Poly(hydroxybutyrate-co-hydroxyvalerate) bilayer skin tissue engineering constructs with improved epidermal rearrangement

Abstract

Bilayer skin substitutes constitute an attractive strategy towards improved skinwound healing.Therefore, solvent casting and freeze-drying methodologies are used to produce polyhydroxybutyrate-co-hydroxyvalerate (PHBV) thin nanoporousmembranes and 3D porous scaffolds thatare combined in bilayer structures to recreate the epidermal and dermal layers, respectively. Thecombination of these methodologies allow attaining a bilayer structure with a high waterretention capability and adequate mechanical properties, susceptible to enzymes degradativeaction. Cultures established with human keratinocytes (hKC) and dermal fibroblasts(hDFb) confirm the suitability of the PHBV structures tosupport cell adhesion and proliferation. Nonetheless,when co-cultured under defined conditions, hKC are ableto grow and rearrange in a multilayer structure withproliferative cells in the basal layer, and cells expressing aterminal differentiation marker in the upper layer.Therefore, PHBV bilayer structures demonstrate propertiesthat favor skin cells performance, thus representing apromising strategy to improve wound healing.