Composite material of functional mesoporous silica loaded drug and siRNA, preparation and application thereof in preparation of anticancer drugs
A technology of mesoporous silica and silica, which is applied in the direction of drug combination, medical preparations without active ingredients, and medical preparations containing active ingredients, etc., can solve the problem that no functionalized mesoporous silica is found Systematic cancer treatment and other issues, to achieve the effect of improving gene silencing efficiency, increasing enrichment, and increasing accumulation
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Embodiment 1
[0038] Example 1: Preparation of functionalized mesoporous silica loaded drug and siRNA composite material
[0039]Preparation of mesoporous silica (MSNs): Dissolve cetyltrimethylammonium bromide (CTAB, 500mg) in distilled water (250mL), then add NaOH aqueous solution (1.75mL, 2Mol / L), and mix the solution in Heat to 80°C with vigorous stirring. When the temperature stabilized, ethyl orthosilicate (TEOS, 2 mL) was slowly added to the mixed solution. After 2 h, the nanoparticles were collected by centrifugation at 10,000 rpm for 3 min, and washed three times with ethanol and distilled water. To remove the template, the collected nanoparticles were resuspended in ethanol (80 mL) containing concentrated hydrochloric acid (4 mL, 37%), refluxed at 80°C for 24 h, collected by centrifugation at 10,000 rpm for 3 minutes, washed three times with ethanol and distilled water, The nanoparticles were vacuum dried overnight at 50°C. Its morphology was observed by TEANAI-10 transmission e...
Embodiment 2
[0042] Example 2: In vitro experiment of functionalized mesoporous silica loaded drug and siRNA composite material as anticancer drug
[0043] In this experiment, human breast cancer cells (MCF-7, Saiqi (Shanghai) Bioengineering Co., Ltd.) were selected as the anti-cancer experimental subjects of the functionalized mesoporous silica drug-loading system.
[0044] MTT test method: Digest the cells with 0.25% trypsin for 5 minutes, blow the cells gently with a pipette to form a suspension in the centrifuge tube of the device, take 10 μL of the cell suspension and count the number of viable cells with a counter, and adjust the concentration of viable cells to 5× 10 3 / mL was added to a 96-well culture plate, 100 μL per well, and after culturing for 24 hours, the composite materials prepared in Example 1 were added at different concentrations, placed at 37°C, and the volume fraction was 5% CO 2 Incubate for 24 hours. Add 20 μL / well of MTT 4 hours before the end, discard the super...
Embodiment 3
[0046] Example 3: Functional Mesoporous Silica Loaded Drug and siRNA Composite Material Enrichment of Drugs in Cells
[0047] MCF-7 cells in the logarithmic phase were digested with 0.25% trypsin for 3-5 minutes, centrifuged, mixed with medium, and inoculated with 1×10 4 Put the cells in the small groove of the laser confocal dish, add 2mL of culture medium after 1h, and add 5% CO 2 , and cultivated in a 37°C incubator for 24h. Add 10 μg / mL of the composite material prepared in Example 1 and an equivalent amount of doxorubicin to treat the cells for 1 h and 3 h, respectively. Then the culture medium in the dish was aspirated, and then washed three times with PBS, then 1 mL of PBS was added to infiltrate the cells, and the laser confocal microscope was used to observe and take pictures. like Figure 4 As shown, compared with free doxorubicin, the doxorubicin in the composite material of the present invention can significantly increase the enrichment amount of doxorubicin in ...
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