间充质干细胞通过调节炎症反应及促进新生物形成治疗盆底器官脱垂的机制研究

Pelvic organ prolapse (POP) has been one of the hotspots and difficulties of gynecological diseases. The surgical therapeutic efficacy based on a mesh is not so satisfaction. The application of tissue engineering offers a new way for the treatment of POP. Studies showed that mesenchymal stem cells implanted into the body, not only could promote new tissue formation for reconstruction of tissue defects, but also could regulate the inflammatory response and the scar tissue hyperplasia. In the present study, we seeded muscle-derived stem cells (MDSCs) and adipose-derived stem cells (ADSCs) labeled with fluorecent protein mediated by adenovirus into Poly(lactic-co-glycolic acid) (PLGA) synthetic and small intestinal submucosa (SIS) biological meshes. After the compounds were induced myogenic differentiation, they were grafted in a model of POP in rats. After implantation, fluorescence microscope was used to measure the prognosis of stem cells, and histological and biomechanical testing was used to measure the repair procedures of POP. ELISA, western blotting, RT-PCR and immunohistochemistry were used to measure the expression level of inflammatory factors and the proliferative and differentiation of inflammatory cells to definite the function of stem cells in regulating the inflammatory response. We also investigated the function of TGF-β/Smad signaling pathway in modulating the proliferative and differentiation of fibroblast and the role of scar tissue hyperplasia induced by stem cells.

盆底器官脱垂（POP）是妇科疾病的热点和难点之一，目前以补片为基础的手术治疗效果尚不满意，组织工程的应用为POP的治疗提供了新的思路。研究表明间充质干细胞植入机体后，不仅可以促进新生物形成、重建组织缺损，还可以调节炎症反应及疤痕组织增生。本课题拟将腺病毒介导荧光蛋白标记的肌源性干细胞（MDSCs）和脂肪源性干细胞（ADSCs）植入聚乳酸-羟基乙酸共聚物（PLGA）及脱细胞小肠粘膜下层（SIS）补片，成肌诱导后植入大鼠POP模型或皮下，荧光显微镜了解干细胞在体内的转归，组织学及生物力学测试明确组织工程补片的修复情况。ELISA, western blotting, RT-PCR及免疫组化等检测植入物及周围组织中炎症因子的表达及炎症细胞的浸润、增殖情况，明确干细胞对补片诱导的炎症反应的调节作用，并进一步探讨TGF-β/Smad信号通路在干细胞调节成纤维细胞增殖、分化及疤痕组织增生中的作用。

本研究项目的结果包括四个方面内容：1）围绕干细胞促进组织再生及调控炎症反应进行深入研究：通过制备脱细胞羊膜支架，并评估其物化性能，负载ADSCs可以重建损伤的子宫内膜。2）制备多孔的支架材料-SMP，并证实该材料可以促进细胞的增殖；3）围绕新技术腹股沟韧带悬吊术治疗盆底器官脱垂的研究：通过尸体解剖，对相关悬挂点进行测量，定位，并应用于临床实践。4）首次探索宫颈癌类器官的构建，为后续研究奠定基础。