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Intra-arterial photodynamic therapy using 5-ALA in a swine model
Abstract
To test the hypothesis that intravascular light could be delivered via a balloon catheter for arterial photodynamic therapy (PDT).
Pig non-injury model.
Clinical catheter equipment.
Large White pigs (15-20 micrograms) were photosensitised with 5-aminolaevulinic acid (5-ALA) induced protoporphyrin IX (PpIX) at a concentration of 120 mg/kg. Arterial biopsies were taken at intervals between 30 mins and 24 h and frozen sections analysed using a CCD camera to give a temporal profile of fluorescence in each arterial layer. PDT was given to normal arterial segments via a 4 mm transparent PTA balloon inflated so as to occlude flow, but not distend the artery. Animals were culled at 3 and 14 days and the above segments harvested.
Fluorescence peaked in the adventitia, intima and medial layers at 1.5, 4 and 6 h respectively. PDT at all time points produced VSMC depletion compared with controls. The degree of depletion mirrored the fluorescence profile of PpIX.
PDT can be delivered via a standard PTA balloon with a transparent channel. This depletes the VSMC population within the arterial wall without complications. Intra-arterial PDT is therefore a potential therapy to reduce the incidence of restenosis post-angioplasty.
... However, only intravascular stents and ionizing irradiation have been proved to clinically reduce this process. 10,11 Photodynamic therapy (PDT) is an effective, alternate experimental approach to favorably modulate this vascular healing response, 12,13 and a first clinical trial for primary atherosclerotic plaque reduction has demonstrated the feasibility of vascular PDT in humans. 14 PDT represents an application of phototherapy in which visible, wavelength-specific light is applied to activate otherwise inert photosensitizer dyes to produce free-radical moieties, without the generation of heat. ...
... 12,17 Because restenosis is essentially a focal obstruction, the precise and strictly local cytotoxic effects of PDT seem to be very well suited for endovascular therapy. 13 However, the clinical use of this promising approach to inhibit restenosis was, in part, limited by the lack of a suitable, readily available, inexpensive, and, therefore, clinically appropriate photosensitizer. ...
... For vascular PDT, photosensitizers have been successfully administered systemically in different animal models. 13,30 Despite the selectivity of confined light irradiation, the site-specific delivery of a photosensitizer directly into a site of vascular intervention has conceptual appeal. 43 First, it would achieve specific local drug concentrations. ...
Photodynamic therapy (PDT), the light activation of photosensitizers to produce free radicals, is known to inhibit experimental intimal hyperplasia (IH). However, its clinical application has been limited by the lack of a suitable approach and a clinically appropriate photosensitizer. The aim of this study was to determine the effectiveness of a clinical approach for PDT, while testing its ability to favorably modulate the vascular wound healing response.
Rat carotid arteries were balloon-injured (BI), and for PDT, the arteries were irradiated with thermoneutral laser light (lambda = 660 nm, 100 J/cm(2)) after the photosensitizer methylene blue (MB) was delivered locally. Control rats included BI alone and MB after BI alone. Arteries were analyzed after 2 weeks with morphometric evaluation (n = 6) and in situ hybridization for versican and procollagen type I gene expression (digitized image pixel analyses, n = 3).
No IH developed in PDT-treated arteries (0 +/- 0 mm(2); compared with BI, 0.192 +/- 0.006 mm(2); P <.0001). The diameters remained unchanged (PDT, 0.95 +/- 0.04 mm; BI, 0.94 +/- 0.05 mm; uninjured artery, 0.91 +/- 0.06 mm). Arterial injury resulted in an increase of versican and procollagen type I messenger RNA (mRNA) in the adventitia and neointima. In the repopulating cells of the adventitia after PDT, there was a significant decrease in versican mRNA (% of positive pixels per high-power field: PDT, 1.13% +/- 0.39%; BI, 2.93% +/- 0.61%; P <.02), but not in procollagen type I mRNA.
The decrease of versican mRNA expression of repopulating cells after PDT reflects favorable healing on a molecular level. Site-specific delivery of MB, a clinically appropriate photosensitizer, followed by PDT represents a suitable method to promote favorable healing after balloon intervention and further supports its role for inhibiting postinterventional restenosis.
... While the biological effects of photoactivation on atheroma are compelling, indeed, the use of currently available photosensitizers is still challenging. The non-specific binding of the photosentizing agent to the vascular structures could potentially cause unexpected damage to the protective barriers, including smooth muscle cells (SMCs) [13,14] and endothelial cells (ECs) [14,15]. Damaged SMCs and endothelial barrier impair the integrity of the fibrous cap overlying the atheroma leading to thrombosis and rupture [16,17]. ...
... While the biological effects of photoactivation on atheroma are compelling, indeed, the use of currently available photosensitizers is still challenging. The non-specific binding of the photosentizing agent to the vascular structures could potentially cause unexpected damage to the protective barriers, including smooth muscle cells (SMCs) [13,14] and endothelial cells (ECs) [14,15]. Damaged SMCs and endothelial barrier impair the integrity of the fibrous cap overlying the atheroma leading to thrombosis and rupture [16,17]. ...
Background
Photoactivation targeting macrophages has emerged as a therapeutic strategy for atherosclerosis, but limited targetable ability of photosensitizers to the lesions hinders its applications. Moreover, the molecular mechanistic insight to its phototherapeutic effects on atheroma is still lacking. Herein, we developed a macrophage targetable near-infrared fluorescence (NIRF) emitting phototheranostic agent by conjugating dextran sulfate (DS) to chlorin e6 (Ce6) and estimated its phototherapeutic feasibility in murine atheroma. Also, the phototherapeutic mechanisms of DS-Ce6 on atherosclerosis were investigated.
Results
The phototheranostic agent DS-Ce6 efficiently internalized into the activated macrophages and foam cells via scavenger receptor-A (SR-A) mediated endocytosis. Customized serial optical imaging-guided photoactivation of DS-Ce6 by light illumination reduced both atheroma burden and inflammation in murine models. Immuno-fluorescence and -histochemical analyses revealed that the photoactivation of DS-Ce6 produced a prominent increase in macrophage-associated apoptotic bodies 1 week after laser irradiation and induced autophagy with Mer tyrosine-protein kinase expression as early as day 1, indicative of an enhanced efferocytosis in atheroma.
Conclusion
Imaging-guided DS-Ce6 photoactivation was able to in vivo detect inflammatory activity in atheroma as well as to simultaneously reduce both plaque burden and inflammation by harmonic contribution of apoptosis, autophagy, and lesional efferocytosis. These results suggest that macrophage targetable phototheranostic nanoagents will be a promising theranostic strategy for high-risk atheroma.
Graphical abstract
... 20 21 5-ALA is attracting particular interest at present. Experiments have shown that there are high concentrations of protoporphyrin IX in the media of pig arteries 4-6 hours after intravenous administration, 21 and also that PDT eVects can be limited to the arterial wall, with no eVect on other adjacent tissues. The drug can be given by mouth, and skin photosensitivity is limited to 24-48 hours. ...
... A laser fibre can be placed within the guide wire channel of a transparent balloon catheter and positioned so that the diffuser at the tip of the fibre is located within the balloon segment (fig 3). 21 Blood is excluded from the lumen of the artery during balloon inflation, and with the balloon inflated the fibre is centred in the arterial lumen, allowing even illumination of the arterial wall. In the coronaries, there is a risk of ischaemia with prolonged balloon inflation and so repeated deflations may be necessary during illumination to minimise the risk of hypoxic injury. ...
The major limitation of the long term success of percutaneous coronary interventions is the development of restenosis, which occurs in 30–50% of patients within six months.1 2 Restenosis is the result of a complex pathophysiological process in the arterial wall in response to balloon dilatation or stent insertion. For many years the hallmark of this response to injury was considered to be the development of intimal hyperplasia resulting from the proliferation and migration of smooth muscle cells from the arterial media.3More recently, it has become clear that this is not the only factor involved. Other key aspects in the development of the restenotic lesion are the extracellular matrix, elastic vessel recoil, and arterial remodelling.4 While our understanding of the processes induced by balloon injury has improved, treatment strategies to limit this have been disappointing in clinical practice.5Stenting, which is now used in up to 80% of all coronary interventional procedures, has almost eliminated problems from elastic recoil and has reduced restenosis rates, but neointimal hyperplasia around stent struts can still lead to in-stent restenosis.6
Intracoronary brachytherapy is one technique that has proved promising in the prevention of restenosis using either catheter based systems for radiation delivery or radioactive stents.7 Concerns have been expressed about late thrombotic occlusion after brachytherapy.8 However, the prolonged use of antiplatelet drugs may control this problem. Brachytherapy is probably the best option for treating restenosis currently available. Nevertheless, it would be attractive to find a method of preventing restenosis after percutaneous intervention that does not involve ionising radiation or prolonged drug treatment. Photodynamic therapy (PDT), which in contrast to brachytherapy uses non-ionising radiation, is emerging as another possible strategy.
PDT involves the interaction of a photosensitising drug, light, and tissue oxygen9 (fig 1). Photosensitising agents, many …
... There were several studies in which PDT was performed through a vessel (Endovascular PDT) for treating different animal experimental disease models [25][26][27][28]: Jenkins et al. performed intra-arterial PDT using 5-ALA in a swine model [25,26]; Edward et al. found that endovascular PDT increases the arterial wall strength in a Wistar rat model [27,28]. Kaplan and other Russian investigators have since reported the performance of systemic PDT on human patients [29]. ...
... There were several studies in which PDT was performed through a vessel (Endovascular PDT) for treating different animal experimental disease models [25][26][27][28]: Jenkins et al. performed intra-arterial PDT using 5-ALA in a swine model [25,26]; Edward et al. found that endovascular PDT increases the arterial wall strength in a Wistar rat model [27,28]. Kaplan and other Russian investigators have since reported the performance of systemic PDT on human patients [29]. ...
Photodynamic therapy (PDT) is attracting attention because of its noticeable inhibitory effects on the growth of dermatological and other solid tumors. Here, we studied the use of PDT in systemic diseases such as leukemia, lymphoma, and metastatic cancer, for which tumor formation areas cannot be clearly compartmentalized. We developed a systemic PDT method and examined its effect in a leukemia mouse model. Growth inhibition of A20 cells (H-2(d), murine B-lymphoma/leukemia, and Balb/c origin) induced by PDT/Photodithazine was evaluated by EZ-Cytox assay. After PDT, changes in cell morphology were assessed by light microscopy. Induction of apoptosis, as well as changes in the cell cycle, were assessed by fluorescence-activated cell sorting (FACS) analysis. A20 cells were injected into Balb/c mice through the tail veins, and PDT was performed. A total of 10 mg kg(-1) body weight of Photodithazine concentration was injected intravenously. After 5 min, micro photofibers (diameter, 200 μm) were inserted into the tail veins and irradiated at 1,200 J with a laser. PDT inhibited growth of A20 cells and resulted in marked morphological changes. PDT also induced apoptosis and G1 arrest. In a leukemia mouse model, systemic PDT increased the survival rate (p < 0.01). This is the first report of the effects of systemic PDT in a leukemia animal model. PDT has been applied only locally in most cases, for example to solid tumors. This study provides experimental evidence that systemic PDT could effectively be applied to systemic and spread tumors, for which tumor formation areas cannot clearly be determined.
... Obviously PCI could be used on superficial lesions or skin disorders. However, by means of optical fibres or other devices many internal sites can also be reached, such as the lungs, the pancreas, the gastrointestinal tract, the urogenital system, joints and blood vessels [Pass, 1993,Jenkins et al., 1998,Trauner et al., 1998,Rockson et al., 2000a. In addition PCI-mediated gene delivery could easily be used as a complement to primary surgical treatment and as an adjuvant treatment to systemic therapy. ...
... restenosis after angioplasty, in graft veins and in coronary stents [Quarck, 2001,Klugherz, 2002, it is crucial to obtain efficient local gene delivery into defined regions of blood vessels. PCI will be very well suited for this, since light can precisely be delivered both through fibre optic catheters [Rockson et al., 2000a] and through standard clinical balloon catheters [Jenkins et al., 1998]. Furthermore, photodynamic treatment on its own has already shown promising results in treating such conditions, both in animal models and in humans [Rockson et al., 2000b]. ...
Numerous gene therapy vectors, both viral and non-viral, are taken into the cell by endocytosis, and for efficient gene delivery the therapeutic genes carried by such vectors have to escape from endocytic vesicles so that the genes can further be translocated to the nucleus. Since endosomal escape is often an inefficient process, release of the transgene from endosomes represents one of the most important barriers for gene transfer by many such vectors. To improve endosomal escape we have developed a new technology, named photochemical internalisation (PCI). In this technology photochemical reactions are initiated by photosensitising compounds localised in endocytic vesicles, inducing rupture of these vesicles upon light exposure. The technology constitutes an efficient light-inducible gene transfer method in vitro, where light-induced increases in transfection or viral transduction of more than 100 and 30 times can be observed, respectively. The method can potentially be developed into a site-specific method for gene delivery in vivo. This article will review the background for the PCI technology, and several aspects of PCI induced gene delivery with synthetic and viral vectors will be discussed. Among these are: (i) The efficiency of the technology with different gene therapy vectors; (ii) use of PCI with targeted vectors; (iii) the timing of DNA delivery relative to the photochemical treatment. The prospects of using the technology for site-specific gene delivery in vivo will be thoroughly discussed, with special emphasis on the possibilities for clinical use. In this context our in vivo experience with the PCI technology as well as the clinical experience with photodynamic therapy will be treated, as this is highly relevant for the clinical use of PCI-mediated gene delivery. The use of photochemical treatments as a tool for understanding the more general mechanisms of transfection will also be discussed.
... Troels et al. utilized ICG in a clinical setting where it was shown to reduce arterial wall thickness upon activation with near-infrared light, illustrating its beneficial role in targeting vascular smooth muscle cells and reducing neointimal hyperplasia. 5-aminolevulinic acid (ALA)-induced protoporphyrin IX is a significant focus of PDT research due to its proclivity to concentrate in atheromas after systemic administration [142,143]. The ease of application and activation using external light sources makes it particularly attractive for clinical use. ...
This review paper examines the evolution of photodynamic therapy (PDT) as a novel, minimally invasive strategy for treating atherosclerosis, a leading global health concern. Atherosclerosis is characterized by the accumulation of lipids and inflammation within arterial walls, leading to significant morbidity and mortality through cardiovascular diseases such as myocardial infarction and stroke. Traditional therapeutic approaches have primarily focused on modulating risk factors such as hypertension and hyperlipidemia, with emerging evidence highlighting the pivotal role of inflammation. PDT, leveraging a photosensitizer, specific-wavelength light, and oxygen, offers targeted treatment by inducing cell death in diseased tissues while sparing healthy ones. This specificity, combined with advancements in nanoparticle technology for improved delivery, positions PDT as a promising alternative to traditional interventions. The review explores the mechanistic basis of PDT, its efficacy in preclinical studies, and the potential for enhancing plaque stability and reducing macrophage density within plaques. It also addresses the need for further research to optimize treatment parameters, mitigate adverse effects, and validate long-term outcomes. By detailing past developments, current progress, and future directions, this paper aims to highlight PDT’s potential in revolutionizing atherosclerosis treatment, bridging the gap from experimental research to clinical application.
... 5-Aminolevulinic acid (5-ALA) is a precursor of protoporphyrin-IX. Photodynamic therapy with 5-ALA has been tested on rabbits and pigs for the treatment of atherosclerotic plaques [207,208]. According to these studies, 5-ALA-based PDT may be an effective method for preventing and treating atherosclerosis [209]. ...
Lipids, together with lipoprotein particles, are the cause of atherosclerosis, which is a pathology of the cardiovascular system. In addition, it affects inflammatory processes and affects the vessels and heart. In pharmaceutical answer to this, statins are considered a first-stage treatment method to block cholesterol synthesis. Many times, additional drugs are also used with this method to lower lipid concentrations in order to achieve certain values of low-density lipoprotein (LDL) cholesterol. Recent advances in photodynamic therapy (PDT) as a new cancer treatment have gained the therapy much attention as a minimally invasive and highly selective method. Photodynamic therapy has been proven more effective than chemotherapy, radiotherapy, and immunotherapy alone in numerous studies. Consequently, photodynamic therapy research has expanded in many fields of medicine due to its increased therapeutic effects and reduced side effects. Currently, PDT is the most commonly used therapy for treating age-related macular degeneration, as well as inflammatory diseases, and skin infections. The effectiveness of photodynamic therapy against a number of pathogens has also been demonstrated in various studies. Also, PDT has been used in the treatment of cardiovascular diseases, such as atherosclerosis and hyperplasia of the arterial intima. This review evaluates the effectiveness and usefulness of photodynamic therapy in cardiovascular diseases. According to the analysis, photodynamic therapy is a promising approach for treating cardiovascular diseases and may lead to new clinical trials and management standards. Our review addresses the used therapeutic strategies and also describes new therapeutic strategies to reduce the cardiovascular burden that is induced by lipids.
... 5-Aminolevulinic acid (5-ALA) is the precursor of protoporphyrin-IX. Studies conducted on both rabbits [50] and pigs [51,52] confirmed that PDT with 5-ALA causes a significant reduction in atherosclerotic plaque and PDT offers a new, promising approach to preventing restenosis after endovascular procedures, as evidenced by a significant reduction in vascular smooth muscle cells (VSMCs) observed 28 days after stenting. A particularly significant reduction in neointimal hyperplasia occurred in the group of rabbits that received PDT before stenting [53]. ...
Cardiovascular diseases are the third most common cause of death in the world. The most common are heart attacks and stroke. Cardiovascular diseases are a global problem monitored by many centers, including the World Health Organization (WHO). Atherosclerosis is one aspect that significantly influences the development and management of cardiovascular diseases. Photodynamic therapy (PDT) is one of the therapeutic methods used for various types of inflammatory, cancerous and non-cancer diseases. Currently, it is not practiced very often in the field of cardiology. It is most often practiced and tested experimentally under in vitro experimental conditions. In clinical practice, the use of PDT is still rare. The aim of this review was to characterize the effectiveness of PDT in the treatment of cardiovascular diseases. Additionally, the most frequently used photosensitizers in cardiology are summarized.
... The study demonstrated that PDT delivered via a standard percutaneous transluminal angioplasty (PTA) balloon effectively depleted the vascular smooth muscle cell population within the arterial wall without complications. This suggests the potential of intravascular PDT to reduce the incidence of restenosis post-angioplasty, showcasing its viability as a minimally invasive intervention for addressing arterial complications [175]. ...
Atherosclerosis, which currently contributes to 31% of deaths globally, is of critical cardiovascular concern. Current diagnostic tools and biomarkers are limited, emphasizing the need for early detection. Lifestyle modifications and medications form the basis of treatment, and emerging therapies such as photodynamic therapy are being developed. Photodynamic therapy involves a photosensitizer selectively targeting components of atherosclerotic plaques. When activated by specific light wavelengths, it induces localized oxidative stress aiming to stabilize plaques and reduce inflammation. The key advantage lies in its selective targeting, sparing healthy tissues. While preclinical studies are encouraging, ongoing research and clinical trials are crucial for optimizing protocols and ensuring long-term safety and efficacy. The potential combination with other therapies makes photodynamic therapy a versatile and promising avenue for addressing atherosclerosis and associated cardiovascular disease. The investigations underscore the possibility of utilizing photodynamic therapy as a valuable treatment choice for atherosclerosis. As advancements in research continue, photodynamic therapy might become more seamlessly incorporated into clinical approaches for managing atherosclerosis, providing a blend of efficacy and limited invasiveness.
... 83 5-Amino-levulinic acid-induced protoporphyrin IX (ALA-PpIX) is a naturally occurring porphyrin precursor, which produces PpIX to sensitize the target cells under irradiation. Based on various animal models [84][85][86][87] and clinical studies, 88 its efficacy in depleting the VSMC population and inhibiting intimal hyperplasia or constrictive remodeling has been demonstrated. However, ALA can cause hemodynamic changes (reductions in systemic and pulmonary pressure and lung resistance), which may limit its use in the cardiovascular field. ...
Atherosclerosis (AS) is a chronic inflammatory disease, which may lead to high morbidity and mortality. Currently, the clinical treatment strategy for AS is administering drugs and performing surgery. However, advanced therapy strategies are urgently required because of the deficient therapeutic effects of current managements. Increased number of energy conversion-based organic or inorganic materials has been used in cancer and other major disease treatments, bringing hope to patients with the development of nanomedicine and materials. These treatment strategies employ specific nanomaterials with specific own physiochemical properties (external stimuli: light or ultrasound) to promote foam cell apoptosis and cholesterol efflux. Based on the pathological characteristics of vulnerable plaques, energy conversion-based nano-therapy has attracted increasing attention in the field of anti-atherosclerosis. Therefore, this review focuses on recent advances in energy conversion-based treatments. In addition to summarizing the therapeutic effects of various techniques, the regulated pathological processes are highlighted. Finally, the challenges and prospects for further development of dynamic treatment for AS are discussed.
... On the other hand, a concern with the balloon methodology of photo-crosslinking may be the irradiation of the parent vessel. However, intravascular light irradiation has already been previously utilized for the treatment of atherosclerosis or vasoconstriction without causing damage to endothelial cells or circulating cells within the bloodstream [43][44][45]. Moreover, histopathological analysis did not reveal damage to the endothelial cells following 5 min of 405 nm light irradiation at this intensity (20 mW). ...
Background: The endovascular treatment of cerebral aneurysms has become widespread but may still be limited by recurrence rates or complications. The discovery of novel embolic strategies may help mitigate these concerns. Methods: We formulated a Photosensitive Hydrogel Polymer (PHP) embolic agent which is low-viscosity, shear-thinning, and radio-opaque. After the filling of an aneurysm with PHP with balloon assistance, we utilized photopolymerization to induce solidification. Different methods of light delivery for photopolymerization were assessed in silicone models of aneurysms and in four acute animal trials with venous anastomosis aneurysms in pigs. Then, balloon-assisted embolization with PHP and photopolymerization was performed in three aneurysms in pigs with a one-month follow-up. Filling volume, recurrence rates, and complications were recorded. Results: The PHP was found to be suitable for the intravascular delivery and treatment of cerebral aneurysms. It was found that light delivery through the balloon catheter, as opposed to light delivery through the injection microcatheter, led to higher rates of filling in the 3D model and acute animal model for cerebral aneurysms. Using the balloon-assisted embolization and light delivery strategy, three wide-necked aneurysms were treated without complication. One-month follow-up showed no recurrence or neck remnants. Conclusions: We demonstrated a novel method of balloon-assisted photosensitive hydrogel polymer embolization and photopolymerization, leading to complete aneurysm filling with no recurrence at 1 month in three wide-necked aneurysms in pigs. This promising methodology will be investigated further with longer-term comparative animal trials.
... The photosensitizer specifically accumulates in the atherosclerotic plaques and, after light activation, triggers a photochemical response that includes the generation of ROS and interference of cell survival and remodeling processes [97][98][99][100]. A number of conventional photosensitizers, namely porphyrins (hematoporphyrin derivative, Verteporfin), phtalocyanine derivatives, chlorins (Talaporfin sodium), 5-aminolevulinic acid and motexafin lutetium showed phototherapeutic properties in atherosclerosis in preclinical studies [101][102][103][104][105][106][107][108][109][110]. However, translation of PDT to clinical use warrants further studies with experimental models similar to human coronary atherosclerosis, clarification of some issues related to the appropriate type, toxicity and dosage of photosensitizer, light hypersensitivity and cutaneous photosensitivity, interference with arterial calcification and lipid-lowering/antiplatelet drugs and selective delivery in the atherosclerotic area [97][98][99]. ...
Despite progress in understanding the pathogenesis of atherosclerosis, the development of effective therapeutic strategies is a challenging task that requires more research to attain its full potential. This review discusses current pharmacotherapy in atherosclerosis and explores the potential of some important emerging therapies (antibody-based therapeutics, cytokine-targeting therapy, antisense oligonucleotides, photodynamic therapy and theranostics) in terms of clinical translation. A chemopreventive approach based on modern research of plant-derived products is also presented. Future perspectives on preventive and therapeutic management of atherosclerosis and the design of tailored treatments are outlined.
... Intraarterial PDT at the time of balloon vessel injury was shown to inhibit intimal hyperplasia in animal model [14]. Despite that PDT is a one-time procedure it can induce biochemical processes leading to long-term inhibition of intimal hyperplasia [15]. ...
In this study we present the porous silica-based material that can be used for in situ drug delivery, offering effective supply of active compounds regardless its water solubility. To demonstrate usability of this new material, three silica-based materials with different pore size distribution as a matrix for doping with Photolon (Ph) and Protoporphyrin IX (PPIX) photosensitizers, were prepared. These matrices can be used for coating cardiovascular stents used for treatment of the coronary artery disease and enable intravascular photodynamic therapy (PDT), which can modulate the vascular response to injury caused by stent implantation—procedure that should be thought as an alternative for drug eluting stent. The FTIR spectroscopic analysis confirmed that all studied matrices have been successfully functionalized with the target photosensitizers. Atomic force microscopy revealed that resulting photoactive matrices were very smooth, which can limit the implantation damage and reduce the risk of restenosis. No viability loss of human peripheral blood lymphocytes and no erythrocyte hemolysis upon prolonged incubations on matrices indicated good biocompatibility of designed materials. The suitability of photoactive surfaces for PDT was tested in two cell lines relevant to stent implantation: vascular endothelial cells (HUVECs) and vascular smooth muscle cells (VSMC). It was demonstrated that 2 h incubation on the silica matrices was sufficient for uptake of the encapsulated photosensitizers. Moreover, the amount of the absorbed photosensitizer was sufficient for induction of a phototoxic reaction as shown by a rise of the reactive oxygen species in photosensitized VSMC. On the other hand, limited reactive oxygen species (ROS) induction in HUVECs in our experimental set up suggests that the proposed method of PDT may be less harmful for the endothelial cells and may decrease a risk of the restenosis. Presented data clearly demonstrate that porous silica-based matrices are capable of in situ delivery of photosensitizer for PDT of VSMC.
... Experimental evidence suggests that PDT, in addition to plaque stabilization and regression, can also be applied to prevent restenosis 42,49,50 . Restenosis is provoked by endothelial or medial injury followed by inflammatory cell infiltration, vascular smooth muscle cell (VSMC) proliferation and migration resulting in intimal hyperplasia and narrowing of vessel lumen. ...
Acute coronary syndromeis a life-threatening condition of utmost clinical importance, which, despite recent progress in the field, is still associated with high morbidity and mortality. Acute coronary syndrome results from a rupture or erosion of vulnerable atherosclerotic plaque with secondary platelet activation and thrombus formation, which leads to partial or complete luminal obstruction of a coronary artery. During the last decade, scientific evidence demonstrated that, when an acute coronary event occurs, several non-culprit plaques are in a “vulnerable” state. Among the promising approaches, several investigations provided evidence of photodynamic therapy (PDT) induced stabilisation and regression of atherosclerotic plaque. Significant development of PDT strategies improved its therapeutic outcome. This review addresses PDT's pertinence and major problems/challenges toward its translation to a clinical reality. This article is protected by copyright. All rights reserved.
... For example, Ahn et al. [4] demonstrated that optical fiber implantation-mediated SPDT resulted in increased survival rates in a leukemia animal model. However, there are several disadvantages associated with the internal irradiation approach, including a small lighting area, less precise lighting times, and unstable PS concentration [5][6][7][8]. While the irradiation of extracorporeal blood could overcome these problems, there have been no reports that have examined the efficacy of this method. ...
Systemic PDT (SPDT) approach is developed to treat a variety of hematological diseases, including cancers and blood-borne infections. We evaluated the efficacy of an SPDT method for treating leukemia using a Brown Norway myeloid leukemia (BNML) rat model with the LT12 cells engineered to express GFP. The survival times of animals receiving SPDT at 5 (early-SPDT) and 10 (mid-SPDT) days post-LT12 injection were prolonged by 2days, the rats in the late-SPDT group (15days) exhibited a 6-day increase in life span (p<0.05). The percentages of GFP-LT12 cells in the bone marrow of the late-SPDT rats decreased from 61.6% to 56.5% on day 17. Likewise, there was a decrease in the serum expression levels of IL-1β, IL-10, TNF-α, and IFN-γ in the late-SPDT rats (p<0.05). Our findings indicate that SPDT could be an effective method for the treatment of leukemia, and that antitumor immunity may play a key role in this process.
... Although motexafin lutetium is well tolerated at lower doses, at higher doses a significant number of patients had paraesthesia (38). The use of deep penetrating red light might be better tolerated in peripheral vessels than in coronary vessels, where PDT with 5-aminolaevulinic acid has shown evidence of myocardial scarring in porcine coronary arteries (39). Although the green light used in the current study requires a blood free field to avoid absorption by circulating red blood cells, the light exposure time using green light (90 s) is significantly less than the time used for red light (12 min) in prior studies. ...
Background:
In humans, rupture of vulnerable plaques is one of the leading causes of
cardiovascular-related mortality. The current therapies for atherosclerosis focus on cholesterol
lowering and reducing inflammation without affecting the proliferative cell types
that cause plaque progression. intravascular Photopoint Photodynamic Therapy (PDT) is
a combination therapy that utilizes a photo activated drug and a light-emitting catheter to
eliminate undesirable cell populations. The aim of the present study is to assess the efficacy
of Photopoint as a therapy for the elimination of atherosclerotic plaque cell populations
with a view to inducing plaque stablizationlregression.
Methods:
New Zealand White rabbits (n=l6) were fed a 1% cholesterol diet followed by
bilateral iliac balloon endothelial denudation. At 5wk post-denudation, rabbits received
photosensitizer MV0611 (3mg/kg I.V.) followed by lkght delivery using a Mwavant catheter-
based diode laser (15J/cm2) at 4, 8 and 24 hrs post-injection. Control animals
received either drug or light alone. Rabbits were sacrificed at 7d post-treatment and iliac
arteries harvested for histopathology and immunohlstochemistry.
Results:
Maximum elimination of plaque cell nuclei was achieved at 6 and 24 hours drug
incubation (78 f 6%, 96i4%; pcO.001). Nuclear depletion was consistent throughout
entire 2cm lengths of plaque. PDT induced complete elimination of macrophage ‘foam’
cells and other inflammatory cells with a corresponding reduction in ‘foam’ cell neutral
lipid. At 7days post PDT, expression of P53 correlated with cell loss. There was no evidence
of Inflammation, necrosis or thrombosis post Photopoint treatment.
Conclusions:
Photopoint PDT may prove to be an effective therapy for the treatment of
vulnerable plaque. An important component of this treatment may be the removal of macrophages
by apoptotic mechanisms. Studies to investigate the chronic effects of PDT are
ongoing.
... The effects of PDT on the intact arterial vessels are also known. It has been demonstrated that PDT of normal vessels causes a unique biological response including endothelial denudation and dose-dependent thinning of medial vascular layer due to depletion of vascular smooth muscle cells [26, 27]. Of note, these striking histological changes occur in the absence of local inflammatory reaction. ...
of cardiovascular diseases are also discussed. Current findings demonstrate that fullerenes may show several potentially physiologically and clinically relevant activities, including antiischemic effect, vasodilatation, inhibition of low-density lipoprotein oxidation, and limitation of proliferative activity of vascular smooth muscle cells. Additional studies will be required to define the molecular mechanisms responsible for the observed effects.
... Although motexafin lutetium is well tolerated at lower doses, at higher doses a significant number of patients had paraesthesia (38). The use of deep penetrating red light might be better tolerated in peripheral vessels than in coronary vessels, where PDT with 5-aminolaevulinic acid has shown evidence of myocardial scarring in porcine coronary arteries (39). Although the green light used in the current study requires a blood free field to avoid absorption by circulating red blood cells, the light exposure time using green light (90 s) is significantly less than the time used for red light (12 min) in prior studies. ...
The purpose of this study was to determine how photodynamic therapy (PDT) promotes stabilization and reduction of regional atherosclerosis.
Photodynamic therapy, a combination of photosensitizer and targeted light to promote cell apoptosis, has been shown to reduce atherosclerotic plaque inflammation.
Forty New Zealand White rabbits were fed with cholesterol. The iliac arteries were balloon denuded and randomized to receive either PhotoPoint PDT treatment (photosensitizer and light) (Miravant Medical Technologies, Santa Barbara, California), photosensitizer (MV0611) alone, or light alone and were then compared at 7 and 28 days. Arteries were examined for evidence of plaque volume, cell number, macrophage and smooth muscle cell (SMC) content, and plaque cell proliferation.
Compared with contralateral iliac artery controls at 7 days, plaque progression was reduced by approximately 35% (p < 0.01); plaque progression was further reduced to approximately 53% (p < 0.01) by 28 days, leading to an increase in lumen patency (p < 0.05). At 7 days after PDT, percent plaque area occupied by macrophages decreased by approximately 98% (p < 0.001) and SMCs by approximately 72% (p < 0.05). At 28 days after PDT, removal of macrophages was sustained (approximately 92% decrease, p < 0.001) and plaques were repopulated with non-proliferating SMCs (approximately 220% increase, p < 0.001). There was no evidence of negative or expansive arterial remodeling, thrombosis, or aneurysm formation.
Photodynamic therapy simultaneously reduces plaque inflammation and promotes repopulation of plaques with a SMC-rich stable plaque cell phenotype while reducing disease progression. These early healing responses suggest that PDT is a promising therapy for the treatment of acute coronary syndromes.
... 10 These free radicals eradicate the entire cell population of the artery wall without inducing inflammation or structural deterioration and thus result in long-term inhibition of experimental IH. 11,12 Vascular PDT has other effects, including inactivation of matrix-associated cytokines and growth factors, which result in alteration of vascular cell function. 13 These matrix effects may influence the observed cellular repopulation of PDTtreated arteries, including reendothelialization and repopulation of the adventitia, but delayed and only sparse repopulation of the media. ...
Photodynamic therapy (PDT) inhibits experimental intimal hyperplasia. PDT results in complete vascular wall cell eradication with subsequent adventitia but minimal media repopulation. This study was designed to test the hypothesis that PDT alters the vascular wall matrix thereby inhibiting invasive cell migration, and as such, provides an important barrier mechanism to favorably alter the vascular injury response. Untreated smooth muscle cells (SMCs) and fibroblasts were seeded on control and PDT-treated (100 J/cm(2); photosensitizer was chloroaluminum-sulfonated phthalocyanine, 5 microg/mL) 3-dimensional collagen matrix gels. Invasive cell migration was temporally quantified by calibrated microscopy. Zymography and ELISA assessed SMC matrix metalloproteinase levels. Molecular changes of gel proteins and their susceptibility to collagenase were analyzed by SDS-PAGE and Western blot. Limited pepsin digestion and histology were used to assess the in vivo relevance of the model, using an established rat carotid artery model at 1 and 4 weeks after balloon injury and PDT. PDT of 3-dimensional matrix of gels led to a 52% reduction of invasive SMCs and to a 59% reduction of fibroblast migration (P<0.001) but did not significantly affect secretion of matrix metalloproteinases. PDT induced collagen matrix changes, including cross-linking, which resulted in resistance to protease digestion. PDT led to a durable 45% reduction in pepsin digestion susceptibility of treated arteries (P<0.001) and inhibition of periadventitial cell migration into the media. These data suggest that PDT of matrix gels generates a barrier to invasive cellular migration. This newly identified effect on matrix proteins underscores its pleiotropic actions on the vessel wall, and as such, PDT may be of considerable potential therapeutic value to inhibit restenosis.
... There has been no evidence of aneurysm formation or thrombosis. [3][4][5] Further studies have shown that PDT can inhibit neointimal hyperplasia following angioplasty to rat carotid, rabbit iliac and pig coronary arteries. [6][7][8][9] Pilot safety studies of its application to peripheral and coronary disease in man have now been reported. ...
Photodynamic therapy (PDT, the combination of light with a photosensitising drug in the presence of oxygen) inhibits restenosis after angioplasty without stenting. This study assesses the potential of PDT for prevention of in-stent re-stenosis.
Normal rabbits were given the photosensitising agent 5-aminolaevulinic acid (ALA) 60 mg/kg, 3 h prior to endovascular illumination of the iliac artery (635 nm at 50 J/cm(2)) either immediately before or after deployment of an oversized (3 mm diameter) stent. PDT treated arteries were retrieved 3 or 28 days later and assessed for cell counts and vascular morphometry. Control arteries (stent but no PDT) were examined at 28 days.
There were no adverse events and all vessels were patent at the end of the study. At 3 days there was almost complete medial cell ablation when light was delivered before stent deployment (17+/-1 cells/hpf), with little effect when illumination followed stent deployment (184+/-17 cells/hpf, p<0.0001). Twenty-eight days after PDT, the neointimal areas were 1.41+/-0.52 mm(2) (stent with no PDT), 1.24+/-0.54 mm(2) (light after stent) and 0.60+/-0.21 mm(2) (light before stent) (p=0.004).
PDT before stent deployment caused almost complete medial cell ablation at 3 days with inhibition of in-stent restenosis at 28 days. PDT is worthy of further study as an adjuvant to percutaneous intervention in patients with vascular disease.
... In addition, phototherapy using ALA administration to generate protoporphyrin IX has been investigated as an approach to treat restenosis (24,28). Studies examining the treatment of animals with ALA for restenosis phototherapy have documented that ALA increases protoporphyrin IX fluorescence levels in the smooth muscle region of the arterial wall (21,28). Treatment of isolated skeletal muscle arterioles with ALA has been used in studies on vascular regulation by the heme oxygenase system as a method of increasing heme availability for the production of carbon monoxide (22). ...
Protoporphyrin IX is an activator of soluble guanylate cyclase (sGC), but its role as an endogenous regulator of vascular function through cGMP has not been previously reported. In this study we examined whether the heme precursor delta-aminolevulinic acid (ALA) could regulate vascular force through promoting protoporphyrin IX-elicited activation of sGC. Exposure of endothelium-denuded bovine pulmonary arteries (BPA) in organoid culture to increasing concentrations of the heme precursor ALA caused a concentration-dependent increase in BPA epifluorescence, consistent with increased tissue protoporphyrin IX levels, associated with decreased force generation to increasing concentrations of serotonin. The force-depressing actions of 0.1 mM ALA were associated with increased cGMP-associated vasodilator-stimulated phosphoprotein (VASP) phosphorylation and increased sGC activity in homogenates of BPA cultured with ALA. Increasing iron availability with 0.1 mM FeSO(4) inhibited the decrease in contraction to serotonin and increase in sGC activity caused by ALA, associated with decreased protoporphyrin IX and increased heme. Chelating endogenous iron with 0.1 mM deferoxamine increased the detection of protoporphyrin IX and force depressing activity of 10 microM ALA. The inhibition of sGC activation with the heme oxidant 10 muM 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ) attenuated the force depressing actions of an NO donor without altering the actions of ALA. Thus control of endogenous formation of protoporphyrin IX from ALA by the availability of iron is potentially a novel physiological mechanism of controlling vascular function through regulating the activity of sGC.
The origins of photodynamic therapy (PDT) date back to 1904. Since then, the amount of research proving PDT and, consequently, its applicability to various disease states has steadily increased. Currently, PDT is mainly used in oncology to destroy cancer cells. It is being worked on for possible use in other medical fields as well, including cardiology. It can be used in the prevention of restenosis, often occurring after vascular surgical interventions, for destroying atherosclerotic plaques and as a new ablative method of ectopic centers in the treatment of atrial fibrillation. The purpose of this review is to summarize the knowledge to date regarding the therapeutic potential of using PDT for various pathological conditions in cardiology. The review also focuses on the current limitations associated with the use of PDT and identifies areas where more research is needed to develop better drug regimens. Materials and methods: The study analyzed 189 medical articles. The articles came from PubMed, Frontiers, Google Scholar, Science Direct and Web of Science databases. Through the excitation of light, a photosensitizer (PS) introduced into the body, the destruction of pathological cells occurs. PTD is widely used in oncology of the central nervous system (CNS). This process is made possible by the production of free oxygen radicals (ROS) and singlet oxygen, which generate oxidative stress that destroys sensitive cancer cells. In recent years, photosensitizers have also been discovered to have a strong affinity for macrophages that fill atherosclerotic plaques, making these compounds suitable for treating atherosclerosis. By inducing apoptosis of smooth muscle cells, inactivating basic fibroblast growth factor (FGF-β) and inhibiting endothelial cell hyperplasia, PDT can be used to prevent restenosis after surgical proceduresPDT appears to be a minimally invasive and highly effective therapeutic method, especially when combined with other therapeutic methods. Unfortunately, the small number of animal model studies and human clinical trials greatly limit the applicability of PDT on a wider scale. Current limitations, such as the depth of penetration, delivery of photosensitizer particles to the direct site of the lesion or the appropriate choice of photosensitizer in relation to the nature of the pathology, unfortunately make it impossible to replace current therapeutic approaches.
Photodynamic therapy (PDT) is a two-step procedure that involves the administration of special drugs, commonly called photosensitizers, followed by the application of certain wavelengths of light. The light activates these photosensitizers to produce reactive molecular species that induce cell death in tissues. There are numerous factors to consider when selecting the appropriate photosensitizer administration route, such as which part of the body is being targeted, the pharmacokinetics of photosensitizers, and the formulation of photosensitizers. While intravenous, topical, and oral administration of photosensitizers are widely used in preclinical and clinical applications of PDT, other administration routes, such as intraperitoneal, intra-arterial, and intratumoral injections, are gaining traction for their potential in treating advanced diseases and reducing off-target toxicities. With recent advances in targeted nanotechnology, biomaterials, and light delivery systems, the exciting possibilities of targeted photosensitizer delivery can be fully realized for preclinical and clinical applications. Further, in light of the growing burden of cancer mortality in low- and middle-income countries and development of low-cost light sources and photosensitizers, PDT could be used to treat cancer patients in low-income settings. This short article introduces aspects of interfaces of intratumoral photosensitizer injections and nano-biomaterials for PDT applications in both high-income and low-income settings but does not present a comprehensive review due to space limitations.
Photodynamic therapy (PDT) is an effective procedure for the treatment of lesions diseases based on the selectivity of a photosensitising compound with the ability to accumulate in the target cell. Atherosclerotic plaque is a suitable target for PDT because of the preferential accumulation of photosensitisers in atherosclerotic plaques. Dendrimers are hyperbranched polymers conjugated to drugs. The dendrimers of ALA hold ester bonds that inside the cells are cleaved and release ALA, yielding PpIX production. The dendrimer 6m-ALA was chosen to perform this study since in previous studies it induced the highest porphyrin macrophage: endothelial cell ratio (Rodriguez et al. in Photochem Photobiol Sci 14:1617–1627, 2015). We transformed Raw 264.7 macrophages to foam cells by exposure to oxidised LDLs, and we employed a co-culture model of HMEC-1 endothelial cells and foam cells to study the affinity of ALA dendrimers for the foam cells. In this work it was proposed an in vitro model of atheromatous plaque, the aim was to study the selectivity of an ALA dendrimer for the foam cells as compared to the endothelial cells in a co-culture system and the type of cell death triggered by the photodynamic treatment. The ALA dendrimer 6m-ALA showed selectivity PDT response for foam cells against endothelial cells. A light dose of 1 J/cm2 eliminate foam cells, whereas less than 50% of HMEC-1 is killed, and apoptosis cell death is involved in this process, and no necrosis is present. We propose the use of ALA dendrimers as pro-photosensitisers to be employed in photoangioplasty to aid in the treatment of obstructive cardiovascular diseases, and these molecules can also be employed as a theranostic agent.
Atherosclerosis is the commonest aetiology in deaths arising from cardiovascular disease. It is characterised by the build up of an intraluminal plaque leading to arterial stenosis. Balloon angioplasty offers a minimally invasive method of dilating such stenoses in both peripheral and coronary arteries. However, despite very favourable immediate results, 15-40% of arteries restenose within 3-6 months following angioplasty with obvious clinical and resource implications. Restenosis is caused by a combination of neointimal hyperplasia (NIH) and negative geometric remodelling, the combined effect of which results in luminal narrowing. The aim of this thesis was to investigate a method of inhibiting restenosis using photodynamic therapy (PDT). PDT involves the interaction of light of a specific wavelength with a pre-administered photosensitiser to produce cell death by oxygen-dependent cytotoxic mediators. Experimental project Preliminary studies established the pharmacokinetics of the chosen photosensitiser 5-aminolaevulinic acid (ALA) in a swine model. From these experiments the optimum drug-light interval was calculated and in a second study, PDT was applied to normal porcine iliac and coronary arteries using an endovascular light source. Depletion of medial vascular smooth muscle cells (VSMC) was seen at 3 and 14 days and was found to be partially dependent on the drug-light interval. Finally, iliac and coronary arteries were balloon injured and then treated with PDT or sham illumination. Histomorphometric studies following harvest at 28 days showed less NIH and less negative remodelling in the group treated with PDT. Clinical project A clinical pilot study of angioplasty with adjuvant PDT was commenced. Patients deemed to be at high risk of restenosis underwent femoral angioplasty followed by endovascular PDT and were followed up by duplex and digital subtraction angiography at 6 months. The results of this small study would suggest that PDT is successful in inhibiting restenosis, but this now needs to be confirmed in a randomised controlled trial.
Aim:
The aim of this article is to summarize and review the use of photodynamic therapy for the treatment of atherosclerotic plaque and the prevention of intimal hyperplasia. Different photosensitizers are discussed and more specifically the role of indocyanine green as a potential photosensitizer.
Methods:
Literature search with focus on the use of photodynamic therapy in atherosclerosis, the mechanism of action and the different photosensitizers for photodynamic therapy.
Results:
In-vitro and in-vivo studies confirm the possibilities of using photodynamic therapy for the treatment of atherosclerosis and the prevention of restenosis. Insufficient specificity in the accumulation of photosensitizer and thus phototoxicity, remains an important problem. Indocyanine green is a photosensitizer with features in favor of photodynamic therapy. Results obtained so far of photodynamic therapy with indocyanine green point towards the potential of indocyanine green as a photosensitizer in photodynamic therapy for atherosclerosis.
Conclusion:
Photodynamic therapy is a promising tool for treating atherosclerosis. Many of the studied photosensitizers have toxic effects. Indocyanine green might be a good photosensitizer for the use of photodynamic therapy in atherosclerosis. These data justify further research to the use of indocyanine green as a photosensitizer in the treatment of atherosclerotic plaque both de novo or in restenotic lesions.
The use of endogenous protoporphyrin IX after administration of 5-aminolaevulinic acid (ALA) has led to many applications in photodynamic therapy (PDT). We have previously reported that the conjugation of ALA dendrimers enhances porphyrin synthesis. The first aim of this work was to evaluate the ability of ALA dendrimers carrying 6 and 9 ALA residues (6m-ALA and 9m-ALA) to photosensitise cancer cells. For this aim, we employed LM3 mammary carcinoma cells. In these tumour cells, porphyrin synthesis from dendrimers was higher as compared to ALA at low concentrations, whereas at high concentrations, porphyrin synthesis was similar from both compounds. Topical application of ALA dendrimers on the skin overlying a subcutaneous LM3 implanted tumour showed no diffusion of the molecules either to distant skin sites or to the adjacent tumour, suggesting a promising use of the ALA macromolecules in superficial cancer models. As a second objective, we proposed the use of ALA-dendrimers in vascular PDT for the treatment of atherosclerosis. Thus, we focused our studies on ALA-dendrimers selectivity towards macrophages in comparison to endothelial cells. For this aim we employed Raw 264.7 macrophages and HMEC-1 microvasculature cells. Porphyrin synthesis induced in macrophages by 6m-ALA and 9m-ALA (3 h, 0.025 mM), was 6 and 4.6 times higher respectively as compared to the endothelial cell line, demonstrating high affinity of ALA dendrimers for macrophages. On the other hand, ALA employed at low concentrations was slightly selective (1.7-fold) for macrophages. Inhibition studies suggested that ALA dendrimers uptake in macrophages is mainly mediated by caveloae-mediated endocytosis. Our main conclusion is that, in addition to being promising molecules in PDT of superficial cancer, ALA dendrimers may also find applications in vascular PDT, since in vitro they showed selectivity for the macrophage component of the atheromatous plaque, as compared to the vascular endothelium.
The Wellman Laboratories of Photomedicine at the Massachusetts General Hospital in
Boston is an internationally known institute for the development and application of laser in medicinco
Besides fundamental research which investigates the interaction of light with cells and molecules,
a great deal of studies arc performed towards clinical application to treat human diseases.
For this reason, all known medical specialties are involved in research programmes examining the
use of laser for their purposes: from ophtalmologists, using photodynamic therapy to treat
choroidal neovascularization, oncologists to photochemically target and remove breast cancer
cells from bone marrow, urologist to detect bladder cancer with fluorescence, gastroenterologists
to treat Barret's oesophagus to dermatologists removing latloes and treating multiple skin diseases
with the laser. The application of laser to treat vascular diseases was started decades ago, but more
recently the concept of photodynamic therapy was introduced by the research group of Dr.
LaMuraglia to prevent restenosis. As a research fellow in surgery I joined this group in 1994 and
our research goal was to better understand how photodynamic therapy affects the vascular wall.
The results from these investigations are presented in tllis thesis. The first part gives an outline of
the problem of restenosis, the concept of photodynamic therapy and aims of the study. The following
chapters describe how photodynamic therapy interacts with biological factors that regulate
the vascular healing process. Finally, considerations for possible clinical use are discussed.
Although the concept photodynamic therapy has been recognised for over a century, it is only over the last 25 years that it has been used in Great Britain. The first applications in the UK were in 1981 by John Carruth, who treated patients with advanced ENT and skin cancers. The following year, he and Stephen Bown set up the British Medical Laser Association (BMLA). Since that time, the use of PDT in the UK has slowly expanded in all fields of medicine and surgery. In 1986, Bown set up the National Medical Laser Centre (NMLC) and later collaborated with Liverpool gastroenterologist, Neville Krasner, in animal studies on rat colon. In 1997, Keyvan Moghissi founded the Yorkshire Laser Centre (YLC) and began treating patients with advanced inoperable bronchial and oesophageal cancers. Stan Brown in Leeds set up the Centre for Photobiology and Photodynamic Therapy at the University of Leeds, working in close collaboration with the Yorkshire Cancer Research Centre. Other pioneers include Hugh Barr in Gloucester, Colin Hopper in London, Grant Fullarton in Glasgow and Roger Ackroyd, Malcolm Reed and Nicky Brown in Sheffield. PDT has now been used in the UK in the treatment of skin, oral, ENT, oesophageal, lung, bladder and gynaecological malignancies.
Photodynamic therapy (PDT) has been used traditionally for oncologic and ophthalmic indications. In addition, the enormous potential for the use of PDT agents in cardiovascular diseases is currently being translated into reality. Preclinical studies with various photosensitizers have demonstrated reduction in atheromatous plaque and prevention of intimal hyperplasia. With recent advances in light-based vascular devices and laser diode technology, the clinical use of cardiovascular photodynamic therapy is even more likely. Two photosensitizers, 5-aminolevulinic acid (ALA) and AntrinR (motexafin lutetium) Injection, are under clinical evaluation with many other agents in preclinical testing. Here, preclinical studies are reviewed and the clinical viability of cardiovascular photodynamic therapy is discussed.© (2000) COPYRIGHT SPIE--The International Society for Optical Engineering. Downloading of the abstract is permitted for personal use only.
Light-dependent therapies, such as photodynamic therapy and extracorporeal photopheresis, are not new, but have remained of interest to chemists and health care professionals since the middle of the twentieth century. While most people link light-dependent therapies only to the treatment of cancer, these therapies may be of use for a diverse set of medical conditions, from acne to AIDS. The techniques arise directly from the application of chemical concepts, such as spectroscopy, MO theory, and organic chemical reactions. Because of its application to health care, the field of photochemistry provides a tool to demonstrate the significance of chemistry to a socially important issue. Keywords (Audience): Upper-Division Undergraduate
IntroductionThe Chemistry of TexaphyrinsTherapeutic Applications of Motexafin Lutetium (a.k.a. Lutetium Texaphyrin, Lu-Tex, 3)Motexafin Gadolinium (Xcytrin®, Gd-Tex, 2) as a Radiation EnhancerTexaphyrins: Future Directions
Purpose:
We investigated hemodynamics in patients receiving delta-Aminolevulinic acid (delta-ALA) to visualize tumor margins prior to radical retro pubic prostatectomy.
Patients:
Twenty patients undergoing elective open radical retro pubic prostatectomy (RRP).
Methods:
Cohort observational study. Ten patients receiving 20 mg/kg of delta-ALA orally prior to surgery (delta-ALA) and 10 patients undergoing RRP without the application of delta-ALA served as a retrospectively matched cohort (CONTROL).
Measurements:
Changes in heart rate (HR), mean arterial blood pressure (MAP), and functional hemodynamic parameters were assessed by electrocardiogram, non-invasive and invasive blood pressure monitoring plus transcardiopulmonary thermodilution.
Results:
Patients of both groups did not differ in means of age, body mass index, or ASA classification. During surgery, HR and MAP did not differ significantly between both groups. Also, the amount of IV crystalloids and colloids did not differ significantly. In contrast, the amount of vasopressor necessary to maintain MAP within the target range of 70-90 mmHg was significantly higher in delta-ALA when compared to CONTROL (0.08 ± 0.04 μg/kg/min (delta-ALA) vs. 0.03 ± 0.02 μg/kg/min (CONTROL); P < 0.01). Immediately after surgery, patients of delta-ALA showed a significantly higher heart rate (82 ± 18 min(-1) vs. 67 ± 9 min(-1); P < 0.05) compared to patients of CONTROL. Cardiac index, global end-diastolic volume index, and extravascular lung water index were significantly higher after surgery, when compared to baseline values (P < 0.05).
Conclusions:
Orally administered delta-ALA prior to open radical prostatectomy induces hemodynamic instability in the perioperative period requiring vasopressor support. Further, an increase of extravascular lung water points toward an increased vascular permeability induced by delta-ALA.
Photodynamic effect was a chance discovery early in the 1900s, demonstrating the lethal effects of light activated chemicals on living cells. Although the application of the principles of photodynamic effect to patients' treatment and what became photodynamic therapy (PDT) was practiced in 1960s and 1970s, clinical trials were only started in the 1980s, following successful synthesis of clinically usable photosensitisers (drugs) and the manufacturing of light sources. We briefly review and highlight some of the landmarks of the development of clinical PDT in Europe.
Photodynamic therapy (PDT) reduces neointimal hyperplasia and negative remodelling following balloon injury in small and large animal models. This clinical study investigated the role of adjuvant PDT following femoral percutaneous transluminal angioplasty (PTA).
Eight PTAs in seven patients (two women) with a median age of 70 (range 59-86) years were performed with adjuvant PDT. All patients had previously undergone conventional angioplasty at the same site which resulted in symptomatic restenosis or occlusion between 2 and 6 months. Each was sensitized with oral 5-aminolaevulinic acid 60 mg/kg, 5-7 h before the procedure. Following a second femoral angioplasty, up to 50 J/cm2 red light (635 nm) was delivered to the angioplasty site via a laser fibre within the angioplasty balloon. Patients were kept in subdued light overnight and discharged the following day. Outcome was assessed by duplex imaging at 24 h, 1, 3 and 6 months and by intravenous digital subtraction angiography at 6 months. A peak systolic velocity ratio (PSVR) of more than 2.0 at the angioplasty site was taken to represent restenosis.
All patients tolerated the procedure well without adverse complications or death. All were rendered asymptomatic which was sustained throughout the study interval. All vessels remained patent and no lesion attained the duplex definition of restenosis. Median (interquartile range) PSVR across stenotic segments was 4.7 (3.7-5.7) before angioplasty, 1.1 (0.9-1.3) at 24 h and 1.4 (1.0-1.8) at 6 months after intervention (P = 0.04 compared with preoperative value).
This pilot study suggests that endovascular PDT is safe and may reduce restenosis follow- ing angioplasty. The data justify a randomized controlled trial.
Photodynamic therapy (PDT) uses red light (non-thermal, non-ionising) to activate a previously administered photosensitizing drug. This inhibits neointimal hyperplasia in injured arteries in small animals where it appears safe and well tolerated. Our aim was to develop a method for percutaneous application of PDT to iliac and coronary arteries in a large animal model and investigate its influence on the remodeling and intimal hyperplastic response to balloon injury.
Studies were undertaken on 13 juvenile Large White-Landrace crossbred pigs (15-20 kg). After intravenous administration of the photosensitizing agent 5-amino laevulinic acid (ALA), the arterial tree was accessed via the left common carotid artery and balloon injuries made by over-distension in both common iliacs (thirteen animals) and one or two main coronary arteries (eight animals). Half the injured sites were then illuminated with red laser light transmitted via the catheter. Animals were culled 28 days later and tissue harvested for histomorphometry.
Compared with control injured vessels, PDT treated, balloon injured coronary arteries had a larger lumen (1.4 vs. 0.8 mm2, P = 0.002), larger area within the external elastic lamina (2.8 vs. 2.2 mm2, P = 0.006) and smaller area of neointimal hyperplasia (0.4 vs. 0.7 mm2, P = 0.06), 28 days after intervention. Less neointimal hyperplasia and the absence of negative remodeling resulted in the lumen of PDT-treated, injured segments being the same as that of adjacent reference segments (1.5 vs. 1.6 mm2). Similar trends, but with smaller differences, were seen in the iliac vessels.
Intra-arterial, trans-catheter PDT favourably influences the arterial response to balloon injury in both the coronary and peripheral circulations. This technique offers a promising new approach to restenosis after endovascular procedures.
Intimal hyperplasia (IH) leading to restenosis is a major complication of arterial revascularization. The purpose of this study was to investigate the effect of photodynamic therapy (PDT) using mono-L-aspartyl chlorin e6 (NPe6) as a photosensitizer and intraluminal radial irradiation for inhibition of IH experimentally.
Study of laser transmission through the blood indicated that exclusion of blood is a prerequisite for intraluminal PDT. For homogeneous radial laser irradiation to the vessel wall, we used a newly developed cylindrical diffusing balloon laser fiber. Injuries were induced by pulling a balloon catheter through the right iliac artery of rabbits. One and 6 hours after the NPe6 injection (5mg/kg i.v.), drug distribution was examined by fluorescence microscopy. Nineteen rabbits received NPe6 at the time of injuries and PDT was performed with 664-nm laser at 30 and 10 J/cm(2) (20, 30, 40 mW/cm(2)) 1 hour after the injuries. The arteries were harvested at 2 days. In a second group of rabbits, PDT was given at 30 mW/cm(2) (30 J/cm(2)). Two weeks after treatment, the arteries were removed and examined histologically.
NPe6 was found to be distributed selectively in the injured media. Endovascular NPe6-PDT showed complete depletion of smooth muscle cells even with 10 J/cm(2) at 2 days. IH was significantly inhibited at 14 days after PDT.
Endovascular PDT of injured artery using NPe6 can prevent IH in this model of arterial wall injury and may become clinically useful for the prophylaxis of IH.
PDT uses non-thermal laser light targeted at the arterial wall to activate previously injected photosensitising agents. The resulting histological changes are associated with a reduction in neointima formation and the absence of negative vessel wall remodelling. It has been used successfully in a pilot clinical trial of peripheral vascular disease and, while further research still needs to be carried out, PDT has promise as a new strategy for the prevention and treatment of restenosis after percutaneous coronary intervention.
Photodynamic therapy (PDT) of tumours is based on a dual selectivity, i.e. preferential uptake and retention of a photosensitiser by tumours and irradiation of the tumour area with light wavelengths specifically absorbed by the photosensitiser. The photoexcited sensitiser generates highly reactive oxygen species that induce irreversible damage to neoplastic cells and vessels. Following sensitiser accumulation and irradiation, damage to sensitive sites within the microvasculature, namely to endothelial cells and the vascular basement membrane, is induced and leads to the establishment of thrombogenic sites within the vessel lumen. This initiates a physiological cascade of responses including platelet aggregation, the release of vasoactive molecules, leukocyte adhesion, increases in vascular permeability and vessel constriction. These result in tumour destruction by vascular collapse, blood flow stasis and tissue hemorrhages. Several photosensitisers are able to induce severe vasculature damage, although by variously different mechanisms. Due to its efficient vascular interactions, photodynamic treatment is also increasingly used for non-cancerous lesions. Successful application of PDT mainly for vessel occlusion and thrombosis in intimal hyperplasia, restenosis, atherosclerotic plaques, corneal and choroidal neovascularisation and port-wine stains is reported.
Cardiovascular interventions to correct arterial occlusive disease have excellent short-term results, but long-term patency is still seriously compromised by the development of restenosis. This review will provide a portrait of the mechanisms following vascular injury, define the clinical scope of the problem and outline developments of different therapeutical strategies to prevent restenosis.
Photodynamic therapy (PDT) based on the photosensitive protoporphyrin IX (PpIX) may prevent restenosis after transluminal angioplasty. PpIX is synthesized in mitochondria, which differ in number and activity among various tissues. Therefore, we questioned whether the course of PpIX concentration after systemic aminolaevulinic acid (ALA) administration differed among various arteries. ALA was administered intravenously (200 mg/kg) to male Wistar rats (n = 21). At varying time intervals (0, 1, 2, 3, 6, 12 and 24 h) both central and peripheral arteries were isolated and homogenized, and the concentration of the various heme intermediates was determined by a fluorometric extraction method. The maximal PpIX concentration was more than two-fold higher in peripheral arteries (20.49 +/- 3.0 to 24.0 +/- 7.5 pmol/mg protein) than in central arteries (0-9.46 +/- 0.01 pmol/mg protein) (P < 0.004). However, the amount of citrate synthase, reflecting the mitochondrial mass, was lower (0.14-0.61 and 1.87-2.32 U/mg protein, respectively). Apparently, the level of PpIX cannot simply be explained by the mitochondrial content of the arteries. The time interval of maximal PpIX accumulation was similar in peripheral and central arteries (2 h and 27 min vs. 2 h and 8 min) (P = 0.13). Thus, if the efficacy of PDT in vivo is directly related to the tissue concentration of PpIX, more effect can be expected in peripheral arteries than in central arteries.
Atherosclerosis and intimal hyperplasia remain obstacles for surgeons to overcome following vascular reconstructions. Even with all of the technical improvements that have occurred in the past several decades, long term patency following intervention is hindered by these inherent adverse developments. Today, the use of light is seen as a potential treatment modality in vascular surgery. Photodynamic therapy (PDT) has been used in the treatment of cancer, and because of its continued success in vascular experimental models it is now being tested in clinical trials for vascular diseases. PDT offers the surgeon many advantages, and it may have unlimited uses in the clinical setting. Is PDT the ultimate treatment modality for the cardiovascular surgeon and will it help to overcome the inherent failures associated with vascular reconstructions? It may be too early to answer these questions, but with the current successes demonstrated by PDT, there is a need for further testing in clinical trials. In the near future, PDT may be used clinically as a treatment modality to inhibit restenosis and intimal hyperplasia following surgical intervention.
Flexible treatments for intimal hyperplasia after angioplasty are still needed. The aim of this study was to demonstrate the long-term effects of vascular photodynamic therapy with talaporfin sodium on intimal hyperplasia following interventional injury. Intimal hyperplasia was induced by balloon distension injury to the carotid artery in 31 rabbits. Talaporfin, 5.0 mg/kg, was delivered systemically immediately after balloon injury. The injury site was irradiated with a diode laser light of wavelength 664 nm using a fluence of 50 J/cm2 after 30 min. At day 3 and weeks 3, 6, 9, 15, and 25 after photodynamic therapy, the treated artery of each rabbit was excised and examined immunohistochemically. Thirty minutes after talaporfin administration, drug fluorescence was found only in the balloon-injured carotid artery wall. At 3 days, no smooth muscle cells were seen in the media of the photodynamic therapy-treated arterial segments. Intimal hyperplasia developed progressively in the balloon-injured and untreated segments; however, in the segments treated with photodynamic therapy, intimal hyperplasia was markedly suppressed until 25 weeks and the media was repopulated by smooth muscle cells without macrophages. Vascular photodynamic therapy with talaporfin may be used to inhibit restenosis after vascular intervention.
To examine the effects of intracoronary PhotoPoint photodynamic therapy (PDT) with a new photosensitiser, MV0611, in the overstretch balloon and stent porcine models of restenosis.
28 pigs were injected with 3 mg/kg of MV0611 systemically 4 h before the procedure. Animals were divided into either the balloon overstretch injury (BI) group (n = 19) or the stented group (n = 9). After BI, a centred delivery catheter was positioned in the artery to cover the injured area, and light (532 nm, 125 J/cm(2)) was applied to activate the drug (n = 10). Control arteries (n = 9) were not activated by light. In the stented group, the drug was light activated before stent deployment. Serial sections of vessels were processed 14 days after treatment in the BI group and 30 days after treatment in the stented group for histomorphometric or immunohistochemical analysis.
Intracoronary PDT significantly reduced intimal thickness in both BI and stented arteries (about 65%: 0.22 (SEM 0.05) mm v 0.62 (0.05) mm, p < 0.01; and about 26%: 0.40 (0.04) mm v 0.54 (0.04) mm, p < 0.01, respectively). PDT increased luminal area by <or= 60% and 50% within BI and stented arteries (3.43 (0.27) mm(2)v 5.51 (0.52) mm(2), p < 0.05; 4.0 (0.02) mm(2)v 6.0 (0.16) mm(2), p < 0.01), respectively. Complete re-endothelialisation was observed by immunohistochemical and gross histological analyses in all PDT and control arteries. There were no cases of aneurysm formation or thrombosis.
Intracoronary PhotoPoint PDT with MV0611 reduces intimal proliferation without suppressing re-endothelialisation in a porcine model of restenosis.
The porphyrin precursor 5‐aminolevulinic acid (ALA) is being widely used in photodynamic therapy of cancer. Improvement in ALA delivery has been sought through the use of ALA derivatives, in particular the esterification of ALA with aliphatic alcohols, which in certain cases can improve cellular penetration and selectivity.
ALA uptake systems appear to be distinctive for each cell type. The LM3 mammary adenocarcinoma cell line takes ALA up by BETA transporters. In this work, we investigated ALA derivative transport systems through the inhibition of radiolabelled ALA uptake in the LM3 cells. We also performed inhibition studies of γ ‐aminobutyric acid (GABA) uptake.
The more lipohilic ALA derivatives hexyl‐ALA and undecanoyl‐ALA inhibit ALA uptake, whereas methyl‐ALA, R , S ‐ALA‐2‐(hydroxymethyl)tetrahydropyranyl ester and the dendron aminomethane tris methyl 5‐ALA does not inhibit ALA uptake. A similar pattern was found for GABA, except that the dendron inhibited GABA uptake. However, hexyl‐ALA and undecanoyl‐ALA are not taken up by BETA transporters, but by simple diffusion, although they still inhibit ALA uptake by binding to the cell membrane.
These results show that different modifications to the ALA molecule lead to different uptake mechanisms. Whereas ALA is taken up by BETA transporters, none of the ALA derivatives shares the same mechanism. Knowledge of the mechanisms of ALA derivatives entry into the cells is essential to understand and improve ALA‐mediated PDT and to the design of new ALA derivatives that may be taken up at a higher rate than ALA.
British Journal of Pharmacology (2006) 147 , 825–833. doi: 10.1038/sj.bjp.0706668
Endogenous porphyrin accumulation after administration of 5-aminolevulinic acid is employed in photodynamic therapy of tumours. Due to its low membrane permeability, esterified 5-aminolevulinic acid derivatives less hydrophilic than the parental compound are under investigation. Knowledge of the mechanisms of 5-aminolevulinic acid derivatives uptake into target cells is essential to understand and improve photodynamic therapy and useful in the design of new derivatives with better affinity and with higher selectivity for tumour cells in specific tissues. The aim of this work was to assess the interaction of 5-aminolevulinic acid derivatives with the intestinal PEPT1 and renal transporter PEPT2 expressed in Pichia pastoris yeasts. We found that Undecanoyl, Hexyl, Methyl and 2-(hydroxymethyl)tetrahydropyranyl 5-aminolevulinic acid esters and the dendron 3m-ALA inhibited (14)C-5-aminolevulinic acid uptake by PEPT2. However, only the Undecanoyl ester inhibited 5-aminolevulinic acid uptake by PEPT1. We have also found through a new developed colorimetric method, that Hexyl and 2-(hydroxymethyl)tetrahydropyranyl 5-aminolevulinic acid esters display more affinity than 5-aminolevulinic acid for PEPT2 whereas none of the compounds surpass 5-aminolevulinic acid affinity for PEPT1. In addition, the Undecanoyl ester binds with high affinity to the membranes of PEPT2 and PEPT1-expressing yeasts and to the control yeasts. The main finding of this work was that some derivatives have the potential to improve 5-aminolevulinic acid-based photodynamic therapy by increased efficiency of transport into cells expressing PEPT2 such as kidney, mammary gland, brain or lung whereas in tissues expressing exclusively PEPT1 the parent 5-aminolevulinic acid remains the compound of choice.
A 77-year-old man presented because of heart enlargement on chest x ray. Transthoracic echocardiography revealed an abnormal mass located in front of the right atrium (RA), with blood flow from the RA to the mass.
Electrocardiogram-gated enhanced, multislice computed tomography …
Protoporphyrin IX (PpIX) is one of the photodynamically active substances that are endogenously synthesized in the metabolic pathway for heme as a precursor. Aminolevulinic acid-esters are more lipophilic than conventional 5-aminolevulinic acid (ALA) and some of them are currently being approved as new drugs for photodynamic diagnosis (PDD) and photodynamic therapy (PDT). In order to investigate the pharmacokinetics of ALA and ALA-ethyl ester (ALA-ethyl) in the atheromatous plaque and normal aortic wall of rabbit postballoon injured artery, each 60 mg kg(-1) of ALA or ALA-ethyl was injected intravenously followed by serial detection of PpIX fluorescence of harvested arteries at 0-48 h post-injection. Maximum PpIX build-up in the atheromatous plaque was seen at 2 h after injecting ALA. In contrast, it occurred at 9 h after injecting ALA-ethyl. In addition, the selective build-up of ALA in the atheromatous plaque compared to normal vessel wall was much higher (10 times) than that of ALA-ethyl. The time of maximum fluorescence intensity of PpIX was employed as drug-light-interval for subsequent PDT treatment of the atheromatous plaque with 50-150 J cm(-1) of light dose. Significant reduction in plaque was observed without damage of the medial wall at both groups, but smooth muscle cell (SMC) was still present in the media region below the PDT-treated atheromatous plaque. In conclusion, ALA may be a more effective compound for endovascular PDT treatment of the atheromatous plaque compared with ALA-ethyl based on their pharmacokinetics, but further optimization of PDT methodology remains to remove completely residual SMC in the media for preventing potential restenosis.
Aminolaevulinic acid (ALA) is the first committed step in haem synthesis. In the presence of excess ALA the natural regulatory feedback system is disrupted allowing accumulation of protoporphyrin IX (PP IX) the last intermediate product before haem, and an effective sensitiser. This method of endogenous photosensitisation of cells has been exploited for photodynamic therapy (PDT). We have studied the fluorescence distribution and biological effect of induced PP IX in normal and tumour tissue in the rat colon. Fluorescence in normal colonic tissue was at a peak of 4 h with a rapid fall off by 6 h. The fluorescence had returned to background levels by 24 h. All normal tissue layers followed the same fluorescence profile but the mucosa showed fluorescent levels six times higher than the submucosa, with muscle barely above background values. At 6 h the ratio of fluorescence levels between normal mucosa and viable tumour was approximately 1:6. At this time laser treatment showed necrosis of normal mucosa and tumour with sparing of normal muscle. There was good correlation between the fluorescence distribution and the biological effect of ALA-induced photosensitisation on exposure to red light. ALA may be superior to conventional sensitisers for tumours that produce haem as the PP IX is synthesised in malignant cells while the other sensitisers mainly localise to the vascular stroma of tumours. There is also a greater concentration difference between the PP IX levels in tumours and in normal mucosa and normal muscle than with the other photosensitisers raising the possibility of more selective necrosis in tumours.
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This paper presents the results of a prospective study of percutaneous transluminal angioplasty (PTA) for the treatment of patients with peripheral arterial occlusive disease and identifies the variables that are predictive of long-term success. The variables believed to be important prognostically were recorded for 984 consecutive PTAs performed between July 1978 and July 1986. Success or failure was defined using a combination of clinical and objective vascular laboratory criteria. The overall long-term success was estimated by the Kaplan-Meier method and differences between curves of success rate versus time for each variable were determined by the Wilcoxon and log-rank statistics. The combination of variables associated with success were determined by the Cox proportional hazards regression model. For all cases, the initial success rate was 88.6 +/- 1.0% and at 5 years was 48.2 +/- 2.3%. The following variables, when considered individually, were associated with success (p less than 0.05): indication for PTA, site of PTA, severity of lesion, runoff, number of sites dilated, diabetes, and the occurrence of a complication. From the Cox model, by using a stepwise multiple regression procedure, the following combination of variables were found to be predictive of success (p less than 0.05): (1) indication (claudication vs. salvage), (2) site (common iliac vs. other), (3) severity of lesion (stenosis vs. occlusion), and (4) runoff (good vs. poor). For all combinations of these four significant variables, curves of the success rate versus time were calculated. In conclusion, this study has identified the combination of four variables that together predict if PTA is likely to be successful in the management of a patient with peripheral arterial occlusive disease.
Photodynamic therapy of cancer exposes adjacent arteries to the risk of injury and the possibility of haemorrhage and thrombosis. The nature of photodynamic injury to normal arteries has not been satisfactorily defined, and the ability of arteries to recover with time is unclear. To clarify these issues, we have investigated the effects of PDT on rat femoral arteries, using a second-generation photosensitiser, disulphonated aluminium phthalocyanine, and a new method of photosensitisation, using endogenous synthesis of protoporphyrin IX following systemic administration of 5-aminolaevulinic acid (ALA). Pharmacokinetic studies of sensitiser fluorescence were carried out to determine peak levels of sensitiser. Subsequently photodynamic therapy at times corresponding to maximal fluorescence was performed using two light doses, 100 and 250 J cm-2. The nature of injury sustained and recovery over a 6 month period was investigated. Three days following PDT, all vessels treated showed complete loss of endothelium, with death of all medial smooth muscle cells, leaving an acellular flaccid artery wall. No vascular occlusion, haemorrhage or thrombosis was found. A striking feature was the lack of inflammatory response in the vessel wall at any time studied. Re-endothelialisation occurred in all vessels by 2 weeks. The phthalocyanine group showed repopulation of the media with smooth muscle cells to be almost complete by 3 months. However, the ALA group failed to redevelop a muscular wall and remained dilated at 6 months. Luminal cross-sectional area of the ALA-treated group was significantly greater than both control and phthalocyanine groups at 6 months. All vessels remained patent. This study indicates that arteries exposed to PDT are not at risk of catastrophic haemorrhage or occlusion, a finding that is of significance for both the local treatment of tumours and the use of PDT as an intraoperative adjunct to surgery for the ablation of microscopic residual malignant disease.
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This study examined the effects of intracoronary irradiation on neointimal proliferation after overstretch balloon angioplasty in a normolipemic swine model of restenosis.
Restenosis after percutaneous transluminal coronary angioplasty represents, in part, a proliferative response of vascular smooth muscle at the site of injury. We have previously shown that ionizing radiation, delivered by means of an intracoronary source, causes focal medial fibrosis. We therefore hypothesized that intracoronary irradiation delivered at the time of balloon angioplasty might impair the restenosis process.
Nineteen juvenile swine underwent coronary angiography; a segment of the coronary artery was chosen as a target for balloon injury. In 10 swine, a ribbon of iridium-192 was positioned at the target segment, and 2,000 cGy was delivered at the vessel wall. Subsequently, overdilation balloon angioplasty was performed at the irradiated segment. In nine control swine, overdilation balloon angioplasty was performed without previous irradiation. Eighteen animals survived and were killed at 30 days. Histopathologic analysis was performed by a pathologist in blinded manner. The area of maximal lumen compromise within the target segment was analyzed by computer-assisted planimetry.
In the control group, mean (+/- SD) neointimal area was 0.84 +/- 0.60 mm2 compared with that in the irradiated group, 0.24 +/- 0.13 mm2 (p = 0.01). In the control group, mean percent area stenosis was 47.6 +/- 20.7%, whereas that in the irradiated group was 17.6 +/- 10.5% (p = 0.001). This represents a 71.4% reduction in neointimal area and a 63.0% reduction in percent area stenosis in the irradiated group. Adjacent coronary segments and surrounding myocardium were unaffected.
Intracoronary irradiation (2,000 cGy) delivered to a target porcine coronary segment before balloon overdilation markedly reduces neointima formation at 30 days and thus significantly impairs the restenosis process.
Although arterial renarrowing after angioplasty has been attributed largely to intimal hyperplasia, there has been no systematic effort to correlate the actual hyperplastic tissue mass with angiographic lumen reduction. Using balloon angioplasty in various animal restenosis models, we quantitatively assessed the separate contributions of intimal hyperplasia and arterial remodeling to angiographic late lumen loss.
Data used for this study were obtained from experiments of conventional and thermal (37 degrees C or 55 degrees to 90 degrees C) balloon angioplasty-treated femoral and iliac arteries in normal rabbits and conventional balloon angioplasty-treated iliac arteries in Yucatan micropigs fed either a normal or an atherogenic diet. Quantitative angiography was performed immediately before and after intervention and at 3 or 8 weeks thereafter, and late loss in lumen diameter was taken as the difference between arterial diameter immediately after treatment and at 3 or 8 weeks of follow-up. Intimal hyperplasia was quantified histologically as the area of tissue mass within the internal elastic lamina. We observed a consistent discrepancy between the actual late loss seen with angiography and the diameter reduction that could be explained by histological intimal thickness alone in both animal models. This discrepancy ranged from 86 +/- 3% of the late loss in the 8 weeks/37 degrees C group to 77 +/- 22% in the conventional group for rabbits and 52 +/- 23% in an atherogenic diet group (n = 10) to 89 +/- 11% in a normal diet group (n = 6) for pigs. This discrepancy appeared to be due predominantly to reduction of the area circumscribed by the internal elastic membrane, a process that is tentatively designated as arterial remodeling. In both the rabbit femoral artery and in the Yucatan iliac artery, remodeling, not intimal hyperplasia, correlated with angiographic late loss.
In both the normal rabbit and the normal and atherosclerotic pig, restenosis after angioplasty results from both intimal hyperplasia and arterial remodeling. The exact etiology of arterial renarrowing after angioplasty has important implications on the design of antirestenosis drugs and new coronary devices.
In animal models of coronary restenosis, intracoronary radiotherapy has been shown to reduce the intimal hyperplasia that is a part of restenosis. We studied the safety and efficacy of catheter-based intracoronary gamma radiation plus stenting to reduce coronary restenosis in patients with previous restenosis.
Patients with restenosis underwent coronary stenting, as required, and balloon dilation and were then randomly assigned to receive catheter-based irradiation with iridium-192 or placebo. Clinical follow-up was performed, with quantitative coronary angiographic and intravascular ultrasonographic measurements at six months.
Fifty-five patients were enrolled; 26 were assigned to the iridium-192 group and 29 to the placebo group. Angiographic studies were performed in 53 patients (96 percent) at a mean (+/-SD) of 6.7+/-2.2 months. The mean minimal luminal diameter at follow-up was larger in the iridium-192 group than in the placebo group (2.43+/-0.78 mm vs. 1.85+/-0.89 mm, P=0.02). Late luminal loss was significantly lower in the iridium-192 group than in the placebo group (0.38+/-1.06 mm vs. 1.03+/-0.97 mm, P=0.03). Angiographically identified restenosis (stenosis of 50 percent or more of the luminal diameter at follow-up) occurred in 17 percent of the patients in the iridium-192 group, as compared with 54 percent of those in the placebo group (P= 0.01). There were no apparent complications of the treatment.
In this preliminary, short-term study of patients with previous coronary restenosis, coronary stenting followed by catheter-based intracoronary radiotherapy substantially reduced the rate of subsequent restenosis.
Photodynamic therapy (PDT) effectively inhibits fibrocellular intimal hyperplasia (FCIH) two and four weeks after arterial traction balloon injury in rat carotid arteries. The aim of the present study was to assess the long term effects of PDT in this rat model of FCIH. 5- aminolaevulinic acid-induced protoporphyrin IX was used to sensitize rats for PDT after traction balloon arterial injury to the whole of the left common carotid artery. Rats were sacrificed at intervals of six to 26 weeks, and perfusion fixed and H and E stained sections were analyzed using computerized morphometry. PDT inhibition of FCIH was only partial at these late times. The amount of FCIH present increased with increasing time after injury. The late occurrence of FCIH appeared to be due to migration of FCIH from balloon injured areas outside the PDT treated field as a result of the traction injury being applied to the whole carotid. We recommend segmental balloon injury rather than the traction injury of the whole common carotid injury for future studies in this model.
Human coronary artery restenosis after percutaneous revascularization is a response to mechanical injury. Smooth muscle cell proliferation is a major component of restenosis, resulting in obstructive neointimal hyperplasia. Because ionizing radiation inhibits cellular proliferation, this study tested in a porcine coronary injury model the hypothesis that the hyperplastic response to coronary artery injury would be attenuated by X-irradiation.Deep arterial injury was produced in 37 porcine left anterior descending coronary artery segments with overexpanded, percutaneously delivered tantalum wire coils. Three groups of pigs were irradiated with 300-kV X-rays after coil injury: Group I (n = 10), 400 cGy at 1 day; Group II (n = 10), 400 cGy at 1 day and 400 cGy at 4 days and Group III (n = 9), 800 cGy at 1 day. Eight pigs in the control group underwent identical injury but received no radiation. Treatment efficacy was histologically assessed by measuring neointimal thickness and percent area stenosis.Mean neointimal thickness in all irradiated groups was significantly higher than in the control groups and thickness was proportional to X-ray dose.X-irradiation delivered at these doses and times did not inhibit proliferative neointima. Rather, it accentuated the neointimal response to acute arterial injury and may have potentiated that injury.
With the high initial success rates for coronary angioplasty that are reported regularly, it has become increasingly difficult to demonstrate methods or techniques that are able to provide more beneficial early results than can be achieved by conventional angioplasty. On the other hand, the incidence of late restenosis has remained much the same over the 10 years that angioplasty has been part of clinical practice, and there is still no proved intervention that modifies the restenosis process. Therefore, the problem of restenosis has assumed increasing relevance in determining the clinical value of coronary angioplasty and, accordingly, studies that address the problem of restenosis need to become more exacting.Although numerous articles have addressed the problem of restenosis in the clinical setting, many defining certain factors associated with restenosis and possible interventions to reduce the incidence of restenosis, there is surprisingly little consensus. Most of the discrepancies can be attributed to three factors: 1) the selection of patients, 2) the method of analysis, and 3) the definition of restenosis employed. This review shows how these three factors influence the outcome and conclusions of restenosis studies.
The author presents an alternative statistical analysis of the results of the University of Toronto series of percutaneous transluminal angioplasty (PTA) of the femoral and popliteal arteries (n = 254). After recalculation of the data with the Kaplan-Meier method, the postprocedure success rate ranged from 88.8% +/- 2.0 at 1 month to 35.7% +/- 4.8 at 6 years. With Cox multiple regression analysis, the type of femoropopliteal lesion and the runoff were the variables that were useful to predict late results. For stenoses with good runoff, the success rate was 53% at 5 years; with poor runoff, 31% at 5 years. For occlusions with good runoff, the success rate was 36% at 5 years; with poor runoff, 16% at 5 years. In initially successful cases, ongoing clinical success at 1, 3, and 5 years was better in patients with good runoff at the time of PTA than in those with poor runoff. Now that more recent studies have documented improved technical success in femoropopliteal PTA, a comparative study of the relative safety, long-term clinical efficacy, and cumulative cost of PTA versus surgery seems warranted.
Photodynamic therapy using photofrin and light energy inhibits human myofibroblast proliferation in cell culture. The purpose of this study is to evaluate its influence on intimal hyperplasia in vivo. Twenty New Zealand White rabbits underwent a standardized intimal injury to both common carotid arteries with a 2 Fr balloon catheter. One week later, half of the animals received photofrin (5 mg/kg) intravenously. The remaining 10 rabbits received no photofrin. Two days later, all neck incisions were reopened and a 1-cm segment of each of the 40 carotid arteries was exposed for 5 min to 80 mW of 630 nm light energy from a continuous wave tunable dye laser (fluence = 7.6 J/cm2). All vessels were harvested 5 weeks post-laser treatment following in vivo fixation with formalin. From each artery, separate cross-sections taken from both the lasered and non-lasered regions of each vessel were mounted and stained for histologic evaluation. Analyzed segments were then divided into four different treatment groups: group I segments consisted of arterial cross-sections which were taken from vessel regions that were injured but received neither photofrin nor laser treatment (group I, n = 20); group II segments also did not receive photofrin but were exposed to light energy (group II, n = 20); group III segments received photofrin but no light energy (group III, n = 20); and cross-sections in group IV were taken from those segments which received both photofrin and laser treatment. Using planimetry, the ratio of the area of intimal hyperplasia (IH) to the area enclosed by the internal elastic lamina (IEL) was measured for each specimen (IH/IEL).(ABSTRACT TRUNCATED AT 250 WORDS)
Photodynamic therapy (PDT) uses light activation of otherwise nontoxic dyes for the production of reactive oxygen species that cause cell injury and death.
The inhibition of intimal hyperplasia (IH) by PDT was studied in the balloon injury model of the rat carotid artery. Chloroaluminum-sulfonated phthalocyanine (CASPc) was the drug chosen for PDT because it does not produce skin photosensitivity and has a high absorption peak of light at 675 nm, a wavelength with good tissue penetration. A pilot study indicated that CASPc administration with laser radiant exposure of 100 J/cm2 resulted in a homogeneous, circumferential effect on the whole artery. Male Sprague-Dawley rats received the balloon catheter injury to the left common carotid artery (day 0) and were equally divided into two groups. Nine rats received either CASPc (5 mg/kg i.v., n = 6) or saline (n = 3) at day 2, before IH was present, and nine rats received CASPc or saline in the same manner on day 7, when IH was already present. Twenty minutes after drug injection, the distal left common carotid artery was irradiated under saline with 675-nm laser light at 100 mW/cm2 for 10(3) seconds (100 J/cm2). At this low laser irradiance, there are no thermal effects, but photoactivation of CASPc occurs. The rats were killed at day 14 after balloon injury when IH reaches a maximum. The arteries were harvested after perfusion-fixation for light microscopy, histological and computerized morphometric evaluation, and transmission electron microscopy (TEM) analysis. The cross-sectional areas of the neointima were measured in the PDT-treated arteries and in the laser-only control arteries. There was a significant mean +/- SD decrease of IH in the PDT-irradiated segments of the arteries (0.06 +/- 0.05 mm2) versus the laser-only control ones (0.17 +/- 0.07 mm2) (t test, p less than 0.001), with no statistical difference between the day 2 and day 7 treated rats. Lack of IH was correlated in 90% of cases with histological absence of medial smooth muscle cells or inflammatory cells, but no other structural injury was identified. TEM analysis showed early evidence of PDT-mediated cytotoxic effects at 4 hours and the absence of collagen or elastic tissue structural alterations.
These data demonstrated that PDT can effectively inhibit the IH response when it is used before or during induction of cellular proliferation in this acute model. Although the long-term implications of PDT in arteries need to be defined, this technique may offer a new method for understanding and treating IH.
The study of potentially effective drug therapies and mechanical devices for the prevention of restenosis after percutaneous coronary revascularization has relied heavily on the use of experimental animal models. To date, greater than 50 experimental studies have been reported and have suggested that at least nine different classes of pharmacologic agents inhibit the intimal proliferative response to arterial injury. However, no pharmacologic intervention has yet been shown to reproducibly reduce the incidence of restenosis after coronary balloon angioplasty in humans. To identify the reasons for the apparent nonspecificity of the animal models and to determine which model should most reliably predict the efficacy of individual therapies in humans, the distinguishing characteristics of the experimental models were compared. Particular attention was paid to the size and morphologic structure of the treated artery, the susceptibility of the species to spontaneous and diet-induced arterial disease, the nature of the stimulus to intimal proliferation and several practical and logistic considerations. Finally, the reported efficacies of specific drug therapies in the respective animal models and in humans were compared. This review suggests that significant interspecies and occasionally intraspecies differences do exist among the respective animal models, particularly in the extent and composition of the neointimal thickening.(ABSTRACT TRUNCATED AT 250 WORDS)
Application of 630 nm light in the presence of blood is often necessary during photodynamic therapy, particularly for proposed intravascular applications. The effect of blood on transmission of 630 nm light was studied using a three dimensional irradiation model and an integrating sphere for measuring light transmitted in any direction through blood layers of different hematocrit (25 to 75) and thickness (.15 to .98 mm). There was an inverse relationship between transmission and hematocrit and transmission and blood thickness, p = .000 for both. At a physiologic hematocrit of 46, transmission through blood layers of .98, .41, .28, and .15 mm were 21%, 33%, 29%, and 58% respectively. These blood thicknesses or more are likely in the clinical environment, and can be expected to result in significant transmission losses. The marked absorption of 630 nm light by blood indicates that removal of the blood or correction for power loss should be employed when 630 nm light is applied in a blood containing environment.
The effect of dihematoporphyrin-ester or -ether (DHE), a photosensitizing porphyrin with different amounts of aggregates, on the growth of cultured smooth muscle cells obtained from nonatherosclerotic arteries and from atheromatous plaques (primary stenosing and restenosing lesions) was examined without photoactivation of the drug. Clinically relevant DHE concentrations ranging from 0.1 to 25 micrograms/ml were used. In all proliferation studies with cells of second and third passage (approximately 10 cumulative population doublings in vitro), the growth rates decreased in a dose-dependent manner. Smooth muscle cells from atherosclerotic lesions were significantly more sensitive than smooth muscle cells from normal arteries. Cells derived from restenosing lesions retained their increased sensitivity even after eight passages in culture (approximately 20 cumulative population doublings). Cell size measurements showed that the decreased proliferative activity mainly occurred in smooth muscle cell subpopulations consisting of small cells. A cytotoxic effect of DHE was observed at concentrations above 5 micrograms/ml, causing cytoplasmic protrusions, vacuoles, and even complete cell lysis. At a DHE concentration of 5 micrograms/ml, the number of viable cells was 73% +/- 14% (means +/- SD) for smooth muscle cells from nonatherosclerotic arterial media and only 38% +/- 11% (means +/- SD) for smooth muscle cells from atherosclerotic plaques. In all experiments, no significant difference in response to DHE treatment was observed between cells from primary stenosing and restenosing lesions, suggesting a specific mechanism for plaque-derived cells causing an increased sensitivity in comparison to smooth muscle cells from nonatherosclerotic arteries. The pronounced sensitivity of plaque cells against DHE treatment suggests that this drug--even without photoactivation--is potentially valuable in vivo as a therapeutic approach to vascular stenoses.
Cultured smooth muscle cells from human nonatherosclerotic arteries and from primary stenosing lesions were labeled with dihematoporphyrinester and ether, a photosensitizing probe used mainly for the detection and photodynamic therapy of tumors. After labeling for 24 h, cells were irradiated with ultraviolet light (wavelength 365 nm, energy densities ranging from 30 to 1,200 mJ/cm2). Twenty-four hours after photoradiation, 80% of smooth muscle cells from nonatherosclerotic arteries and only 20% of smooth muscle cells from atherosclerotic plaques were viable and still adherent. Moreover, dynamic cell and cytoskeletal alterations in response to irradiation are described. The differential sensitivity of smooth muscle cells from nonatherosclerotic arteries and from atherosclerotic plaques provides evidence that a photodynamic treatment might be a valuable therapeutic approach to vascular stenosis.
To further understand the temporal mode and mechanisms of coronary restenosis, 229 patients were studied by prospective angiographic follow-up on day 1 and at 1, 3 and 6 months and 1 year after successful percutaneous transluminal coronary angioplasty. Quantitative measurement of coronary stenosis was achieved by cinevideodensitometric analysis. Actuarial restenosis rate was 12.7% at 1 month, 43.0% at 3 months, 49.4% at 6 months and 52.5% at 1 year. In 219 patients followed up for greater than or equal to 3 months, mean stenosis diameter was 1.91 +/- 0.53 mm immediately after coronary angioplasty, 1.72 +/- 0.52 mm on day 1, 1.86 +/- 0.58 mm at 1 month and 1.43 +/- 0.67 mm at 3 months. In 149 patients followed up for greater than or equal to 6 months, mean stenosis diameter was 1.66 +/- 0.58 mm at 3 months and 1.66 +/- 0.62 mm at 6 months. In 73 patients followed up for 1 year, mean stenosis diameter was 1.65 +/- 0.56 mm at 6 months and 1.66 +/- 0.57 mm at 1 year. Thus, stenosis diameter decreased markedly between 1 month and 3 months after coronary angioplasty and reached a plateau thereafter. In conclusion, restenosis is most prevalent between 1 and 3 months and rarely occurs beyond 3 months after coronary angioplasty.
The pathologic changes in the coronary arteries of three patients who died 5, 17 and 62 days, respectively, after percutaneous transluminal coronary angioplasty were studied. Changes in the vessel wall seen early after angioplasty included focal denudation of the endothelium, splits in the intima extending to and along the inner aspect of the media, focal intimal necrosis and adventitial hemorrhage. Extensive medial dissections were seen in the coronary arteries of the two patients who died 5 and 17 days after coronary angioplasty. Fibrin was deposited on the surface of the intima, within intimal cracks and in areas of intimal and medial necrosis. Focal proliferation of smooth muscle cells was prominent on neointimal surfaces of the coronary artery from the patient who died 17 days after angioplasty. The previously dilated coronary segment from the patient who died 62 days after angioplasty was stenosed by an extensive recent proliferation of smooth muscle cells that were distributed over the entire circumference of the intimal surface as well as within gaps in the old atherosclerotic plaques. This type of intimal proliferation would appear to be responsible for the recurrent coronary artery stenosis that develops in some patients after coronary angioplasty.
The results of follow-up angiography in patients from 27 clinical centers enrolled in the PTCA Registry were analyzed to evaluate restenosis after PTCA. Of 665 patients with successful PTCA, 557 (84%) had follow-up angiography (median follow-up 188 days). Restenosis, defined as an increase of at least 30% from the immediate post-PTCA stenosis to the follow-up stenosis or a loss of at least 50% of the gain achieved at PTCA, was seen in 187 patients (33.6%). The incidence of restenosis in patients who underwent follow-up angiography was highest within the first 5 months after PTCA. Restenosis was found in 56% of patients with definite or probable angina after PTCA and in 14% of patients without angina after PTCA. Twenty-four percent of patients with restenosis did not have either definite or probable angina. Multivariate analysis selected 4 factors associated with increased rate of restenosis: male sex, PTCA of bypass graft stenosis, severity of angina before PTCA and no history of MI before PTCA.
Neointima formation contributing to recurrent stenosis remains a major limitation of percutaneous transluminal angioplasty. Endovascular low-dose gamma-irradiation has been shown to reduce intimal thickening (hyperplasia) after balloon overstretch injury in pig coronary arteries, a model of restenosis. The objective of this study was to determine whether the use of a beta-emitting radioisotope for this application would have similar effects and to examine the dose-response relations with this approach.
Normal domestic pigs underwent balloon overstretch injury in the left anterior descending and left circumflex and coronary arteries. A flexible catheter was introduced by random assignment into one of these arteries and was afterloaded with a 2.5-cm ribbon of encapsulated 90Strontium/90Yttrium sources (90Sr/Y, a pure beta-emitter). It was left in place for a period of time sufficient to deliver one of four doses: 7, 14, 28, or 56 Gy, to a depth of 2 mm. Animals were killed 14 days after balloon injury, the coronary vasculature was pressure-perfusion fixed, and histomorphometric analysis of arterial cross sections was performed. All arteries treated with radiation demonstrated significantly decreased neointima formation compared with control arteries. The ratio of intimal area to medial fracture length was inversely correlated with increasing radiation dose: control (no radiation), 0.47; 7 Gy, 0.34; 14 Gy, 0.20; 28 Gy, 0.08; and 56 Gy, 0.02 (r = -.78, P < .000001). Scanning electron microscopy demonstrated a confluent layer of endothelium-like cells both in control and in 14 Gy-irradiated arteries. There was neither evidence of significant necrosis nor excess fibrosis in the media, adventitia, or perivascular space of the coronary arteries or adjacent myocardium in the irradiated groups. Furthermore, the exposure to the staff and the total body exposure to the pig with the beta source was a small fraction of the dose previously measured and calculated with 192Ir, a gamma-emitting radioisotope.
Administration of endovascular beta-radiation to the site of coronary arterial overstretch balloon injury in pigs with 90Sr/Y is technically feasible and safe. Radiation doses between 7 and 56 Gy showed evidence of inhibition of neointima formation. A dose-response relation was demonstrated, but no further inhibitory effect was seen beyond 28 Gy. These data suggest that intracoronary beta-irradiation is practical and feasible and may aid in preventing clinical restenosis.
Photodynamic therapy (PDT) for tumor ablation is effective in the treatment of superficial cancers. Adjunctive intraoperative PDT has been proposed for the “sterilization” of tumor beds after the resection of malignancies. Arteries in photosensitized animal models exposed to appropriate light receive characteristic injury. This study was conducted to determine whether photodynamic injury to the rabbit carotid artery results in thrombotic occlusion or weakening of the vessel wall.
PDT of the carotid arteries of New Zealand white rabbits, using either disulphonated aluminum phthalocyanine or 5-aminolevulinic-acid-induced protoporphyrin IX as the photosensitizer, was performed with a light dose of 100 J/cm2. Histologic examination of the carotids treated with either agent demonstrated typical full-thickness loss of cellularity 3 days after PDT. All vessels remained patent, and no inflammatory infiltrate was evident. Elastin van Gieson staining showed preservation of inner and medial elastic laminae and medial and adventitial collagen.
Additional rabbits were similarly treated with PDT to 1-cm segments of both common carotid arteries. The animals were sacrificed at 3, 7, and 21 days. The carotids were exposed, and both control and treated segments were subjected to intraluminal hydrostatic distention until the vessels burst. No reduction in the pressure required to burst the vessels was evident in the treated vessels as compared with the control vessels. The authors of the study concluded that despite full-thickness cell death, PDT-treated arteries are not at risk for thrombotic occlusion or hemorrhage.
Although the management of atherosclerotic disease by the use of balloon angioplasty is widespread, the treatment is limited by restenosis in 30% to 50% of cases. Fibrocellular intimal hyperplasia, the main cause of restenosis, arises from proliferation and migration of medial smooth muscle cells (SMC) into the intimal layer. Factors leading to intimal hyperplasia are incompletely understood, and drugs have universally failed to influence clinical restenosis. Photodynamic therapy (PDT), the light activation of photosensitizing drugs to generate cytotoxic mediators, may have potential as prophylaxis for intimal hyperplasia. 5-Amino-levulinic acid-induced protoporphyrin IX (ALA-PPIX), a naturally occurring porphyrin precursor, and its product, -PPIX, offers a novel method of sensitization for PDT. We have investigated the pharmacokinetics of ALA in arteries and the effects of ALA-PPIX-sensitized PDT on normal and balloon-injured arteries.
ALA (20 to 200 mg/kg) was injected into healthy rats, and PPIX fluorescence was measured in the carotid arteries. In a second group of rats, the exposed carotid artery was laser illuminated (50 J/cm2, 630 nm) 30 to 90 minutes after sensitization. Three and 14 days after PDT, histological sections from treated arteries were analyzed by light microscopy. Subsequently, two new groups of rats underwent PDT (ALA, 100 mg/kg; laser, 50 J/cm2, 630 nm [at 60 to 90 minutes]). The left carotid arteries underwent balloon angioplasty by intraluminal passage of a Fogarty FG2 catheter immediately before irradiation. These rats were killed at 14 and 28 days together with laser-only, ALA-only, and untreated control rats. The arteries were perfusion-fixed in vivo. ALA-PPIX induced arterial media fluorescence in a dose-dependent manner. In the normal arteries, PDT produced a dose-dependent cellular depletion in the treated arterial segment at 3 days, and this was complete with 100 and 200 mg/kg of ALA. At 14 days, the media remained acellular, although the endothelial lining had regenerated. In the balloon-injured arteries, PDT produced complete inhibition of intimal hyperplasia at both 14 and 28 days (0%). This was significantly greater than that produced by any of the control rats (34% to 69% and 37% to 66% at the two times, respectively). Significance was at 99% using ANOVA and Fisher's PLSD test. No hemorrhage, thrombosis, or aneurysm formation was seen.
ALA-PPIX-sensitized PDT applied at the time of angioplasty effectively inhibits experimental intimal hyperplasia development in rats. This may offer a new approach to the management of angioplasty restenosis in patients.
This study was performed to define the evolution of lesion morphology and its relation to thrombus formation and smooth muscle cell proliferation after experimental coronary stent placement.
Restenosis after percutaneous revascularization may develop because of thrombus accumulation and smooth muscle cell proliferation. In animal models of restenosis, thrombus may assume a significant role in neointimal formation by providing an absorbable matrix into which smooth muscle cells proliferate.
Twenty-eight oversized stents were placed in the coronary arteries of 23 juvenile domestic pigs. The histologic degree of vessel injury, lesion morphometry and smooth muscle cell proliferation measured by immunolocalization with a monoclonal antibody to proliferating cell nuclear antigen (PCNA) were assessed at 24 h and 7, 14 and 28 days after stent placement.
The area of thrombus was minimal at 24 h ([mean +/- SE] 0.44 +/- 0.12 mm2). Neointimal area at 7 days (0.72 +/- 0.20 mm2) was similar to the area of thrombus, followed by a significant increase at 14 days (3.15 +/- 0.39 mm2) and 28 days (3.30 +/- 0.28 mm2) (p < 0.0036, 24 h and 7 days vs. 14 and 28 days). At 14 and 28 days, neointimal thickness correlated with the histologic degree of vessel injury (p < 0.003). In arteries with severe injury, the increase in neointimal thickness is accounted for by replacement of the damaged media. The smooth muscle cell proliferation index was 18.6 +/- 3.5% at 7 days compared with 9.6 +/- 1.3% by 14 days (p = 0.0247) and declined to 1.1 +/- 0.97% by 28 days (p < 0.008, 7 and 14 days vs. 28 days).
Early thrombus formation is minimal, and thrombus accounts for a small portion of subsequent neointimal formation. Smooth muscle cell proliferation and matrix formation are the major factors relating to neointimal formation in this proliferative model of restenosis. The evolution of neointimal formation after coronary stenting shows maximal smooth muscle cell proliferation at 7 days, with a decline to low levels by 28 days. Therefore, these data may be useful for developing effective therapies for restenosis.
Removal of the carotid artery adventitia from rabbits induced the formation of an intimal hyperplastic lesion. In rabbits fed a normal diet, the lesion (measured as the intimal:medial ratio) was maximal by day 14 (0.456 +/- 0.079, n = 5, P < 0.01) and thereafter, regressed towards control dimensions (0.037 +/- 0.003, n = 14) by day 28 (0.080 +/- 0.025, n = 7, P = 0.14). In rabbits fed a high cholesterol diet, the lesion was again maximal by day 14 (0.376 +/- 0.056, n = 8, P < 0.01). Although some regression was seen, the lesion persisted to day 42 (0.272 +/- 0.052, n = 8, P < 0.01). Electron microscopy and immunocytochemistry showed two types of lesion, (a) smooth muscle cell predominant on normal diet and, (b) macrophage predominant on high cholesterol diet. Smooth muscle cell predominant lesions underwent almost complete regression, whereas macrophage predominant lesions persisted. We propose that lesion formation may be initiated following the development of arterial wall hypoxia, secondary to excision of the adventitial vasa vasorum. Furthermore, we have devised a novel method to restore a highly vascular 'neoadventitia' to an artery whose adventitia has previously been removed, using loosely placed PVC tubing. We suggest this 'neoadventitia' was able to inhibit the formation of an intimal hyperplastic lesion and to promote regression of an already established lesion by restoring arterial wall oxygenation.
In this editorial, the problem of restenosis after coronary balloon angioplasty and other transluminal interventions is reviewed from the perspective of quantitative coronary angiography. The review is largely based on the experience of the Thoraxcentre in the application of quantitative angiography to the study of restenosis over the past decade, with incorporation and discussion of relevant and significant contributions from other groups. Current discrepancies in the angiographic definition of restenosis are highlighted and the use of percent diameter stenosis or MLD as the measurement parameter of choice is objectively addressed. Perspectives on the pathologic paradigm of restenosis are briefly reviewed as a basis from which to evaluate quantitative angiographic information provided by various studies. Particular attention is then paid, in chronologic fashion, to discussion and elaboration of insights to the restenosis process provided by quantitative angiographic studies, which have led to the introduction of some new methodological approaches to the comparison of short- and long-term angiographic luminal changes after various interventions. A word of caution on the potential pitfalls of quantitative angiographic studies is provided and counterbalanced with a discussion of clinical correlations of quantitative angiographic measurements. Finally, a proposal is made for the application of quantitative angiographic measurements to randomized clinical trials for the purpose of comparing new interventional devices.
Restenosis complicates a significant proportion of endovascular and open vascular procedures such as carotid endarterectomy. In contrast to the primary atheroma, restenosis is characterized by intimal hyperplasia of vascular smooth muscle cells. We hypothesized that gamma radiation would reduce restenosis by limiting intimal hyperplasia after arterial injury.
To demonstrate the effect of gamma radiation on smooth muscle hyperplasia in vivo, a standardized bilateral carotid balloon catheter arterial injury was produced in 37 rats. A single dose of 750, 1500, or 2250 cGy (1cGy = 1 rad) gamma radiation was delivered to the right carotid artery at either 1 or 2 days after injury; the shielded contralateral carotid artery served as matched control. At 21 days after injury, vessels were perfusion-fixed in situ, and cross-sectional area of neointima was determined from axial sections using image analysis.
Marked reductions in neointimal cross-sectional area were demonstrated in vessels subjected to 1500- and 2250-cGy radiation at both 1 and 2 days after injury. A less prominent effect was noted for 750 cGy, reaching statistical significance only at 2 days after injury. By two-way ANOVA, radiation dose (P = .0002), timing of radiation delivery (P = .003), and an interaction between timing and dose (P = .0278) were significantly associated with reduction in neointimal cross-sectional area. At 1500 cGy, delivery of radiation 1 day after injury inhibited neointimal hyperplasia more prominently than the same dose 2 days after injury; a dose-response relation was evident at 1 day.
Radiation may be an important adjunctive therapy for reducing the incidence of restenosis after angioplasty or endarterectomy.
Intraluminal thrombus formation and medial smooth muscle (SM) cell proliferation are recognized responses of the arterial system to injury. In contrast to these well-characterized processes during vascular repair, changes involving the adventitia have been largely neglected in previous studies. Hence, the goal of this investigation was to assess the response of the adventitia to coronary arterial injury.
Adventitial changes in porcine coronary arteries subjected to medial injury were characterized by immunohistochemistry, histochemistry, and microscopic morphometry. The rapid development of a hypercellular response in the adventitia was evident 3 days after balloon-induced medial injury. Cell proliferation, as assessed by proliferating cell nuclear antigen immunostaining, reached the maximum level in the adventitia at 3 days, whereas at 14 and 28 days, the number of replicating cells reverted toward the baseline. The proliferating activity in the adventitia exceeded that seen in the media at all times after injury. To further define the changes in the phenotype of adventitial cells, the expression of three cytoskeletal proteins (vimentin, alpha-SM actin, and desmin) was characterized. Fibroblasts in normal adventitia expressed vimentin but no alpha-SM actin or desmin. After injury, these cells acquired characteristics of myofibroblasts expressing alpha-SM actin, which peaked at 7 and 14 days. Desmin expression was patchy in the adventitia, as opposed to its homogeneous distribution in medial SM cells. The modulation of fibroblast phenotype was transient, inasmuch as alpha-SM actin immunostaining declined at 28 days after injury, when dense, collagen-rich scar was evident within the adventitia. The above-described changes involving hypercellularity of the adventitia, myofibroblast formation, and fibrosis were associated with a significant focal adventitial thickening at 3, 7, 14, and 28 days after injury (P < .01 versus uninjured coronary arteries).
This study demonstrates the involvement of the adventitia in the vascular repair process after medial injury. The hypercellularity of the adventitial layer, proliferation of fibroblasts, and modulation of their phenotype to myofibroblasts are associated with the development of the thickened adventitia. It is postulated that these phenomena affect vascular remodeling and may provide an important insight into the mechanisms of vascular disorders.
We investigated the effects of Photodynamic therapy (PDT) using Aluminium disulphonated phthalocyanine (AlS2Pc) on experimental intimal hyperplasia (FCIH).
(a) Pharmacokinetics: Normal rats were injected with Als2Pc and carotid artery fluorescence was measured. (b) Normal artery PDT: Sensitised rats underwent carotid artery laser irradiation (50J/cm2, 675nm) and were assessed after 3 and 14 days and 1-6 months. (c) PDT: Rats underwent standard carotid artery balloon injury immediately prior to PDT and arteries were assessed at 2 to 26 weeks, together with laser, AlS2Pc, and untreated controls.
(a) Fluorescence intensity in different arterial layers. (b) Medial smooth muscle cell counts per high power field (light microscopic). (c) Percentage amount of FCIH (area of intimal hyperplasia) as a ratio of the IEL (area enclosed by the internal elastic lamina).
(a) AlS2Pc fluorescence intensity increased with increasing dosage, with maximal fluorescence in the arterial media at 30 min. (b) PDT produced medial cell depletion at 3 days and persisted over 6 months without loss of vessel integrity. (c) PDT completely inhibited FCIH at 2 and 4 weeks. This was partial at 6 to 26 weeks (51% of untreated level). PDT inhibition of FCIH was significantly greater than in any of the control groups. p < 0.0001. Mann-Whitney Test.
Adjunctive AlS2Pc sensitised photodynamic therapy inhibits experimental intimal hyperplasia, by causing medial smooth muscle cell depletion. This offers a new approach to the management of angioplasty restenosis in patients.
Oversized balloon dilatation of normal porcine coronary arteries usually heals without stenosis formation.
With the purpose of developing a stenotic model and examining the mechanisms of luminal narrowing after angioplasty, we produced a circumferential deep vessel wall injury by inflating and withdrawing an oversized chain-encircled angioplasty balloon in the left anterior descending coronary artery (LAD) of 20 pigs. Three pigs died and did not complete the study. In 8 pigs (group 1), serial coronary arteriography was performed. The lumen diameter (mean+/-SD) before dilation was 3.4 +/- .4 mm; after dilation, 4.2 +/- 0.6 mm; and at follow-ups 2 and 4 weeks later, 1.6 +/- 0.4 mm (P<.0001). In 9 pigs (group 2) examined postmortem 3 weeks after dilatation, histology revealed that the injury was deep (out to adventitia) in all arteries and completely circumferential (360 degrees) in all but two arteries. Adventitia was markedly thickened as a result of neoadventitial formation. Injury correlated strongly with neointimal formation (middle LAD, r=.71, P=.00001, but neither injury nor neointima correlated with lumen size (r=.14, P=.46 and r=.34, P=.07, respectively); ie, neointimal formation did not explain late luminal narrowing. Lumen size, however, did correlate strongly with vessel size (r=.74, P=000005). The late loss in lumen diameter observed angiographically in group 1 substantially exceeded that caused by neointimal formation seen by histology in group 2.
The chain-encircled angioplasty balloon produced a circumferential deep vessel wall injury that healed by luminal narrowing. In this porcine model, arterial remodeling was more important than neointimal formation in late luminal narrowing.
Studies have suggested that restenosis within Palmaz-Schatz stents results from neointimal hyperplasia or chronic stent recoil and occurs more frequently at the articulation.
Serial intravascular ultrasound (IVUS) was performed after intervention and at follow-up in 142 stents in 115 lesions. IVUS measurements (external elastic membrane [EEM], stent, and lumen cross-sectional areas [CSAs] and diameters) were performed, and plaque CSA (EEM lumen in reference segments and stent lumen in stented segments), late lumen loss (delta lumen), remodeling (delta EEM in reference segments and delta stent in stented segments), and tissue growth (delta plaque) were calculated. After intervention, the lumen tended to be smallest at the articulation because of tissue prolapse. At follow-up, tissue growth was uniformly distributed throughout the stent; the tendency for greater neointimal tissue accumulation at the central articulation reached statistical significance only when normalized for the smaller postintervention lumen CSA. In stented segments, late lumen area loss correlated strongly with tissue growth but only weakly with remodeling. Stents affected adjacent vessel segments; remodeling progressively increased and tissue growth progressively decreased at distances from the edge of the stent. These findings were similar in native arteries and saphenous vein grafts and in lesions treated with one or two stents. There was no difference in the postintervention or follow-up lumen (at the junction of the two stents) when overlapped were compared with nonoverlapped stents.
Late lumen loss and in-stent restenosis were the result of neointimal tissue proliferation, which tended to be uniformly distributed over the length of the stent.
Restenosis occurs after 30% to 50% of transcatheter coronary procedures; however, the natural history and pathophysiology of restenosis are still incompletely understood.
Serial (postintervention and follow-up) intravascular ultrasound imaging was used to study 212 native coronary lesions in 209 patients after percutaneous transluminal coronary angioplasty, directional coronary atherectomy, rotational atherectomy, or excimer laser angioplasty. The external elastic membrane (EEM) and lumen cross-sectional areas (CSA) were measured; plaque plus media (P+M) CSA was calculated as EEM minus lumen CSA. The anatomic slice selected for serial analysis had an axial location within the target lesion at the smallest follow-up lumen CSA. At follow-up, 73% of the decrease in lumen (from 6.6+/-2.5 to 4.0+/-3.7 mm2, P<.0001) was due to a decrease in EEM (from 20.1+/-6.4 to 18.2+/-6.4 mm2, P<.0001); 27% was due to an increase in P+M (from 13.5+/-5.5 to 14.2+/-5.4 mm2, P<.0001). Delta Lumen CSA correlated more strongly with delta EEM CSA (r=.751, P<.0001) than with delta P+M CSA (r=.284, P<.0001). Delta EEM was bidirectional; 47 lesions (22%) showed an increase in EEM. Despite a greater increase in P+M (1.5+/-2.5 versus 0.5+/-2.0 mm2, P=.0009), lesions exhibiting an increase in EEM had (1) no change in lumen (-0.1+/-3.3 versus 3.6+/-2.3 mm2, P<.0001), (2) a reduced restenosis rate (26% versus 62%, P<.0001), and (3) a 49% frequency of late lumen gain (versus 1%, P<.0001) compared with lesions with no increase in EEM.
Restenosis appears to be determined primarily by the direction and magnitude of vessel wall remodeling (delta EEM). An increase in EEM is adaptive, whereas a decrease in EEM contributes to restenosis.
Vessel remodeling after angioplasty: comparative anatomic studies
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