Loading of plasmid DNA into PLGA microparticles using TROMS (Total Recirculation One-Machine System): evaluation of its integrity and controlled release properties

• Published in: Journal of controlled release 2003-01, Vol.86 (1), p.123-130
• Main Authors: García del Barrio, G, Novo, F.J, Irache, J.M
• Format: Article
• Language: English
• Subjects: Animals; Biological and medical sciences; Delayed-Action Preparations - administration & dosage; Delayed-Action Preparations - chemical synthesis; Delayed-Action Preparations - pharmacokinetics; DNA - administration & dosage; DNA - chemical synthesis; DNA - pharmacokinetics; Female; General pharmacology; Lactic Acid - administration & dosage; Lactic Acid - chemical synthesis; Lactic Acid - pharmacokinetics; Medical sciences; Microspheres; Pharmaceutical technology. Pharmaceutical industry; Pharmacology. Drug treatments; Plasmid DNA, muscle gene transfer; Plasmids - administration & dosage; Plasmids - chemical synthesis; Plasmids - pharmacokinetics; PLGA microparticles, controlled release; Polyglycolic Acid - administration & dosage; Polyglycolic Acid - chemical synthesis; Polyglycolic Acid - pharmacokinetics; Polymers - administration & dosage; Polymers - chemical synthesis; Polymers - pharmacokinetics; Rats; Rats, Wistar; Technology, Pharmaceutical - instrumentation; Technology, Pharmaceutical - methods; Transfection - methods; TROMS (Total Recirculation One-Machine System)
• ISSN: 0168-3659; 1873-4995
• DOI: 10.1016/S0168-3659(02)00371-1

Loading plasmid DNA into poly(ester) microparticles usually involves the formation of a multiple emulsion, using homogenisation techniques such as sonication or Ultra-Turrax®. These procedures may negatively affect the integrity of the macromolecule and consequently its activity. The aim of this study was to prepare and evaluate DNA-loaded microparticles by TROMS (Total Recirculation One-Machine System), a new procedure that is based on the formation of a multiple emulsion by the injection of the phases under a turbulent regime. Microparticles were prepared with either Resomer® RG 502 (MP 502) or RG 756 (MP 756) and DNA loading was quantified fluorimetrically. DNA loading in MP 756 was almost twice as high as in MP 502 (510 vs. 285 ng/mg, respectively). Under both formulations, the loaded plasmid was released while maintaining its integrity for at least 24 days (MP 502) and 40 days (MP 756). Finally, the transfection efficiency was studied after injection of the microparticles (MP 502) into rat skeletal muscle and compared with naked DNA injection. Injection of naked DNA (150 μg DNA per muscle) achieved higher but variable expression levels that decreased after 3 weeks. In contrast, the muscles injected with microparticles (6.8 μg DNA per muscle) showed lower but homogeneous expression values, which were maintained for at least 3 weeks.