Traumatic brain injury (TBI) is a major risk factor for disabilities globally with no effective treatment thus far. Recently, homogenous population of clonal mesenchymal stem cells (cMSC) and their ...derived extracellular vesicles (cMSC-EVs) have been proposed as a promising TBI treatment strategy. We herein investigated possible therapeutic effect of cMSC-EVs in TBI treatment and the underlying mechanisms considering cis p-tau as an early hallmark of TBI.
We examined the EVs morphology, size distribution, marker expression, and uptake. Moreover, the EVs neuroprotective effects were studied in both in-vitro and in-vivo model. We also examined the anti-cis p-tau antibody-loading characteristics of the EVs. We treated TBI mouse model with EVs; prepared from cMSC-conditioned media. TBI mice were given cMSC-EVs intravenously and their cognitive functions were analyzed two months of the treatment. We employed immunoblot analysis to study the underlying molecular mechanisms.
We observed a profound cMSC-EVs uptake by primary cultured neurons. We found a remarkable neuroprotective effect of cMSC-EVs upon nutritional deprivation stress. Furthermore, cMSC-EVs were effectively loaded with an anti-cis p-tau antibody. There was a significant improvement in cognitive function in TBI animal models treated with cMSC-EVs compared to the saline-treated group. There was a decreased cis p-tau and cleaved caspase3 as well as increased p-PI3K in all treated animals.
The results revealed that cMSC-EVs efficiently improved animal behaviors after TBI by reducing cistauosis and apoptosis. Moreover, the EVs can be employed as an effective strategy for antibody delivery during passive immunotherapy.
•Mesenchymal stem cell-derived extracellular vesicles (MSC-EVs) improved behavioral recovery after traumatic brain injury.•MSC-EVs reduced cistauosis and apoptosis after traumatic brain injury.•MSC-EVs ameliorated traumatic brain injury as effective as anti-cis p-tau antibody.
The therapeutic potential of naturally secreted micro- and nanoscale extracellular vesicles (EVs) makes them attractive candidates for regenerative medicine and pharmaceutical science applications. ...To date, the results of numerous publications have shown the practicality of using EVs to replace mesenchymal stromal cells (MSCs) or liposomes. This article presents a systematic review of pre-clinical studies conducted over the past decade of MSC-derived EVs (MSC-EVs) used in animal models of disease. The authors searched the relevant literature in the PubMed and Scopus databases (9358 articles), and 690 articles met the inclusion criteria. The eligible articles were placed in the following disease categories: autoimmune, brain, cancer, eye, gastrointestinal, heart, inflammation/transplantation, liver, musculoskeletal, pancreas, spinal cord and peripheral nervous system, respiratory system, reproductive system, skin, urinary system and vascular-related diseases. Next, the eligible articles were assessed for the rate of publication and global distribution, methodology of EV isolation and characterization, route of MSC-EV administration, length of follow-up, source of MSCs and animal species. The current review classifies and critically discusses the technical aspects of these MSC-EV animal studies and discusses potential relationships between methodological details and the effectiveness of MSC-EVs as reported by these pre-clinical studies.
Extracellular vesicles (EVs) are nanosized vesicles with a lipid bilayer that are secreted by cells and play a critical role in cell‐to‐cell communication. Despite the promising reports regarding ...their diagnostic and therapeutic potential, the utilization of EVs in the clinical setting is limited due to insufficient information about their cargo and a lack of standardization in isolation and analysis methods. Considering protein cargos in EVs as key contributors to their therapeutic potency, we conducted a tandem mass tag (TMT) quantitative proteomics analysis of three subpopulations of mesenchymal stem cell (MSC)‐derived EVs obtained through three different isolation techniques: ultracentrifugation (UC), high‐speed centrifugation (HS), and ultracentrifugation on sucrose cushion (SU). Subsequently, we checked EV marker expression, size distribution, and morphological characterization, followed by bioinformatic analysis. The bioinformatic analysis of the proteome results revealed that these subpopulations exhibit distinct molecular and functional characteristics. The choice of isolation method impacts the proteome of isolated EVs by isolating different subpopulations of EVs. Specifically, EVs isolated through the high‐speed centrifugation (HS) method exhibited a higher abundance of ribosomal and mitochondrial proteins. Functional apoptosis assays comparing isolated mitochondria with EVs isolated through different methods revealed that HS‐EVs, but not other EVs, induced early apoptosis in cancer cells. On the other hand, EVs isolated using the sucrose cushion (SU) and ultracentrifugation (UC) methods demonstrated a higher abundance of proteins primarily involved in the immune response, cell‐cell interactions and extracellular matrix interactions. Our analyses unveil notable disparities in proteins and associated biological functions among EV subpopulations, underscoring the importance of meticulously selecting isolation methods and resultant EV subpopulations based on the intended application.
Amyotrophic lateral sclerosis (ALS) is a fatal, severely debilitating and rapidly progressing disorder affecting motor neurons in the brain, brainstem, and spinal cord. Unfortunately, there are few ...effective treatments, thus there remains a critical need to find novel interventions that can mitigate against its effects. Whilst the aetiology of ALS remains unclear, ageing is the major risk factor. Ageing is a slowly progressive process marked by functional decline of an organism over its lifespan. However, it remains unclear how ageing promotes the risk of ALS. At the molecular and cellular level there are specific hallmarks characteristic of normal ageing. These hallmarks are highly inter-related and overlap significantly with each other. Moreover, whilst ageing is a normal process, there are striking similarities at the molecular level between these factors and neurodegeneration in ALS. Nine ageing hallmarks were originally proposed: genomic instability, loss of telomeres, senescence, epigenetic modifications, dysregulated nutrient sensing, loss of proteostasis, mitochondrial dysfunction, stem cell exhaustion, and altered inter-cellular communication. However, these were recently (2023) expanded to include dysregulation of autophagy, inflammation and dysbiosis. Hence, given the latest updates to these hallmarks, and their close association to disease processes in ALS, a new examination of their relationship to pathophysiology is warranted. In this review, we describe possible mechanisms by which normal ageing impacts on neurodegenerative mechanisms implicated in ALS, and new therapeutic interventions that may arise from this.
This study aimed to investigate the therapeutic potential of a homogenous clonal population of mesenchymal stem cells (cMSC) and their extracellular vesicles (cMSC-EV) subpopulations on isolated rat ...islets in vitro and in inflammatory-mediated type 1 diabetes (T1D) non-human primate models.
EV subpopulations were isolated from human bone marrow-derived cMSC supernatant by low- and high-speed ultracentrifuge (EV-20K and EV-U110K) and sucrose density gradient (EV-S110K). The EVs were characterized generally and for the level of albumin, acetylcholinesterase (AChE) activity, co-isolate apoptotic markers, and expression of CD63+/annexin V+. Rat islet-derived single cells (iSCs) proliferation was measured using a Ki-67 proliferation assay. Diabetes was induced by multiple low-dose administrations of streptozotocin in rhesus monkeys. The diabetic monkeys were divided into three groups: the cMSC group, received two injections of 1.5 × 106 cMSC/kg body weight; the EV group received two injections of EVs isolated from 1.5 × 106 cMSC/kg, and the vehicle group received phosphate-buffered saline.
EV-S110K showed higher AChE activity, lower expression of CD63+/annexin V+, and lower apoptotic co-isolates. EV-S110K induced β-cell proliferation in vitro in a dose-dependent manner. The administration of EV-S110K and/or cMSC in diabetic monkeys demonstrated no significant changes in general diabetic indices and β-cell mass in the pancreas of the monkeys. Both treatments demonstrated a lowering trend in blood glucose levels and reduced pro-inflammatory cytokines. In contrast, regulatory T cells and anti-inflammatory cytokines were increased.
cMSC and cMSC-EV provided initial evidence to attenuate clinical symptoms in inflammatory-mediated T1D non-human primates through immunomodulation.
Background: Mesenchymal stem cells (MSCs) have been increasingly used in treatment of type 1 diabetes (T1D). In attention to disadvantages of cell therapy versus cell-free therapy, extracellular ...vesicles (EVs) released from MSCs have drawn wide attention as a promising alternative in cell therapy. In this study, we investigated the effect of human bone marrow mesenchymal stem cells derived EVs (hBMSC-EVs) on the function of dispersed rat islet cells in vitro. Methods: we used supernatant derived from the dynamic expansion of hBMSCs to isolate EVs through gradient ultracentrifugation. EVs were measured for their protein content using a BCA Protein Assay Kit and then characterized by electron microscopy and the size distribution of EVs was measured by dynamic light scattering (DLS) in order to measure the cytotoxicity of the dose-dependent manner of EVs, MTS assay was tested. Also, we tested if cells could uptake hBMSC-EVs labelled with red fluorescent PKH-26 to follow their functional assay on dispersed rat pancreatic islet cells. To evaluate the effect of hBMSC-derived EVs on single cell viability we assayed dispersed islet cells using fluorescein diacetate (FDA) and propidium iodide (PI) staining. Results: We quantified that according to the amount of 36 x 106 hBMSC could produce approximately 1218 µg exosomes and 1190 µg microvesicle. DLS and electron microscopy also have been done for the collected EVs. Cells were plated at 30,000 cells/well and incubated with exosomes at different concentration (0, 10,100 µg/ml) and the control (PBS) for 48 h. The nanoparticle have been shown to interact with MTS reagent and caused false positive results against the bright field microscopy images after co-culture of islet cells with PKH-26 labelled EVs at different time points (2, 24 and 48 h), the results showed that EVs could be internalized by islet cells. FDA-PI staining also showed the effect of hBMSC-EVs on the viability of dispersed rat islet cell. Summary/Conclusion: In this study, we have worked on the characterization of hBMSC-EVs and a cytotoxicity assays on dispersed rat islet cells in vitro.
Background: Mesenchymal stem cells (MSC) therapy is one of the most commonly employed cellular therapy in human clinical trials. Since MSCs secrete extracellular vesicles (EVs) to mediate in ...regeneration, EVs are undergoing extensive evaluation as a replacement or adjutant to cells in cellular therapy in pre-clinical studies. To date, there has been no meta-analysis of studies using MSC-EV therapy in animal studies. Methods: By searching systematically in PubMed and Scopus databases, more than 1000 reports were identified. After screening for eligibility, a total of approximately 100 studies are found to report MSC-EV therapy in animal disease models. Results: All the found pre-clinical studies reporting the therapeutic potential of MSC-derived EVs underwent comprehensive review, quality assessment and data extraction. Most of these studies employed animal models for kidney, heart, skin and lung disease as well as cancer. Although culture conditions of the EV-producing cells have overlapping characteristics, we discussed many different technical aspects, the methods of EV isolation and downstream characterizations that can make a significant difference in the overall results of animal studies. In addition, we explored the effect of clinically relevant in vivo animal administration parameters on functional outcome to better design future clinical studies utilizing MSC-EVs. Summary/conclusion: Our study provides new insight regarding parameters affecting the therapeutic potential of MSC-derived EVs in preclinical studies. Although there is significant improvement in animal disease model after MSC-derived EVs therapy, most of reports are still far from transparency required for designing a clinical trial.
