J Korean Soc Hypertens.  2010 Jun;16(2):28-38.

Establishment of a eNOS Gene Therapy Model Using PLGA Nanosphere in Spontaneous Hypertensive Rat

• Shim TJ
• Choi EJ
• Park KJ
• Kim YJ
• Kim SJ
• Lee SY
• Kim SM
• Bae JW
• Hwang KK
• Kim DW
• Cho MC

Abstract

BACKGROUND
Poly(lactic-co-glycolic acid) (PLGA) has been studied for many years as a suitable drug delivery material, because of its chemical biocompatibility, total biodegradability, and non-toxic degradation products. The aim of this study was to establish the eNOS (endothelial Nitric Oxide Synthase) plasmid DNA-loaded PLGA nanosphere gene delivery system to control blood pressure in spontaneous hypertensive rat.
METHODS
To conjugate eNOS plasmid DNA to PLGA, we used the Water in Oil in Water method to formulate DNA-nanosphere mixture. For physicochemical characterization, the particle size and zetapotential were measured by a light scattering spectrophotometer. Release kinetics of PLGA nanosphere was investigated in rat vascular smooth muscle cells with FITC (Fluorescein isothiocyanate) and DAPI (4',6-diamidino-2-phenylindole). Also, the blood pressure control capability of the eNOS/PLGA gene delivery system was evaluated in spontaneous hypertensive rats with 2 times intravenous injection over 10 weeks.
RESULTS
The average size of PLGA nanosphere was 398+/-37 nm, and DNA loading efficiency within PLGA nanosphere was about 30~40%. With 2 times of intravenous injection, systolic blood pressure was controlled in spontaneous hypertensive rats in 10 weeks (eNOS/PLGA group 198+/-14mmHg, control group 215+/-15 mmHg, p<0.05). Nitric oxide concentrations in serum (eNOS/PLGA group 45.7+/-3.2 nM, control group 42.7+/-4.2 nM) and urine (eNOS/PLGA group 35.7+/-2.9 nM, control group 29.9+/-4.9 nM) were increased in the eNOS/PLGA delivery group. Carotid artery and aortic relaxation was observed after the eNOS/PLGA gene delivery.
CONCLUSION
In this study, the optimal condition to prepare PLGA nanosphere eNOS gene therapy was set up for a non-viral gene delivering system and the effectiveness of this model was tested in spontaneous hypertensive rats.

Keyword

PLGA; eNOS; Nanospheres; Blood pressure

MeSH Terms

Animals
Blood Pressure
Carotid Arteries
DNA
Fluorescein-5-isothiocyanate
Gene Transfer Techniques
Genetic Therapy
Indoles
Injections, Intravenous
Kinetics
Lactic Acid
Light
Muscle, Smooth, Vascular
Nanospheres
Nitric Oxide
Particle Size
Plasmids
Polyglycolic Acid
Rats
Relaxation
Water
DNA
Fluorescein-5-isothiocyanate
Indoles
Lactic Acid
Nitric Oxide
Polyglycolic Acid
Water