Development of porous nano/microfibrous layered skin regenerating scaffold supporting human skin equivalent formation

摘要

Grafting is preferable for the healing of a full-thickness wound of more than one cm in diameter due to lack of dermal tissue. Autografts, allografts, xenografts and cell therapies are the most common therapeutic options to heal full-thickness skin wounds. However, antigenicity, post-operative infections, and limitation of donor sites, restricts the use of the skin substitutes mentioned above. Therefore, tissue engineering approach to promote tissue repair, regeneration, and recovery of their functions is appreciable. Collagen, the structural and functional component of dermal extracellular matrix, plays a vital role in wound healing process. This fostered collagen as the most favoured material for dermal substitute Collagen as healing matrix is used in different forms, namely nanofiber, sponge, and hydrogel. In this study, emulsion based electrospinning was done to develop highly porous nano/microfibrous scaffold of PCL/chitosan. The scaffolds were then layered with collagen, extracted from fish scale . The microstructures of samples were evaluated by scanning electron microscopy The collagen layer had the natural d- spacing as seen in dermal collagen. FTIR spectroscopy, mechanical properties, swelling behavior, biodegradation kinetics of matrices was studied for their suitability as skin regenerating scaffold. Normal fibroblast cells were grown to check the scaffold's cytocompatibility and SEM images revealed cell infiltration from day 1 Normal human fibroblast and keratinocyte were co-cultured and a stratified human skin equivalent was developed. Acute full thickness wound healing rate was tested on rat on time dependent scale and better wounding healing kinetics was reported.