Effect of laminin 5 on the formation of the lamina densa in vitro and in vivo

层粘连蛋白5对体外和体内致密层形成的影响

基本信息

批准号：
07807003
负责人：
ADACHI Eijiro
金额：
$ 1.28万
依托单位：
KITASATO UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1995
资助国家：
日本
起止时间：
1995 至 1996
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07807003/
关键词：
Lamina densa Laminin 5 Collagen IV Nude mouse Keratinocytes 人口皮膚線維芽細胞ゲル収縮

项目摘要

This study includes the development of supporting tissues, collagen IV for example, on which epidermal cells can adhere preferencially, and administering laminin 5, preferably located on the epidermal-dermal junction, thereby promoting the lamina densa formation.We tried to form the skin equivalents by seeding keratinocytes on the dermal equivalents, contracted collagen I gels containing fibroblasts, and continued to culture for additional one week in the culture medium containing laminin 5 at the concentration of 0,1,5, and 20mug/ml. After 2 weeks culture, the epidermal-dermal junction was examined by electronmicroscopy.The recognizable laminae densae were rarely detected in the absence of laminin 5. By contrast to the control culture, the laminae densae were frequently observed along the epidermal-dermal junction of the skin equivalents cultured in the presence of laminin 5 (1,5,20mug/ml). Morphometrical data of the experiments were confirmed by measuring the total length of the lamina densa found in the unit length of the epidermal-dermal junction. The data indicated that the lamina densa was developed approximately 3 times as long as that of control in the skin equivalents cultured in the presence of laminin 5 at the concentration of 5mug/ml, and 6 times at 20mug/ml. By immuno-electronmicroscopy, laminin 5 was detected at the lamina densa in the skin equivalents.Keratinocytes sheets were grafted to the panniculus carnosus of the nude mice. The samples were taken and assessed by the electronmicroscopy. At day 3 after grafting, the lamina densa of grafts supplemented with laminin 5 were more continuous than that found in the control transplantation. The supplement of laminin 5 promoted the formation of the lamina densa resulting in the improvement of Keratinocytes coverage.In conclusion, exogenous laminin 5 being present at a sufficient concentration allowed to form the lamina densa and promote the keratinocytes coverage.

This study includes the development of supporting tissues, collagen IV for example, on which epidermal cells can adhere preferencially, and administering laminin 5, preferably located on the epidermal-dermal junction, thereby promoting the lamina densa formation.We tried to form the skin equivalents by seeding keratinocytes on the dermal equivalents, contracted collagen I gels containing成纤维细胞，并在含有层粘连蛋白5的培养基中继续培养一个星期，浓度为0,1,5和20mug/ml。 After 2 weeks culture, the epidermal-dermal junction was examined by electronmicroscopy.The recognizable laminae densae were rarely detected in the absence of laminin 5. By contrast to the control culture, the laminae densae were frequently observed along the epidermal-dermal junction of the skin equivalents cultured in the presence of laminin 5 (1,5,20mug/ml).通过测量表皮 - dermal连接的单位长度中发现的椎板densa的总长度来证实实验的形态图数据。数据表明，在5mug/ml浓度下层粘连蛋白5的情况下，在存在层粘连蛋白5的情况下，在皮肤等效物中开发了薄片的长度约为3倍，而在20mug/ml的浓度为6倍。通过免疫电子显微镜检查，在皮肤当量的层片上检测到层粘连蛋白5。通过电子显微镜检查并评估样品。在嫁接后的第3天，补充层粘连蛋白5的移植物的椎板比对照移植中发现的更连续。层粘连蛋白5的补充促进了椎板densa的形成，从而改善了角质形成细胞覆盖率。结论，以足够的浓度存在外源性层粘连蛋白5，可以形成层片并促进角质形成细胞覆盖。