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目的:拟以C57糖尿病小鼠为研究对象,建立糖尿病小鼠OPN-/-MSCs组、糖尿病小鼠MSCs组及溶剂组对照组,采用小鼠创面愈合率检测、HE染色、免疫组化等试验方法,观察骨髓间充质干细胞(MSCs)及OPN基因敲除的MSCs对糖尿病小鼠创面愈合的影响。方法:原代提取MSCs及OPN-/-MSCs细胞;利用流式细胞仪检测细胞表面抗原;利用STZ溶液腹腔注射的方法建立糖尿病小鼠模型;将MSCs及OPN-/-MSCs分别注射至小鼠尾静脉,并观察创面愈合情况。结果:成功提取和培养了两组细胞,并成功建立了糖尿病小鼠模型;溶剂组小鼠完全愈合用了19.90±0.55天,OPN-/-MSCs注射组完全愈合用了18.52±0.75天,MSCs尾静脉注射糖尿病组小鼠用了13.70±0.35天。尾静脉注射MSCs组创面愈合时间明显短于普通糖尿病小鼠组。结论:通过观察三组糖尿病小鼠创面愈合情况,证实MSCs尾静脉注射糖尿病组小鼠比OPN-/-MSCs注射组小鼠和溶剂组小鼠的创面愈合速度快,说明OPN具有调控骨髓间充质干细胞促进创面愈合的作用,为临床治疗糖尿病足提供新的理论依据。
OBJECTIVE: To investigate the effects of wound healing rate, HE staining and immunohistochemistry in mice with C57 diabetic mice as the research object, to establish diabetic mice OPN - / - MSCs group, diabetic mice MSCs group and solvent group control group Methods The effects of bone marrow mesenchymal stem cells (MSCs) and OPN knockout MSCs on wound healing in diabetic mice were observed. Methods: Primary cultured MSCs and OPN - / - MSCs were harvested. Cell surface antigens were detected by flow cytometry. Diabetic mouse models were established by intraperitoneal injection of STZ solution. MSCs and OPN - / - MSCs were injected into mice Tail vein, and observe the wound healing. RESULTS: Two groups of cells were successfully extracted and cultured. The diabetic mice model was established successfully. The mice in the solvent group completely healed for 19.90 ± 0.55 days and the OPN - / - MSCs injected group completely healed for 18.52 ± 0.75 days. MSCs The tail vein injection of diabetic mice used 13.70 ± 0.35 days. The wound healing time of tail vein injection of MSCs was significantly shorter than that of normal diabetic mice. Conclusion: The healing of wounds in three groups of diabetic mice was observed, and it was confirmed that the wound healing rate of mice injected with tail vein of MSCs was higher than that of mice injected with OPN - / - MSCs and solvent group, indicating that OPN has the ability to regulate bone marrow The role of mesenchymal stem cells in promoting wound healing provides a new theoretical basis for the clinical treatment of diabetic foot.