Nahid Heidari Hafshejani
1 
, Azam Rahimpour
1, Hojjat Allah Abbaszadeh
2,3, Hojatollah Montazeri
4, Sepideh Ghani
5, Simzar Hosseinzadeh
4,6* , Bahram Kazemi
1,5,7 1 Department of Medical Biotechnology, School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran
2 Laser Applications in Medical Sciences Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran
3 Hearing Disorders Research Center, Loghman Hakim Hospital, Shahid Beheshti University of Medical Sciences, Tehran, Iran
4 Department of Tissue Engineering and Applied Cell Sciences, School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran
5 Cellular and Molecular Biology Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran
6 Medical Nanotechnology and Tissue Engineering Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran
7 Parasitology Department, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran
Abstract
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.