Feasibility of biolistic gene therapy in burns.
Skin is an especially attractive target for genetic manipulation because it is readily accessible and easily monitored for both the presence and the expression of inserted genes. This study was designed to assess the feasibility of particle mediated gene transfer to burned skin and to compare the transfection efficiency, anatomic distribution, and duration of transgene expression achievable in normal versus burned skin. Two days following scald injury of varying depths in 60 degrees C water (10 s: superficial partial; 20 s: deep partial; 40 s: full thickness) reporter gene (beta-galactosidase) constructs were delivered using a gene gun at various helium pressures (200-600 psi) to normal and burned skin. A time course study was performed to examine the kinetics of transgene expression. Animals received a superficial partial thickness burn and were sacrificed 12 h, 1, 3, 5, 7, 14, or 21 days after gene transfer. India Ink injection and immunohistochemistry were used to assess the depth of the scald injury. Transfection efficiency was measured in skin homogenates 24 h after gene transfer by morphometric and chemoluminescent assays. We found that the extent of tissue damage was directly related to the duration of heat source exposure. Reporter gene activity was significantly higher in superficial partial thickness burns compared to normal controls and gradually declined with increasing tissue injury. No activity was seen in the full thickness burn group. Beta-galactosidase activity reached a maximum level 12 h after gene transfer in both normal and superficial partial thickness burned skin with no levels seen after 5 days post-transfection. These findings indicate that particle-mediated gene transfer in thermally injured skin is feasible and may provide a means of introducing biologic agents into injured tissue capable of enhancing bacterial clearance and improving wound healing.
Steinstraesser
L
L
Department of Surgery, University of Michigan, 48109-0666, USA.
Föhn
M
M
Klein
R D
RD
Aminlari
A
A
Remick
D G
DG
Su
G L
GL
Wang
S C
SC
D000818:Animals; D019470:Biolistics; D002056:Burns / Q000473:pathology / Q000628:therapy*; D004274:DNA, Recombinant / Q000008:administration & dosage; D005240:Feasibility Studies; D015870:Gene Expression; D017930:Genes, Reporter; D015316:Genetic Therapy; D005822:Genetic Vectors / Q000008:administration & dosage; D006046:Gold; D007763:Lac Operon; D008163:Luminescent Measurements; D008297:Male; D008863:Microspheres; D011897:Random Allocation; D051381:Rats; D017207:Rats, Sprague-Dawley; D011993:Recombinant Fusion Proteins / Q000032:analysis; D013047:Specific Pathogen-Free Organisms; D014162:Transfection; D019076:Transgenes; D001616:beta-Galactosidase / Q000032:analysis
D004274:DNA, Recombinant; D011993:Recombinant Fusion Proteins; D006046:Gold; D001616:beta-Galactosidase