Preparation of Antibacterial Epidermal Skin Graft

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1. Application antibacterial peptide-presenting skin sheetAntibacterial activity of epitherial cell which was genetically modified with sapecin gene was examined. After transplanting of eptherial cells into mouse, E. coli or S. aureus was infected. Moreover, we constructed new gene which had sapecin gene, linker containing a thrombin digesting motif and the PDGF (blood platelet growth factor) receptor. After transfection by lipofection method, the cells were dyed with the fluorescence antibody to the Myc tag which is a marker. However, the antibacterial activity was not observed, because peptide production was very low. Then, BAM-antibacterial peptide which was able to anchor on the cell surface was prepared, and its antibacterial activity was evaluated. When BAM-antibacterial peptide was added to the culture medium of mouse fibroblast cell, antibacterial peptide was able to be shown to the cell surface in several minutes. When E coli. culture medium was contacted into this cell, antibacterial peptide was released gradually and. On the other hand, BAM-antibacterial peptide was easily anchored on mouse outer skin and also antibacterial activity was observed against E. coli.2. Establishment of the preparation method of cultured skin cell sheetThe biodegradable hydrogels were prepared though crosslinking of gelatin with transglutaminase (TGase). We found that the concentration of 5wt5 gelatin and 1 unit/ml TGase were optimum for the proliferation of NIH/3T3 fibroblast. The cell proliferation was enhanced by incorporation of cell adhesion factors in to gelatin hydorgels. Synthetic peptide, RGDLLQ, were added to the gelatin solution where LLQ motif is a substrate of TGase by virtue of a glutamine residue. These results suggest that TGase-mediated incorporation of cell adhesion factors into gelatin matrices enhanced cell proliferation and this novel biomaterial is a potent tool for wound dressing or tissue engineering.