Yang Cheng, Ni Jiangdong, Zhang Shou, Fan Zhongcheng
Department of Orthopedics, Second Xiangya Hospital, Central South University, Changsha 410011, China.
Orthopedics Center, Haikou Hospital Affiliated to Xiangya School of Medicine, Central South University, Haikou 570100, China.
Zhong Nan Da Xue Xue Bao Yi Xue Ban. 2017 Aug 28;42(8):919-926. doi: 10.11817/j.issn.1672-7347.2017.08.008.
To investigate the feasibility of construction of tissue engineered cartilage by co-culture of bone marrow mesenchymal stem cells (BMSCs) and costal chondrocytes (CCs), and to provide theoretical basis and experimental basis for clinical repair of articular cartilage defects by Wuzhishan miniature pig knee cartilage defects with co-cultured cells. Methods: Density gradient centrifugation method was used to isolate BMSCs from Wuzhishan miniature pig. The double enzyme digestion method was used to isolate CCs. The passage 3 generation of BMSCs and passage 2 generation of CCs were randomly divided into 3 groups: a co-culture group of BMSCs:CCs for 1:2 (Group A), a simple CCs (Group B), and a simple BMSCs (Group C). The cell growth curve was drawn, and the content of glycosaminoglycan (GAG) of external separation in chondrocytes was determined. The 12 Wuzhishan miniature pigs were randomly divided into a co-culture cells/collagen membrane experimental group, a collagen membrane control group and the blank group. In the co-culture cells/collagen membrane experimental group, the co-cultured cells/collagen membrane were implanted into the cartilage defects of the mandibular condyle; in the collagen membrane control group, only collagen membrane was implanted; while in the blank group, nothing was implanted. Six animals were sacrificed at 8 and 16 weeks after surgery respectively (2 animals in each group). General observation, cartilage histological score and histopathological examination were carried out. Results: The BMSCs and co-culture cells grew well. The biological activity of CCs was good. After 16 weeks of operation, the repair tissues in the co-cultured cells/collagen membrane experimental group showed hyaline cartilage features: smooth, flat, and integrated well with the surrounding cartilage and subchondral bone. The collagen membrane in the collagen membrane control group was fibrously repaired. Repair tissue gross score in the co-culture cells/collagen membrane experimental group was significantly better than that in the collagen membrane control group and the blank group (both P<0.05), but there was no significant difference between the collagen membrane control group and the blank group (P>0.05). Conclusion: BMSCs, CCs and co-cultured cells can function as the seed cells for cartilage tissue engineering, and the co-culture cells (BMSCs:CCs=1:2) possess more advantages; the short-term effect of co-culture cells with collagen membrane on repairing cartilage defects is satisfied.
探讨骨髓间充质干细胞（BMSCs）与肋软骨细胞（CCs）共培养构建组织工程软骨的可行性，为五指山小型猪膝关节软骨缺损采用共培养细胞进行临床修复提供理论依据和实验依据。方法：采用密度梯度离心法从五指山小型猪分离BMSCs，采用双酶消化法分离CCs。将第3代BMSCs和第2代CCs随机分为3组：BMSCs与CCs按1∶2共培养组（A组）、单纯CCs组（B组）、单纯BMSCs组（C组）。绘制细胞生长曲线，检测软骨细胞外泌糖胺聚糖（GAG）含量。将12只五指山小型猪随机分为共培养细胞/胶原膜实验组、胶原膜对照组和空白组。共培养细胞/胶原膜实验组将共培养细胞/胶原膜植入下颌髁突软骨缺损处；胶原膜对照组仅植入胶原膜；空白组不植入任何材料。术后8周和16周分别处死6只动物（每组2只），进行大体观察、软骨组织学评分及组织病理学检查。结果：BMSCs及共培养细胞生长良好，CCs生物学活性良好。术后16周，共培养细胞/胶原膜实验组修复组织呈透明软骨特征：表面光滑、平整，与周围软骨及软骨下骨整合良好。胶原膜对照组胶原膜呈纤维性修复。共培养细胞/胶原膜实验组修复组织大体评分显著优于胶原膜对照组和空白组（均P<0.05），但胶原膜对照组与空白组比较差异无统计学意义（P>0.05）。结论：BMSCs、CCs及共培养细胞均可作为软骨组织工程种子细胞，共培养细胞（BMSCs∶CCs=1∶2）具有更多优势；共培养细胞与胶原膜修复软骨缺损的短期效果满意。
