聚己内酯-透明质酸静电纺丝膜联合间充质干细胞修复子宫内膜损伤  

Mesenchymal stem cells combined with polycaprolactone-hyaluronic acid electrospinning membrane in repair of endometrial injury

作　　者：安江茹 张锦艺 王秋花 杨杨杨 王文双[2] 张晓晴 An Jiangru;Zhang Jinyi;Wang Qiuhua;Yang Yangyang;Wang Wenshuang;Zhang Xiaoqing(School of Basic Medical Sciences,Binzhou Medical University,Department of Human Anatomy and Histoembryology,Joint Laboratory for International Cooperation in Biomaterials and Tissue Regeneration,Yantai 264003,Shandong Province,China;Department of Gynecology,Yantai Yuhuanding Hospital,Yantai 264000,Shandong Province,China)

机构地区：[1]滨州医学院基础医学院,人体解剖与组织胚胎学教研室,生物材料与组织再生国际合作联合实验室,山东省烟台市264003 [2]烟台毓璜顶医院妇科,山东省烟台市264000

出　　处：《中国组织工程研究》2025年第16期3369-3379,共11页Chinese Journal of Tissue Engineering Research

基　　金：山东省自然科学基金-青年基金项目(ZR2021QC034),项目负责人:张晓晴;泰山学者青年项目(tsqn202103111),项目负责人:张晓晴。

摘　　要：背景:人子宫内膜间充质干细胞能够直接修复受损的子宫内膜,促进血管生成、恢复子宫形态结构,然而将干细胞直接注入受损子宫内膜后的细胞存活率低、滞留时间短,修复效果有限。目的:观察聚己内酯-透明质酸静电纺丝膜复合人子宫内膜间充质干细胞修复大鼠子宫内膜损伤的效果。方法:①细胞实验:采用胶原酶消化法提取人子宫内膜间充质干细胞,静电纺丝技术制备聚己内酯-透明质酸静电纺丝膜。将人子宫内膜间充质干细胞分别接种于聚苯乙烯培养板与聚己内酯-透明质酸静电纺丝膜上,通过DNA定量分析、WST-1细胞活性实验、鬼笔环肽染色、扫描电镜观察细胞的增殖与黏附能力,qRT-PCR检测静电纺丝膜上细胞CD90、Meflin的mRNA表达。②动物实验:取27只处于动情期的雌性SD大鼠,通过机械搔刮法建立宫腔粘连模型后随机分为3组,每组9只:空白对照组不进行任何治疗,对照组将聚己内酯-透明质酸静电纺丝膜植入宫腔损伤部位,实验组将聚己内酯-透明质酸静电纺丝膜/人子宫内膜间充质干细胞补片植入宫腔损伤部位。术后第3,7,14天取材,采用苏木精-伊红染色观察子宫形态结构及腺体数量,qRT-PCR和免疫荧光染色观察子宫组织CD31、血管内皮生长因子的表达。结果与结论:①细胞实验:与聚苯乙烯培养板相比,聚己内酯-透明质酸静电纺丝膜可促进人子宫内膜间充质干细胞的增殖与黏附,并且聚己内酯-透明质酸静电纺丝膜支持人子宫内膜间充质干细胞基因CD90和Meflin的表达;②动物实验:苏木精-伊红染色显示,聚己内酯-透明质酸静电纺丝膜/人子宫内膜间充质干细胞补片可促进子宫内膜损伤后形态结构的恢复,术后第14天的内膜厚度与腺体数量均多于空白对照组、对照组(P<0.05);qRT-PCR和免疫荧光染色检测显示,实验组术后第7,14天的CD31、血管内皮生长因子mRNA与蛋白表达均高BACKGROUND:Human endometrial mesenchymal stem cells can directly repair the damaged endometrium,promote angiogenesis,and restore the morphological structure of the uterus.However,after the direct injection of stem cells into the damaged endometrium,the cell survival rate is low,the retention time is short,and the repair effect is limited.OBJECTIVE:To observe the effect of polycaprolactone-hyaluronic acid electrospinning membrane combined with human endometrial mesenchymal stem cells on endometrial injury in rats.METHODS:(1)Cell experiment:Human endometrial mesenchymal stem cells were extracted by collagenase digestion method.Polycaprolactone-hyaluronic acid electrospinning membrane was prepared by electrospinning technology.The human endometrial mesenchymal stem cells were inoculated on polystyrene culture plate and polycaprolactone-hyaluronic acid electrospinning membrane.The proliferation and adhesion of the cells were observed by DNA quantitative analysis,WST-1 cell activity test,phalloidin staining,and scanning electron microscopy.The mRNA expressions of CD90 and Meflin in electrospun membrane were detected by qRT-PCR.(2)Animal experiments:27 female SD rats in estrus were selected to establish uterine adhesion model by mechanical scratching method and randomly divided into three groups with nine rats in each group:The blank control group did not receive any treatment;the control group was implanted with polycaprolacton-hyaluronic acid electrospinning membrane;the experimental group was implanted with polycaprolacton-hyaluronic acid electrospinning membrane/human endometrial mesenchymal stem cell mesh.Samples were collected at 3,7,and 14 days after surgery.Hematoxylin-eosin staining was used to observe the morphological structure of uterus and the number of glands.qRT-PCR and immunofluorescence staining were used to observe the expression of CD31 and vascular endothelial growth factor in uterine tissue.RESULTS AND CONCLUSION:(1)Cell experiment:Compared with polystyrene culture plate,polycaprolactone-hyaluronic

关 键 词：子宫内膜 宫腔粘连 人子宫内膜间充质干细胞 静电纺丝膜 聚己内酯 透明质酸 

分 类 号：R459.9[医药卫生—治疗学] R318.08[医药卫生—临床医学] R713