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微流控芯片上胰岛素样生长因子-1(IGF-1)、碱性成纤维细胞生长因子(bFGF)对兔关节软骨细胞体外增殖的影响
The Effects of Insulin-Like Growth Factor-1 and Basic Fibroblast Growth Factor on Proliferation of Rabbit Articular Chondrocytes Embedded in Collagen Gel I on Microfluidic Chips
【作者】 李元城;
【导师】 张卫国;
【作者基本信息】 大连医科大学 , 骨科学, 2010, 博士
【摘要】 目的:微流控芯片平台上探索三维培养环境下胰岛素样生长因子-1(Insulin-Like Growth Factor-1, IGF-1)、碱性成纤维细胞生长因子(basic Fibroblast Growth Factor, bFGF)对软骨细胞体外增殖的影响;通过验证微流控平台的有效性将微流控芯片技术引入软骨组织工程领域。方法:微流控芯片上,分别在IGF-1、bFGF以及二者组合浓度梯度下三维培养软骨细胞,两周后计算软骨细胞增殖率并与96孔板传统方法所获得的数据进行比较。结果:IGF-1在57.14ng/ml产生最大增殖效应(平均倍增2.38倍),bFGF在5.72ng/ml产生最大增殖效应(平均倍增3.85倍),85.71ng/ml的IGF-1与1.43ng/ml bFGF的组合使软骨细胞产生最大的增殖效应(平均倍增4.76倍);芯片与传统96孔板所得增殖率不存在统计学显著性差异。结论:一定浓度组合的IGF-1和bFGF可协同产生最大的增殖效应;微流控芯片技术可有效应用于软骨组织工程领域,其在软骨组织工程领域存在广阔的应用前景。
【Abstract】 Objective:The effects of the insulin-like growth factor-1 (IGF-1) and basic fibroblast growth factor (bFGF) on the proliferation of chondrocytes in 3D were investigated. By validating the application of microfluidic chip, we introduced it for cartilage tissue engineering.Methods:Rabbit articular chondrocytes embedded in collagen gel were cultured on chip and 96-well plate for two weeks and their proliferation rate was investigated in the presence of IGF-1, bFGF and their combination.Results:The cell proliferation rate revealed to be highest in 57.14ng/ml of IGF-1 (up to 2.38-fold), highest in 5.72ng/ml of bFGF (up to 3.58-fold) and highest in the combination of 85.73ng/ml IGF-1 and 1.43ng/ml bFGF (up to 4.76-fold). Moreover, there was no statistically significant difference between microfluidic platform and 96-well plate with the proliferation rate.Conclusion:A certain combination of IGF-1 and bFGF could synergistically increases the proliferation of chondrocytes in 3D while maintaining their morphology. Microfluidic chips could be applied in cartilage tissue engineering efficiently.
【Key words】 Cartilage; Tissue Engineering; Microfluidic Chip; Insulin-Like Growth Factor-1; basic Fibroblast Growth Factor; Concentration Gradient;