Development of chitosan/fish scales collagen/glycerin/ nano-ZnO 3D porous scaffolds for skin tissue engineering and regeneration applications

This study was aimed to develop three-dimensional (3D) porous scaffold from blending of chitosan/fish scales collagen/glycerin/nano zinc oxide for skin tissue engineering and skin regenerating template. 3D porous scaffolds were fabricated through freeze dry technique. The 3D porous scaffolds were subjected to phosphate buffer solution (PBS) washing while dehydrothermal (DHT) treatment and chemical treatment using 1-ethyl- 3-(3-dimethylaminopropyl)carbodiimide (EDAC)-N-hydroxysuccinimide (NHS) were used to create crosslinks. The mechanical properties, porosity and swelling ratio of the 3D porous scaffolds have been determined using general procedures. Crosslink density was calculated using the Fourier Transform Infrared (FTIR) technique while the crosslinking degree has been determined using 2,4,6-trinitrobenzenasulfonic acid (TNBS). Field emission scanning electron microscopy (FESEM) analysis was used to examine the morphology of 3D porous scaffolds. The biodegradation rate and cytocompatibility tests were performed using common procedure. The results showed that the 3D scaffolds were porous with interconnected pores with average pore sizes between 100 and 200 microns while mechanical properties and biostability fulfilled the requirements of the tissue engineering scaffold excepted for PBS washing treated scaffolds. In vitro analysis using human fibroblast and keratinocyte indicates that all 3D porous scaffolds were good cytocompatibility but scaffold containing nano zinc oxide and scaffold containing high fish scales collagen content were excellently facilitated cell proliferation and adhesion. In conclusions, the 3D porous scaffolds treated with DHT and EDAC-NHS and also the addition of zinc oxide nanoparticles and various chitosan/collagen/glycerin ratios showed excellent biostability. Furthermore, 3D scaffolds were also cytocompatible with mechanical strengths suitable for skin tissue engineering applications. The implication of this study indicate that porous 3D scaffolds produced most suitable to be used as skin tissue engineering and skin regeneration template applications.