Design of Novel 3D-Scaffold as a Potential Material to Induct Epidermal-Dermal Keratinocytes of Human-Adipose-Derived Stem Cells and Promote Fibroblast Cells Proliferation for Skin Regeneration

Abstract

Dermal lesions and chronic wounds associated with burns or some diseases like diabetes are the more important public health concerns which can affect the quality of life. Currently, tissue engineering is considered as the most effective therapeutic method although the design of polymeric substrates for epidermal-dermal differentiation and wound healing (scar-free) is the main challenge. For this purpose, we designed a hybrid three-dimensional scaffold (CPCP) based on collagen/chitosan modified by PEG/PCL composite that can imitate differentiation pattern of both epidermis/dermis cells, via mimicking the structure and function of human skin. The physicochemical, mechanical and biological properties of designed scaffolds were evaluated to study their function for skin tissue engineering applications. Comparison of FTIR analysis showed a chemical similarity between CPCP and decellularized dermal matrix (DDM). Our results showed that combination of two natural/two synthetic polymers led to the formation of stronger 3D-network together with higher modulus (-18), water absorption (4-fold), porosity (-92) and consequently lower pores size (-54 um), compared to natural, synthetic and natural/ synthetic copolymer-based scaffolds. The observation of human skin fibroblast cells proliferation and morphology showed that CPCP was more beneficial to cell adhesion, proliferation, and extension than that of other designed scaffolds due to its hydrophilicity and higher wettability (WCA=60°). According to the results of RT-PCR, the more expression of epidermal-dermal keratinocytes induced by human-adipose-derived stem cells was observed on the CPCP along with a pattern similar to skin. The results demonstrate CPCP can act as a super-absorbent substrate/dressing for continuous absorption of wound exudates. Furthermore, it can potentially be effective for re-epithelialization of skin together with its derivative (hair follicles, sebaceous/sweat glands). This study indicates new insights into the design of skin- engineered scaffolds.