RESUMO
Calcium phosphate (CaP) has been widely used as the vector for gene transfection in the past three decades. However, clinical application is still not popular due to the poor-controlling of DNA/CaP complexes preparation, cytotoxicity and its low transfection efficiency. In this study, a novel amphipathic octadecyl-quatemized carboxymethyl chitosan (OQCMC) derivative from chitosan was combined with calcium phosphate to synthesize CaP/OQCMC nanoparticles (CaP/OQCMC NPs). The nanoparticles were 122-177 nm in diameter exhibited neutral zeta potential (from -0.115 mV to 0.216 mV), and they were applied as DNA vectors for DNA loading and in vitro transfection. The results showed that CaP/OQCMC NPs displayed high DNA loading capacity and enhanced transfection efficiency with extremely low cytotoxicity. In addition, both CaP and OQCMC are biocompatible and biodegradable, thus the as-prepared CaP/OQCMC NPs are promising in gene delivery.
Assuntos
Fosfatos de Cálcio/química , Quitosana/análogos & derivados , Quitosana/química , Nanopartículas/química , Nanotecnologia/métodos , Transfecção , Animais , Materiais Biocompatíveis/química , Linhagem Celular , DNA/química , Técnicas de Transferência de Genes , Vetores Genéticos , Luz , Camundongos , Microscopia Eletrônica de Transmissão , Modelos Genéticos , Nanocompostos/química , Polímeros/química , Espalhamento de RadiaçãoRESUMO
The main purpose of this study was to evaluate the targeting effect of cyclic arginine-glycine-aspartic peptide (cRGD)-modified monomethoxy (polyethylene glycol)-poly (D, L-lactide-co-glycolide)-poly (L-lysine) nanoparticles (mPEG-PLGA-PLL-cRGD NPs) for gastric cancer SGC-7901 cells. We prepared the 5-Fulorouracil (5Fu)-loaded mPEG-PLGA-PLL-cRGD (5Fu/mPEG-PLGA-PLL-cRGD) NPs that had an average particle size of 180 nm and a zeta potential 2.77 mV. The results of cytotoxicity demonstrated the mPEG-PLGA-PLL-cRGD NPs showed the ignorable cytotoxicity and the 5Fu/mPEG-PLGA-PLL-cRGD NPs could significantly enhance the cytotoxicity of 5Fu. In vitro drug release experiments showed that the release of drug was effectively prolonged and sustained. The results of confocal laser scanning microscope (CLSM) and flow cytometer analysis demonstrated that the fluorescence intensity of the SGC-7901 gastric cancer cells treated with Rb/mPEG-PLGA-PLL-cRGD NPs was significantly higher than that treated with Rb, this suggested that Rb/mPEG-PLGA-PLL-cRGD NPs could effectively be internalized by SGC-7901 gastric cancer cells. In summary, the above experimental results illustrate that mPEG-PLGA-PLL-cRGD NPs have great potential to be used as an effective delivery carriers.