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Methods: An ethanol extract of Origanum vulgare L. (in further text: OE), exhibiting exceptional antioxidant (100%) and antibacterial activity (&gt;99% against E.coli and S.aureus), was employed to biofunctionalize cotton, wool, and silk fabrics. Results: All biofunctionalized fabrics demonstrated strong antioxidant activity (&gt;99%), while antibacterial efficacy varied by fabric: cotton &gt; 54%, wool &gt; 99%, and silk &gt; 89%. OE-biofunctionalized wool possessed the highest release of OE\u2019s bioactive compounds, followed by silk and cotton, indicating substrate-dependent release behavior. This tunable fabrics\u2019 OE release profile, along with their unique bioactivity, supports targeted applications: OE-functionalized silk for luxury or prolonged therapeutic use (skin-care textiles, post-surgical dressings, anti-aging products), cotton for disposable or short-term use (protective wipes, minor wound coverings), and wool for wound dressings. The biocompatibility and cytotoxicity of OE-biofunctionalized wool were evaluated via in vitro assays using healthy human keratinocytes and in vivo testing in Wistar albino male rats. The obtained results revealed that OE-functionalized wool significantly accelerated wound closure (97.8% by day 14), enhanced collagen synthesis (6.92 \u00b5g\/mg hydroxyproline), and improved tissue and systemic antioxidant defense while reducing oxidative stress markers in skin and blood samples of rats treated with OE-biofunctionalized wool. Conclusions: OE-biofunctionalized wool demonstrates strong potential as an advanced natural solution for managing chronic wounds. Further clinical validation is recommended to confirm its performance in real-world healthcare settings. This work introduces an entirely new application of OE in textile biofunctionalization, offering alternatives for healthcare and medical textiles.<\/jats:p>","DOI":"10.3390\/pharmaceutics17070856","type":"journal-article","created":{"date-parts":[[2025,7,2]],"date-time":"2025-07-02T06:46:12Z","timestamp":1751438772000},"page":"856","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":11,"title":["Development of Bioactive Cotton, Wool, and Silk Fabrics Functionalized with Origanum vulgare L. for Healthcare and Medical Applications: An In Vivo Study"],"prefix":"10.3390","volume":"17","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-6846-9583","authenticated-orcid":false,"given":"Aleksandra","family":"Ivanovska","sequence":"first","affiliation":[{"name":"Innovation Center of the Faculty of Technology and Metallurgy, University of Belgrade, Karnegijeva 4, 11000 Belgrade, Serbia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3048-4264","authenticated-orcid":false,"given":"Anica","family":"Petrovi\u0107","sequence":"additional","affiliation":[{"name":"Department of Pharmacy, Faculty of Medical Sciences, University of Kragujevac, Svetozara Makovi\u0107a 69, 34000 Kragujevac, Serbia"},{"name":"Center of Excellence for Redox Balance Research in Cardiovascular and Metabolic Disorders, Svetozara Makovi\u0107a 69, 34000 Kragujevac, Serbia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6220-2079","authenticated-orcid":false,"given":"Tamara","family":"Lazarevi\u0107-Pa\u0161ti","sequence":"additional","affiliation":[{"name":"VIN\u010cA Institute of Nuclear Sciences\u2013National Institute of the Republic of Serbia, University of Belgrade, Mike Petrovica Alasa 12-14, 11000 Belgrade, Serbia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7198-2855","authenticated-orcid":false,"given":"Tatjana","family":"Ilic-Tomic","sequence":"additional","affiliation":[{"name":"Institute of Molecular Genetics and Genetic Engineering, University of Belgrade, Vojvode Stepe 444a, 11042 Belgrade, Serbia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6397-7711","authenticated-orcid":false,"given":"Katarina","family":"Dimi\u0107-Mi\u0161i\u0107","sequence":"additional","affiliation":[{"name":"Department of Chemical and Metallurgical Engineering, School of Chemical Engineering, Aalto University, 02150 Espoo, Finland"},{"name":"Institute of General and Physical Chemistry, Studentski Trg 12\/V, 11158 Belgrade, Serbia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5554-7295","authenticated-orcid":false,"given":"Jelena","family":"La\u0111arevi\u0107","sequence":"additional","affiliation":[{"name":"Faculty of Technology and Metallurgy, University of Belgrade, Karnegijeva 4, 11000 Belgrade, Serbia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5388-6085","authenticated-orcid":false,"given":"Jovana","family":"Bradi\u0107","sequence":"additional","affiliation":[{"name":"Department of Pharmacy, Faculty of Medical Sciences, University of Kragujevac, Svetozara Makovi\u0107a 69, 34000 Kragujevac, Serbia"},{"name":"Center of Excellence for Redox Balance Research in Cardiovascular and Metabolic Disorders, Svetozara Makovi\u0107a 69, 34000 Kragujevac, Serbia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,6,30]]},"reference":[{"key":"ref_1","unstructured":"Chassagne, F. 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