Cell Imprint‐Mediated Differentiation of Adipose‐Derived Stem Cells Into Keratinocytes Enhances Wound Healing on Collagen‐Based Scaffolds: An Ovine Model Study

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Full-thickness skin wound management remains a significant clinical challenge, necessitating innovative approaches that combine cellular therapy with biomaterial scaffolds. One of the promising approaches in regenerative medicine is the recellularisation of wound dressings. A polydimethylsiloxane (PDMS) substrate imprinted with ovine foetal keratinocytes was used as a template to induce the keratinocyte differentiation of adipose-derived mesenchymal stem cells (ADSCs) in this study. The therapeutic efficacy on full-thickness cutaneous wound regeneration was evaluated by employing keratinocyte-differentiated ADSCs (KC-ADSCs) in combination with collagen scaffolds in an ovine model. The successful differentiation of ADSCs into keratinocyte-like cells through the imprinted PDMS substrate was confirmed via immunocytochemical analysis of specific keratinocyte markers. The study implemented a randomised controlled design comparing four treatment groups: KC-ADSCs seeded on a collagen scaffold, undifferentiated ADSCs on a collagen scaffold, acellular collagen scaffold, and untreated controls. Wound healing was evaluated with and without polypropylene wound isolation chambers to prevent keratinocyte migration and wound contraction. Healing outcomes were assessed through standardised macroscopic documentation and comprehensive histopathological analysis over 3 weeks. The KC-ADSC/collagen scaffold combination demonstrated significantly superior wound healing characteristics (p < 0.05), including enhanced re-epithelialisation, advanced granulation tissue maturation, reduced inflammatory infiltrate, and improved neovascularisation compared to control groups. This therapeutic superiority was particularly evident in chambered wounds, where the KC-ADSC/collagen construct promoted substantial epithelial regeneration despite restricted wound contraction and cell migration. Ultimately, this method can be introduced as a growth factor-independent approach for cell differentiation and a clinically applicable therapeutic strategy for skin tissue engineering and regenerative medicine.

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