Self Organized Skin Equivalent by Epithelial and Fibroblast Cells on Polycaprolactone Electrospun Mat with Porous Fibers

Abstract

Tissue engineered skin substitutes are used in chronic, partial or full thickness wounds, and serve to augment the growth and repair of the skin at the wound site, by acting as a temporary or permanent skin replacement. In this study Polycaprolactone a biodegradable polymer was electrospun in a binary solvent system to form nonwoven mats with porous fibers (PPCL). These mats were hydrolysed with alkali to make them more hydrophilic and promote cell adhesion. The electrospun mat served as a scaffold for culture of human keratinocyte (HaCaT) and rabbit hair follicle dermal fibroblast cells (HFDF) cells. F-actin staining and SEM analysis showed that the cells exhibited their characteristic morphology, with good adhesion and spreading. Live dead staining confirmed the viability of the cells on the PPCL mat. A bioengineered construct of HaCaT and HFDF cells cultured on either side of the PPCL mat was generated in vitro. Light microscopy of Toluidine blue stained cryo sections of the construct reveal that the HaCaT cells formed intact multilayers, while the fibroblasts showed a more distributed growth pattern with migration into the mat, hence emulating the epidermis and dermis of the skin. The results were substantiated in the SEM micrographs and fluorescent images of DiI and DiO labeled cocultures. The PPCL mat cocultured with HaCaT and HFDF cells thus finds potential as a self organized tissue engineered skin substitute.