Annual Meeting of the Society for Cutaneous Ultrastructure Research (33rd Annual Meeting of SCUR) 8–10 June 2006 WARSZAWA, POLAND

Abstract

Unsupported keratinocyte grafts, while fragile, provide excellent clinical benefits and wound coverage. We aim to overcome the fragile nature of these grafts by using keratinocytes and fibroblast grafts grown on biocompatible, biodegradable, hexagonalpacked porous poly-calprolactone membranes. To confirm the feasibility of this proposal, human epidermal keratinocytes and dermal fibroblasts were cultured on membranes with controlled-sized pores (ranging from 3–20 microns). We examined the effects of growing monocultures on these porous membranes and on flat (pore less) polymer films. Cell adhesion, growth and migration rates and basic morphology were examined using confocal laser scanning microscopy. Cell ultrastructure was subsequently examined using scanning and transmission electron microscopy. Human keratinocytes and fibroblasts attached to all porous membranes. Furthermore, small pore (3 and 5 micron) membranes provided the best surfaces, on which fibroblasts could attach, adhere, divide and grow. However, keratinocytes, the most discerning of the two cell types adhered and grew best on 3 micron pore membranes. In addition, cells were prevented from undergoing transmembrane migration (to the opposing membrane surface) by the 3 micron porous membrane. Larger pores (45microns) allowed migration of both fibroblasts (and to a lesser extent keratinocytes) on to the opposite side of the membrane. Our data show that keratinocytes and fibroblasts attached and grew optimally on small pore membranes due to greater pores numbers and a larger surface area over which cells adhere. The small pore size also inhibits cells from traversing the membrane while, at the same time allowing cell adhesion, matrix communication and diffusion of soluble nutrients/factors from the environment. These characteristics are important in development of graft technology, in the treatment of human skin wounds and in the study of cell-substrate adhesion on novel patterned structures