| Budget Amount *help |
¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 2002: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 2001: ¥1,000,000 (Direct Cost: ¥1,000,000)
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| Research Abstract |
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.
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