Department of Biology and Anatomical Sciences, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
Department of Stem Cells and Developmental Biology, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran.
Stem Cell Res Ther. 2023 Apr 3;14(1):61. doi: 10.1186/s13287-023-03279-7.
Asherman syndrome (AS), or intrauterine adhesions, is a main cause of infertility in reproductive age women after endometrial injury. Mesenchymal stem cells (MSCs) and their extracellular vesicles (EVs) are promising candidates for therapies that repair damaged endometria. However, concerns about their efficacy are attributed to heterogeneity of the cell populations and EVs. A homogenous population of MSCs and effective EV subpopulation are needed to develop potentially promising therapeutic options in regenerative medicine.
AS model was induced by mechanical injury in adult rat uteri. Then, the animals were treated immediately with homogeneous population of human bone marrow-derived clonal MSCs (cMSCs), heterogenous parental MSCs (hMSCs), or cMSCs-derived EV subpopulations (EV20K and EV110K). The animals were sacrificed two weeks post-treatment and uterine horns were collected. The sections were taken, and hematoxylin-eosin was used to examine the repair of endometrial structure. Fibrosis was measured by Masson's trichrome staining and α-SMA and cell proliferation by Ki67 immunostaining. The function of the uteri was explored by the result of mating trial test. Expression changes of TNFα, IL-10, VEGF, and LIF were assayed by ELISA.
Histological analysis indicated fewer glands, thinner endometria, increased fibrotic areas, and decreased proliferation of epithelial and stroma of the uteri in the treated compared with intact and sham-operated animals. However, these parameters improved after transplantation of both types of cMSCs and hMSCs and/or both cryopreserved EVs subpopulations. The cMSCs demonstrated more successful implantation of the embryos in comparison with hMSCs. The tracing of the transplanted cMSCs and EVs showed that they migrated and localized in the uteri. Protein expression analysis results demonstrated downregulation of proinflammatory factor TNFα and upregulation of anti-inflammatory cytokine IL-10, and endometrial receptivity cytokines VEGF and LIF in cMSC- and EV20K-treated animals.
Transplantation of MSCs and EVs contributed to endometrial repair and restoration of reproductive function, likely by inhibition of excessive fibrosis and inflammation, enhancement of endometrial cell proliferation, and regulation of molecular markers related to endometrial receptivity. Compared to classical hMSCs, cMSCs were more efficient than hMSCs in restoration of reproductive function. Moreover, EV20K is more cost-effective and feasible for prevention of AS in comparison with conventional EVs (EV110K).
Asherman 综合征（AS）或宫腔粘连是子宫内膜损伤后生殖年龄妇女不孕的主要原因。间充质干细胞（MSCs）及其细胞外囊泡（EVs）是修复受损子宫内膜的有前途的治疗候选物。然而，对其疗效的关注归因于细胞群体和 EVs 的异质性。需要同种 MSC 群体和有效的 EV 亚群来开发再生医学中潜在有前途的治疗选择。
通过机械损伤诱导成年大鼠子宫中的 AS 模型。然后，立即用同种异体人骨髓来源的克隆 MSC（cMSC）、异质亲本 MSC（hMSC）或 cMSC 衍生的 EV 亚群（EV20K 和 EV110K）处理动物。在治疗后两周处死动物并收集子宫角。取切片，用苏木精-伊红染色观察子宫内膜结构的修复。通过 Masson 三色染色和 Ki67 免疫染色测量纤维化程度和细胞增殖。通过交配试验的结果探索子宫的功能。通过 ELISA 测定 TNFα、IL-10、VEGF 和 LIF 的表达变化。
组织学分析表明，与完整和假手术动物相比，处理后的子宫中腺体减少、子宫内膜变薄、纤维化区域增加、上皮和基质细胞增殖减少。然而，在移植两种类型的 cMSC 和 hMSC 以及/或两种冷冻保存的 EV 亚群后，这些参数得到了改善。与 hMSC 相比，cMSC 显示出胚胎植入的成功率更高。移植的 cMSC 和 EV 的示踪表明它们迁移并定位于子宫。蛋白质表达分析结果表明，cMSC 处理的动物中促炎因子 TNFα下调，抗炎细胞因子 IL-10、子宫内膜容受性细胞因子 VEGF 和 LIF 上调。
MSC 和 EV 的移植有助于子宫内膜修复和生殖功能的恢复，可能通过抑制过度纤维化和炎症、增强子宫内膜细胞增殖以及调节与子宫内膜容受性相关的分子标志物来实现。与经典的 hMSC 相比，cMSC 在恢复生殖功能方面比 hMSC 更有效。此外，与传统 EV（EV110K）相比，EV20K 在预防 AS 方面更具成本效益和可行性。
