Efficient Non-Invasive Plasmid-DNA Administration into Tibialis Cranialis Muscle of “Little” Mice

Title:Efficient Non-Invasive Plasmid-DNA Administration into Tibialis Cranialis Muscle of “Little” Mice

Volume: 17 Issue: 3

Author(s): C. R. Cecchi*, E. Higuti, E. R. Lima, D. P. Vieira, P. L. Squair, C. N. Peroni and P. Bartolini

Affiliation:

• Biotechnology Center, Instituto de Pesquisas Energeticas e Nucleares (IPEN-CNEN), Av. Prof. Lineu Prestes 2242, 05508–000, Sao Paulo, SP,Brazil

Keywords: Electrotransfer, gene therapy, human growth hormone (hGH), immunodeficient “little” mice, quadriceps, serum mIGF-1 levels, tibialis.

Abstract: Background: An alternative treatment for growth hormone deficiency based on hGH-DNA administration, followed by electro gene transfer, was investigated by injecting the plasmid into surgically exposed or non-exposed quadriceps or tibialis muscle of immunodeficient “little” mice.

Methods: An optimization of electrotransfer conditions via a new combination of high/low voltage pulses is presented. After 3 days, serum hGH was determined and in a 28-day assay, the relative growth parameters were compared.

Results: Both groups exhibited similar results: 5.0 ± 2.2 (SD) and 3.5 ± 0.9 ng hGH/ml (P>0.05; n=7) for the exposed quadriceps and non-exposed tibialis treatments, respectively. The final body weight increases were 16.1% for the quadriceps and 18.9% for the tibialis group. The tail and nose-to-tail length increases were 4.5% and 7.1% for the quadriceps and 4.8 and 4.6% for the tibialis group. The right and left femur length increases, obtained from radiographic measurements, were 16.9% and 12.7% for the quadriceps and 19.4% and 12.3% for the tibialis, respectively. A non-significant difference between exposed quadriceps and non-exposed tibialis treatments (P=0.48) was confirmed via a completely integrated statistical analysis. Circulating mIGF-1 levels were 126 ± 47, 106 ± 93 (P>0.05) and 38 ± 15 ng/ml for the quadriceps, tibialis and saline treatments, respectively.

Conclusion: These results show that hGH-DNA administration into non-exposed tibialis muscle followed by the new HV/LV electrotransfer protocol was an equally efficient, less traumatic treatment, much more suitable for pre-clinical testing than administration into exposed quadriceps.

Cite this article as:

Cecchi R. C.*, Higuti E. , Lima R. E., Vieira P. D., Squair L. P., Peroni N. C. and Bartolini P. , Efficient Non-Invasive Plasmid-DNA Administration into Tibialis Cranialis Muscle of “Little” Mice, Current Molecular Medicine 2017; 17 (3) . https://dx.doi.org/10.2174/1566524017666170721152119