title: Calcium phosphate embedded PLGA nanoparticles: a promising gene delivery vector with high gene loading and transfection efficiency.
abstract: In the purpose of increasing incorporation efficiency and improving the release kinetics of plasmid DNA (pDNA) from poly(D,L-lactide-co-glycolide) (PLGA) nanoparticles, a facile method for the fabrication of calcium phosphate (CaPi) embedded PLGA nanoparticles (CaPi-pDNA-PLGA-NPs) was developed. The effect of several preparation factors on the particle size, incorporation efficiency, pDNA release and transfection efficiency in vitro was studied by Single Factor Screening Method. These preparation factors included the molecular weight (MW), hydrolysis degree (HD) of polyvinyl alcohol (PVA), sonication power and time, composition of organic phase, initial concentration of calcium phosphate and calcium (Ca) to phosphate ion (P) ratio (Ca/P ratio), etc. The CaPi-pDNA-PLGA-NPs made according to the optimal formulation were spherical in shape observed by transmission electron microscopy (TEM) with a mean particle size of 207±5 nm and an entrapment efficiency of 95.7±0.8%. Differential scanning calorimetry (DSC) suggested that there existed interaction between the DNA-calcium-phosphate (CaPi-pDNA) complexes and the polymeric matrices of PLGA. X-ray diffractometry (XRD) further proved the conclusion and indicated that the CaPi-pDNA was in weak crystallization form inside the nanoparticles. The Brunauer-Emmett-Teller (BET) surface area measurement demonstrated that the CaPi-pDNA-PLGA-NPs are mesoporous with specific surface area of 57.5m(2)/g and an average pore size of 96.5 Å. The transfection efficiency of the CaPi-pDNA-PLGA-NPs on human embryonic kidney 293 (HEK 293) cells in vitro was 22.4±1.2%, which was much higher than those of both the pDNA loaded PLGA nanoparticles (pDNA-PLGA-NPs) and the CaPi-pDNA embedded PLGA microparticles (CaPi-pDNA-PLGA-MPs). The CaPi-pDNA-PLGA-NPs are promising vectors for gene delivery.
authors.affiliation: College of Pharmacy, State Key Laboratory of Biotherapy, Sichuan University, No. 17, Section 3, Renmin Nan Road, Chengdu 610041, PR China.
authors.forename: Jie
authors.initials: J
authors.lastname: Tang
authors.forename: Jin-Ying
authors.initials: JY
authors.lastname: Chen
authors.forename: Jing
authors.initials: J
authors.lastname: Liu
authors.forename: Min
authors.initials: M
authors.lastname: Luo
authors.forename: Ying-Jing
authors.initials: YJ
authors.lastname: Wang
authors.forename: Xia-wei
authors.initials: XW
authors.lastname: Wei
authors.forename: Xiang
authors.initials: X
authors.lastname: Gao
authors.forename: Bi-lan
authors.initials: BL
authors.lastname: Wang
authors.forename: Yi-Bo
authors.initials: YB
authors.lastname: Liu
authors.forename: Tao
authors.initials: T
authors.lastname: Yi
authors.forename: Ai-Ping
authors.initials: AP
authors.lastname: Tong
authors.forename: Xiang-Rong
authors.initials: XR
authors.lastname: Song
authors.forename: Yong-Mei
authors.initials: YM
authors.lastname: Xie
authors.forename: Yinglan
authors.initials: Y
authors.lastname: Zhao
authors.forename: Mingli
authors.initials: M
authors.lastname: Xiang
authors.forename: Yuan
authors.initials: Y
authors.lastname: Huang
authors.forename: Yu
authors.initials: Y
authors.lastname: Zheng
mesh_terms: D002130:Calcium Phosphates / Q000008:administration & dosage* / Q000737:chemistry; D002152:Calorimetry, Differential Scanning; D004247:DNA / Q000008:administration & dosage / Q000235:genetics*; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000235:genetics; D057809:HEK293 Cells; D006801:Humans; D019344:Lactic Acid / Q000008:administration & dosage* / Q000737:chemistry; D046529:Microscopy, Electron, Transmission; D053758:Nanoparticles / Q000008:administration & dosage* / Q000737:chemistry / Q000648:ultrastructure; D010316:Particle Size; D011100:Polyglycolic Acid / Q000008:administration & dosage* / Q000737:chemistry; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D016062:Porosity; D013499:Surface Properties; D014162:Transfection / Q000379:methods*; D014961:X-Ray Diffraction
chemical_list: D002130:Calcium Phosphates; D049452:Green Fluorescent Proteins; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D011100:Polyglycolic Acid; D019344:Lactic Acid; D004247:DNA; C020243:calcium phosphate