William Edward Hotham, Charlotte Thompson, Lin Szu-Ting, Frances Margaret Daphne Henson
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However, it is hypothesised that mesenchymal stem cells (MSC) exert their effects via paracrine factors including the production of extracellular vesicles that can themselves recapitulate the MSC effects in the joint.</p><p><strong>Objectives: </strong>To isolate extracellular vesicles from bone marrow MSC and investigate their anti-inflammatory effects on chondrocytes.</p><p><strong>Study design: </strong>An in vitro assessment of the effect of direct culturing extracellular vesicles on artificially inflamed chondrocytes.</p><p><strong>Methods: </strong>Extracellular vesicles were isolated from bone marrow MSC using differential sequential ultracentrifugation. Vesicles were characterised using electron microscopy, nanoparticle tracing analysis and protein analysis. Vesicle internalisation was carried out via vesicles being pre-stained and co-cultured with equine chondrocytes before analysis using confocal microscopy. The effects of vesicles on artificially inflamed chondrocytes was examined using quantitative PCR.</p><p><strong>Results: </strong>To the best of the authors' knowledge, this is the first study to isolate and characterise extracellular vesicles from equine bone MSC. Vesicles were taken up by autologous chondrocytes and had anti-inflammatory effects on gene expression following chondrocyte exposure to tumour necrosis factor α and Interleukin 1β.</p><p><strong>Main limitations: </strong>Only three independent biological repeats were performed and the work was done in vitro.</p><p><strong>Conclusion: </strong>Extracellular vesicles can be isolated from equine bone marrow MSC; they may be taken up by chondrocytes and have an anti-inflammatory action.</p>","PeriodicalId":23565,"journal":{"name":"Veterinary Record Open","volume":null,"pages":null},"PeriodicalIF":1.3000,"publicationDate":"2021-11-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8580791/pdf/","citationCount":"0","resultStr":"{\"title\":\"The anti-inflammatory effects of equine bone marrow stem cell-derived extracellular vesicles on autologous chondrocytes.\",\"authors\":\"William Edward Hotham, Charlotte Thompson, Lin Szu-Ting, Frances Margaret Daphne Henson\",\"doi\":\"10.1002/vro2.22\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><strong>Background: </strong>Osteoarthritis (OA) in the horse is an economic and welfare issue and there are no current disease modifying drugs available. 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引用次数: 0
摘要
背景:马的骨关节炎(OA)是一个经济和福利问题,目前还没有改变疾病的药物。干细胞被认为是治疗 OA 的干预措施,最初是基于其再生能力。然而,有一种假设认为,间充质干细胞(MSC)是通过旁分泌因子(包括产生细胞外囊泡)发挥其作用的,而细胞外囊泡本身可在关节中重现间充质干细胞的作用:从骨髓间充质干细胞中分离出细胞外囊泡,并研究它们对软骨细胞的抗炎作用:研究设计:体外评估直接培养细胞外囊泡对人工发炎软骨细胞的影响:采用差序超速离心法从骨髓间充质干细胞中分离出细胞外小泡。利用电子显微镜、纳米颗粒追踪分析和蛋白质分析对囊泡进行表征。在使用共聚焦显微镜进行分析之前,先对囊泡进行预染色并与马软骨细胞共同培养,从而实现囊泡内化。使用定量 PCR 检测囊泡对人工发炎软骨细胞的影响:据作者所知,这是首次从马骨间质干细胞中分离出细胞外囊泡并对其进行定性的研究。囊泡被自体软骨细胞吸收,并在软骨细胞暴露于肿瘤坏死因子α和白细胞介素1β后对基因表达产生抗炎作用:主要局限性:只进行了三次独立的生物重复,而且工作是在体外完成的:结论:可从马骨髓间充质干细胞中分离出细胞外囊泡;它们可被软骨细胞吸收并具有抗炎作用。
The anti-inflammatory effects of equine bone marrow stem cell-derived extracellular vesicles on autologous chondrocytes.
Background: Osteoarthritis (OA) in the horse is an economic and welfare issue and there are no current disease modifying drugs available. Stem cells have been suggested as a therapeutic intervention for OA, originally on the basis of their regenerative capacity. However, it is hypothesised that mesenchymal stem cells (MSC) exert their effects via paracrine factors including the production of extracellular vesicles that can themselves recapitulate the MSC effects in the joint.
Objectives: To isolate extracellular vesicles from bone marrow MSC and investigate their anti-inflammatory effects on chondrocytes.
Study design: An in vitro assessment of the effect of direct culturing extracellular vesicles on artificially inflamed chondrocytes.
Methods: Extracellular vesicles were isolated from bone marrow MSC using differential sequential ultracentrifugation. Vesicles were characterised using electron microscopy, nanoparticle tracing analysis and protein analysis. Vesicle internalisation was carried out via vesicles being pre-stained and co-cultured with equine chondrocytes before analysis using confocal microscopy. The effects of vesicles on artificially inflamed chondrocytes was examined using quantitative PCR.
Results: To the best of the authors' knowledge, this is the first study to isolate and characterise extracellular vesicles from equine bone MSC. Vesicles were taken up by autologous chondrocytes and had anti-inflammatory effects on gene expression following chondrocyte exposure to tumour necrosis factor α and Interleukin 1β.
Main limitations: Only three independent biological repeats were performed and the work was done in vitro.
Conclusion: Extracellular vesicles can be isolated from equine bone marrow MSC; they may be taken up by chondrocytes and have an anti-inflammatory action.
期刊介绍:
Veterinary Record Open is a journal dedicated to publishing specialist veterinary research across a range of topic areas including those of a more niche and specialist nature to that considered in the weekly Vet Record. Research from all disciplines of veterinary interest will be considered. It is an Open Access journal of the British Veterinary Association.