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Tissue engineering for higher take rate of cultured epidermis in human

组织工程提高人体培养表皮的利用率

基本信息

批准号：
12470378
负责人：
YAMAUCHI Toshihiko
金额：
$ 1.28万
依托单位：
KURUME UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2000
资助国家：
日本
起止时间：
2000 至 2002
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12470378/
关键词：
Cultured human epidermis basal membrane anchoring fibrin matrix complex of cultured epidermis artificial dermis take rate

项目摘要

Human epidermic cells and feeder cells were co-cultured on fibrin matrix (gel) in order to maintain the components of basal membrane during culture. The cultured human epidermis was then combined with absorbable mesh that functions as an artificial anchor to graft bed, and this complex-sheet was grafted to the full-thickness skin defect on the back of nude rats. Take rate was comparatively examined in the 3 groups that received grafting of conventionally cultured human epidermis (Control), the above-mentioned complex-sheet (Complex group), or the complex-sheet and artificial dermis (Complex + Dermis group). As a result, take rate was significantly higher in the Complex group (71.4%, n = 14) and the Complex + Dermis group (66.8%, n = 30) than in the Control (16.7%, n = 12) (Px0.002). Histological examinations were conducted in the Control and the complex-sheet component of the Complex + Dermis. Tissues on the graft area were surgically resected in the 2nd, 3rd and 4th week after grafting for the monitoring under electron microscope, hematoxylin and eosin (HE) staining, immunohistochemical staining and fluorochrome staining HE staining showed the complex-sheet was taken by the graft bed The other 2 stainings demonstrated the expression of adhesive factors to basal membrane such as Type IV collagen, laminin and fibronectin. On the electron microscopy in the 3rd week, structure of basal membrane was not found in the Control, whereas the Complex + Dermis group had lamina densa, lamina lucida, hemidesmosome and anchoring fibrin. These results showed that our complex-sheet and artificial dermis were better taken than the conventionally cultured epidermis.

将人表皮细胞和饲养层细胞在纤维蛋白基质（凝胶）上共培养，以在培养过程中保持基底膜的成分。然后将培养的人表皮与可吸收网片结合，可吸收网片作为移植床的人工锚，并将这种复合片移植到裸大鼠背部的全层皮肤缺损处。在接受常规培养的人表皮（对照组）、上述复合片（复合组）或复合片和人工真皮（复合+真皮组）的3组中比较检查了移植率。结果，复合物组（71.4%，n = 14）和复合物+真皮组（66.8%，n = 30）的取用率显著高于对照组（16.7%，n = 12）（P <0.002）。在对照组和复合物+真皮的复合物-片材组分中进行组织学检查。术后2、3、4周分别取移植区组织行电镜观察、HE染色、免疫组化及荧光染色，HE染色显示移植床周围有复合片形成，免疫组化及荧光染色显示基底膜粘附因子Ⅳ型胶原、层粘连蛋白及纤维连接蛋白的表达。3周电镜观察，对照组未见基底膜结构，复合物+真皮组可见致密层、透明层、半桥粒和锚定纤维蛋白。这些结果表明，我们的复合片和人工真皮比传统培养的表皮更好。