骨髓基质细胞修复山羊髁突软骨缺失的绿色荧光蛋白示踪

In vivo tracing of bone marrow stromal cells differentiating into chondrocytes to repair condylar articular cartilage defects by green fluorescent protein gene transfection:An experimental study in goats

目的:采用绿色荧光蛋白(green fluorescent protein,GFP)对骨髓基质细胞体内修复髁突软骨全层缺失进行示踪观察。方法:扩增PG13细胞株,获得大量含GFP基因的假病毒液,并直接转染骨髓基质细胞(bone marrow stromal cells,BMSCs)。将含有GFP基因转染标记的BMSCs和少量软骨细胞与生物可降解材料复合后,植入山羊髁突软骨全层缺失处,1个月后应用激光共聚焦显微镜检测修复组织中GFP标记的BMSCs的分布。结果:标记的BMSCs植入关节软骨缺损1个月后,修复组织仍能高效表达GFP。激光共聚焦显微镜下显示:多数新生软骨陷窝内有GFP标记的细胞。结论:标记的BMSCs可在髁突软骨全层缺失区分化为成熟的软骨细胞,并在髁突软骨缺失的修复中发挥重要作用。

PURPOSE： To trace directly the distribution and differentiation of bone marrow stromal cells in temporoman- dibular joint condylar articular cartilage defects by green fluorescent protein gene transfection. METHODS： Recombinant GFP expression vector transfected cell PGI3 were obtained and in vitro expanded.The virus supernatant from infected PGI3 cell culture was used to infect BMSCs （bone marrow stromal cells） directly. Autologous GFP labeled BMSCs combined with chondrocytes were mixed with Pluronic F-127 and implanted into goat temporomandibular joint condylar articular cartilage defects. One month later, the repaired tissue was evaluated by laser confocal microscope.RESULTS： Laser confocal microscope revealed that GFP labeled cells existed in many neocartilage defects after 1 month. CONCLUSIONS： The recombinant GFP expression vector transfected cells can be used to trace the distribution and differentiation of bone marrow stromal cells in temporomandibular joint condylar articular cartilage defects in vivo. BMSCs play an important role in repairing articular cartilage defects.