Department of Obstetrics and Gynecology, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai, 200233, China.
School of Biomedical Engineering, Shanghai Jiao Tong University, Shanghai, 200240, China.
Stem Cell Res Ther. 2022 Jul 23;13(1):331. doi: 10.1186/s13287-022-03003-x.
Due to the large area and deep width of the artificial neovagina after vaginoplasty, it takes a considerable amount of time to achieve complete epithelization of the neovagina. Currently, the clinical therapies for vaginal epithelization after vaginoplasty are still dissatisfactory. Recent studies showed that small extracellular vesicles (sEVs) derived from stem cells could accelerate wound epithelization. The sustained release of sEVs from optimized hydrogels may be a promising strategy to accelerate vaginal epithelization after vaginoplasty.
The efficacy of phototriggered imine crosslink hydrogels (piGEL) containing sEVs derived from human urine-derived stem cells (hUSC-sEVs, piGEL-sEVs) on vaginal mucosa defects in rabbits was assessed by wound closure rates, histological analysis and immunofluorescence staining analysis. Cell counting kit-8, 5-ethynyl-2'-deoxyuridine and scratch wound assays were performed to assess the effects of hUSC-sEVs on the proliferation and migration ability of vaginal epithelial cells (VK2/E6E7). Quantitative real-time polymerase chain reaction (qRT-PCR) was carried out to test the expression of epithelial differentiation markers in VK2 cells. Moreover, a microRNA (miRNA) microarray was used to find hUSC-sEVs-specific miRNAs that potentially affected the proliferation, migration and differentiation ability of VK2 cells.
The in vitro release profile revealed that the piGEL could ensure sustained release of hUSC-sEVs. The in vivo results showed that piGEL-sEVs effectively promoted epithelization and angiogenesis of vaginal mucosa defects in rabbits. According to miRNA microarray and qRT-PCR results, miR-126-3p might be the crucial molecule among the various miRNAs contained in hUSC-sEVs. The data showed that hUSC-sEVs promoted the migration and differentiation of VK2 cells by delivering miR-126-3p to suppress the expression of Spred1 and PIK3R2, thereby activating the ERK1/2 and ATK signaling pathways.
The results indicated that piGEL-sEVs could be a novel promising approach for enhancing the epithelization of the neovagina after vaginoplasty and provided useful data for understanding the underlying mechanism of the effect of hUSC-sEVs on epithelization.
由于阴道成形术后人工阴道的面积大、宽度深，阴道上皮化需要相当长的时间。目前，阴道成形术后阴道上皮化的临床治疗仍不尽如人意。最近的研究表明，来源于干细胞的小细胞外囊泡（sEVs）可以加速创面上皮化。从优化的水凝胶中持续释放 sEVs 可能是加速阴道成形术后阴道上皮化的一种很有前途的策略。
通过伤口闭合率、组织学分析和免疫荧光染色分析，评估了含有人尿液来源的干细胞（hUSC-sEVs）衍生的 sEVs 的光触发亚胺交联水凝胶（piGEL-sEVs）对兔阴道黏膜缺损的疗效。细胞计数试剂盒-8、5-乙炔基-2'-脱氧尿苷和划痕伤口试验用于评估 hUSC-sEVs 对阴道上皮细胞（VK2/E6E7）增殖和迁移能力的影响。实时定量聚合酶链反应（qRT-PCR）用于检测 VK2 细胞中上皮分化标志物的表达。此外，还使用 miRNA 微阵列寻找可能影响 VK2 细胞增殖、迁移和分化能力的 hUSC-sEVs 特异性 miRNAs。
体外释放曲线表明，piGEL 可以确保 hUSC-sEVs 的持续释放。体内结果表明，piGEL-sEVs 可有效促进兔阴道黏膜缺损的上皮化和血管生成。根据 miRNA 微阵列和 qRT-PCR 结果，miR-126-3p 可能是 hUSC-sEVs 中各种 miRNAs 中的关键分子。研究数据表明，hUSC-sEVs 通过递送 miR-126-3p 来抑制 Spred1 和 PIK3R2 的表达，从而激活 ERK1/2 和 ATK 信号通路，促进 VK2 细胞的迁移和分化。
结果表明，piGEL-sEVs 可能是一种增强阴道成形术后新阴道上皮化的新方法，为了解 hUSC-sEVs 对上皮化影响的潜在机制提供了有用的数据。
