A textile mesh-collagen hybrid matrix and cell sheet technology for skin tissue engineering

Abstract

A number of natural and synthetic materials have been explored for the purpose of skin tissue engineering. However, each of these materials used has its limitations. The ideal matrix for skin tissue engineering, from both functional and structural perspectives, has not been found. We present a novel concept of using a knitted textile mesh-collagen hybrid matrix, in combination with cell sheet culture technology, which has the potential to improve some limitations in currently available skin equivalent products. Poly(lactic-co-glycolic acid) (PLGA, 10:90) fibers of ca. 20μm were knitted into a stretchable mesh and freeze-dried with extracted rat tail collagen to produce a hybrid matrix. Human dermal fibroblasts were cultured with this matrix and the resulting cell sheet was folded around the matrix to form a 3D dermal substitute. Human keratinocytes were subsequently seeded onto the dermal substitutes and a bilayered skin equivalent was obtained by established protocols. The cell-matrix constructs were evaluated for cell viability, proliferation, distribution and protein expression. Preliminary results showed that this hybrid matrix, used in combination with a cell sheet technique, allows for the development of a dermal substitute with mature extracellular matrix formation. Undesirable effects of repeated trypsinization cycles on cellular functions could also be minimised. This novel technology platform offers the potential to culture a functionally sound and structurally stable skin equivalent.