{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T01:19:31Z","timestamp":1760059171807,"version":"build-2065373602"},"reference-count":115,"publisher":"MDPI AG","issue":"11","license":[{"start":{"date-parts":[[2025,5,25]],"date-time":"2025-05-25T00:00:00Z","timestamp":1748131200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Funda\u00e7\u00e3o para a Ci\u00eancia e Tecnologia (FCT)","award":["2023.00246.BD","2021.05265.BD","2023.00544.BD"],"award-info":[{"award-number":["2023.00246.BD","2021.05265.BD","2023.00544.BD"]}]},{"name":"Animal Science Studies Centre (CECA)","award":["2023.00246.BD","2021.05265.BD","2023.00544.BD"],"award-info":[{"award-number":["2023.00246.BD","2021.05265.BD","2023.00544.BD"]}]},{"name":"Technologies and Sciences Institute (ICETA)","award":["2023.00246.BD","2021.05265.BD","2023.00544.BD"],"award-info":[{"award-number":["2023.00246.BD","2021.05265.BD","2023.00544.BD"]}]},{"name":"Porto University (UP)","award":["2023.00246.BD","2021.05265.BD","2023.00544.BD"],"award-info":[{"award-number":["2023.00246.BD","2021.05265.BD","2023.00544.BD"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>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\u2122 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.<\/jats:p>","DOI":"10.3390\/ijms26115081","type":"journal-article","created":{"date-parts":[[2025,5,25]],"date-time":"2025-05-25T20:26:50Z","timestamp":1748204810000},"page":"5081","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Rat Hair Follicle Stem Cell-Derived Exosomes: Isolation, Characterization and Comparative Analysis of Their In Vitro Wound Healing Potential"],"prefix":"10.3390","volume":"26","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-2531-3196","authenticated-orcid":false,"given":"Patr\u00edcia","family":"Sousa","sequence":"first","affiliation":[{"name":"Departamento de Cl\u00ednicas Veterin\u00e1rias, Instituto de Ci\u00eancias Biom\u00e9dicas de Abel Salazar (ICBAS), Universidade do Porto (UP), Rua de Jorge Viterbo Ferreira, No. 228, 4050-313 Porto, Portugal"},{"name":"Centro de Estudos de Ci\u00eancia Animal (CECA), Instituto de Ci\u00eancias, Tecnologias e Agroambiente da Universidade do Porto (ICETA), Rua D. Manuel II, Apartado 55142, 4051-401 Porto, Portugal"},{"name":"Associate Laboratory for Animal and Veterinary Science (AL4AnimalS), 1300-477 Lisboa, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8168-2415","authenticated-orcid":false,"given":"Bruna","family":"Lopes","sequence":"additional","affiliation":[{"name":"Departamento de Cl\u00ednicas Veterin\u00e1rias, Instituto de Ci\u00eancias Biom\u00e9dicas de Abel Salazar (ICBAS), Universidade do Porto (UP), Rua de Jorge Viterbo Ferreira, No. 228, 4050-313 Porto, Portugal"},{"name":"Centro de Estudos de Ci\u00eancia Animal (CECA), Instituto de Ci\u00eancias, Tecnologias e Agroambiente da Universidade do Porto (ICETA), Rua D. Manuel II, Apartado 55142, 4051-401 Porto, Portugal"},{"name":"Associate Laboratory for Animal and Veterinary Science (AL4AnimalS), 1300-477 Lisboa, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8800-2195","authenticated-orcid":false,"given":"Ana Catarina","family":"Sousa","sequence":"additional","affiliation":[{"name":"Departamento de Cl\u00ednicas Veterin\u00e1rias, Instituto de Ci\u00eancias Biom\u00e9dicas de Abel Salazar (ICBAS), Universidade do Porto (UP), Rua de Jorge Viterbo Ferreira, No. 228, 4050-313 Porto, Portugal"},{"name":"Centro de Estudos de Ci\u00eancia Animal (CECA), Instituto de Ci\u00eancias, Tecnologias e Agroambiente da Universidade do Porto (ICETA), Rua D. Manuel II, Apartado 55142, 4051-401 Porto, Portugal"},{"name":"Associate Laboratory for Animal and Veterinary Science (AL4AnimalS), 1300-477 Lisboa, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1347-8789","authenticated-orcid":false,"given":"Al\u00edcia","family":"de Sousa Moreira","sequence":"additional","affiliation":[{"name":"Departamento de Cl\u00ednicas Veterin\u00e1rias, Instituto de Ci\u00eancias Biom\u00e9dicas de Abel Salazar (ICBAS), Universidade do Porto (UP), Rua de Jorge Viterbo Ferreira, No. 228, 4050-313 Porto, Portugal"},{"name":"Centro de Estudos de Ci\u00eancia Animal (CECA), Instituto de Ci\u00eancias, Tecnologias e Agroambiente da Universidade do Porto (ICETA), Rua D. Manuel II, Apartado 55142, 4051-401 Porto, Portugal"},{"name":"Associate Laboratory for Animal and Veterinary Science (AL4AnimalS), 1300-477 Lisboa, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4319-2404","authenticated-orcid":false,"given":"Alexandra","family":"R\u00eama","sequence":"additional","affiliation":[{"name":"Departamento de Cl\u00ednicas Veterin\u00e1rias, Instituto de Ci\u00eancias Biom\u00e9dicas de Abel Salazar (ICBAS), Universidade do Porto (UP), Rua de Jorge Viterbo Ferreira, No. 228, 4050-313 Porto, Portugal"},{"name":"Centro de Estudos de Ci\u00eancia Animal (CECA), Instituto de Ci\u00eancias, Tecnologias e Agroambiente da Universidade do Porto (ICETA), Rua D. Manuel II, Apartado 55142, 4051-401 Porto, Portugal"},{"name":"Associate Laboratory for Animal and Veterinary Science (AL4AnimalS), 1300-477 Lisboa, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1945-6377","authenticated-orcid":false,"given":"Rui","family":"Alvites","sequence":"additional","affiliation":[{"name":"Departamento de Cl\u00ednicas Veterin\u00e1rias, Instituto de Ci\u00eancias Biom\u00e9dicas de Abel Salazar (ICBAS), Universidade do Porto (UP), Rua de Jorge Viterbo Ferreira, No. 228, 4050-313 Porto, Portugal"},{"name":"Centro de Estudos de Ci\u00eancia Animal (CECA), Instituto de Ci\u00eancias, Tecnologias e Agroambiente da Universidade do Porto (ICETA), Rua D. Manuel II, Apartado 55142, 4051-401 Porto, Portugal"},{"name":"Associate Laboratory for Animal and Veterinary Science (AL4AnimalS), 1300-477 Lisboa, Portugal"},{"name":"Instituto Universit\u00e1rio de Ci\u00eancias da Sa\u00fade (IUCS), Instituto Universit\u00e1rio de Ci\u00eancias da Sa\u00fade (CESPU), Avenida Central de Gandra 1317, 4585-116 Paredes, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6962-831X","authenticated-orcid":false,"given":"Stefano","family":"Geuna","sequence":"additional","affiliation":[{"name":"Centre for Rapid and Sustainable Product Development, Polytechnic of Leiria, 2430-028 Marinha Grande, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5016-0868","authenticated-orcid":false,"given":"Nuno","family":"Alves","sequence":"additional","affiliation":[{"name":"Department of Clinical and Biological Sciences, Cavalieri Ottolenghi Neuroscience Institute, University of Turin, Ospedale San Luigi, 10043 Turin, Italy"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0018-9363","authenticated-orcid":false,"given":"Ana Colette","family":"Maur\u00edcio","sequence":"additional","affiliation":[{"name":"Departamento de Cl\u00ednicas Veterin\u00e1rias, Instituto de Ci\u00eancias Biom\u00e9dicas de Abel Salazar (ICBAS), Universidade do Porto (UP), Rua de Jorge Viterbo Ferreira, No. 228, 4050-313 Porto, Portugal"},{"name":"Centro de Estudos de Ci\u00eancia Animal (CECA), Instituto de Ci\u00eancias, Tecnologias e Agroambiente da Universidade do Porto (ICETA), Rua D. Manuel II, Apartado 55142, 4051-401 Porto, Portugal"},{"name":"Associate Laboratory for Animal and Veterinary Science (AL4AnimalS), 1300-477 Lisboa, Portugal"}]}],"member":"1968","published-online":{"date-parts":[[2025,5,25]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Sousa, P., Lopes, B., Sousa, A.C., Moreira, A., Coelho, A., Alvites, R., Alves, N., Geuna, S., and Maur\u00edcio, A.C. (2023). Advancements and Insights in Exosome-Based Therapies for Wound Healing: A Comprehensive Systematic Review (2018\u2013June 2023). Biomedicines, 11.","DOI":"10.20944\/preprints202306.1736.v1"},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Sousa, P., Lopes, B., Sousa, A.C., Coelho, A., de Sousa Moreira, A., R\u00eama, A., Gon\u00e7alves-Maia, M., Amorim, I., Alvites, R., and Alves, N. (2024). Isolation, Expansion, and Characterization of Rat Hair Follicle Stem Cells and Their Secretome: Insights into Wound Healing Potential. 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