Pre-clinical evaluation of liposomal gene transfer to improve dermal and epidermal regeneration.
Liposomal gene transfer effectively enhances dermal and epidermal regeneration in burned rodents. To advance this treatment to clinical studies, we investigated the efficacy of liposomal gene transfer in a clinically relevant porcine wound model. Mimicking the clinical scenario, six female Yorkshire pigs (40-50 kg) received up to 12 burns of 50 cm(2) area that were fully excised and covered with skin autograft meshed at 4:1 ratio 24 h post-burn. Animals received control injections (empty liposomes), liposomes (DMRIE-C) containing 1 mg LacZ-cDNA, or liposomes (DMRIE-C) with 1 mg of platelet-derived growth factor (PDGF)-cDNA, or the naked PDGF gene. Serial biopsies were taken from different wound sites at multiple time points up to 12 days post-wounding. Transfection efficacy and transfection rate of LacZ and localization of beta-gal were determined by immunohistochemical and immunofluorescent techniques. RT-PCR and multiplex protein analysis (ELISA) were used to measure levels of growth factor mRNA transcribed and growth factor protein translated. Wound re-epithelialization and graft adhesion was evaluated using planimetric analysis and clinical scores. We found that peak transfection of liposomal beta-galactosidase occurred on day 2, with a fluorescence increase of 154% to baseline (P<0.001). Transfection intensity dropped to 115% above baseline on day 4 (P<0.001) and 109% on day 7. Immunohistochemistry showed a maximum transfection rate of 34% of cells in wound tissue. Gene transfer of liposomal PDGF-cDNA resulted in increased PDGF-mRNA and protein expression on days 2 and 4, and accelerated wound re-epithlialization as well as graft adhesion on day 9 (P<0.05). In this study, we showed that liposomal cDNA gene transfer is possible in a porcine wound model, and by using PDGF-cDNA we further showed that dermal and epidermal regeneration can be improved. These data indicate that liposomal gene transfer can be a new therapeutic approach to improve wound healing in humans.
Branski
L K
LK
Shriners Hospitals for Children and University of Texas Medical Branch, Galveston, TX 77550, USA.
Masters
O E
OE
Herndon
D N
DN
Mittermayr
R
R
Redl
H
H
Traber
D L
DL
Cox
R A
RA
Kita
K
K
Jeschke
M G
MG
D000818:Animals; D002056:Burns / Q000628:therapy*; D004817:Epidermis; D005260:Female; D018014:Gene Transfer Techniques; D008081:Liposomes; D023421:Models, Animal; D010982:Platelet-Derived Growth Factor / Q000235:genetics*; D012038:Regeneration; D012867:Skin / Q000293:injuries*; D016038:Skin Transplantation / Q000379:methods*; D013552:Swine; D014162:Transfection; D014945:Wound Healing / Q000235:genetics
D008081:Liposomes; D010982:Platelet-Derived Growth Factor