Development and evaluation of a new composite Laserskin graft

Background: Tremendous effort has been made to improve the graft take rate of cultured epidermal autograph. The purpose of this study is to develop an d evaluate a new composite Laserskin graft (CLSG) as a human skin substitut e for wound resurfacing, Methods: The seeding efficacy of cultured keratinocytes on plain Laserskin was compared with the 3T3 cell-seeded Laserskin and allogenic fibroblast-po pulated Laserskin, Three different types of CLSG, 2 cm in diameter each, we re prepared and tested in rats. Type A CLSG consisted of proliferative allo genic rat fibroblasts on both sides of the Laserskin with autologous kerati nocytes also on the upper side. Fibroblasts and keratinocytes were seeded o nly on the upper side of the Laserskin in type B CLSG, Keratinocytes alone mere seeded on plain Laserskin in type C CLSG, Type B CLSG consisting of au tologous keratinocytes and autologous dermal fibroblasts was tested on five selected wounds (5 x 5 cm each) of a patient with full-thickness burn. In another burn patient, type B CLSG consisting of autologous keratinocytes an d allogenic dermal fibroblasts was grafted onto three wounds (5 x 5 cm each ). Results: The seeding efficacy of human keratinocytes on plain Laserskin inc reased from 75% to 95% when proliferative allogenic fibroblasts were grown as a feeder layer on the Laserskin, The seeding efficacy of rat keratinocyt es increased from 36% to 88% in the presence of a proliferative allogenic f ibroblast feeder layer, whereas human/rat keratinocytes had respective seed ing efficacy of 98%/91% on Laserskin preseeded with mitomycin C-treated 3T3 cells, Skin biopsies of grafted type A CLSG on day 14 after grafting showe d complete epithelialization without severe inflammation in 16 of 20 (80%) grafted surgical wounds in rats. There were eight (40%) and seven (35%) "ta kes" of the CLSG in types B and C, respectively. The infection rate in type 11 CLSG was two (10%). There was one (5%) infection in types A and C, The respective take rates on the two patients grafted with type B CLSG were 60% and 100%. Conclusion: The animal experiment and the preliminary clinical data showed that the CSLGs consisting of autologous keratinocytes and of autologous/all ogenic fibroblasts are good human skin substitutes in terms of durability, biocompatibility, high seeding efficacy for keratinocytes, high graft take rate, and low infection rate.