Gene delivery to the epidermal cells of human skin explants using microfabricated microneedles and hydrogel formulations.
OBJECTIVE
Microneedles disrupt the stratum corneum barrier layer of skin creating transient pathways for the enhanced permeation of therapeutics into viable skin regions without stimulating pain receptors or causing vascular damage. The cutaneous delivery of nucleic acids has a number of therapeutic applications; most notably genetic vaccination. Unfortunately non-viral gene expression in skin is generally inefficient and transient. This study investigated the potential for improved delivery of plasmid DNA (pDNA) in skin by combining the microneedle delivery system with sustained release pDNA hydrogel formulations.


METHODS
Microneedles were fabricated by wet etching silicon in potassium hydroxide. Hydrogels based on Carbopol polymers and thermosensitive PLGA-PEG-PLGA triblock copolymers were prepared. Freshly excised human skin was used to characterise microneedle penetration (microscopy and skin water loss), gel residence in microchannels, pDNA diffusion and reporter gene (beta-galactosidase) expression.


RESULTS
Following microneedle treatment, channels of approximately 150-200 microm depth increased trans-epidermal water loss in skin. pDNA hydrogels were shown to harbour and gradually release pDNA. Following microneedle-assisted delivery of pDNA hydrogels to human skin expression of the pCMVbeta reporter gene was demonstrated in the viable epidermis proximal to microchannels.


CONCLUSIONS
pDNA hydrogels can be successfully targeted to the viable epidermis to potentially provide sustained gene expression therein.
Pearton
Marc
M
Gene Delivery Research Group, Welsh School of Pharmacy, Cardiff University, Cardiff, CF10 3XF, UK.
Allender
Chris
C
Brain
Keith
K
Anstey
Alexander
A
Gateley
Chris
C
Wilke
Nicolle
N
Morrissey
Anthony
A
Birchall
James
J
D018014:Gene Transfer Techniques / Q000295:instrumentation; D015316:Genetic Therapy / Q000379:methods*; D006801:Humans; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate; D008845:Microinjections / Q000295:instrumentation*; D009339:Needles; D012867:Skin / Q000378:metabolism*
D020136:Hydrogel, Polyethylene Glycol Dimethacrylate