Combined use of novel chitosan-grafted N-hydroxyethyl acrylamide polyurethane and human dermal fibroblasts as a construct for in vitro-engineered skin

Abstract

A rich plethora of information about grafted chitosan (CS) for medical use has been reported. The capability of CS-grafted poly(N-hydroxyethyl acrylamide) (CS-g-PHEAA) to support human dermal fibroblasts (HDFs) in vitro has been proven. However, CS-grafted copolymers lack good stiffness and the characteristic microstructure of a cellular matrix. In addition, whether CS-g-PHEAA can be used to prepare a scaffold with a suitable morphology and mechanical properties for skin tissue engineering (STE) is unclear. This study aimed to show for the first time that step-growth polymerizations can be used to obtain polyurethane (PU) platforms of CS-g-PHEAA, which can also have enhanced microhardness and be suitable for in vitro cell culture. The PU prepolymers were prepared from grafted CS, polyethylene glycol, and 1,6-hexamethylene diisocyanate. The results proved that a poly(saccharide-urethane) [(CS-g-PHEAA)-PU] could be successfully synthesized with a more suitable microarchitecture, thermal properties, and topology than CS-PU for the dynamic culturing of fibroblasts. Cytotoxicity, proliferation, histological and immunophenotype assessments revealed significantly higher biocompatibility and cell proliferation of the derivative concerning the controls. Cells cultured on (CS-g-PHEAA)-PU displayed a quiescent state compared to those cultured on CS-PU, which showed an activated phenotype. These findings may be critical factors in future studies establishing wound dressing models.