Tabriz University of Medical Sciences BioImpacts 2228-5652 16 1 2026 01 04 Design and evaluation of a multifunctional electrospun scaffold loaded with amniotic membrane and allantoin for dermal tissue engineering 33538 33538 10.34172/bi.33538 EN Nahid Heidari Hafshejani https://orcid.org/0009-0000-1093-0904 Azam Rahimpour Hojjat Allah Abbaszadeh Hojatollah Montazeri Sepideh Ghani Simzar Hosseinzadeh https://orcid.org/0000-0001-8749-2447 Bahram Kazemi https://orcid.org/0000-0002-3072-8831 Journal Article 10.34172/bi.33538 2026 02 10 2026 09 16 Introduction: Chronic wounds remain difficult to treat because healing is impaired, inflammation persists, and infection risk is high. Whether a spatially organized three-layer electrospun scaffold containing amniotic membrane powder and allantoin can combine mechanical support with sustained bioactive delivery during full-thickness dermal regeneration remains unclear. Methods: The scaffold was electrospun in 3 layers: polyurethane, polyethylene oxide containing amniotic membrane powder and allantoin, and a gelatin/carbomer blend. Physicochemical properties and fibroblast responses were assessed, followed by wound healing in a rat full-thickness excisional model (n = 12 per group; n = 3 per time point) over 21 days. Results: All layers displayed uniform nanofibers. The 3L/Amnion/Allantoin scaffold reached an ultimate tensile strength of 38.19 MPa. Superhydrophilic behavior, with a contact angle of 0°, was observed for 3L/Allantoin. The 3L/Amnion/Allantoin scaffold increased fibroblast viability (P < 0.0001 versus controls at day 14) and upregulated VEGF (2-fold) and DDR2 (1.7-fold) expression. On day 7, wound closure reached 50.0 ± 1.0% in the 3L/Fibroblast/Amnion/Allantoin group versus 15.0 ± 1.0% in the untreated control (P < 0.0001). By day 14, this group reached 90.0 ± 1.0% closure versus 50.0 ± 1.0% in controls (P < 0.0001), accompanied by accelerated re-epithelialization and organized collagen deposition. By day 21, wound closure reached ≥ 99% in all groups, with no significant intergroup differences. Conclusion: The three-layer scaffold combined structural reinforcement with local bioactive delivery and accelerated wound closure through day 14, supporting further evaluation in dermal tissue engineering.