Multilayered 3-D nanofibrous scaffold with chondroitin sulfate sustained release as dermal substitute

Electrospun nanofibers for skin tissue engineering applications face two main challenges. The low thickness of electrospun mats is the main reason for their weak load-bearing performance at clinical applications and limited cell penetration due to their small pore sizes. We have developed multi-layered nanofibrous 3D (M3DN) scaffolds comprising gelatin, polyvinyl alcohol, and chondroitin sulfate (CS) by an electrospinning method and attaching three electrospun layers via ethanol to cause interface fibers to come in contact with each other. Prepared M3DN scaffolds revealed a sustained CS release profile. The improved mechanical performance, stable release of CS, and penetration capability of the cells and blood vessels through the spaces between layers in the prepared multi-layered nanofibrous scaffolds demonstrate their potential applications in response to the increasing demand for replacement of damaged dermis. The results of animal studies on the dorsal skin of Rat with full-thickness wounds have shown that the reconstruction of full-thickness skin lesions is significantly higher for M3DN scaffolds than a control group (treated with sterile gauze). The amount of epithelization, collagen arrangement, and inflammatory cells (acute and chronic) has been investigated, and their associated results demonstrated that M3DN scaffolds have great potential for full-thickness wound restoration.