Figure - available from: Histochemistry and Cell Biology
This content is subject to copyright. Terms and conditions apply.
Mouse pericyte-like vascular cells (PVC) associate with cords formed by endothelial cells (HUVEC) on matrigel. HUVEC alone (a), HUVEC and PVC (ratio 5:1) (b) or PVC alone (c) were plated on matrigel, and cord formation was followed by time-lapse video microscopy. PVC are labelled by the fluorescent vital cells marker CFDA. Representative areas are shown after 0 and 6 h. HUVEC cords (d) or PVC aggregates (e) were preformed for 24 h, and PVC or HUVEC were added for 2 h, respectively. Interactions of labelled PVC and HUVEC were detected by fluorescence microscopy. Bars 500 µm. Cocultures of HUVEC and CFDA-labelled pericyte-like cells (PVC/MII) (5:1) were cultured on matrigel for 2 h (f) or 24 h (g). HUVEC were identified by immunodetection of CD31 (red), PVC were identified by the CFDA label (green), nuclei were stained (blue) and merged pictures are shown. HUVEC cords (h) or PVC aggregates (i) were preformed for 24 h on matrigel before CFDA-labelled PVC or HUVEC were added for 2 h, respectively, and identified by immunodetection of CD31 (red), PVC by their CFDA label (green), nuclei were stained (blue). Interacting PVC are marked in merged images (arrows; in b, f). Bars 200 µm
Source publication
Activation of endothelial cells and recruitment of mural cells define critical steps during the formation of stable vascular elements. Both events are reflected by cocultures of endothelial cells and isolated murine pericyte-like cells and define a versatile platform for the analysis of distinct steps during the angiogenic process in vitro. Isolate...
Similar publications
Recent documentation shows that a curcumin-induced growth arrest of renal cell carcinoma (RCC) cells can be amplified by visible light. This study was designed to investigate whether this strategy may also contribute to blocking metastatic progression of RCC. Low dosed curcumin (0.2 µg/mL; 0.54 µM) was applied to A498, Caki1, or KTCTL-26 cells for...
Prostate adenocarcinoma (PCa) is the most common cause of death due to malignancy among men, and bone metastasis is the leading cause of mortality in patients with PCa. Therefore, identifying the causes and molecular mechanism of bone metastasis is important for early detection, diagnosis, and personalized therapy. In this study, we systematically...
Many different diseases are associated with fibrosis of the skin. The clinical symptoms can vary considerably with a broad range from isolated small areas to the involvement of the entire integument. Fibrosis is triggered by a multitude of different stimuli leading to activation of the immune and vascular system that then initiate fibroblast activa...
Citations
... This indicates that reconstruction of the segmental defects with RP, cells & AVB resulted in greatest blood vessel proliferation and the most mature vasculature. This finding is in agreement with the fact that pericytes induce the production of the collagen IV matrix, which initiates migration of endothelial cells, formation of cell-cell contacts, maturation and vascular stabilization (Zhou et al., 2016). CD31 is a transmembrane glycoprotein located at the cell-cell junction of endothelial cells which are homotypic contacts and associated with cell adhesion. ...
Introduction: Recently, efforts towards the development of patient-specific 3D printed scaffolds for bone tissue engineering from bioactive ceramics have continuously intensified. For reconstruction of segmental defects after subtotal mandibulectomy a suitable tissue engineered bioceramic bone graft needs to be endowed with homogenously distributed osteoblasts in order to mimic the advantageous features of vascularized autologous fibula grafts, which represent the standard of care, contain osteogenic cells and are transplanted with the respective blood vessel. Consequently, inducing vascularization early on is pivotal for bone tissue engineering. The current study explored an advanced bone tissue engineering approach combining an advanced 3D printing technique for bioactive resorbable ceramic scaffolds with a perfusion cell culture technique for pre-colonization with mesenchymal stem cells, and with an intrinsic angiogenesis technique for regenerating critical size, segmental discontinuity defects in vivo applying a rat model. To this end, the effect of differing Si-CAOP (silica containing calcium alkali orthophosphate) scaffold microarchitecture arising from 3D powder bed printing (RP) or the Schwarzwalder Somers (SSM) replica fabrication technique on vascularization and bone regeneration was analyzed in vivo. In 80 rats 6-mm segmental discontinuity defects were created in the left femur.
Methods: Embryonic mesenchymal stem cells were cultured on RP and SSM scaffolds for 7d under perfusion to create Si-CAOP grafts with terminally differentiated osteoblasts and mineralizing bone matrix. These scaffolds were implanted into the segmental defects in combination with an arteriovenous bundle (AVB). Native scaffolds without cells or AVB served as controls. After 3 and 6 months, femurs were processed for angio-µCT or hard tissue histology, histomorphometric and immunohistochemical analysis of angiogenic and osteogenic marker expression.
Results: At 3 and 6 months, defects reconstructed with RP scaffolds, cells and AVB displayed a statistically significant higher bone area fraction, blood vessel volume%, blood vessel surface/volume, blood vessel thickness, density and linear density than defects treated with the other scaffold configurations.
Discussion: Taken together, this study demonstrated that the AVB technique is well suited for inducing adequate vascularization of the tissue engineered scaffold graft in segmental defects after 3 and 6 months, and that our tissue engineering approach employing 3D powder bed printed scaffolds facilitated segmental defect repair.
... Given the abundant AnxA5 levels in many tissues, several studies utilized the AnxA5 KO-mice to address possible roles in development. Indeed, the targeted disruption and replacement with an Anxa5-lacZ fusion gene enabled detailed AnxA5 expression analysis during embryonal mouse development, identifying Anxa5-lacZ expression specifically in populations of perivascular cells of non-skeletal mouse tissues, with critical roles in initial steps of angiogenesis differentiation and maturation of endothelial cells by pericytes [234,235]. However, and despite high AnxA5 levels in cartilaginous tissues and bone, the AnxA5 KO-mouse lacked any defects in the calcification process during skeletal development [236] (Table 5). ...
Routine manipulation of the mouse genome has become a landmark in biomedical research. Traits that are only associated with advanced developmental stages can now be investigated within a living organism, and the in vivo analysis of corresponding phenotypes and functions advances the translation into the clinical setting. The annexins, a family of closely related calcium (Ca2+)- and lipid-binding proteins, are found at various intra- and extracellular locations, and interact with a broad range of membrane lipids and proteins. Their impacts on cellular functions has been extensively assessed in vitro, yet annexin-deficient mouse models generally develop normally and do not display obvious phenotypes. Only in recent years, studies examining genetically modified annexin mouse models which were exposed to stress conditions mimicking human disease often revealed striking phenotypes. This review is the first comprehensive overview of annexin-related research using animal models and their exciting future use for relevant issues in biology and experimental medicine.
... Pericytes regulate capillary integrity through stabilization of the basement membrane and direct interactions with endothelial cells. Indeed, whereas endothelial cells do synthesize and deposit collagen IV in the basement membrane [32,46], pericyte recruitment to capillaries further stimulates assembly of basement membrane proteins [34,47,48]. We here show that -i) the endothelial culture medium could induce collagen IV deposition by SHED, and -ii) co-culture with endothelial cells was further increasing collagen IV deposition by pericytes, suggesting mutual stimulation of synthesis and deposition of the vascular basement membrane. ...
Blood perfusion of grafted tissue constructs is a hindrance to the success of stem cell-based therapies by limiting cell survival and tissue regeneration. Implantation of a pre-vascularized network engineered in vitro has thus emerged as a promising strategy for promoting blood supply deep into the construct, relying on inosculation with the host vasculature. We aimed to fabricate in vitro tissue constructs with mature microvascular networks, displaying perivascular recruitment and basement membrane, taking advantage of the angiogenic properties of dental pulp stem cells and self-assembly of endothelial cells into capillaries. Using digital scanned light-sheet microscopy, we characterized the generation of dense microvascular networks in collagen hydrogels and established parameters for quantification of perivascular recruitment. We also performed original time-lapse analysis of stem cell recruitment. These experiments demonstrated that perivascular recruitment of dental pulp stem cells is driven by PDGF-BB. Recruited stem cells participated in deposition of vascular basement membrane and vessel maturation. Mature microvascular networks thus generated were then compared to those lacking perivascular coverage generated using stem cell conditioned medium. Implantation in athymic nude mice demonstrated that in vitro maturation of microvascular networks improved blood perfusion and cell survival within the construct. Taken together, these data demonstrate the strong potential of in vitro production of mature microvasculature for improving cell-based therapies.
... As described in the previous section, the collagen-dense ECM is tightly regulated at the transcriptional level to facilitate endothelial cell migration during angiogenesis. The vascular basement membrane, upon which endothelial cells reside, is composed predominantly of collagen IV, which has six genetically distinct α chains that selfassemble into heterotrimers and interact with laminins and perlecan (Mundel & Kalluri, 2007;Zhou, Pausch, Schlötzer-Schrehardt, Brachvogel, & Pöschl, 2016). Readers are referred to a review by Brown et al. (Brown, Cummings, Vanacore, & Hudson, 2017) to understand the molecular mechanisms that dictate collagen IV scaffold assembly. ...
... Readers are referred to a review by Brown et al. (Brown, Cummings, Vanacore, & Hudson, 2017) to understand the molecular mechanisms that dictate collagen IV scaffold assembly. Although it is not well understood how it functions during angiogenesis, the secretion of collagen IV is speculated to support blood vessel formation and maturation in vivo (Bonanno, Iurlaro, Madri, & Nicosia, 2000;Mundel & Kalluri, 2007;Zhou et al., 2016). In vitro, collagen IV was secreted in cultures of microvascular endothelial cells derived from human myocardium and skin after 10 days with sheet-like networks by Day 25 (Bahramsoltani, Slosarek, De Spiegelaere, & Plendl, 2014). ...
Engineering an angiogenic material for regenerative medicine requires knowledge of native extracellular matrix remodeling by cellular processes in angiogenesis. Vascularization remains a key challenge in the field of tissue engineering, one that can be mitigated by developing platforms conducive to guiding dynamic cell‐matrix interactions required for new vessel formation. In this review, we highlight nuanced processes of angiogenesis and demonstrate how materials engineering is being used to interface with dynamic type I collagen remodeling, Notch and VEGF signaling, cell migration, and tissue morphogenesis. Because α1(I)‐collagen is secreted by endothelial tip cells during sprouting angiogenesis and required for migration, collagen is a very useful natural biomaterial and its angiogenic modifications are described. The balance between collagen types I and IV via secretion and degradation is tightly controlled by proteinases and other cell types that are capable of internalizing collagen to maintain tissue integrity. Thus, we provide examples in skin and cardiac tissue engineering of collagen tailoring in diverse cellular microenvironments for tissue regeneration. As our understanding of how to drive collagen remodeling and cellular phenotype through angiogenic pathways grows, our capabilities to model and manipulate material systems must continue to expand to develop novel applications for wound healing, angiogenic therapy, and regenerative medicine.
... In addition, pericytes and brain microvascular endothelial cells communicate through gap junctions, tight junctions, focal adhesions, and some soluble factors, which in turn secrete growth factors to regulate the formation of tight junctions and permeability of endothelial cells that are important for the remodeling and maintenance of the vascular system . At the same time, pericytes can also regulate the proliferation, migration, and differentiation of endothelial cells (Zhou et al., 2016). Moreover, type IV collagen, polysaccharides, and laminin synthesized by the mouse kidney pericytes have been reported to have an effect on the formation of basement membrane (Wnuk et al., 2017). ...
Pericytes are functional components of the neurovascular unit (NVU) that are located around the blood vessels, and their roles in the regulation of cerebral health and diseases has been reported. Currently, the potential properties of pericytes as emerging therapeutic targets for cerebrovascular diseases have attracted considerable attention. Nonetheless, few reviews have comprehensively discussed pericytes and their roles in cerebrovascular diseases. Therefore, in this review, we not only summarized and described the basic characteristics of pericytes but also focused on clarifying the new understanding about the roles of pericytes in the pathogenesis of cerebrovascular diseases, including white matter injury (WMI), hypoxic–ischemic brain damage, depression, neovascular insufficiency disease, and Alzheimer’s disease (AD). Furthermore, we summarized the current therapeutic strategies targeting pericytes for cerebrovascular diseases. Collectively, this review is aimed at providing a comprehensive understanding of pericytes and new insights about the use of pericytes as novel therapeutic targets for cerebrovascular diseases.
... However, their functional characteristics are more homogeneous within the tissues. Functional characteristics of pericytes include migration toward endothelial platelet-derived growth factor bb (PDGFbb) signaling and the ability to mediate tubule formation in co-culture with endothelial cells (Chen et al. 2015;Zhou et al. 2016). Cardiac pericytes are pro-angiogenic and procoagulatory (Juchem et al. 2010). ...
... However, their functional characteristics are more homogeneous within the tissues. Functional characteristics of pericytes include migration toward endothelial platelet-derived growth factor bb (PDGFbb) signaling and the ability to mediate tubule formation in co-culture with endothelial cells Zhou et al. 2016). Cardiac pericytes are pro-angiogenic and procoagulatory (Juchem et al. 2010). ...
This review chapter describes the current knowledge about the nature of pericytes in the gut, their interaction with endothelial cells in blood vessels, and their pathophysiological functions in the setting of chronic liver disease. In particular, it focuses on the role of these vascular cell types and related molecular signaling pathways in pathological angiogenesis associated with liver disease and in the establishment of the gut-vascular barrier and the potential implications in liver disease through the gut-liver axis.
... However, their functional characteristics are more homogeneous within the tissues. Functional characteristics of pericytes include migration toward endothelial platelet-derived growth factor bb (PDGFbb) signaling and the ability to mediate tubule formation in co-culture with endothelial cells (Chen et al. 2015;Zhou et al. 2016). Cardiac pericytes are pro-angiogenic and procoagulatory (Juchem et al. 2010). ...
Mural cells known as pericytes envelop the endothelial layer of microvessels throughout the body and have been described to have tissue-specific functions. Cardiac pericytes are abundantly found in the heart, but they are relatively understudied. Currently, their importance is emerging in cardiovascular homeostasis and dysfunction due to their pleiotropism. They are known to play key roles in vascular tone and vascular integrity as well as angiogenesis. However, their dysfunctional presence and/or absence is critical in the mechanisms that lead to cardiac pathologies such as myocardial infarction, fibrosis, and thrombosis. Moreover, they are targeted as a therapeutic potential due to their mesenchymal properties that could allow them to repair and regenerate a damaged heart. They are also sought after as a cell-based therapy based on their healing potential in preclinical studies of animal models of myocardial infarction. Therefore, recognizing the importance of cardiac pericytes and understanding their biology will lead to new therapeutic concepts.
... Collagen IV (ColIV or Col4) is an exclusively distributed collagen within the basement membrane zone, of which the basement membrane is an extracellular matrix (ECM) separating connective tissue from epithelia, endothelia, muscle fibers and the entire nervous system (6,7). Mutations of the gene coding for ColIV may cause thinning and splitting ...
Involvement of collagen IV (ColIV) and fibronectin (FN) in the occurrence and development of diabetic nephropathy (DN) and the effects of telmisartan and Salvia miltiorrhiza injection in the treatment of the patients were investigated. Two hundred and fifty-eight patients with stage IV DN were selected as the case group, and another 110 normal healthy subjects were incorporated as the control group. Involved patients were subdivided into different groups according to different treatment therapies; patients in the telmisartan group (T group) were given oral telmisartan; patients in the Salvia miltiorrhiza injection + telmisartan (S + T group) were administered with Salvia miltiorrhiza injection combined with telmisartan treatment, and there was a group of patients who received no intervention as the placebo group. After intervention, levels of glycemic indexes and renal damage indexes indicated downwards trends both in the T group and the S + T group when compared to the placebo group; besides, levels in the S + T group were much lower than those in the T group (all P<0.05). Additionally, in comparison among the above three intervention groups, differences in the fasting blood glucose, 2 h post-prandial blood glucose, glycosylated hemoglobin, blood urea nitrogen, serum creatinine and urinary albumin excretion rate were significant after treatment (all P<0.05). Further, before intervention, both Co1IV and FN in the urine were increased in the case group compared to the control group (all P<0.05). After intervention, both levels were apparently decreased. There were remarkable differences of Co1IV and FN levels in the urine when compared among three different intervention groups after treatment (P<0.05). Increased ColIV and FN levels may be partially responsible for the development of DN. Salvia miltiorrhiza injection with telmisartan have beneficial synergistic effects for DN patients through attenuating the increase in ColIV and FN, reversing hyperglycemia state and postponing ultrastructure changes of glomerular basement membrane.
... Human umbilical vein endothelial cells (HUVECs) were obtained from TCS Cellworks (Buckingham, UK) and murine MII perivascular cells (M2) were isolated as previously described [27]. For all experiments, HUVECs were used between passages 5 and 9, M2 was used between passages 35 and 40. ...
Sulforaphane (SFN) exhibits chemopreventive effects through various mechanisms. However, few studies have focused on the bioactivities of its metabolites. Here, three metabolites derived from SFN were studied, known as sulforaphane glutathione, sulforaphane cysteine and sulforaphane-N-acetylcysteine. Their effects on cell viability, DNA damage, tumorigenicity, cell migration and adhesion were measured in human hepatoma HepG2 cells, and their anti-angiogenetic effects were determined in a 3D co-culture model of human umbilical vein endothelial cells (HUVECs) and pericytes. Results indicated that these metabolites at high doses decreased cancer cell viability, induced DNA damage and inhibited motility, and impaired endothelial cell migration and tube formation. Additionally, pre-treatment with low doses of SFN metabolites protected against H2O2 challenge. The activation of the nuclear factor E2-related factor 2 (Nrf2)-antioxidant response element (ARE) pathway and the induction of intracellular glutathione (GSH) played an important role in the cytoprotective effects of SFN metabolites. In conclusion, SFN metabolites exhibited similar cytotoxic and cytoprotective effects to SFN, which proves the necessity to study the mechanisms of action of not only SFN but also of its metabolites. Based on the different tissue distribution profiles of these metabolites, the most relevant chemical forms can be selected for targeted chemoprevention.
