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尼古丁对藻酸盐珠培养中大鼠骨髓间充质干细胞软骨分化的影响。

Effect of nicotine on chondrogenic differentiation of rat bone marrow mesenchymal stem cells in alginate bead culture.

作者信息

Deng Yu, Li Tian-Qi, Yan You-E, Magdalou Jacques, Wang Hui, Chen Liao-Bin

机构信息

Department of Orthopaedic Surgery, Zhongnan Hospital of Wuhan University, Wuhan, China.

出版信息

Biomed Mater Eng. 2012;22(1-3):81-7. doi: 10.3233/BME-2012-0692.

Abstract

This study was carried out to explore environmental compound such as nicotine can cause adverse effect on chondrogenic differentiation of bone marrow mesenchymal stem cells (BMSCs). Rat BMSCs were capsulated in alginate beads incubated with a chondrogenic differentiation medium and while chondrogenic differentiation of rat BMSCs were cultured for 4 weeks treated with nicotine at concentrations of 25, 50 and 100 μM. The effect of nicotine on BMSCs viability was tested using MTT assay. After chondrogenic differentiation, alginate beads sections were stained for glycosaminoglycan (GAG) with alcian blue and safranin-O. The mRNA expression of chondrogenesis related genes, including collagen type 2 alpha 1 (Col2A1), aggrecan, insulin-like growth factor-1 (IGF-1) were determined by RT-PCR. Nicotine did not affect viability of BMSCs at any indicated concentration. Continuous exposure to nicotine for 4 weeks resulted in significant decrease of the area stained with alcian blue and safranin-O in a concentration-dependent manner compared with the control (P<0.05). After 4 weeks in chondrogenic medium, nicotine dose-dependently decreased the expression of aggrecan, Col2A1 and IGF-1 genes in rat BMSCs chondrogenesis compared with the control (P<0.05). It turned out that nicotine suppresses chondrogenic differentiation potential of BMSCs, leading to a poorly differentiated cartilage.

摘要

本研究旨在探讨尼古丁等环境化合物是否会对骨髓间充质干细胞(BMSCs)的软骨分化产生不利影响。将大鼠BMSCs封装在藻酸盐珠中,与软骨分化培养基一起孵育,同时将大鼠BMSCs的软骨分化培养4周,用浓度为25、50和100μM的尼古丁处理。使用MTT法检测尼古丁对BMSCs活力的影响。软骨分化后,用阿尔新蓝和番红O对藻酸盐珠切片进行糖胺聚糖(GAG)染色。通过RT-PCR测定软骨生成相关基因的mRNA表达,包括Ⅱ型胶原α1(Col2A1)、聚集蛋白聚糖、胰岛素样生长因子-1(IGF-1)。在任何所示浓度下,尼古丁均不影响BMSCs的活力。与对照组相比,连续4周暴露于尼古丁导致阿尔新蓝和番红O染色面积显著降低,且呈浓度依赖性(P<0.05)。在软骨分化培养基中培养4周后,与对照组相比,尼古丁剂量依赖性地降低了大鼠BMSCs软骨生成中聚集蛋白聚糖、Col2A1和IGF-1基因的表达(P<0.05)。结果表明,尼古丁抑制了BMSCs的软骨分化潜能,导致软骨分化不良。

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