In vitro reconstructed tissues on hyaluronan-based temporary scaffolding

Tissue engineering offers the possibility to reconstruct tissue substitutes in order to replace lost or damaged tissues. The availability of appropriate biomaterial devices is essential to allow in vitro cultured cells to behave as in the original tissues in vivo. In our studies we utilized a seminatural biomaterial made up by the benzyl ester of hyaluronan to grow keratinocytes, fibroblasts and chondrocytes. Keratinocytes and fibroblasts were isolated from human foreskin. Cells were separately cultured on two different hyaluronan based biomaterial devices for the first 15 days and then co-cultured for an additional period of 2 weeks. Keratinocytes gave rise to a well-differentiated epithelial layer, while fibroblasts were able to synthesize all the main extracellular molecules inside the biomaterial spaces, forming dermal-like tissues. When these two tissues were co-cultured, a skin equivalent was formed with a dermal-epidermal junction. Chondrocytes were obtained from chick-embryo sterna and cultured for 21 days inside a non-woven scaffolding made up of a hyaluronan-based biomaterial. Cells were able to organize themselves into nodules embedded in a dense metachromatic substance in which type II collagen was present. Data from this study suggest that this novel class of hyaluronan derived biomaterials is suitable for different cell culture and in vitro tissue reconstruction.