Objective: Bone marrow mesenchymal stem cells (BMSCs) have the potential to differentiate into chondrocytes, it is the ideal seed cells for treating cartilage injury. The purpose of this study was to investigate the i...Objective: Bone marrow mesenchymal stem cells (BMSCs) have the potential to differentiate into chondrocytes, it is the ideal seed cells for treating cartilage injury. The purpose of this study was to investigate the inducible factors that stimulate the differentiation of bone marrow mesenchymal stem cells into chondrocytes. Methods: Rat BMSCs were isolated and subcultured, and then inoculated on PLGA scaffolds after successfully identificating of BMSCs. They were divided into 5 groups. Blank serum group, TGF-β1 + Yougui Yin drug-containing serum induction group, TGF-β1 + mechanical stimulation induction group, Yougui Yin drug-containing serum + mechanical stimulation induction group, TGF-β1 + Yougui Yin drug-containing serum + mechanical stimulation induction group, 4 weeks later, the adhesion status of cells and scaffolds was observed by Scanning Electron Microscopy, and then chondrocyte induction was performed on different treatment groups. After induction, the expression of Aggrecan and Collagen II was detected by immunofluorescence, and the mRNA expression of Aggrecan and Collagen II was detected by qPCR. Results: BMSCs were identified as rat bone marrow mesenchymal stem cells by immunofluorescence CD34 and CD44 cells. Scanning Electron Microscopy observed that BMSCs grew well on PLGA. Compared with PLGA group, the scaffold space of BMSCS-PLGA composite was significantly reduced, and the growth of cell adhesion scaffold was significantly increased. Compared with the blank serum group, the expression of Aggrecan in TGF-β1 + Yougui Yin drug-containing serum group and Yougui Yin drug-containing serum + mechanical stimulation group was significantly increased. The expression of Collagen II in TGF-β1 + Yougui Yin medicated serum group, Yougui Yin medicated serum + mechanical stimulation group, TGF-β1 + mechanical stimulation group and TGF-β1 + Yougui Yin medicated serum + mechanical stimulation group were significantly increased;PCR results showed that compared with blank serum group, the expression of Aggrecan in other groups was significantly increased, except for Collagen II expression in Yougui Yin drug-containing serum + mechanical stimulation group. Conclusion: BMSCs can differentiate into chondrocytes on PLGA scaffold. Compared with blank serum group, BMSCs can be induced to differentiate into chondrocytes in all induction groups, among all induction groups, TGF-β1 + Yougui-yin drug-containing serum group and TGF-β1 + mechanical stimulation group were more significant, and the difference was statistically significant.展开更多
目的探讨TGF-β1及其受体TGF-β1受体-Ⅰ(TGF-beta typeⅠreceptors,TβRⅠ)和受体-Ⅱ(TGF-beta type II receptors,TβRⅡ)mRNA在PCOS大鼠卵巢中的表达及作用。方法采用皮下注射脱氢表雄酮的方法建立PCOS动物模型,运用免疫组化方法检...目的探讨TGF-β1及其受体TGF-β1受体-Ⅰ(TGF-beta typeⅠreceptors,TβRⅠ)和受体-Ⅱ(TGF-beta type II receptors,TβRⅡ)mRNA在PCOS大鼠卵巢中的表达及作用。方法采用皮下注射脱氢表雄酮的方法建立PCOS动物模型,运用免疫组化方法检测卵巢中TGF-β1蛋白表达,并进行图像分析;采用RT-PCR方法检测TGF-β1受体TβRⅠ和TβRⅡmRNA在卵巢中的表达。结果与对照组相比,TGF-β1在PCOS各阶段卵泡卵母细胞、颗粒细胞中的表达强度差异无统计学意义(P>0.05)。而在PCOS间质细胞、窦状卵泡膜细胞的表达显著高于对照组(P<0.05,P<0.01)。凝胶图像分析显示TβRⅠ和TβRⅡmRNA相对含量显著高于对照组(P<0.05)。结论PCOS组TGF-β1表达异常是导致卵巢间质纤维化、包膜增厚的主要原因;TGF-β1也参与PCOS卵泡发育、闭锁的调控。PCOS大鼠TGF-β1受体表达的上调,是使TGF-β1致纤维化功能增强的原因之一。展开更多
文摘Objective: Bone marrow mesenchymal stem cells (BMSCs) have the potential to differentiate into chondrocytes, it is the ideal seed cells for treating cartilage injury. The purpose of this study was to investigate the inducible factors that stimulate the differentiation of bone marrow mesenchymal stem cells into chondrocytes. Methods: Rat BMSCs were isolated and subcultured, and then inoculated on PLGA scaffolds after successfully identificating of BMSCs. They were divided into 5 groups. Blank serum group, TGF-β1 + Yougui Yin drug-containing serum induction group, TGF-β1 + mechanical stimulation induction group, Yougui Yin drug-containing serum + mechanical stimulation induction group, TGF-β1 + Yougui Yin drug-containing serum + mechanical stimulation induction group, 4 weeks later, the adhesion status of cells and scaffolds was observed by Scanning Electron Microscopy, and then chondrocyte induction was performed on different treatment groups. After induction, the expression of Aggrecan and Collagen II was detected by immunofluorescence, and the mRNA expression of Aggrecan and Collagen II was detected by qPCR. Results: BMSCs were identified as rat bone marrow mesenchymal stem cells by immunofluorescence CD34 and CD44 cells. Scanning Electron Microscopy observed that BMSCs grew well on PLGA. Compared with PLGA group, the scaffold space of BMSCS-PLGA composite was significantly reduced, and the growth of cell adhesion scaffold was significantly increased. Compared with the blank serum group, the expression of Aggrecan in TGF-β1 + Yougui Yin drug-containing serum group and Yougui Yin drug-containing serum + mechanical stimulation group was significantly increased. The expression of Collagen II in TGF-β1 + Yougui Yin medicated serum group, Yougui Yin medicated serum + mechanical stimulation group, TGF-β1 + mechanical stimulation group and TGF-β1 + Yougui Yin medicated serum + mechanical stimulation group were significantly increased;PCR results showed that compared with blank serum group, the expression of Aggrecan in other groups was significantly increased, except for Collagen II expression in Yougui Yin drug-containing serum + mechanical stimulation group. Conclusion: BMSCs can differentiate into chondrocytes on PLGA scaffold. Compared with blank serum group, BMSCs can be induced to differentiate into chondrocytes in all induction groups, among all induction groups, TGF-β1 + Yougui-yin drug-containing serum group and TGF-β1 + mechanical stimulation group were more significant, and the difference was statistically significant.
基金Supported by The Program of Population and Family Planning Commission of Guangdong Province(No.2006002)
文摘目的探讨TGF-β1及其受体TGF-β1受体-Ⅰ(TGF-beta typeⅠreceptors,TβRⅠ)和受体-Ⅱ(TGF-beta type II receptors,TβRⅡ)mRNA在PCOS大鼠卵巢中的表达及作用。方法采用皮下注射脱氢表雄酮的方法建立PCOS动物模型,运用免疫组化方法检测卵巢中TGF-β1蛋白表达,并进行图像分析;采用RT-PCR方法检测TGF-β1受体TβRⅠ和TβRⅡmRNA在卵巢中的表达。结果与对照组相比,TGF-β1在PCOS各阶段卵泡卵母细胞、颗粒细胞中的表达强度差异无统计学意义(P>0.05)。而在PCOS间质细胞、窦状卵泡膜细胞的表达显著高于对照组(P<0.05,P<0.01)。凝胶图像分析显示TβRⅠ和TβRⅡmRNA相对含量显著高于对照组(P<0.05)。结论PCOS组TGF-β1表达异常是导致卵巢间质纤维化、包膜增厚的主要原因;TGF-β1也参与PCOS卵泡发育、闭锁的调控。PCOS大鼠TGF-β1受体表达的上调,是使TGF-β1致纤维化功能增强的原因之一。
