应用旋转生物反应器开展软骨组织工程研究

Tissue - engineered biological neocartilage utilizing rotary cell culture system

目的 应用旋转生物反应器(RCCS)在体外条件下培育具有一定形状的组织工程化人工软骨,以便为软骨组织工程产品产业化生产奠定基础。方法 将兔关节软骨细胞接种到圆形或方形可降解聚合物材料上,然后在RCCS内进行培养,并以在培养瓶内培养复合体为对照组。在体外培养期间,对复合体培育产物氨基糖胺聚糖(GAG)及DNA含量进行定量测量。经体外培养后,将复合体植入到裸鼠背部皮下,术后不同时间取材,进行大体、组织学等检查。结果 经RCCS培养的复合体在体外培养期间以及体内植入后均有明显软骨形成,而对照组则仅见少量软骨形成。复合体体外培育产物检测结果表明,在RCCS内培育的复合体GAG和DNA含量明显高于对照组。结论 RCCS与以单层生长方式为主的传统细胞培养装置—培养瓶相比,能够提供更适宜的外部环境,从而有利于软骨细胞在支架内形成人工软骨组织。

Objective To evaluate the possibility of growing tissue - engineered cartilage in a environment of rotary cell culture system. Methods Polymer templates were formed in the shape of laminate or round using foams of PLGA. Chondrocytes were isolated enzymatically from rabbit articular cartilage. Each PLGA template was seeded with chondrocytes and then kept in RCCS or flask for 4 weeks prior to subcutaneous transplantation into athymic mice. The chondrocyte - polymer constructs in vitro or in vivo were harvested and examined macroscopically, histologically and immunohistochemically. Results Tissue constructs grown in RCCS had higher GAG and DNA contents than control constructs grown in flasks. Specimens implanted into athymic mice from experimental groups exhibited more neocartilage formation than that from the control groups. Conclusions These findings demonstrate that RCCS can provide a suitable culture environment that supports tissue assemblies. RCCS has great potential for producing tissue engineered cartilage.