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Extracellular vesicles (EVs), such as microvesicles and exosomes, are membranous structures containing bioactive material released by several cells types, including mesenchymal stem/stromal cells (MSCs). Increasing lines of evidences point to EVs as paracrine mediators of the beneficial effects on tissue remodeling associated with cell therapy. Adm...
Citations
... However, the clinical translation of MSC exosomes faces challenges due to a lack of effective methods to label and track the biodistribution of exosomes or small EVs. Establishing a spatiotemporal biodistribution congruent with a proposed model is essential for elucidating their mechanism of action, as previously outlined [20][21][22]. Radiolabeling currently stands out as the most sensitive and effective method for directly labeling and tracking EVs in vivo, allowing for deep tissue penetration in biodistribution studies [23,24]. ...
Mesenchymal stem/stromal cells (MSCs) are an extensively studied cell type in clinical trials due to their easy availability, substantial ex vivo proliferative capacity, and therapeutic efficacy in numerous pre-clinical animal models of disease. The prevailing understanding suggests that their therapeutic impact is mediated by the secretion of exosomes. Notably, MSC exosomes present several advantages over MSCs as therapeutic agents, due to their non-living nature and smaller size. However, despite their promising therapeutic potential, the clinical translation of MSC exosomes is hindered by an incomplete understanding of their biodistribution after administration. A primary obstacle to this lies in the lack of robust labels that are highly sensitive, capable of directly and easily tagging exosomes with minimal non-specific labeling artifacts, and sensitive traceability with minimal background noise. One potential candidate to address this issue is radioactive iodine. Protocols for iodinating exosomes and tracking radioactive iodine in live imaging are well-established, and their application in determining the biodistribution of exosomes has been reported. Nevertheless, the effects of iodination on the structural or functional activities of exosomes have never been thoroughly examined. In this study, we investigate these effects and report that these iodination methods abrogate CD73 enzymatic activity on MSC exosomes. Consequently, the biodistribution of iodinated exosomes may reflect the biodistribution of denatured exosomes rather than functionally intact ones.
... Exosomes, as biologically derived nanoparticles, offer efficient drug delivery and excellent biocompatibility with minimal side effects. They can elicit robust cellular responses both in vitro and in vivo, making them promising therapeutic agents [7,8]. NF-kB exists in an inactive state within the cytoplasm of nearly all mammalian cells, and it forms associations with inhibitory proteins collectively known as IκB (inhibitory κB proteins). ...
Background
This study aims to investigate the potential anti-inflammatory effects of exosomes engineered to carry super-repressor IκB (Exo-srIκB), an exosome-based NF-κB inhibitor, in the context of RA.
Methods
Peripheral blood mononuclear cells (PBMCs) and synovial fluid mononuclear cells (SFMCs) were collected from patients diagnosed with RA and treated with Exo-srIκB to test the therapeutic potential. Flow cytometry analysis was performed to assess the production of inflammatory cytokines (IL-17A and GM-CSF) by the cells. ELISA was utilized to measure the levels of TNF-α, IL-17A, IL-6, and GM-CSF. Arthritis was induced in SKG mice by intraperitoneal injection of curdlan. DBA/1 J mice were used in collagen-induced arthritis (CIA) experiments. After the development of arthritis, mice were injected with either Exo-Naïve (control exosome) or Exo-srIκB. Arthritis scores were recorded biweekly, and histological observations of the ankle joint were conducted using H&E and safranin-O staining. Additionally, bone erosion was evaluated using micro-CT imaging.
Results
In the ex vivo study involving human PBMCs and SFMCs, treatment with Exo-srIκB demonstrated a notable reduction in inflammatory cytokines. Furthermore, in both the SKG and CIA models, Exo-srIκB treatment exhibited significant reductions in inflammation, cartilage destruction, and bone erosion within the joint tissues when compared to the Exo-Naive control group. Additionally, the radiographic score assessed through microCT showed a significant decrease compared to the Exo-Naive control group.
Conclusion
Overall, these findings suggest that Exo-srIκB possesses anti-inflammatory properties in human RA cells and animal models, making it a promising therapeutic candidate for the treatment of RA.
... We obtained similar results to those of a few studies investigating the biodistribution of large EVs administrated by intravenous injection, showing that EVs rapidly localized to lungs, liver, spleen and kidneys [61]. Here, we demonstrated the presence of EVs in the same organs and in addition, were detected in the stomach and intestine, probably due to the intraperitoneal route of administration. ...
The discovery of extracellular vesicles (EVs) as efficient exogenous biotransporters of therapeutic agents into cells across biological membranes is an exciting emerging field. Especially the potential of EVs as targeted delivery systems for diseases with selective treatments, such as fibrosis, whose treatment causes side effects in other organs not involved in the disease. Methods: In this study, we collected embryonic fibroblast-derived EVs from two different centrifugation fractions, 10 K g and 100 K g fractions from a NIH-3T3 cell line loaded with an experimental drug. Mice with fibrotic hearts and lungs were obtained by administration of angiotensin II. We generated fluorescent EVs and bioluminescent drug to observe their accumulation by colocalization of their signals in fibrotic heart and lung. The biodistribution of the drug in various organs was obtained by detecting the Au present in the drug nanostructure. Results: The drug-loaded EVs successfully reduced fibrosis in pathological fibroblasts in vitro, and modified the biodistribution of the experimental drug, enabling it to reach the target organs in vivo. We described the pre-analytical characteristics of EVs related to physical variables, culture and harvesting conditions, crucial for their in vivo application as nanotransporters using a previously validated protein-based antifibrotic drug. The results showed the colocalization of EVs and the experimental drug in vivo and ex vivo and the efficient reduction of fibrosis in vitro. This work demonstrates that 10K-EVs and 100K-EVs derived from fibroblasts can act as effective biotransporters for targeted drug delivery to profibrotic fibroblasts, lungs, or heart.
Conclusion: We observed that fibroblast-derived 10K-EVs and 100K-EVs are useful biotransporters encapsulating a new generation drug leading to a reduction of fibrosis in profibrotic fibroblasts in vitro. In addition, drug containing EVs were shown to reach fibrotic heart and lungs in vivo, enhancing free drug biodistribution.
... Except for the membrane composition, various factors can affect the absorption, distribution, metabolism, and excretion (ADME) of MSC-EV either directly or indirectly. The choice of dosing regimen, including dosages, frequency, routes, and timing of administration, may also affect the survival time, and therapeutic effects of MSC-EV (Wiklander et al., 2015;Di Rocco et al., 2016). Besides, the size of MSC-EV also affects its elimination in vivo. ...
Liver disease has emerged as a significant worldwide health challenge due to its diverse causative factors and therapeutic complexities. The majority of liver diseases ultimately progress to end-stage liver disease and liver transplantation remains the only effective therapy with the limitations of donor organ shortage, lifelong immunosuppressants and expensive treatment costs. Numerous pre-clinical studies have revealed that extracellular vesicles released by mesenchymal stem cells (MSC-EV) exhibited considerable potential in treating liver diseases. Although natural MSC-EV has many potential advantages, some characteristics of MSC-EV, such as heterogeneity, uneven therapeutic effect, and rapid clearance in vivo constrain its clinical translation. In recent years, researchers have explored plenty of ways to improve the therapeutic efficacy and rotation rate of MSC-EV in the treatment of liver disease. In this review, we summarized current strategies to enhance the therapeutic potency of MSC-EV, mainly including optimization culture conditions in MSC or modifications of MSC-EV, aiming to facilitate the development and clinical application of MSC-EV in treating liver disease.
... Numerous studies have suggested that purified BMSC-EXOs offer several unique advantages over BMSCs in repairing damaged tissues. They have stable biological activity, long-term preservation, fast transportation, strong permeability, good biocompatibility, ease of engineering and the ability to avoid immune responses and tumorigenesis (8,14,125). Therefore, BMSC-EXO-based decellularized therapy is a promising therapeutic approach for maintaining the regenerative properties of BMSCs, while avoiding the potential downsides associated with cell therapy (126,127). ...
In sports medicine, injuries related to the insertion of tendons into bones, including rotator cuff injuries, anterior cruciate ligament injuries and Achilles tendon ruptures, are commonly observed. However, traditional therapies have proven to be insufficient in achieving satisfactory outcomes due to the intricate anatomical structure associated with these injuries. Adult bone marrow mesenchymal stem cells possess self-renewal and multi-directional differentiation potential and can generate various mesenchymal tissues to aid in the recovery of bone, cartilage, adipose tissue and bone marrow hematopoietic tissue. In addition, extracellular vesicles derived from bone marrow mesenchymal stem cells known as exosomes, contain lipids, proteins and nucleic acids that govern the tissue microenvironment, facilitate tissue repair and perform various biological functions. Studies have demonstrated that bone marrow mesenchymal stem cell-derived exosomes can function as natural nanocapsules for drug delivery and can enhance tendon-bone healing strength. The present review discusses the latest research results on the role of exosomes released by bone marrow mesenchymal stem cells in tendon-bone healing and provides valuable information for implementing these techniques in regenerative medicine and sports health.
... Furthermore, administration of mesenchymal stem cell EVs to mouse models of induced liver fibrosis improved the fibrotic phenotype (Li et al., 2012). As EVs can travel throughout the body via the circulatory and lymphatic systems, there is significant interest in where they ultimately distribute to and, from a therapeutic perspective, whether they can be used to target specific organs or tumours for drug delivery (Di Rocco et al., 2016;Lai et al., 2014). ...
Extracellular vesicles (EVs) are lipid-bound vesicles released from cells that play a crucial role in many physiological processes and pathological mechanisms. As such, there is great interest in their biodistribution. One currently accessible technology to study their fate in vivo involves fluorescent labelling of exogenous EVs followed by whole-animal imaging. Although this is not a new technology, its translation from studying the fate of whole cells to subcellular EVs requires adaptation of the labelling techniques, excess dye removal and a refined experimental design. In this Review, we detail the methods and considerations for using fluorescence in vivo and ex vivo imaging to study the biodistribution of exogenous EVs and their roles in physiology and disease biology.
... advantages over organic or inorganic nanoparticles in that they have inherent biocompatibility, elevate physicochemical resilience, have low immunogenicity, and have long-distance information, including an inherent aiming potential to intermingle along cells through membrane fusion with signal transduction [10,11]. The lack of knowledge of ECVs' in vivo bio function in real stint, despite efforts and advancements in the field, poses a significant obstacle to their usage in diagnostic or therapeutic settings [12]. For ECVs to be used effectively as a curative or drug deployment framework in bioscience utilizations, it is necessary to determine their circulation kinetics and biodistribution profile, as well as their aiming potency to exact cells or tissues and uptake route, including cargo transport reliability to inheritor cells [13]. ...
Extracellular vesicles (ECVs) have been abandoned as bio-inspired drug delivery systems (DDS) in the biomedical field. ECVs have a natural ability to cross over extracellular and intracellu-lar barriers, making them superior to manufactured nanoparticles. Additionally, they have the ability to move beneficial biomolecules among far-flung bodily cells. These advantages and the accomplishment of favorable in vivo results convincingly show the value of ECVs in medication delivery. The usage of ECVs is constantly being improved, as it might be difficult to develop a consistent biochemical strategy that is in line with their useful clinical therapeutic uses. Extracellular vesicles (ECVs) have the potential to enhance the therapy of diseases. Imaging technologies, particularly radiolabelled imaging, have been exploited for non-invasive tracking to better understand their in vivo activity.
... The demonstration of EVs pro-osteogenic capacity in vitro has led researchers to investigate their potential use for the treatment of systematic skeletal diseases such as OP. Although EV administration via intravenous injection for the treatment of OP has shown promise [160], the nanoparticles rapid clearance from systemic circulation hinders its therapeutic efficacy [182][183][184]. Moreover, natural unmodified EVs often exhibit insufficient tropism to the tissue of interest [185], hindering the desired therapeutic response of these nanoparticles. ...
In the past decade, extracellular vesicles (EVs) have emerged as key regulators of bone development, homeostasis and repair. EV-based therapies have the potential to circumnavigate key issues hindering the translation of cell-based therapies including functional tissue engraftment, uncontrolled differentiation and immunogenicity issues. Due to EVs’ innate biocompatibility, low immunogenicity, and high physiochemical stability, these naturally-derived nanoparticles have garnered growing interest as potential acellular nanoscale therapeutics for a variety of diseases. Our increasing knowledge of the roles these cell-derived nanoparticles play, has made them an exciting focus in the development of novel pro-regenerative therapies for bone repair. Although these nano-sized vesicles have shown promise, their clinical translation is hindered due to several challenges in the EV supply chain, ultimately impacting therapeutic efficacy and yield. From the biochemical and biophysical stimulation of parental cells to the transition to scalable manufacture or maximising vesicles therapeutic response in vivo, a multitude of techniques have been employed to improve the clinical efficacy of EVs. This review explores state of the art bioengineering strategies to promote the therapeutic utility of vesicles beyond their native capacity, thus maximising the clinical potential of these pro-regenerative nanoscale therapeutics for bone repair.
Graphical Abstract
... CD206 is a marker of M2 macrophage, an anti-inflammatory macrophage that recognizes the foreign matter as phagocyted. 12 The M2 macrophage, an anti-inflammatory macrophage, has been reported to have higher phagocytic affinity than the M1 macrophage under low and high density of cell target. 13 The phagocytosis ability of macrophages is diverse depending on the stimulation. ...
Background : Secretomes have been gaining interest in treating several diseases due to their pharmaceutical effects, such as the immunomodulatory effect. This study aimed to determine the immunomodulatory effect of secretomes derived from human umbilical cord mesenchymal stem cells (MSCs) and their safety.
Methods : We conducted an in vivo immunomodulatory study using a carbon clearance assay. The safety of single-dose administration of secretome was done using fixed-dose methods of acute toxicity test.
Results : The phagocytic index was higher in mice treated with secretome than in untreated mice. The acute toxicity study also showed that the administration of secretome derived from human umbilical cord MSCs did not change the mice’s body weight, physical examination results, organ weight, and gross anatomy examination.
Conclusions : This study presents the potential of secretome derived from MSCs as a safe immunomodulatory agent.
... Assays that lead to co-precipitation of these vesicles with soluble proteins and supplemental chemical methods are also used to isolate EVs from conditioned media. Immunoaffinity methods and sEV extraction kits have also been employed to enable the isolation of these EVs in a shorter period of time (Raposo and Stoorvogel, 2013;Schageman et al., 2013;Rocco et al., 2016;Sarvar et al., 2016;Silva et al., 2016). Each method presents different advantages that should be evaluated according to application, prioritizing processing time, purity, cost, ease of FIGURE 2 Extracellular vesicles are a generic name for many kinds of vesicles released by cells; their origin and size are determinants in their classification. ...
Mesenchymal stem cells (MSCs) are crucial for tissue homeostasis and repair, secreting vesicles to the extracellular environment. Isolated exosomes were shown to affect angiogenesis, immunomodulation and tissue regeneration. Numerous efforts have been dedicated to describe the mechanism of action of these extracellular vesicles (EVs) and guarantee their safety, since the final aim is their therapeutic application in the clinic. The major advantage of applying MSC-derived EVs is their low or inexistent immunogenicity, prompting their use as drug delivery or therapeutic agents, as well as wound healing, different cancer types, and inflammatory processes in the neurological and cardiovascular systems. MSC-derived EVs display no vascular obstruction effects or apparent adverse effects. Their nano-size ensures their passage through the blood–brain barrier, demonstrating no cytotoxic or immunogenic effects. Several in vitro tests have been conducted with EVs obtained from different sources to understand their biology, molecular content, signaling pathways, and mechanisms of action. Application of EVs to human therapies has recently become a reality, with clinical trials being conducted to treat Alzheimer’s disease, retina degeneration, and COVID-19 patients. Herein, we describe and compare the different extracellular vesicles isolation methods and therapeutic applications regarding the tissue repair and regeneration process, presenting the latest clinical trial reports.
