应用旋转式生物反应器体外构建组织工程软骨具优越性

The advantage of cultivating tissue-engineered cartilage in rotating bioreactor

目的:应用HFB-40型旋转式生物反应器体外培养一定形状的人工软骨组织,为组织工程软骨应用于更广泛的领域打下基础。方法:将第3代兔软骨细胞接种于2mm×4mm×4mm的长方体聚乳酸聚羟基乙酸聚合物(PLGA)上,随机分为A、B两组。A组为实验组,培养于生物反应器内;B组为对照组,培养于普通培养瓶内。4周后定量检测支架中氨基糖胺聚糖(GAG)、DNA含量,分析Ⅱ型胶原免疫组织化学染色面积百分比差异。结果:实验组支架-细胞复合体体积较大,较厚,表面光滑,有弹性,有光泽,其GAG、DNA含量以及Ⅱ型胶原免疫组织化学染色面积百分比明显高于对照组(P<0.01)。结论:生物反应器相对于普通的培养瓶,能够提供更加有利于软骨细胞在PLGA中生长的外部环境,更有利于组织工程软骨的形成。

Objective：To evaluate the possibility of cultivating tissue-engineered cartilage in a rotating bioreactor（HFB-40）. Methods：The PLGA constructs were formed in the shape of rectangular parallelopiped （2 mm ×4 mm × 4 mm）. Each PLGA template was seeded with chondrocytes and then cultivated in the bioreactor or flasks for 4 weeks. The chondrocyte-PLGA constructs were harvested and detected by the content of GAG, DNA and the percentage of the type II collagen dyeing area. Results：The chondrocytes-PLGA constructs grew much better in the bioreactor than in the flasks：The constructs grown in the bioreactor had much higher content of GAG,DNA and percentage of the type II collagen dyeing area than those in the flasks. Conclusion：It is advantageous for the tissue-engineered cartilage cultivated in the bioreactor,which can provide more suitable culture environment for chondrocyte-PLGA constructs.