Stomatology Hospital, School of Stomatology, Zhejiang University School of Medicine, Zhejiang Provincial Clinical Research Center for Oral Diseases, Key Laboratory of Oral Biomedical Research of Zhejiang Province, Cancer Center of Zhejiang University, Hangzhou, 310000, China.
Stem Cell Res Ther. 2024 Nov 28;15(1):455. doi: 10.1186/s13287-024-04081-9.
Increasing evidence suggests that mesenchymal stem cells (MSCs) repair traumatized tissues primarily through paracrine secretion and differentiation into specific cell types. However, the role of epithelial differentiation of MSCs in cutaneous wound healing is unclear. This study aimed to investigate the epithelial differentiation potential of gingival tissue-derived MSCs (GMSCs) in epithelial cell growth medium and the mechanisms underlying their differentiation into an epithelial-like cell phenotype.
We used scanning electron microscopy to examine GMSCs for epithelial differentiation. Quantitative real-time polymerase chain reaction and Western blotting were respectively used to measure genes and proteins related to epithelial differentiation. Immunofluorescence was used to examine subcellular localization of KLF4, KRT19, and β-catenin proteins. Transcriptome sequencing was used to enrich the mechanisms underlying epithelial differentiation in GMSCs. An MSAB inhibitor was used to validate the Wnt signaling pathway further. The wound healing rate and re-epithelialization were assessed through macroscopical observation and hematoxylin and eosin staining.
GMSCs cultured in epithelial cell growth medium from days 3 to 15 exhibited decreased expression of mesenchymal-epithelial transition and stemness-related proteins (N-cadherin, Vimentin, KLF4, and SOX2), increased expression of epithelial-related proteins (KRT12, KRT15, KRT19, and E-cadherin), and exhibited epithelial-like morphology. Mechanistically, high-throughput sequencing revealed that the Wnt and TGF-beta signaling pathways were inhibited during epithelial differentiation of GMSCs (Epi-GMSCs). MSAB-induced Wnt signaling pathway inhibition promoted epithelial-related gene and protein expression. Furthermore, we demonstrated the ability of Epi-GMSCs to facilitate wound healing by improving re-epithelialization in a full-thickness skin defect model.
Collectively, this study uncovers that GMSCs have the ability to differentiate into epithelia and highlights a promising strategy for using Epi-GMSCs to improve cutaneous wound healing.
越来越多的证据表明，间充质干细胞（MSCs）主要通过旁分泌和分化为特定细胞类型来修复受损组织。然而，MSCs 的上皮分化在皮肤伤口愈合中的作用尚不清楚。本研究旨在探讨牙龈组织来源的间充质干细胞（GMSCs）在上皮细胞生长培养基中的上皮分化潜能及其向上皮样细胞表型分化的机制。
我们使用扫描电子显微镜观察 GMSCs 的上皮分化情况。实时定量聚合酶链反应和 Western blot 分别用于测量与上皮分化相关的基因和蛋白。免疫荧光用于检测 KLF4、KRT19 和 β-catenin 蛋白的亚细胞定位。转录组测序用于富集 GMSCs 上皮分化的机制。MSAB 抑制剂进一步验证 Wnt 信号通路。通过宏观观察和苏木精-伊红染色评估伤口愈合率和再上皮化。
GMSCs 在上皮细胞生长培养基中培养 3 至 15 天，间充质-上皮转化和干细胞相关蛋白（N-钙黏蛋白、波形蛋白、KLF4 和 SOX2）表达下调，上皮相关蛋白（KRT12、KRT15、KRT19 和 E-钙黏蛋白）表达上调，呈现上皮样形态。机制上，高通量测序显示，GMSCs 上皮分化过程中 Wnt 和 TGF-β 信号通路被抑制（Epi-GMSCs）。MSAB 诱导的 Wnt 信号通路抑制促进了上皮相关基因和蛋白的表达。此外，我们在全层皮肤缺损模型中证明了 Epi-GMSCs 能够通过改善再上皮化来促进伤口愈合的能力。
总之，本研究揭示了 GMSCs 具有分化为上皮细胞的能力，并强调了使用 Epi-GMSCs 改善皮肤伤口愈合的有前途的策略。
