A preliminary in vitro study on the fabrication and tissue engineering applications of a novel chitosan bilayer material as a scaffold of human neofetaf dermal fibroblasts

The bilayer structure of chitosan him and sponge was designed as a scaffold of skin tissue engineering and a dermis substitute. It was processed succe ssively via the formation of a dense chitosan him by casting method and a p orous chitosan sponge by lyophilization. The dry thickness of the him layer was 19.6 mum and that of the sponge layer was controlled at 60-80 mum. Por ogens such as sodium chloride, glucose, and sucrose were used to create lar ge pores of the chitosan sponge layer. Human neofetal dermal fibroblasts we re seeded in the chitosan sponge layer and cultured for 4 weeks. It was fou nd that the cells could grow and proliferate well in an extended shape on t he flat bottom of large pores with 15-100 mum width and in spherical form o n the rough pore walls or at the edges of micropores less than 5 mum. Fibro blasts after the culture could bind tightly with the sponge layer via newly formed extracellular matrices to give a living cell-matrix-chitosan compos ite. The bilayer chitosan material remained stable in shape and size during the cell culture. The results suggested that the bilayer chitosan material would be an alternative of collagen materials which was obviously contract ed during cell culture. (C) 2001 Elsevier Science Ltd. All rights reserved.