(1) Background: Meshed split skin grafting (SSG) still remains the gold standard of care for deep wound treatment. The aim of this study was to evaluate an autologous bioengineered dermal–epidermal composite skin for treating full-thickness wounds in a porcine model using a clinically relevant two-stage grafting approach. (2) Methods: Multiple full-thickness wounds were created on the dorsal sites of animals. Wounds were initially grafted with a polyurethane dermal template, followed three weeks later by either Platelet hydroGel–Pig Skin Equivalent (PG-PSE) or meshed SSG control. Human platelet lysate was also assessed as a substitute for foetal calf serum during skin maturation. (3) Results: By two weeks post-grafting, epidermisation of PG-PSE-treated wounds were comparable to SSG-treated wounds. EdU labelling confirmed early engraftment and contribution of graft-derived cells to both neo-epidermis and neo-dermis. Cytokeratin 5+ immunofluorescence staining, showed a hyperproliferative neo-epidermis, enriched in progenitor keratinocytes, typical of early-stage skin grafts in pigs. Neo-dermis was marked with abundance of vimentin⁺ fibroblasts and rete ridges indicating a mature dermal–epidermal junction. Robust neovascularisation was observed in both superficial and deep neo-dermis, with platelet lysate-matured grafts showing greater microvessel formation in superficial neo-dermis in comparison with native skin. Picrosirius red staining of the superficial neo-dermis revealed a lower abundance of thick, red collagen fibres in PG-PSE grafts compared to SSGs, consistent with ongoing collagen deposition and tissue integration in PG-PSE. However, the ratio of thin green collagen fibres to thick red collagen fibres in the deep neo-dermis was largely comparable between group conditions, suggesting that bioengineered grafting does not alter dermal template maturation. (4) Conclusion: Overall, autologous dermal–epidermal bioengineered skin effectively vascularises and repairs wounds over a dermal template. Platelet lysate is a promising alternative to foetal calf serum, although PG-PSE may require longer time for complete dermal maturation and warrants further long-term evaluation.

(1) Background: Meshed split skin grafting (SSG) still remains the gold-standard of care for deep wound treatment. The aim of this study was to evaluate an autologous bioengineered dermal–epidermal composite skin for treating full-thickness wounds in a porcine model using a clinically relevant two-stage grafting approach. (2) Methods: Multiple full thickness wounds were created on the dorsal sites of animals. Wounds were initially grafted with a polyurethane dermal template, followed three weeks later by either Platelet hydroGel–Pig Skin Equivalent (PG-PSE) or meshed SSG control. Human platelet lysate was also assessed as a substitute for foetal calf serum during skin maturation. (3) Results: By two weeks post grafting, epidermisation of PG-PSE–treated wounds was comparable to SSG-treated wounds. EdU labelling confirmed early engraftment and contribution of graft-derived cells to both neo-epidermis and neo-dermis. Immunofluorescence staining for vimentin+ fibroblasts and cytokeratin 5+ keratinocytes demonstrated formation of a neo-dermis and a hyperproliferative neo-epidermis enriched in progenitor keratinocytes, with rete ridge architecture indicative of dermal–epidermal maturation. Robust neovascularisation was observed in both papillary and reticular neo-dermis, with platelet lysate–matured grafts showing greater microvessel formation than foetal calf serum–matured grafts. Collagen deposition in PG-PSE grafts, particularly collagen I in the papillary neo-dermis, remained lower than in SSG-treated wounds, consistent with ongoing integration. However, collagen type I/type III composition in the reticular neo-dermis was largely comparable between PG-PSE and SSG, suggesting that bioengineered grafting does not alter dermal template maturation. (4) Conclusion: Overall, autologous dermal–epidermal bioengineered skin effectively vascularises and repairs wounds over a dermal template. Platelet lysate is a promising alternative to foetal calf serum, although PG-PSE may require longer for complete dermal maturation and warrants further long-term evaluation.