Department of Otolaryngology, Eye and ENT Hospital of Shanghai Medical School, Fudan University, Shanghai, China.
J Biomed Mater Res A. 2014 Apr;102(4):1071-8. doi: 10.1002/jbm.a.34763. Epub 2013 Jun 11.
To explore the feasibility of constructing a functional biomaterial complex with regenerated silk fibroin membrane and immortalized chondrocytes in vivo. Rat auricular chondrocytes (RACs) were transfected with the lentivirus vector pGC-FU-hTERT-3FLAG or pGC-FU-GFP-3FLAG, encoding the human telomerase reverse transcriptase (hTERT) or GFP gene. The effects of regenerated silk fibroin film on the adhesion, growth of immortalized chondrocytes and expression of collagen II in vitro were analyzed with immunofluorescent histochemistry. Immortalized RACs were transformed. Induction by nutrient medium promoted higher expression levels of collagen II in transformed chondrocytes. The regenerated silk fibroin film was not cytotoxic to immortalized chondrocytes and had no adverse influence on their adhesion. Collagen II expression was good in the immortalized chondrocytes in vivo. The construction of a silk-based biomaterial complex with immortalized chondrocytes may provide a feasible kind of functional biomaterial for the repair of cartilage defects in clinical applications.
探讨构建具有再生丝素蛋白膜和永生化软骨细胞的功能性生物材料复合体的可行性。用携带人端粒酶逆转录酶（hTERT）或 GFP 基因的慢病毒载体 pGC-FU-hTERT-3FLAG 或 pGC-FU-GFP-3FLAG 转染大鼠耳软骨细胞（RAC）。采用免疫荧光组织化学方法分析再生丝素蛋白膜对永生化软骨细胞黏附、生长和Ⅱ型胶原表达的影响。永生化 RAC 转化。诱导营养培养基可促进转化软骨细胞中Ⅱ型胶原的高表达。再生丝素蛋白膜对永生化软骨细胞无细胞毒性，对其黏附也无不良影响。体内永生化软骨细胞Ⅱ型胶原表达良好。构建具有永生化软骨细胞的丝基生物材料复合体可为临床软骨缺损修复提供一种可行的功能性生物材料。
