Development and characterization of pectinate micro/nanoparticles for gene delivery.
The aim of this study was to investigate the possibility of using pectinate micro/nanoparticles as gene delivery systems. Pectinate micro/nanoparticles were produced by ionotropic gelation. Various factors were studied for their effects on the preparation of pectinate micro/nanoparticles: the pH of the pectin solution, the ratio of pectin to the cation, the concentration of pectin and the cation, and the type of cation (calcium ions, magnesium ions and manganese ions). After the preparation, the size and charge of the pectin micro/nanoparticles and their DNA incorporation efficiency were evaluated. The results showed that the particle sizes decreased with the decreased concentrations of pectin and cation. The type of cations affected the particle size. Sizes of calcium pectinate particles were larger than those of magnesium pectinate and manganese pectinate particles. The DNA loading efficiency showed that Ca-pectinate nanoparticles could entrap DNA up to 0.05 mg when the weight ratio of pectin:CaCl(2):DNA was 0.2:1:0.05. However, Mg-pectinate could entrap only 0.01 mg DNA when the weight ratio of pectin:MgCl(2):DNA was 1:100:0.01 The transfection efficiency of both Ca-pectinate and Mg-pectinate nanoparticles yielded relatively low levels of green fluorescent protein expression and low cytotoxicity in Huh7 cells. Given the negligible cytotoxic effects, these pectinate micro/nanoparticles can be considered as potential candidates for use as safe gene delivery carriers.
Opanasopit
Praneet
P
Nanotechnology for Drug/Gene Delivery Systems Group, Faculty of Pharmacy, Silpakorn University, Nakhon Pathom, Thailand. praneet@email.pharm.su.ac.th
Apirakaramwong
Auayporn
A
Ngawhirunpat
Tanasait
T
Rojanarata
Theerasak
T
Ruktanonchai
Uracha
U
D006528:Carcinoma, Hepatocellular / Q000473:pathology / Q000503:physiopathology; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D003460:Crystallization / Q000379:methods; D004247:DNA / Q000008:administration & dosage / Q000737:chemistry*; D004337:Drug Carriers / Q000737:chemistry*; D004339:Drug Compounding / Q000379:methods*; D004353:Drug Evaluation, Preclinical; D018390:Gene Targeting / Q000379:methods*; D006801:Humans; D008113:Liver Neoplasms / Q000473:pathology / Q000503:physiopathology; D008863:Microspheres; D053758:Nanoparticles / Q000737:chemistry* / Q000648:ultrastructure*; D010316:Particle Size; D010368:Pectins / Q000737:chemistry*; D014162:Transfection / Q000379:methods*
D004337:Drug Carriers; D010368:Pectins; D004247:DNA