Preparation method of a cationic lipid-mesoporous silicon composite nanocarrier co-loading siRNA and chemotherapy drugs
A technology of cationic lipids and chemotherapeutic drugs, applied in drug combinations, pharmaceutical formulas, genetic material components, etc., can solve the problem of poor treatment effect of malignant tumors
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Embodiment 1
[0031] (1) Preparation of mesoporous silicon nanoparticles: Add a magnet rotor and 1.0 g of CTAB into a three-neck flask, then add 100 ml of deionized water, and stir at 95° C. for 30 min. Add 160μL DEA solution to the bottle and continue to stir for 15min. Take 8ml of TEOS and add it dropwise slowly (dropping for 1 hour). After the dropwise addition is completed, reflux for 3 hours, centrifuge, and discard the supernatant. The template was removed by cross-washing three times with methanol-HCl solution (4:1) and deionized water. Dry in a drying oven at 50° C. for 12 hours, and then vacuum-dry for 12 hours. That is, mesoporous silicon nanoparticles (MSNs) with an average particle diameter of about 80 nm were obtained.
[0032] (2) Take mesoporous silicon particles in a 10ml centrifuge tube, add an appropriate amount of PBS to disperse, add dropwise 5mg / ml doxorubicin (DOX) solution, mix and make up to volume. Stir for 24 hours, centrifuge, and wash with PBS until the washin...
Embodiment 2
[0039] (1) Chemically synthesize two complementary single-stranded DNAs corresponding to siRNA 1, with Hind III and BglII restriction sites at both ends. After annealing, it was ligated with the double-enzyme-digested plasmid pDual, and the product was transformed into Escherichia coli competent DH5a, single-clonal colonies were picked for amplification and culture, and the plasmid was extracted and sequenced for identification.
[0040] (2) Loading of siRNA 1: Centrifuge the dry powdered siRNA for 10 minutes, dissolve it in 375 μl of DEOC water, mix the obtained solution with the above-mentioned CLMSNs, and let it stand still, and record it as siRNA-CLMSNS. The particle size, PDI and zeta potential of siRNA-CLMSNS were measured with a Malvern particle size analyzer. The results are shown in Table 1. Compared with CLMSNS, the particle size of siRNA-CLMSNS was significantly larger, and the zeta potential was significantly lower. This is due to the negatively charged The siRNA n...
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