Jeyakumar Vivek, Halbwirth Florian, Niculescu-Morzsa Eugenia, Bauer Christoph, Zwickl Hannes, Kern Daniela, Nehrer Stefan
1 Centre for Regenerative Medicine and Orthopedics, Danube University Krems, Krems, Austria.
Cartilage. 2017 Apr;8(2):191-198. doi: 10.1177/1947603516657641. Epub 2016 Jul 7.
Objective The purpose of the current study was to compare the donor age variation of chondrocytes from non-OA (osteoarthritic) trauma joints in patients of young to middle age (20.5 ± 3.7, 31.8 ± 1.9, 41.9 ± 4.1 years) embedded in matrix-associated autologous chondrocyte transplantation (MACT) grafts (CaReS). The chondrocyte-specific gene expression of CaReS grafts were then compared to chondrocytes from OA joints (in patients aged 63.8 ± 10 years) embedded in a collagen type I hydrogel. Design OA chondrocytes and articular chondrocyte-laden grafts were cultured over 14 days in chondrogenic growth medium. We performed reverse transcriptase quantitative polymerase chain reaction (RT-qPCR) to evaluate the mRNA expression levels of chondrocyte-specific and hypertrophic markers. Results Gene expression analysis with RT-qPCR revealed no significant difference in chondrocyte-specific genes ( COL2A1, ACAN, SOX9, SOX5, SOX6) among 3 different age group of patients with CaReS grafts. In a comparative analysis of OA chondrocytes to articular chondrocytes, chondrogenic markers ( COL2A1, SOX6) exhibited higher expression in OA chondrocytes ( P < 0.05). Hypertrophic or OA cartilage pathogenesis marker ( MMP3, MMP13) expression was higher and COL1A1 had significantly lower expression ( P < 0.05) in OA chondrocytes than articular chondrocytes when cultivated in collagen type I hydrogels. Conclusion In summary, we identify that donor age variation does not influence the chondrogenic gene expression of the CaReS system. We also identified that freshly isolated OA chondrocytes embedded in collagen type I hydrogels can exhibit chondrogenic gene expression as observed in articular chondrocytes on the CaReS grafts. Transforming OA chondrocytes to articular chondrocytes can be regarded as an alternative option in the MACT technique.
目的 本研究旨在比较年轻至中年患者(20.5±3.7岁、31.8±1.9岁、41.9±4.1岁)非骨关节炎(OA)创伤关节软骨细胞嵌入基质相关自体软骨细胞移植(MACT)移植物(CaReS)中的供体年龄差异。然后将CaReS移植物的软骨细胞特异性基因表达与嵌入I型胶原水凝胶中的OA关节软骨细胞(63.8±10岁患者)进行比较。设计 将OA软骨细胞和负载关节软骨细胞的移植物在软骨生成生长培养基中培养14天。我们进行逆转录定量聚合酶链反应(RT-qPCR)以评估软骨细胞特异性和肥大标志物的mRNA表达水平。结果 RT-qPCR基因表达分析显示,CaReS移植物的3个不同年龄组患者的软骨细胞特异性基因(COL2A1、ACAN、SOX9、SOX5、SOX6)无显著差异。在OA软骨细胞与关节软骨细胞的比较分析中,软骨生成标志物(COL2A1、SOX6)在OA软骨细胞中表达较高(P<0.05)。在I型胶原水凝胶中培养时,OA软骨细胞中的肥大或OA软骨发病机制标志物(MMP3、MMP13)表达较高,而COL1A1表达显著较低(P<0.05)。结论 总之,我们发现供体年龄差异不影响CaReS系统的软骨生成基因表达。我们还发现,嵌入I型胶原水凝胶中的新鲜分离的OA软骨细胞可表现出与CaReS移植物上关节软骨细胞中观察到的软骨生成基因表达。将OA软骨细胞转化为关节软骨细胞可被视为MACT技术中的一种替代选择。