Liu Minjie, Ye Ganqin, Li Ruiyang, Gao Tenghui, Zhuang Caiwei, Huang Shiyun, Wang Sheng, Hu Jinhua, Xiang Andy Peng, Jiang Mei Hua
Program of Stem Cells and Regenerative Medicine, Affiliated Guangzhou Women and Children's Hospital and Department of Anatomy, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, China.
Center for Stem Cell Biology and Tissue Engineering, Key Laboratory for Stem Cells and Tissue Engineering, Ministry of Education, Sun Yat-sen University, Guangzhou, China.
Cell Biol Int. 2025 Jul 29. doi: 10.1002/cbin.70063.
Fibroblasts are pivotal cellular components in cutaneous wound healing and are regarded as promising therapeutic candidates. However, their functional heterogeneity within tissue microenvironments significantly limits their clinical application. In contrast, whether fibroblast-derived extracellular vesicles (EVs) can overcome fibroblast heterogeneity while retaining the bioactivity and regenerative potential of homeostatic fibroblasts remains unclear. In this study, we systematically analyzed and compared the therapeutic potential and functional advantages of human dermal fibroblast-derived EVs (hDF-EVs) in promoting cutaneous wound healing. Our findings highlight the translational potential of fibroblast-derived EVs as a novel strategy to improve clinical outcomes for skin injuries. hDF-EVs were internalized by fibroblasts and keratinocytes at the wound margins, thereby attenuating early inflammatory responses and accelerating tissue repair following dermal excisional injuries. hDF-EVs significantly enhanced the proliferation and migration of both fibroblasts and keratinocytes in a coculture system. Transcriptomic analysis revealed that hDF-EVs upregulated genes involved in cell proliferation and cytokine regulation. Integrated miRNA profiling revealed a subset of hDF-EVs-enriched miRNAs that mechanistically orchestrate fibroblast activation through coordinated MAPK, Wnt, and Ras signaling axis engagement, consequently reprogramming inflammatory mediator secretion dynamics in wound microenvironments. Furthermore, cytokine array analysis demonstrated that hDF-EVs enhanced the expression of various cytokines, including Amphiregulin, GCSF, IL-7, and IL-2, while activating Ras, Rap1, PI3K-Akt, and MAPK signaling pathways during the early stage of wound healing. Collectively, hDF-EVs promote wound healing by modulating early growth factor dynamics and enhancing fibroblast-keratinocyte crosstalk, presenting a novel cell-free strategy for skin regeneration.
成纤维细胞是皮肤伤口愈合过程中的关键细胞成分，被视为有前景的治疗候选者。然而，它们在组织微环境中的功能异质性显著限制了其临床应用。相比之下，成纤维细胞衍生的细胞外囊泡（EVs）能否克服成纤维细胞的异质性，同时保留稳态成纤维细胞的生物活性和再生潜力仍不清楚。在本研究中，我们系统地分析和比较了人真皮成纤维细胞衍生的EVs（hDF-EVs）在促进皮肤伤口愈合方面的治疗潜力和功能优势。我们的研究结果突出了成纤维细胞衍生的EVs作为一种改善皮肤损伤临床结果的新策略的转化潜力。hDF-EVs被伤口边缘的成纤维细胞和角质形成细胞内化，从而减轻早期炎症反应并加速皮肤切除损伤后的组织修复。在共培养系统中，hDF-EVs显著增强了成纤维细胞和角质形成细胞的增殖与迁移。转录组分析显示，hDF-EVs上调了参与细胞增殖和细胞因子调节的基因。综合miRNA谱分析揭示了一组hDF-EVs富集的miRNA，它们通过协调的MAPK、Wnt和Ras信号轴参与机制性地协调成纤维细胞活化，从而重新编程伤口微环境中炎症介质的分泌动态。此外，细胞因子阵列分析表明，hDF-EVs在伤口愈合早期增强了包括双调蛋白、粒细胞集落刺激因子、白细胞介素-7和白细胞介素-2在内的多种细胞因子的表达，同时激活了Ras、Rap1、PI3K-Akt和MAPK信号通路。总体而言，hDF-EVs通过调节早期生长因子动态和增强成纤维细胞-角质形成细胞相互作用来促进伤口愈合，为皮肤再生提供了一种新的无细胞策略。
