Extracellular Vesicle Preparation and Analysis: A State‐of‐the‐Art Review

Advanced Science, Год журнала: 2024, Номер 11(30)

Опубликована: Июнь 14, 2024

Abstract In recent decades, research on Extracellular Vesicles (EVs) has gained prominence in the life sciences due to their critical roles both health and disease states, offering promising applications diagnosis, drug delivery, therapy. However, inherent heterogeneity complex origins pose significant challenges preparation, analysis, subsequent clinical application. This review is structured provide an overview of biogenesis, composition, various sources EVs, thereby laying groundwork for a detailed discussion contemporary techniques preparation analysis. Particular focus given state‐of‐the‐art technologies that employ microfluidic non‐microfluidic platforms EV processing. Furthermore, this discourse extends into innovative approaches incorporate artificial intelligence cutting‐edge electrochemical sensors, with particular emphasis single proposes current outlines prospective avenues future research. The objective motivate researchers innovate expand methods analysis fully unlocking biomedical potential.

Язык: Английский

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Clinical-grade extracellular vesicles derived from umbilical cord mesenchymal stromal cells: preclinical development and first-in-human intra-articular validation as therapeutics for knee osteoarthritis

Journal of Nanobiotechnology, Год журнала: 2025, Номер 23(1)

Опубликована: Янв. 13, 2025

Osteoarthritis (OA) is a joint disease characterized by articular cartilage degradation. Persistent low-grade inflammation defines OA pathogenesis, with crucial involvement of pro-inflammatory M1-like macrophages. While mesenchymal stromal cells (MSC) and their small extracellular vesicles (sEV) hold promise for treatment, achieving consistent clinical-grade sEV products remains significant challenge. This study aims to develop fully characterized, reproducible, batches derived from umbilical cord (UC)-MSC the treatment while assessing its efficacy safety. Initially, standardized, research-grade manufacturing protocol was established ensure production. UC-MSC-sEV characterization under non-cGMP conditions showed miRNA protein profiles, suggesting potential standardized manufacturing. In vitro studies evaluated efficacy, safety, potency sEV; animal confirmed effectiveness vitro, polarized macrophages an anti-inflammatory M2b-like phenotype, through STAT1 modulation, indicating create environment in affected joints. silico sEV's immunosuppressive signature proteome analysis. mouse model, injected intra-articularly (IA) induced hyaline regeneration, validated histological μCT analyses. The unique detection signals within knee over time highlights safety profile confirming retention joint. product development involved refining, standardizing, validating processes compliance GMP standards. initial assessment via IA administration first-in-human no adverse effects after 12 month follow-up period. These results support progress this sEV-based therapy early-phase clinical trial, details which are presented discussed work. provides data on using as local OA, highlighting regenerative properties preclinical proof-of-principle application.

Язык: Английский

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Therapeutic role of hucMSC-sEV-enriched miR-13896 in cisplatin-induced acute kidney injury through M2 macrophage polarization

Cell Biology and Toxicology, Год журнала: 2025, Номер 41(1)

Опубликована: Фев. 24, 2025

Human umbilical cord mesenchymal stem cell-derived small extracellular vesicles (hucMSC-sEV) have recently garnered attention as a potential therapeutic approach for kidney diseases with anti-inflammatory effects. Infiltrated macrophages play an important role in facilitating tissue regeneration. However, the intricate regulatory effects of hucMSC-sEV on during cisplatin-induced acute injury (AKI) remain unknown. In this study, we uncovered that exhibited potent anti-inflammation and effectively inhibited polarization M1 phenotype macrophages. Mechanically, miRNA sequencing analysis qRT-PCR indicated novel miRNA, named miR-13896, was enriched hucMSC-sEV. When transfected miR-13896 mimic, displayed M2 elevated levels Arg1 IL-10, while inhibitor promoted phenotype. Furthermore, firstly established repressed Tradd expression by targeting its 3' untranslated region subsequently NF-κB signaling pathway Additionally, to improve effects, were engineered through electroporation, which resulted promoting macrophages, inhibiting inflammatory factors, enhancing repair. Conclusively, our findings provide insights into mechanisms underlying AKI, also highlighting electroporation promising strategy treating AKI.

Язык: Английский

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Engineering exosomes derived from TNF-α preconditioned IPFP-MSCs enhance both yield and therapeutic efficacy for osteoarthritis

Journal of Nanobiotechnology, Год журнала: 2024, Номер 22(1)

Опубликована: Сен. 11, 2024

Язык: Английский

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ECM stiffness affects cargo sorting into MSC-EVs to regulate their secretion and uptake behaviors

Journal of Nanobiotechnology, Год журнала: 2024, Номер 22(1)

Опубликована: Март 21, 2024

Abstract Mesenchymal stem cell-derived extracellular vesicles (MSC-EVs) have garnered extensive attention as natural product-based nanomedicines and potential drug delivery vehicles. However, the specific mechanism for regulating MSC-EVs secretion remains unclear. Here, we demonstrate that matrix (ECM) stiffness regulates of EVs by affecting MSCs' cargo sorting mechanically. Using multi-omics analysis, found a decrease in ECM impeded vesicular transport-related proteins autophagy-related lipids into MSC-EVs, impairing their subsequent uptake macrophages. Hence, with different behaviors can be produced changing culture substrates. This study provides new insights MSC-EV biology establishes connection between from biophysical perspective, providing basis rational design biomedical materials. Graphical

Язык: Английский

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Engineered MSC‐sEVs as a Versatile Nanoplatform for Enhanced Osteoarthritis Treatment via Targeted Elimination of Senescent Chondrocytes and Maintenance of Cartilage Matrix Metabolic Homeostasis

Advanced Science, Год журнала: 2025, Номер unknown

Опубликована: Янв. 4, 2025

Abstract Chondrocyte senescence is an important pathogenic factor causing osteoarthritis (OA) progression through persistently producing pro‐inflammatory factors. Mesenchymal stem cells‐derived small extracellular vesicles (MSC‐sEVs) have shown anti‐inflammatory effects in OA models, while persistent existence of senescent chondrocytes still promotes cartilage destruction. Therefore, improving the targeted elimination ability on required to facilitate translation MSC‐sEVs treatment. In this study, versatile engineered are developed targetedly clear and maintain metabolic homeostasis. Specifically, loaded with siRNA mouse double minute 2 homologue (siMDM2) modified cartilage‐targeting peptide WYRGRL‐PEG 2K ‐DSPE (WPD), named WPD‐sEVs siMDM2 . The results demonstrate modification improves cellular uptake chondrocytes, thus antiaging effects. Importantly, multifunctional enhances penetration extends joint retention time MSC‐sEVs. both post‐traumatic mice naturally aged mice, more effectively eliminates maintained matrix By using P53 phosphorylation inhibitor, essential role MDM2‐P53 pathway function verified. ex vivo cultured human explants, it confirmed that alleviates phenotype. Altogether, findings suggest promising translational potential for

Язык: Английский

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Efficacy and safety of mesenchymal stromal cell transplantation in the treatment of autoimmune and rheumatic immune diseases: a systematic review and meta-analysis of randomized controlled trials

Stem Cell Research & Therapy, Год журнала: 2025, Номер 16(1)

Опубликована: Фев. 11, 2025

This study aims to assess the effectiveness and safety of mesenchymal stem cell (MSC) transplantation in treatment autoimmune rheumatic immune diseases through randomized controlled trials (RCTs). Two researchers conducted a comprehensive search Chinese English databases from their inception until Dec. 2023. The literature screening data extraction were then performed. Statistical analysis was carried out using RevMan 5.4 software. A total 42 relevant RCTs, involving 2,183 participants, ultimately included this study. These RCTs encompassed four types bone diseases, namely rheumatoid arthritis (RA), osteoarthritis (OA), spondyloarthritis, systemic sclerosis arthritis, lupus erythematosus (SLE), inflammatory bowel disease, multiple sclerosis, primary Sjögren's syndrome (PSS). systematic review indicates that MSC may improve RA, PSS. meta-analysis reveals significantly improved symptoms patients with OA [VAS (visual analogue scale): marrow: SMD = − 0.95, 95% CI 1.55 0.36, P 0.002; umbilical cord: 1.25, 2.04 0.46, adipose tissue: -1.26, -1.99 0.52, 0.0009)], SLE [Systemic disease activity index (SLEDAI): 2.32, 3.59 1.06, 0.0003], [clinical efficacy: RR 2.02, 1.53 2.67, < 0.00001]. However, not (Ssc). Importantly, did increase incidence adverse events (OA: 1.23, 0.93 1.65, 0.15; SLE: 0.83, 0.28 2.51, 0.76; Inflammatory disease: 0.99, 0.81 1.22, 0.96; Multiple sclerosis: 1.12, 1.53, 0.50), supporting its profile across studies. findings suggest holds promise for several while highlighting areas where further research is warranted. have potential treat diseases. Moreover. appears be relatively safe could considered as viable alternative option

Язык: Английский

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Therapeutics of the future: Navigating the pitfalls of extracellular vesicles research from an osteoarthritis perspective

Journal of Extracellular Vesicles, Год журнала: 2024, Номер 13(7)

Опубликована: Июнь 28, 2024

Abstract Extracellular vesicles have gained wide momentum as potential therapeutics for osteoarthritis, a highly prevalent chronic disease that still lacks an approved treatment. The membrane‐bound are secreted by all cells carrying different cargos can serve both biomarkers and modifiers. Nonetheless, despite significant peak in research regarding EVs OA therapeutics, clinical implementation seems distant. In addition to scalability standardization challenges, researchers often omit focus on consider the proper tropism of vesicles, practicality relevance their source, low native therapeutic efficacy, whether they address whole. These considerations necessary better understand light been comprehensively discussed ultimately summarized this review into conceptualized framework termed nanodiamond concept. Future perspectives also discussed, alternatives presented some challenges concerns.

Язык: Английский

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Allogenic bone marrow–derived mesenchymal stromal cell–based therapy for patients with chronic low back pain: a prospective, multicentre, randomised placebo controlled trial (RESPINE study)

Annals of the Rheumatic Diseases, Год журнала: 2024, Номер 83(11), С. 1572 - 1583

Опубликована: Окт. 11, 2024

Objectives

To assess the efficacy of a single intradiscal injection allogeneic bone marrow mesenchymal stromal cells (BM-MSCs) versus sham placebo in patients with chronic low back pain (LBP).

Methods

Participants were randomised prospective, double-blind, controlled study to receive either or 20 million BM-MSC, between April 2018 and December 2022. The first co-primary endpoint was rate responders defined by improvement Visual Analogue Scale (VAS) for at least 20% mm, Oswestry Disability Index (ODI) baseline month 12. secondary structural assessed disc fluid content measured quantitative MRI T2, Secondary endpoints included VAS, ODI, Short Form (SF)-36 minimal clinically important difference all timepoints (1, 3, 6, 12 24 months). We determined immune response associated cell 6 months. Serious adverse events (SAEs) recorded.

Results

114 (n=58, BM-MSC group; n=56, group). At months, primary outcome not reached (74% group vs 69% p=0.77). groups did differ outcomes. No SAE related intervention occurred.

Conclusions

While our conclusively demonstrate BM-MSCs LBP, procedure safe. Long-term outcomes MSC therapy LBP are still being studied.

Trial registration number

EudraCT 2017-002092-25/ClinicalTrials.gov: NCT03737461.

Язык: Английский

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Aberrant Activation of Immune and Non-Immune Cells Contributes to Joint Inflammation and Bone Degradation in Rheumatoid Arthritis

International Journal of Molecular Sciences, Год журнала: 2023, Номер 24(21), С. 15883 - 15883

Опубликована: Ноя. 1, 2023

Abnormal activation of multiple immune and non-immune cells proinflammatory factors mediate the development joint inflammation in genetically susceptible individuals. Although specific environmental like smoking infections are associated with disease pathogenesis, until now, we did not know autoantigens arthritogenic that trigger initiation clinical disease. Autoantibodies recognizing post-translationally modified unmodified antigens generated circulation before onset disease, could serve as diagnostic prognostic markers. The characteristic features autoantibodies change regarding sub-class, affinity, glycosylation pattern, epitope spreading onset. Some these antibodies were proven to be pathogenic using animal cell-culture models. However, all them can induce animals. This review discusses aberrant major contributing inflammation. Recent studies explored protective effects extracellular vesicles from mesenchymal stem bacteria on joints by targeting pathways. Current therapeutics clinics target inflammatory pathways attenuate protect cartilage bones degradation, but none cure Hence, more basic research is needed investigate triggers mechanisms involved initiating relapses prevent chronic damaging architecture.

Язык: Английский

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