Facile Construction of Chloroquine Containing PLGA-Based pDNA Delivery System for Efficient Tumor and Pancreatitis Targeting in Vitro and in Vivo

Abstract

Chloroquine diphosphate (CQ) was ingeniously used to take place of phosphate salt in traditional calcium phosphate coprecipitation method for pDNA transfection. With multiple roles of CQ in the novel Ca-CQ-pDNA complex including pDNA compaction and assistance in lysosome escape, the transfection efficiency of the pDNA was significantly increased relative to the traditional method. CQ did not intercalate into the DNA double helix as free CQ did, which was probably ascribed to the prior mixing of the pDNA with high concentration of calcium chloride. In order to construct efficacious vector for in vivo gene delivery, Ca-CQ-pDNA-PLGA-NPs was designed and prepared. With entrapment efficiency, particle size and pDNA integrity as screening conditions, the optimal prescription was obtained and CaPi-pDNA-PLGA-NPs made with classic calcium phosphate coprecipitation method after optimization was also prepared as control to systematically study the role of CQ in the novel vector. Physical characters of the vectors were comprehensively studied using TEM, DSC, and XRD. The safety of the vector both in vitro and in vivo was evaluated using MTT, hemolysis test, and histological sections. The Ca-CQ-pDNA-PLGA-NPs dramatically enhanced the gene tranfection efficiency in Human Embryonic kidney HEK293 cells compared with the CaPi-pDNA-PLGA-NPs and presented an increasing gene transfection for up 144 h. The relative fast release of the CQ compared with pDNA from the nanoparticles was responsive for the increased transfection. The Did-labeled-Ca-CQ-pDNA-PLGA-NPs exhibited excellent tumor targeting efficiency and sustained circulation time in CT26 mouse model. The Ca-CQ-pDNA-PLGA-NP loaded with the plasmid pVITRO2 expressing mSurvivin-T34A protein gave 70% tumor inhibition rate, which was partially ascribed to CQ. The Ca-CQ-pDNA-PLGA-NPs showed high targeting efficiency in C57 acute pancreatitis model. In all, the Ca-CQ-pDNA-PLGA-NP was a promising candidate for targeted gene delivery to both tumor and pancreatitis.

Ultrasound-Mediated Microbubble Destruction (UMMD) Facilitates the Delivery of CA19-9 Targeted and Paclitaxel Loaded mPEG-PLGA-PLL Nanoparticles in Pancreatic Cancer

“…The uptake of RN-Ab was evidently greater than RN because the targeted NPs with the anti-CA19-9 antibody could specifically bind to the capan-1 cells and increase the local concentration of RB. The uptake of RN-Ab was similar to that of free RB before 8 h but kept increasing until 48 h. These results were consistent with the previous observations that the RB delivered via NPs was internalized by endocytosis, accumulated inside the cells, and could avoid degradation 37 - 40 .…”

Hyaluronic acid hydrogel scaffolds loaded with cationic niosomes for efficient non-viral gene delivery

“… 25 The use of chloroquine has also been reported to enhance gene delivery both in vitro and in vivo . 26 Chloroquine prevents the acidification of endosomes, fusion of endosomes and lysosomes, and inhibits the lysosomal enzymes. 27 , 28 Therefore, the selection of the components, and their concentrations used to prepare the niosome formulations, as well as the cationic lipid/DNA mass ratios (w/w) to form the corresponding nioplexes were based on available data and previous studies developed in our laboratory.…”

Integrated in silico and experimental methods revealed that Arctigenin inhibited angiogenesis and HCT116 cell migration and invasion through regulating the H1F4A and Wnt/β-catenin pathway

“…[26][27][28][29][30] Briefly, zebrafish embryos at the 13-somite stage (30 embryos per group) were incubated overnight with vehicle, curcumin or Arctigenin. [26][27][28][29][30] Briefly, zebrafish embryos at the 13-somite stage (30 embryos per group) were incubated overnight with vehicle, curcumin or Arctigenin.…”