Sun Hao, Huang Yue, Zhang Lei, Li Biao, Wang Xudong
Department of Oral and Cranio-Maxillofacial Surgery, Shanghai Ninth People's Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 200011, P.R. China.
Department of Oral and Maxillofacial Surgery, The First Affiliated Hospital of Fujian Medical University, Fuzhou, Fujian 350005, P.R. China.
Exp Ther Med. 2018 Oct;16(4):2969-2977. doi: 10.3892/etm.2018.6551. Epub 2018 Aug 1.
This study explored the feasibility of inducing the differentiation of BMSCs into chondrocytes through co-culture with chondrocytes in hydrogel constructs (Pluronic F-127 gel) for the repair of goat mandibular condylar cartilage defects. Chondrocytes and BMSCs were isolated from goat auricular cartilage and bone marrow, respectively, and were mixed at a ratio of 3:7. BMSCs were labelled with green fluorescence protein (GFP) using a retrovirus vector for tracing. Mixed cells were re-suspended in 30% Pluronic F-127 at a concentration of 5×10 cells/ml to form a gel-cell complex. The gel-cell complex was implanted into the temporomandibular joint condylar articular cartilage defects. The whole temporomandibular joint and adjacent tissues were harvested at 4, 8, and 12 weeks after surgery, and gross observation, histology and collagen II expression were evaluated. In the co-culture group, cartilage-like tissues were formed, and abundant type II collagen could be detected by immunohistochemistry in the condylar cartilage defects. Confocal microscopy revealed that implanted GFP-labelled BMSCs were embedded in cartilage-like tissues. The co-culture system described herein provides a chondrogenic microenvironment to induce the chondrogenic differentiation of BMSCs without any additional cellular factors.
本研究探讨了通过在水凝胶构建体(普朗尼克F-127凝胶)中与软骨细胞共培养诱导骨髓间充质干细胞(BMSCs)向软骨细胞分化以修复山羊下颌髁突软骨缺损的可行性。分别从山羊耳廓软骨和骨髓中分离软骨细胞和BMSCs,并以3:7的比例混合。使用逆转录病毒载体用绿色荧光蛋白(GFP)标记BMSCs以进行追踪。将混合细胞以5×10个细胞/ml的浓度重悬于30%的普朗尼克F-127中以形成凝胶-细胞复合物。将凝胶-细胞复合物植入颞下颌关节髁突关节软骨缺损处。在术后4、8和12周采集整个颞下颌关节及相邻组织,进行大体观察、组织学检查和Ⅱ型胶原表达评估。在共培养组中,形成了软骨样组织,在髁突软骨缺损处通过免疫组织化学可检测到丰富的Ⅱ型胶原。共聚焦显微镜显示,植入的GFP标记的BMSCs嵌入软骨样组织中。本文所述的共培养系统提供了一种软骨生成微环境,可在无任何额外细胞因子的情况下诱导BMSCs的软骨生成分化。