权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Preparation of Antibacterial Epidermal Skin Graft

抗菌表皮植皮片的制备

基本信息

批准号：
13650855
负责人：
SHINKAI Masashige
金额：
$ 2.3万
依托单位：
THE UNIVERSITY OF TOKYO
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2001
资助国家：
日本
起止时间：
2001 至 2002
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13650855/
关键词：
Epidermal Skin Graft Tissue Transplantation Cell Engineering Tissue Engineering Antibacterial peptide Gene Therapy Medical Materials Cell Surface Engineering

项目摘要

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.

1.应用抗菌肽呈递皮肤片检测转基因sapecin基因的上皮细胞的抗菌活性。将上皮细胞移植到小鼠体内后，大肠杆菌或金黄色葡萄球菌被感染。此外，我们构建了具有 sapecin 基因、含有凝血酶消化基序的接头和 PDGF（血小板生长因子）受体的新基因。通过脂转染法转染后，用针对作为标记物的Myc标签的荧光抗体对细胞进行染色。然而，由于肽产量非常低，因此没有观察到抗菌活性。然后，制备了能够锚定在细胞表面的BAM抗菌肽，并评价其抗菌活性。当将BAM抗菌肽添加到小鼠成纤维细胞的培养基中时，抗菌肽能够在几分钟内显现到细胞表面。当大肠杆菌。将培养基接触到该细胞中，抗菌肽逐渐释放出来。另一方面，BAM抗菌肽很容易锚定在小鼠外皮上，并且还观察到对大肠杆菌的抗菌活性。2。培养皮肤细胞片制备方法的建立通过明胶与转谷氨酰胺酶(TGase)交联制备可生物降解的水凝胶。我们发现5wt5明胶和1单位/ml TGase的浓度最适合NIH/3T3成纤维细胞的增殖。通过将细胞粘附因子掺入明胶水凝胶中来增强细胞增殖。将合成肽 RGDLLQ 添加到明胶溶液中，其中 LLQ 基序通过谷氨酰胺残基成为 TGase 的底物。这些结果表明，TGase 介导的细胞粘附因子与明胶基质的结合增强了细胞增殖，这种新型生物材料是伤口敷料或组织工程的有效工具。