GET protocols: Hyaluronidase is an effective adjuvant in the permeabilization of muscle fibers

N OLAIZ; E SIGNORI; L GUERRA; F MAGLIETTI; C SUAREZ; M VILLVERDE; L COLOMBO; S MICHINSKI; G MARSHALL

Portoroz

Congreso; 1st World Congress on Electroporation and Pulsed Electric Fields in Biology, Medicine and Food & Environmental Technologies; 2015

p { margin-bottom: 0.1in; direction: ltr; line-height: 120%; text-align: left; widows: 2; orphans: 2; }p.western { font-family: "Calibri",serif; font-size: 11pt; }p.cjk { font-family: "Calibri"; font-size: 11pt; }p.ctl { font-size: 11pt; } p { text-indent: 0.16in; margin-bottom: 0in; direction: ltr; line-height: 120%; text-align: justify; widows: 2; orphans: 2; }p.western { }p.cjk { }a:link { color: rgb(0, 0, 10); text-decoration: none; } ince 1950, hyaluronidase has been known as a ?spreading factor? favoring better diffusion of traditional vaccines in the site of inoculation. Nowadays, hyaluronidaseis used in the clinical practice as adjuvant therapy to increase the absorption and dispersion of injected drugs. In our laboratory we are evaluating the optimal conditions for effective gene electrotransfer (GET) protocols for the delivery of plasmid vectors. We have theoretically predicted by in silico modeling that optimal efficiency in GET protocols could be achieved when the applied electric field is near158 V/cm, using hyaluronidase as an adjuvant (Olaiz N et al, Bioelectrochemistry, 100: 105-11, 2014). Here, we evaluated in vivo our previous theoretical predictions. Our results show that: a) the presence of pH fronts in mice skins when electric fields are applied is experimentally corroborated; b) enhanced level of the expression vector in tissues pretreated withhyaluronidase and electric fields >150 V/cm is experimentally corroborated; c) evidence that electric fields between 150 and 200 V/cm would be optimal for GET protocols based on pretreatment with hyaluronidase is experimentally corroborated; d) the presence of tissue damage when the electric field applied is near 200V/cm is experimentally corroborated; e) theoretical predictions from (1) are experimentally corroborated.