Sousa Patrícia, Lopes Bruna, Sousa Ana Catarina, de Sousa Moreira Alícia, Rêma Alexandra, Alvites Rui, Geuna Stefano, Alves Nuno, Maurício Ana Colette
Departamento de Clínicas Veterinárias, Instituto de Ciências Biomédicas de Abel Salazar (ICBAS), Universidade do Porto (UP), Rua de Jorge Viterbo Ferreira, No. 228, 4050-313 Porto, Portugal.
Centro de Estudos de Ciência Animal (CECA), Instituto de Ciências, Tecnologias e Agroambiente da Universidade do Porto (ICETA), Rua D. Manuel II, Apartado 55142, 4051-401 Porto, Portugal.
Int J Mol Sci. 2025 May 25;26(11):5081. doi: 10.3390/ijms26115081.
Stem cell-derived secretome and exosomes present a promising cell-free strategy for tissue repair and wound healing. This study aimed to isolate and characterize, for the first time, exosomes derived from rat hair follicle stem cells (rHFSCs) and to evaluate their wound-healing potential alongside rHFSC secretome. Exosomes were isolated via ultracentrifugation and characterized using Reverse Transcriptase Polymerase Chain Reaction (RT-PCR), biomarker profiling and protein quantification. Scanning electron microscopy (SEM) with energy-dispersive X-ray spectroscopy (EDS) confirmed their spherical morphology, diameter and elemental composition. Protein quantification showed higher protein content in the secretome than in exosomes. RT-PCR and biomarker profiling highlighted the therapeutic relevance of the exosomal cargo compared to parent rHFSCs. Functional analysis of 30 wound-healing biomolecules validated their pro-regenerative potential. Cytocompatibility was confirmed via the PrestoBlue™ viability assay, while scratch assays demonstrated significant wound closure in the treated groups, both with and without mitomycin C. These findings highlight the potential of rHFSC-derived exosomes and secretome as innovative, cell-free therapeutic agents for cutaneous regeneration. This study advances our understanding of their role in wound healing and underscores their broader applicability in regenerative medicine.
干细胞来源的分泌组和外泌体为组织修复和伤口愈合提供了一种很有前景的无细胞策略。本研究旨在首次分离并鉴定源自大鼠毛囊干细胞（rHFSCs）的外泌体，并将其与rHFSC分泌组一同评估其伤口愈合潜力。通过超速离心分离外泌体，并使用逆转录聚合酶链反应（RT-PCR）、生物标志物分析和蛋白质定量进行表征。采用能量色散X射线光谱（EDS）的扫描电子显微镜（SEM）证实了它们的球形形态、直径和元素组成。蛋白质定量显示分泌组中的蛋白质含量高于外泌体。RT-PCR和生物标志物分析突出了外泌体货物与亲代rHFSCs相比的治疗相关性。对30种伤口愈合生物分子的功能分析验证了它们的促再生潜力。通过PrestoBlue™活力测定法确认了细胞相容性，而划痕试验表明，无论有无丝裂霉素C，治疗组的伤口均有显著闭合。这些发现突出了rHFSC来源的外泌体和分泌组作为用于皮肤再生的创新无细胞治疗剂的潜力。本研究推进了我们对它们在伤口愈合中作用的理解，并强调了它们在再生医学中更广泛的适用性。
