基于器械的膝关节滑膜细胞富集以驱动 MSC 软骨形成,无需体外预先培养扩增:更接近一步法骨科手术。
Device-Based Enrichment of Knee Joint Synovial Cells to Drive MSC Chondrogenesis Without Prior Culture Expansion In Vitro: A Step Closer to 1-Stage Orthopaedic Procedures.
机构信息
Leeds Institute of Rheumatic and Musculoskeletal Medicine, University of Leeds, Leeds, UK.
Leeds Biomedical Research Centre, National Institute for Health Research, Leeds, UK.
出版信息
Am J Sports Med. 2022 Jan;50(1):152-161. doi: 10.1177/03635465211055164. Epub 2021 Nov 15.
BACKGROUND
Synovial fluid (SF) mesenchymal stem cells (MSCs) are derived from the synovial membrane and have cartilage repair potential. Their current use in clinical practice is largely exploratory. As their numbers tend to be small, therapeutic procedures using MSCs typically require culture expansion. Previous reports indicate that the stem cell-mobilizing device (STEM device) intraoperatively increases SF-MSCs.
PURPOSE
This study evaluated the chondrogenic potential of non-culture expanded synovium-mobilized MSCs and SF-microfragments obtained after enrichment using the STEM device and ascertained if device-mediated synovial membrane manipulation facilitated ongoing MSC release.
STUDY DESIGN
Controlled laboratory study.
METHODS
Two samples of aspiration fluid were collected intraoperatively before and after STEM device utilization from patients (n = 16) undergoing diagnostic or therapeutic knee arthroscopy. Human knee synovium (n = 5) was collected during total knee replacement, and a suspended culture was performed to assess the effect of the STEM device on ongoing MSC release. Colony forming unit-fibroblastic assays were used to determine the number of MSCs. Additionally, cytometric characterization of stromal and immune cells and chondrogenesis differentiation assay were performed without culture expansion. Filtered platelet concentrates were prepared using the HemaTrate system.
RESULTS
After STEM device use, a significant increase was evident in SF-MSCs ( = .03) and synovial fluid-resident synovial tissue microfragments ( = .03). In vitro-suspended synovium released significantly more MSCs following STEM device use than nonstimulated synovium ( = .01). The STEM device-released total cellular fraction produced greater in vitro chondrogenesis with significantly more glycosaminoglycans (GAGs; < .0001) when compared with non-STEM device synovial fluid material. Nonexpanded SF-MSCs and SF-microfragments combined with autologous filtered platelet concentrate produced significantly more GAGs than the complete chondrogenic media ( < .0001). The STEM device-mobilized cells contained more M2 macrophage cells and fewer M1 cells.
CONCLUSION
Non-culture expanded SF-MSCs and SF-microfragments had the potential to undergo chondrogenesis without culture expansion, which can be augmented using the STEM device with increased MSC release from manipulated synovium for several days. Although preliminary, these findings offer proof of concept toward manipulation of the knee joint environment to facilitate endogenous repair responses.
CLINICAL RELEVANCE
Although numbers were small, this study highlights 3 factors relevant to 1-stage joint repair using the STEM device: increased SF-MSCs and SF-microfragments and prolonged synovial release of MSCs. Joint repair strategies involving endogenous MSCs for cartilage repair without the need for culture expansion in a 1-stage procedure may be possible.
背景
滑液(SF)间充质干细胞(MSCs)来源于滑膜,具有软骨修复潜力。它们目前在临床实践中的应用主要是探索性的。由于其数量往往较少,使用 MSC 的治疗程序通常需要培养扩增。先前的报告表明,手术中使用的滑膜动员装置(STEM 装置)可增加 SF-MSCs。
目的
本研究评估了非培养扩增的滑膜动员 MSC 和使用 STEM 装置富集后的 SF-微碎片的软骨形成潜力,并确定装置介导的滑膜处理是否促进了 MSC 的持续释放。
研究设计
对照实验室研究。
方法
从接受诊断或治疗性膝关节镜检查的患者(n=16)术中在使用 STEM 装置前后采集 2 份抽吸液样本。在全膝关节置换期间收集人膝关节滑膜(n=5),并进行悬浮培养以评估 STEM 装置对持续 MSC 释放的影响。集落形成单位-成纤维细胞测定用于确定 MSC 的数量。此外,在没有培养扩增的情况下,还进行了间充质和免疫细胞的流式细胞术特征分析以及软骨分化测定。使用 HemaTrate 系统制备血小板浓缩物。
结果
使用 STEM 装置后,SF-MSCs(=.03)和滑膜液驻留滑膜组织微碎片(=.03)明显增加。与未受刺激的滑膜相比,使用 STEM 装置后悬浮培养的滑膜释放出更多的 MSC(=.01)。与非 STEM 装置滑膜液材料相比,STEM 装置释放的总细胞部分产生了更多的体外软骨形成,且糖胺聚糖(GAGs)明显更多(<.0001)。未扩增的 SF-MSCs 和 SF-微碎片与自体过滤血小板浓缩物结合可产生比完整软骨形成培养基更多的 GAGs(<.0001)。STEM 装置动员的细胞含有更多的 M2 巨噬细胞和更少的 M1 细胞。
结论
未经培养扩增的 SF-MSCs 和 SF-微碎片具有在不进行培养扩增的情况下进行软骨形成的潜力,而使用 STEM 装置可增加 MSC 释放,并可通过几天内对受处理滑膜的 MSC 释放进行多次操作来增强。尽管这是初步研究,但这些发现为通过操纵膝关节环境促进内源性修复反应来实现 1 期关节修复提供了概念验证。
临床相关性
尽管数量较少,但本研究强调了使用 STEM 装置进行 1 期关节修复的 3 个相关因素:SF-MSCs 和 SF-微碎片增加以及滑膜 MSC 释放时间延长。涉及软骨修复的内源性 MSC 的关节修复策略可能无需在 1 期手术中进行培养扩增即可实现。