Accelerated wound healing by in vivo application of keratinocytes overexpressing KGF.
Epidermal regeneration is a complex process, strongly influenced by growth factors, including keratinocyte growth factor (KGF). The objective of this study was to establish immortalized HaCaT keratinocytes and KMST-6-fibroblasts stably expressing KGF. Transfection efficiency, genomic integration, and functionality of the transgene were determined by ELISA and PCR, and KGF-expressing clones were selected using an air-liquid interface test system. HaCaT cells displayed stronger transgene expression compared to transfected fibroblasts, and the most effective HaCaT clone was incubated on a membrane carrier to form a "membrane cell graft." Twenty-one superficial second-degree burn wounds were created in each of three pigs, and wound healing capacity of the generated "polypeptide cell delivery system" after grafting was examined. Untransfected HaCaT keratinocytes and membrane-covered and untreated burn wounds served as controls. Histological and macroscopical follow-up revealed that grafting of transfected HaCaT cells resulted in complete reepithelialization within 5 days, while wounds covered with untransfected cells needed 2 days longer. At untreated sites, a thin epithelium was detectable after 10 days. The results indicate that wound healing processes can be stimulated distinctly by growth factors secreted from HaCaT cells, with a prominent role for transgenic KGF.
Kopp
Jürgen
J
Department of Plastic and Hand Surgery, University Medical Center, Friedrich-Alexander-University, 91054 Erlangen-Nürnberg, Germany. juergen.kopp@chir.imed.uni-erlangen.de
Wang
Guang Y
GY
Kulmburg
Peter
P
Schultze-Mosgau
Stefan
S
Huan
Jing N
JN
Ying
Kang
K
Seyhan
Harun
H
Jeschke
Mark D
MD
Kneser
Ulrich
U
Bach
Alexander D
AD
Ge
Sheng D
SD
Dooley
Steven
S
Horch
Raymund E
RE
D000818:Animals; D002056:Burns / Q000473:pathology / Q000628:therapy; D002460:Cell Line; D017690:Cell Transplantation / Q000379:methods*; D004817:Epidermis / Q000502:physiology*; D051523:Fibroblast Growth Factor 7; D005346:Fibroblast Growth Factors / Q000235:genetics* / Q000378:metabolism; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D015603:Keratinocytes / Q000378:metabolism* / Q000637:transplantation*; D010957:Plasmids / Q000235:genetics; D013552:Swine; D014162:Transfection; D014945:Wound Healing
C496453:FGF7 protein, human; D051523:Fibroblast Growth Factor 7; D005346:Fibroblast Growth Factors