Limits...
Topical gene electrotransfer to the epidermis of hairless guinea pig by non-invasive multielectrode array.

Guo S, Israel AL, Basu G, Donate A, Heller R - PLoS ONE (2013)

Bottom Line: Gene expression was observed exclusively in the epidermis.A low level of human Factor IX was detected in the plasma of hairless guinea pig after gene electrotransfer with the MEA, although a significant increase of Factor IX was obtained in the skin of animals.These results suggest gene electrotransfer with the MEA can be a safe, efficient, non-invasive skin delivery method for skin disorders, vaccinations and potential systemic diseases where low levels of gene products are sufficient.

View Article: PubMed Central - PubMed

Affiliation: Frank Reidy Research Center for Bioelectrics, Old Dominion University, Norfolk, Virginia, United States of America.

ABSTRACT
Topical gene delivery to the epidermis has the potential to be an effective therapy for skin disorders, cutaneous cancers, vaccinations and systemic metabolic diseases. Previously, we reported on a non-invasive multielectrode array (MEA) that efficiently delivered plasmid DNA and enhanced expression to the skin of several animal models by in vivo gene electrotransfer. Here, we characterized plasmid DNA delivery with the MEA in a hairless guinea pig model, which has a similar histology and structure to human skin. Significant elevation of gene expression up to 4 logs was achieved with intradermal DNA administration followed by topical non-invasive skin gene electrotransfer. This delivery produced gene expression in the skin of hairless guinea pig up to 12 to 15 days. Gene expression was observed exclusively in the epidermis. Skin gene electrotransfer with the MEA resulted in only minimal and mild skin changes. A low level of human Factor IX was detected in the plasma of hairless guinea pig after gene electrotransfer with the MEA, although a significant increase of Factor IX was obtained in the skin of animals. These results suggest gene electrotransfer with the MEA can be a safe, efficient, non-invasive skin delivery method for skin disorders, vaccinations and potential systemic diseases where low levels of gene products are sufficient.

Show MeSH

Related in: MedlinePlus

Kinetic of gene expression in HLGP skin after intradermal DNA injection and non-invasive GET.A, Time course of luciferase expression in HLGP skin after delivery by different electric fields at day 0. 50 µL DNA and 1 pulse delivery for all delivery groups, IO: no pulse delivery; GET: applied voltage of 30, 40, 50, 60 or 70. *: p<0.05 for GET-50, GET-60 or GET-70 vs IO. B, Time course of luciferase expression in HLGP skin after delivery to different sizes of skin. Delivery groups, 50-IO: 50 µL DNA without pulse delivery; 50-GET: 50 µL DNA with 1 pulse delivery on the injection site; 50 µLx4-IO: 4 injections with 50 µL DNA without pulse delivery; 50 µLx4-GET: 4 injections with 50 µL DNA and each pulse delivery on the injection site. Bars represent mean ± SD. 4–6 sites were analyzed for each delivery parameter, p/s  =  photons/second. * p<0.05 for 4×50-GET vs 50-GET and 4×50-GET vs 4×50-IO.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC3756005&req=5

pone-0073423-g001: Kinetic of gene expression in HLGP skin after intradermal DNA injection and non-invasive GET.A, Time course of luciferase expression in HLGP skin after delivery by different electric fields at day 0. 50 µL DNA and 1 pulse delivery for all delivery groups, IO: no pulse delivery; GET: applied voltage of 30, 40, 50, 60 or 70. *: p<0.05 for GET-50, GET-60 or GET-70 vs IO. B, Time course of luciferase expression in HLGP skin after delivery to different sizes of skin. Delivery groups, 50-IO: 50 µL DNA without pulse delivery; 50-GET: 50 µL DNA with 1 pulse delivery on the injection site; 50 µLx4-IO: 4 injections with 50 µL DNA without pulse delivery; 50 µLx4-GET: 4 injections with 50 µL DNA and each pulse delivery on the injection site. Bars represent mean ± SD. 4–6 sites were analyzed for each delivery parameter, p/s  =  photons/second. * p<0.05 for 4×50-GET vs 50-GET and 4×50-GET vs 4×50-IO.

Mentions: Because electrogene delivery had not been previously performed in hairless guinea pig skin, we first optimized the delivery condition by adopting the parameters from our previous work in Hartley guinea pigs [12], while testing a wider range of electric fields. As shown in Fig 1A, the expression level of luciferase positively correlated to the applied voltage up to 50 V. Although all delivery groups with above 50 V showed significantly increased gene expression compared to DNA injection only (p<0.05 for all 3 groups vs DNA injection only), the level of gene expression was no longer enhanced when the applied voltage was further increased up to 70 V. In contrast to intradermal DNA injection only, GET remarkably enhanced gene expression 2 to 4 logs and prolonged gene expression up to 12 to 15 days. While the expression level of non-GET-treated groups was relatively low at day one and rapidly dropped at day two, the gene expression of GET-treated groups maintained high levels until peak expression was reached at day eight then slowly decreased to background level after day 19 or later.


Topical gene electrotransfer to the epidermis of hairless guinea pig by non-invasive multielectrode array.

Guo S, Israel AL, Basu G, Donate A, Heller R - PLoS ONE (2013)

Kinetic of gene expression in HLGP skin after intradermal DNA injection and non-invasive GET.A, Time course of luciferase expression in HLGP skin after delivery by different electric fields at day 0. 50 µL DNA and 1 pulse delivery for all delivery groups, IO: no pulse delivery; GET: applied voltage of 30, 40, 50, 60 or 70. *: p<0.05 for GET-50, GET-60 or GET-70 vs IO. B, Time course of luciferase expression in HLGP skin after delivery to different sizes of skin. Delivery groups, 50-IO: 50 µL DNA without pulse delivery; 50-GET: 50 µL DNA with 1 pulse delivery on the injection site; 50 µLx4-IO: 4 injections with 50 µL DNA without pulse delivery; 50 µLx4-GET: 4 injections with 50 µL DNA and each pulse delivery on the injection site. Bars represent mean ± SD. 4–6 sites were analyzed for each delivery parameter, p/s  =  photons/second. * p<0.05 for 4×50-GET vs 50-GET and 4×50-GET vs 4×50-IO.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3756005&req=5

pone-0073423-g001: Kinetic of gene expression in HLGP skin after intradermal DNA injection and non-invasive GET.A, Time course of luciferase expression in HLGP skin after delivery by different electric fields at day 0. 50 µL DNA and 1 pulse delivery for all delivery groups, IO: no pulse delivery; GET: applied voltage of 30, 40, 50, 60 or 70. *: p<0.05 for GET-50, GET-60 or GET-70 vs IO. B, Time course of luciferase expression in HLGP skin after delivery to different sizes of skin. Delivery groups, 50-IO: 50 µL DNA without pulse delivery; 50-GET: 50 µL DNA with 1 pulse delivery on the injection site; 50 µLx4-IO: 4 injections with 50 µL DNA without pulse delivery; 50 µLx4-GET: 4 injections with 50 µL DNA and each pulse delivery on the injection site. Bars represent mean ± SD. 4–6 sites were analyzed for each delivery parameter, p/s  =  photons/second. * p<0.05 for 4×50-GET vs 50-GET and 4×50-GET vs 4×50-IO.
Mentions: Because electrogene delivery had not been previously performed in hairless guinea pig skin, we first optimized the delivery condition by adopting the parameters from our previous work in Hartley guinea pigs [12], while testing a wider range of electric fields. As shown in Fig 1A, the expression level of luciferase positively correlated to the applied voltage up to 50 V. Although all delivery groups with above 50 V showed significantly increased gene expression compared to DNA injection only (p<0.05 for all 3 groups vs DNA injection only), the level of gene expression was no longer enhanced when the applied voltage was further increased up to 70 V. In contrast to intradermal DNA injection only, GET remarkably enhanced gene expression 2 to 4 logs and prolonged gene expression up to 12 to 15 days. While the expression level of non-GET-treated groups was relatively low at day one and rapidly dropped at day two, the gene expression of GET-treated groups maintained high levels until peak expression was reached at day eight then slowly decreased to background level after day 19 or later.

Bottom Line: Gene expression was observed exclusively in the epidermis.A low level of human Factor IX was detected in the plasma of hairless guinea pig after gene electrotransfer with the MEA, although a significant increase of Factor IX was obtained in the skin of animals.These results suggest gene electrotransfer with the MEA can be a safe, efficient, non-invasive skin delivery method for skin disorders, vaccinations and potential systemic diseases where low levels of gene products are sufficient.

View Article: PubMed Central - PubMed

Affiliation: Frank Reidy Research Center for Bioelectrics, Old Dominion University, Norfolk, Virginia, United States of America.

ABSTRACT
Topical gene delivery to the epidermis has the potential to be an effective therapy for skin disorders, cutaneous cancers, vaccinations and systemic metabolic diseases. Previously, we reported on a non-invasive multielectrode array (MEA) that efficiently delivered plasmid DNA and enhanced expression to the skin of several animal models by in vivo gene electrotransfer. Here, we characterized plasmid DNA delivery with the MEA in a hairless guinea pig model, which has a similar histology and structure to human skin. Significant elevation of gene expression up to 4 logs was achieved with intradermal DNA administration followed by topical non-invasive skin gene electrotransfer. This delivery produced gene expression in the skin of hairless guinea pig up to 12 to 15 days. Gene expression was observed exclusively in the epidermis. Skin gene electrotransfer with the MEA resulted in only minimal and mild skin changes. A low level of human Factor IX was detected in the plasma of hairless guinea pig after gene electrotransfer with the MEA, although a significant increase of Factor IX was obtained in the skin of animals. These results suggest gene electrotransfer with the MEA can be a safe, efficient, non-invasive skin delivery method for skin disorders, vaccinations and potential systemic diseases where low levels of gene products are sufficient.

Show MeSH
Related in: MedlinePlus