Vinod Elizabeth, Parasuraman Ganesh, J Jeya Lisha, Varghese Jithu James, Livingston Abel, Rebekah Grace, Francis Deepak Vinod, Sathishkumar Solomon, Daniel Alfred Job, Ramasamy Boopalan
Department of Physiology, Christian Medical College, Vellore, Tamil Nadu, 632002, India.
Centre for Stem Cell Research (A Unit of InStem, Bengaluru), Christian Medical College, Vellore, India.
Biotechnol Lett. 2025 Jan 8;47(1):17. doi: 10.1007/s10529-024-03558-0.
Cartilage repair necessitates adjunct therapies such as cell-based approaches, which commonly use MSCs and chondrocytes but is limited by the formation of fibro-hyaline cartilage. Articular cartilage-derived chondroprogenitors(CPs) offer promise in overcoming this, as they exhibit higher chondrogenic and lower hypertrophic phenotypes. The study aimed to compare the efficacy of various cell types derived from adult and foetal cartilage suspended in platelet-rich plasma(PRP) in repairing chondral defects in an Ex-vivo Osteochondral Unit(OCU) model.
In-vitro characterization of the cells included growth kinetics, FACS, qRT-PCR, and multilineage differentiation potential using histology and GAG analysis. Ex-vivo human OCUs with chondral defects containing the different cells in PRP were cultured and subjected to analysis for matrix and collagen staining.
The ex-vivo OCU analysis, in terms of defect repair, showed that adult chondrocytes, sorted-CPs, and foetal MCPs displayed better host integration and filling. The In-vitro analysis of adult chondrocytes displayed greater chondrogenic genes ACAN and COL2A1 expression, with sorted-CPs also showing higher levels of ACAN. In terms of accumulation of extracellular matrix uptake evident by Safranin O staining and collagen type II fibrillar uptake, the AD-MSCs, BM-MSCs, and sorted CPs outperformed the other groups. BM-MSCs also showed corroborative higher CD146 levels, however, the gene analysis of the AD-MSCs showed a high hypertrophic tendency in terms of its COL1A1 and RUNX2 expression.
Sorted chondroprogenitors outperformed both in terms of filling and hyaline-like repair, with AD-MSC and BM-MSC groups also achieving functional cartilage of a hyaline nature, warranting further evaluation using in-vivo and clinical studies.
软骨修复需要辅助治疗，如基于细胞的方法，该方法通常使用间充质干细胞（MSCs）和软骨细胞，但受纤维透明软骨形成的限制。关节软骨来源的软骨祖细胞（CPs）有望克服这一问题，因为它们表现出更高的软骨生成和更低的肥大表型。本研究旨在比较悬浮于富血小板血浆（PRP）中的成人和胎儿软骨来源的各种细胞类型在体外骨软骨单元（OCU）模型中修复软骨缺损的效果。
细胞的体外特性包括生长动力学、流式细胞术（FACS）、定量逆转录聚合酶链反应（qRT-PCR），以及使用组织学和糖胺聚糖（GAG）分析的多谱系分化潜能。对含有PRP中不同细胞的体外人OCU软骨缺损进行培养，并进行基质和胶原染色分析。
就缺损修复而言，体外OCU分析表明，成人软骨细胞、分选的CPs和胎儿间充质软骨祖细胞（MCPs）表现出更好的宿主整合和填充。成人软骨细胞的体外分析显示软骨生成基因ACAN和COL2A1表达更高，分选的CPs也显示出更高水平的ACAN。就番红O染色和II型胶原纤维摄取所显示的细胞外基质摄取积累而言，脂肪来源的MSCs（AD-MSCs）、骨髓来源的MSCs（BM-MSCs）和分选的CPs优于其他组。BM-MSCs也显示出更高的CD146水平，然而，AD-MSCs的基因分析显示其COL1A1和RUNX2表达具有较高的肥大倾向。
分选的软骨祖细胞在填充和类透明软骨修复方面均表现出色，AD-MSC和BM-MSC组也实现了透明软骨性质的功能性软骨，值得通过体内和临床研究进行进一步评估。
