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目的探讨骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs)促进皮肤创伤愈合作用及细胞因子在愈合过程中的表达变化。方法本研究于2013年7月至2014年10月在沈阳万类生物技术实验室完成。体外分离、培养大鼠BMSCs,镜下观察细胞生长形态,取第3代BMSCs进行实验。流式细胞仪检测细胞表面标记物鉴定BMSCs。选择健康雄性SD大鼠48只,随机分成2组。于大鼠尾静脉注射PKH-26标记的BMSCs,1周后进行皮肤创伤模型制备,实验组于大鼠背部正中线划长3 cm的纵行切口,对照组不做创伤处理。荧光染色观察不同时间点正常皮肤及创面组织BMSCs数量、分布情况,HE染色观察不同时间点正常皮肤及创面组织病理变化情况。ELISA检测大鼠尾静脉血中不同时间点基质细胞衍生因子(SDF-1)、肿瘤坏死因子(TNF-α)、白细胞介素(IL-1β)、碱性成纤维细胞生长因子(b FGF)、血小板衍生生长因子(PDGF)的含量变化。结果镜下观察培养第三代的BMSCs细胞形态为均一梭形,贴壁生长。BMSCs移植能够加速创面愈合,ELISA结果显示,BMSCs移植后血清中SDF-1α、TNF-α、IL-1β、b FGF和PDGF的含量随着创伤愈合进程呈先上升后下降的趋势。结论BMSCs能促进大鼠皮肤创伤愈合。
Objective To investigate the effects of bone marrow mesenchymal stem cells (BMSCs) on wound healing and the changes of cytokines in the process of healing. Methods The study was performed at Shenyang Wanchu Biotechnology Laboratory from July 2013 to October 2014. BMSCs were isolated and cultured in vitro. The morphology of the cells was observed under the microscope. The 3rd generation BMSCs were used for the experiment. BMSCs were identified by flow cytometry using cell surface markers. Forty-eight healthy male SD rats were randomly divided into two groups. PKH-26-labeled BMSCs were injected into the caudal vein of rats and skin wounds were prepared after 1 week. The experimental group was given a longitudinal incision with a length of 3 cm in the midline of the back of the rats, while the control group was not treated with wounds. Fluorescent staining was used to observe the number and distribution of BMSCs in normal skin and wound tissues at different time points. The pathological changes of normal skin and wound tissues at different time points were observed by HE staining. The levels of SDF-1, TNF-α, IL-1β and bFGF were measured by ELISA at different time points in tail vein of rats. , Platelet-derived growth factor (PDGF) content changes. RESULTS: The third generation of BMSCs cultured under microscope showed a uniform spindle shape and adherent growth. BMSCs transplantation can accelerate the healing of wounds. The results of ELISA showed that the levels of SDF-1α, TNF-α, IL-1β, b FGF and PDGF in sera of BMSCs initially increased and then decreased with the progress of wound healing. Conclusion BMSCs can promote wound healing in rats.