高糖环境下人骨髓间充质干细胞复合兔脱细胞真皮支架体外成骨能力检测

Detection of human bone marrow mesenchymal stem cells integrate rabbit acellular dermal matrix stenting of in vitro osteogenesis ability in high glucose environment

目的探讨脱细胞真皮支架在高糖环境下对骨髓间充质干细胞(h BMSC)的保护作用,以及促进h BMSC向成骨细胞分化的能力。方法建立脱细胞真皮基质和含300 mmol/L葡萄糖的细胞高糖模型,在高糖环境下,培养h BMSC为高糖对照组,在高糖条件下,h BMSC种植在脱细胞真皮基质上为基质实验组,正常条件下培养的h BMSC为正常对照组,每组均进行成骨细胞诱导实验。利用MTT实验检测h BMSC的增殖情况;利用骨形成蛋白-1(BMP-1)免疫荧光染色检测h BMSC成骨细胞分化情况,ELISA实验检测各组h BMSC的BMP-1、碱性磷酸酶(ALP)的含量。结果与高糖对照组比较,基质实验组h BMSC的细胞增殖光密度(OD值)升高,差异有统计学意义(P<0.01);h BMSC在脱细胞真皮基质的生长状态良好,基质实验组BMP-1染色阳性的细胞数目明显高于高糖对照组(P<0.01);基质实验组BMP-1、ALP等蛋白的含量明显增高(P<0.01)。结论高糖环境下,脱细胞真皮基质有利于促进h BMSC分化为成骨细胞。

Objective To investigate the protective effect of human bone marrow mesenchymal stem cells（h BMSC） with the acellular dermal matrix stenting in the high glucose environment, and the differentiation ability with the promotion form human bone marrow mesenchymal stem cells to osteoblasts. Methods Acellular dermal matrix and the high glucose model of containing 300 mmol/L glucose were established. With the high glucose environment, h BMSC was cultured to be the control group. With the high glucose condition, h BMSC was planted to be the acellular dermal matrix experimental group. With the normal condition, h BMSC was planted to be the normal control group. All groups had the osteoblasts induction experiment, the proliferation of h BMSC was detected by using MTT. the osteoblasts differentiation of h BMSC was detected by using fluorescence staining, and the content of BMP-1 and ALP in the experiment groups was detected by using ELISA. Results Compared with the high glucose control group, cell proliferation and matrix optical density（OD） of the h BMSC matrix experiment group was increased, with significant statistic difference（P 0.01）; h BMSC was in good condition in the acellular dermal matrix. The number of BMP-1 positive cells in the culture group was significantly higher than that in the high glucose group（P 0.01）; The contents of BMP-1, ALP proteins were significantly increased（P 0.01）. Conclusion In the high glucose environment, the acellular dermal matrix has the significant promotion of differentiation from h BMSC to osteoblasts.