Chitosan-DNA microparticles as mucosal delivery system: synthesis, characterization and release in vitro.
BACKGROUND
Mucosal immunity is important to defense against dental caries. To enhance mucosal immunity, a DNA vaccine mucosal delivery system was prepared by encapsulating anticaries DNA vaccine (plasmid pGJA-P/VAX) in chitosan under optimal conditions and the characteristics of the microparticles was investigated. Furthermore, the release properties and protective action of microparticles for plasmid were studied in vitro.


METHODS
Plasmid loaded chitosan microparticles were prepared by complex coacervation. Three factors, concentration of DNA, sodium sulfate, and the chitosan/DNA ratios in complexes [better expressed as N/P ratio: the number of poly nitrogen (N) per DNA phosphate (P)] influencing preparation were optimized by orthogonal test. The characteristics of microparticles were evaluated by scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM). DNA release rate of microparticles in similar gastro fluid (SGF) or similar intestinal fluid (SIF) at 37 degrees C was determined by ultraviolet spectrophotometry.


RESULTS
High encapsulation efficiency (96.8%) was obtained with chitosan microparticles made under optimal conditions of 50 mmol/L Na2SO4, 200 microg/ml DNA and N/P ratio of 4. The size of particles was about 4 to 6 microm. The encapsulation process did not destroy the integrity of DNA. When incubated with SIL, after a release of about 10% in the first 60 minutes, no further DNA was released during the following 180 minutes. When incubated with SGL, the microparticles released a small burst (about 11%) in the first 60 minutes, and then slowly released at a constant, but different rate.


CONCLUSIONS
These chitosan microparticles showed suitable characteristics in vitro for mucosal vaccination and are therefore a promising carrier system for DNA vaccine mucosal delivery.
Li
Yu-hong
YH
Key Laboratory for Oral Biomedical Engineering Ministry of Education, School of Stomatology, Wuhan University, Wuhan 430079, China.
Fan
Min-wen
MW
Bian
Zhuan
Z
Chen
Zhi
Z
Zhang
Qi
Q
Yang
Hai-rui
HR
D000818:Animals; D016466:CHO Cells; D002470:Cell Survival; D048271:Chitosan / Q000008:administration & dosage*; D006224:Cricetinae; D016503:Drug Delivery Systems; D004587:Electrophoresis, Agar Gel; D018928:Immunity, Mucosal; D010316:Particle Size; D019444:Vaccines, DNA / Q000008:administration & dosage* / Q000378:metabolism
D019444:Vaccines, DNA; D048271:Chitosan