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纸质出版:2016
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杨化超,李茂,黄文.人脐带间充质干细胞对小鼠皮肤创面愈合及VEGF表达的影响[J].陆军军医大学学报(原第三军医大学学报),2016,38(05):456-462.
Yang Huachao,Li Mao,Huang Wen.Effect of human umbilical cord mesenchymal stem cells on skin wound healing and expression of VEGF in mice[J].J Amry Med Univ (J Third Mil Med Univ),2016,38(05):456-462.
目的 研究人脐带间充质干细胞(human umbilical cord mesenchymal stem cells,hUC-MSCs)对小鼠全层皮肤切除创面愈合及血管内皮细胞生长因子(vascular endothelial growth factor,VEGF)表达的影响。 方法 采用组织块贴壁法分离培养hUC-MSCs,流式细胞术检测细胞免疫表型,并做向成骨、成脂细胞方向分化。48只雄性BALB/c小鼠采用完全随机分组方法分为hUC-MSCs组和PBS组,每组24只,在其背部形成一全层皮肤缺损创面模型,分别于创周注射hUC-MSCs悬液和PBS液。于3、7、14 d观察创面愈合情况,HE染色观察病理学改变,免疫组织化学及Western blot检测VEGF蛋白的表达。 结果 流式细胞术检测显示所培养的细胞为hUC-MSCs,体外诱导其能向成骨、成脂细胞方向分化。hUC-MSCs组创面愈合率在各时间点均较PBS组显著增高(P0.05)。HE染色显示hUC-MSCs组创面愈合质量优于PBS组。免疫组化显示hUC-MSCs组VEGF在3 d时表达最多,7、14 d逐渐减少,但均较PBS组增高。Western blot结果显示hUC-MSCs组3、7、14 d创面VEGF蛋白表达显著高于PBS组(P0.05)。 结论 hUC-MSCs可增强皮肤创面VEGF的表达,促进皮肤创面的修复。
Objective To investigate the effect of human umbilical cord mesenchymal stem cells (hUC-MSCs) on wound healing and the expression of vascular endothelial growth factor (VEGF) after full-thickness excision dermal wounds in mice. Methods hUC-MSCs were isolated from the human umbilical cord tissues from a full-term neonate by tissue explants adherent method
and then cultured and identified with flow cytometry for cellular immune phenotypes. Then the obtained hUC-MSCs were induced to differentiate into osteoblasts and adipocytes. Forty-eight male BALB/c mice were randomly divided into hUC-MSCs group and PBS group
with 24 animals in each group. The mice were injected with hUC-MSCs suspension and PBS fluid respectively around the wounds which were made as a full-thickness excision skin wounds in size of 1.5 cm on the dorsum in mice. The wound healing condition were examined in 3
7
and 14 d after operation
and pathological changes were observed with HE staining. The protein expression of VEGF were detected with immunohistochemical assay and Western blotting. Results Flow cytometry results showed that the cultured cells was hUC-MSCs
and the obtained cells could be induced to osteogenic and adipogenic cells after differentiation. The wound healing rate was obviously higher at all time points in the hUC-MSCs group than the PBS group (P0.05). HE staining indicated that the wound healing quality was better in the former than the latter group. Immunohistochemical assay displayed that the expression level of VEGF reached the highest in wound at 3 d
and then reduced gradually at 7 and 14 d in the hUC-MSCs group
but still significantly higher than in the PBS group. Western blot assay showed that the protein expression level of VEGF was obviously higher in hUC-MSCs group than in the PBS group at 3
7
and 14 d after operation (P0.05). Conclusion hUC-MSCs promote the expression of VEGF and facilitate the healing of skin wound.
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