基底膜组织特异性支架修复子宫内膜缺损的实验研究

Scaffold based regeneration showed great potential in repairing the endometrium defects. The ideal scaffold should be uterus endometrium basement membrane(UEBM) or tissue-specific scaffold with basement membrane structure. However, related researches haven't been found yet. In this aspect, we have separated the UEBM from the uterus whole organ acellualr matrix and prepared urinary bladder matrix(UBM) with similar contents and micro-archtecture. UBM was chosen as scaffold in our research for its clinical feasibility. UBM combined with sustained-release vascular endothelial growth factor (VEGF-UBM) will be manufactured. The amounts of VEGF will be tailored to obtain satisfied in vitro release behavior. Its effects on cell metabolism, proliferation and differentiation endometrial stem cells will be seeded on the VEGF-UBM to investigate its effects on intracutaneous stimulation, blood compatibility, pyrogen test are carried out to assess the bio-safety of the ERM. In vivo, the microstructure, vessel density, scar formation and intrauterine adhesions of regenerated tissue will be evaluated in endometrial defects in canine models repaired with VEGF-UBM. Recovery of pregnancy and menstruation will be analyzed as well. The current research aims to discover appropriate scaffold materials for endometrium repair. The applicant focused on the research concerning acellular matrix and sustained release of bio-active factors, and have published 2 articles (IF 8.557 & IF 2.472) and applied 1 patent.

基于支架材料的诱导再生技术是子宫内膜缺损修复的重要研究方向，最佳的修复材料理论上应是子宫内膜基底膜（UEBM）或基底膜组织特异性支架，但至今未见相关报道。这方面申请者前期制备出UEBM和其类似物-高生物活性膀胱粘膜基底膜（UBM）并证实二者结构与成分均相近，但UBM拥有更佳的临床实用性。在此基础上，结合自主研发的载药缓释技术，本课题拟①制备可缓释血管内皮生长因子的UBM，体外鉴定释放曲线（≈21d）和对子宫内膜干细胞趋化、分化、增殖的作用及生物安全性；②体内修复犬重度子宫内膜缺损模型，设立UEBM、非组织特异性支架对照，评价组织再生、血供、修复区妊娠功能恢复和宿主-材料免疫反应。预期找到适合子宫内膜缺损修复的新支架材料，课题成果具有良好临床转化前景。申请者一直从事脱细胞基质生物材料及载药缓释相关研究，已发表生物材料学顶级期刊《Biomaterials》论著等SCI论文2篇、申请发明专利1项

基于支架材料的诱导再生技术是子宫内膜缺损修复的重要研究方向，最佳的修复材料理论上应是子宫内膜基底膜（UEBM）或基底膜组织特异性支架，但至今未见相关报道。这方面申请者前期制备出UEBM和其类似物-高生物活性膀胱粘膜基底膜（UBM）并证实二者结构与成分均相近，但UBM拥有更佳的临床实用性。在此基础上，结合自主研发的载药缓释技术，本课题在项目期内完成了以下研究：①构建负载VEGF的UBM，并完成相关检测。体外释放实验证实其可以实现VEGF的缓释，显著促进细胞的趋化、增殖且生物安全性良好。②构建大鼠子宫内膜缺损模型，以载VEGF的UBM修复缺损，发现与未载VEGF的UBM对照相比，VEGF-UBM显著促进组织再生、新生血管化、降低纤维化和提升子宫受容性，妊娠试验证实其可以帮助子宫一定程度上的恢复功能。③分析UBM的降解产物成分，确定其富含小分子多肽组成且具有促进组织再生的功能成分；分析其免疫原性、植入炎症反应，发现其体内外免疫原性极低，炎症反应轻微；完成UBM促进组织生长的机理的研究。课题发现UBM是适合子宫内膜缺损修复的新型支架材料，课题成果计划与上海长征医院妇产科联合申报多中心前瞻性非劣效性随机单盲对照研究临床研究，证实其临床疗效和应用前景。