人脐带间充质干细胞条件培养基脂质体修复大鼠皮肤创面损伤

Liposome prepared from human umbilical cord mesenchymal stem cell conditioned medium for repair of skin wound in rats

背景:干细胞条件培养基中含有大量有助于伤口修复的细胞分泌因子,用脂质体包裹有助于细胞因子在伤口处的缓释作用。目的:研究人脐带间充质干细胞条件培养基脂质体对大鼠皮肤创面愈合的影响。方法:采用组织块贴壁法分离培养人脐带间充质干细胞并进行相关因子检测。取20只SD大鼠(购自山西医科大学实验动物中心)用于制备直径1 cm的皮肤创面模型。薄膜分散制备条件培养基脂质体囊泡用于大鼠皮肤缺损伤口,取修复部位皮肤及周围组织进行苏木精-伊红染色病理切片观察,通过组织学评分统计分析判断其对创伤的修复情况。结果与结论:(1)人脐带间充质干细胞表面标志物CD73、CD90、CD105阳性率分别为99.7%、95.7%、99.4%,CD45、CD34阳性率分别为1.1%、0.1%,且能诱导分化为骨细胞、脂肪细胞;(2)条件培养基中血管内皮生长因子、白细胞介素8、单核细胞趋化蛋白1的质量浓度分别为558.60 ng/L、5 236.48 ng/L、2 972.55 ng/L;(3)用薄膜分散法制备得到人脐带间充质干细胞条件培养基脂质体平均粒径为262.3 nm;(4)人脐带间充质干细胞条件培养及其脂质体均可促进大鼠皮肤缺损处的毛囊、腺体发生,且表皮层加厚,组织学评分显著升高,第2天的条件培养基与条件培养基脂质体差异无显著性意义(P=0.730),其他各组之间有显著差异(P <0.05);(5)结果提示,人脐带间充质干细胞条件培养基及其脂质体可以促进大鼠缺损伤口的修复。

BACKGROUND:The stem cell conditioned medium contains a large number of cell secretory factors contributing to wound repair.Liposome encapsulation facilitates the sustained release of cytokines at the wound site.OBJECTIVE:To study the effect of liposomes prepared from the conditioned medium of human umbilical cord mesenchymal stem cells on the healing of skin wounds in rats.METHODS:Human umbilical cord mesenchymal stem cells were isolated and cultured by tissue explant method,and cytokine detection was then performed.Twenty Sprague-Dawley rats were provided by the Experimental Animal Center of Shanxi Medical University in China to make skin wound models of1cm in diameter.Membrane dispersion technique was used to prepare the conditioned medium liposomal vesicles used for skin wounds in the rats.The skin and surrounding tissues of the repair site were taken for hematoxylin-eosin staining.The histological scores were used to judge the repair of wounds.RESULTS AND CONCLUSION:(1)The human umbilical cord mesenchymal stem cells were positive for CD73(99.7%),CD90(95.7%),CD105(99.4%),CD45(1.1%)and CD34(0.1%).The cells could be induced to differentiate into osteocytes and adipocytes.(2)The concentrations of vascular endothelial growth factor,interleukin-8,and monocyte chemotactic protein1in the conditioned medium were558.60ng/L,5236.48ng/L,and2972.55ng/L,respectively.(3)The average particle size of liposome in the conditioned medium prepared by membrane dispersion method was262.3nm.Conditional culture of human umbilical cord mesenchymal stem cells and its liposome could increase the hair follicles and glands at the site of rat skin defects,the epidermis was thickened,and the histological score was markedly increased.There was no difference between the conditioned medium and conditioned medium liposome groups at2days of culture(P=0.730),but significant differences existed among the other groups(P<0.05).To conclude,the human umbilical cord mesenchymal stem cell conditioned medium and its liposomes can promote the repair of skin defects in rats.