Further biocompatibility testing of silica-chitosan complex membrane in the production of tissue plasminogen activator by epithelial and fibroblast cells.

The effects of the physicochemical characteristics of a silica-chitosan complex membrane (SiCM) on the expression of tissue plasminogen activator (tPA) by contacting cells were investigated with the aim of improving the biocompatibility of the novel implant biomaterial. Expression of tPA is considered to be effective in wound healing by preventing thrombus formation, which causes inflammatory responses and rejection of implant materials. Inducing the epithelial cells surrounding implant materials to secrete tPA, which serves as an early signaling system to proliferate cells underlying connective tissues, would be further effective in accelerating wound healing. An epithelial 293 cell line derived from human embryonic kidney and a fibroblast IMR-90 cell line from human lung possessing the ability to secrete tPA were cultured on SiCMs, whose composition was stepwise controlled by adjusting the mixing ratio between silica and chitosan to give silica contents of 20, 33, 43, and 50wt%. Both strains showed strong adhesion on chitosan (0%-SiCM) and 50%-SiCM. The cell proliferation rates were also accelerated in a manner that was dependent on the increase in the adhesion strength of the cells cultured on the SiCMs. Furthermore, the tPA activity in the culture medium increased in accordance with the cell density, while the cellular specific activity of IMR-90 cells to secrete tPA was synergistically enhanced by strong adhesion and a high cell density on the surface of chitosan and 50%-SiCM. Analysis of the physico-chemical effects of the SiCMs revealed that the cells were dominantly affected by the surface hydrophobicity rather than by the zeta potential, as well as by the mixing ratio between chitosan and silica. The wet contact angles of 50%-SiCM and chitosan, which were 68 degrees and 65 degrees , respectively, were found to be suitable for adhesion and growth of both the epithelial 293 cells and fibroblast IMR-90 cells. A hydrophobic surface at 65 degrees -68 degrees was also effective for the production of tPA by IMR-90 cells, whereas the tPA activity of 293 cells reached its highest level on the SiCM with a wet contact angle of 63 degrees . These results suggest that a suitable adhesion strength is a significant factor in the expression of tPA by cells contacting an implant biomaterial.