β1转化生长因子浓度对体外诱导骨髓间充质细胞构建软骨的影响  

Influence of TGF-β1 concentration on in vitro cartilage engineering with bone marrow stromal cells

作　　者：刘天一[1] 周广东[1] 陈瑾君[1] 崔磊[1] 刘伟[1] 曹谊林[1] 

机构地区：[1]上海交通大学医学院附属第九人民医院整复外科、上海市组织工程重点实验室,上海200011

出　　处：《中华医学美学美容杂志》2007年第5期288-291,共4页Chinese Journal of Medical Aesthetics and Cosmetology

基　　金：国家973计划项目（编号：2005CB522702）;国家863计划重大专项（编号：2006AA02A126）;国家自然科学基金（编号：303003531）

摘　　要：目的 探讨β1转化生长因子（TGF-β1）浓度对体外诱导猪骨髓间充质细胞（BMSCs）构建组织工程化软骨的影响，明确TGF-β1诱导剂量对细胞分化的作用，为体外软骨构建提供适宜的诱导因子应用浓度参数。方法 抽取8周龄猪髂嵴骨髓，应用贴壁法分选单个核细胞，体外培养扩增后获得BMSCs，收集第2代细胞，以5×10^7个／cm。细胞的密度接种到聚羟基乙酸（PGA）制成的圆柱形三维支架材料上（直径5mm，厚度2mm），7d后应用不同浓度TGFβ1（A组：5ng／ml、B组：10ng／ml、C组：20ng／ml、D组：50ng／m1）与IGF-1（50ng／m1）及地塞米松（40ng／m1）组成诱导剂，分别进行体外诱导培养。8周后取材行大体观察，体积、湿重及聚合蛋白多糖（GAG）定量，组织学及Ⅱ型胶原免疫组织化学等检测。结果 B、C、D组形成细胞材料复合物组织学结构较为类似，有明显的软骨陷窝，分布有大量Ⅱ型胶原及GAG；A组软骨陷窝结构较少，胞外基质染色较浅。B、C、D组的组织湿重、体积和GAG含量均明显高于A组。结论 诱导三维支架上的BMSCs体外构建组织工程化软骨过程中，10ng／ml的TGF-β1诱导浓度具有良好的促分化效能，TGF-1的促分化作用并未表现出明显的剂量依赖性。Objective To explore the relationship between TGF-β1 concentration and in-vitro induced chondrogenesis of BMSCs, and the effective TGF-β1 dose for induced BMSCs differentiation. Methods Bone marrow was aspirated from cristae iliaca of 8-week-old pigs to select mononuclear cells by cell culture. The cells were cultured and expended in vitro, and passage 2 BMSCs were seeded at the density of 5×10^7 cells/cm^2 to disc-shaped scaffolds with 5 mm in diameter and 2 cm in thickness. At 7 days, the scaffolds were induced in vitro with 50ng/ml of IGF-Ⅰ, 40ng/ml of dexomethasone, and different concentrations of TGFβ1 5ng/ml in group A, 10 ng/ml in group B, 20 ng/ml in group C, and 50 ng/ml in group D. And specimens were collected at 8 weeks for gross observation, size evaluation, wet weight, glycoaminoglycan （GAG） content, histology, and immunohistochemistry of type Ⅱ collagen. Results The constructs of groups B, C, and D showed similar histological features with typical lacuna structures and abundant type II collagen and GAG, while those constructs in group A showed few lacunas and light ECM staining. Wet weight, size, and GAG content in groups B, C, and D were significantly higher than those in group A. Conclusion TGF-β1 can induce differentiation toward cartilage at a concentration of 10 ng/ml during in vitro culture in a dose-in dependent manner.

关 键 词：骨髓间充质细胞 Β1转化生长因子 软骨 组织工程 

分 类 号：R6[医药卫生—外科学]