IGF-Ⅰ与TGF-β_2单独及联合应用对软骨细胞培养的影响  

The effects of IGF-Ⅰ and TGF-β_2 alone and combination on chondrocytes culture

作　　者：张帆[1] 刘兴龙[1] 周广东[2] 杨彪炳[3] 

机构地区：[1]潍坊医学院,山东潍坊261041 [2]上海交通大学组织工程国家研究中心,上海200240 [3]潍坊医学院整形外科研究所,山东潍坊261041

出　　处：《现代医药卫生》2014年第2期161-163,166,共4页Journal of Modern Medicine & Health

基　　金：山东省高等学校科技计划项目(J09LF20)

摘　　要：目的探讨胰岛素样生长因子-1(IGF-Ⅰ)与转化生长因子-β2(TGF-β2)单独及联合应用对软骨细胞形态、增殖速度、黏多糖含量、Ⅱ型胶原含量及表型的影响。方法采用酶消化法取兔软骨细胞并培养,于第2代时分别加IGF-Ⅰ及TGF-β2。分为A(空白对照)、B(含IGF-Ⅰ)、C(含TGF-β2)、D(含IGF-Ⅰ及TGF-β2)组。用苏木精-伊红染色法、甲苯胺蓝染色观察软骨细胞形态,MTT法检测增殖速度,阿尔新蓝比色法检测黏多糖含量,免疫组化法检测Ⅱ型胶原含量及表型。结果在维持细胞形态、细胞增殖速度、Ⅱ型胶原含量及表型方面,D组优于B组,C组次之,A组最差,差异均有统计学意义(P<0.05)。在促进细胞外基质及黏多糖分泌方面,D组优于C组,B组次之,A组最差(P<0.05)。结论 IGF-Ⅰ、TGF-β2能维持软骨细胞形态,促进其增殖及Ⅱ型胶原的分泌,并维持其表型,增加细胞外基质及黏多糖的分泌,联合应用有协同作用,可抑制软骨细胞在体外培养过程中的老化。Objective To investigate the effects of insulin-like growth factor - I （IGF- I ） and transforming growth factor β2（TGF-β2） alone and combined application on the morphology ,proliferation ratio ,mueopolysaccharides content,collagen type 11 content and phenotype of chondrocytes. Methods Rabbit chondrocytes were obtained using enzyme digestion method,and the second passage of them were added with IGF- I and TGF-β2 for culturen, and clivided into group A （blank control group）, group B （with IGF- I ） ,group C（with TGF-β2） and group D（with IGF- I and TGF-β2）. HE and toluidine blue staining were used to detect chondrocytes morphology. MTT and Alcian blue colorimetric method were used to measure the proliferation ratio and mucopolysaccharides content respectively. Immunohistochemistry was applied to detect the collagen type II content and phenotype of chondrocytes. Results In the respect of maintaining the chondrocytes morphology,proliferation ratio, the content of collagen type II and retaining the phenotype of chondrocytes, the group D was superior to the group C, followed by group B, and the group A was the worst. The difference had statistical significance （P〈0.05）. The promotion of extracellular matriss and secretion of glycosaminoglycan in group D was better than those in group C ,fallowed by group B, and group A was the worst （P〈0.05）. Conclusion IGF- I and TGF-β2 can maintain the morphology and phenotype of chondrocytes, promote the proliferation ratio and the secretion of collagen type II as well as the secretion of mucopolysaccharides, and the combined application of IGF- I and TGF-β2 has synergistic effect. Therefore, the combined application of these two factors can inhibit chondrocytes aging in vitro culture.

关 键 词：胰岛素样生长因子Ⅰ 转化生长因子Β 软骨细胞 组织工程 联合应用 

分 类 号：R329-33[医药卫生—人体解剖和组织胚胎学]