A colon-targeted nanofibrous membrane containing quercetin and prebiotic factors and its preparation method and application
A nanofiber membrane, probiotic factor technology, applied in the directions of medical preparations containing active ingredients, fiber treatment, medical preparations without active ingredients, etc., can solve the problems of poor stability, low bioavailability, etc., and achieve high porosity. , the fiber diameter can be controlled, and the effect of improving the release rate
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Embodiment 1
[0048] A method for preparing a colon-targeted nanofiber membrane containing quercetin and prebiotics prepared based on an electrospinning method, the specific steps are as follows:
[0049] (1) Prepare the chitosan nanoparticle solution containing the active substance by ion cross-linking: at first chitosan (160kDa, deacetylation degree 87%) is dissolved in (1%, v / v) acetic acid solution, stirred to obtain 9mL of chitosan solution with a concentration of 4mg / ml, then adjust the pH to 5.3 and filter with a 0.45μm membrane; then add 0.2g of poloxamer F127 and 1mL of 1mg / ml quercetin-containing ethanol aqueous solution; finally add 1mg The sodium tripolyphosphate solution of / ml is added dropwise in the above-mentioned solution with the speed of 400 μ l / min and realizes that the mass ratio of chitosan and sodium tripolyphosphate is 5:1, obtains the pale yellow solution that opalescence is arranged, and this is that contains quercetin Chitosan nanoparticle solution of corticoster...
Embodiment 2
[0054] The X-ray diffraction pattern of the nanoparticles containing quercetin, the specific steps are as follows:
[0055] (1) prepare the chitosan nanoparticle solution containing quercetin according to the method of step (1) in Example 1;
[0056] (2) Centrifuge (20000rpm, 10oC, 20min) the obtained quercetin-containing chitosan nanoparticle solution with a high-speed centrifuge, remove the supernatant, and freeze-dry the lower layer to obtain the quercetin-containing nanoparticle solution. particle.
[0057] (3) performing X-ray diffraction scanning on the chitosan, quercetin powder and the quercetin-containing nanoparticles obtained in (2).
[0058] image 3 It is the X-ray diffraction pattern of quercetin-containing nanoparticles. As can be seen from the figure, the absorption peaks of quercetin at 2θ=10.56, 12.45, 15.78, 23.94, and 27.48 and chitosan at 2θ=10.13 and 20.14 indicate respectively Both crystal structures. However, in the quercetin-containing nanoparticle...
Embodiment 3
[0060] (1) Cell culture: Caco-2 cells were cultured with 10% fetal bovine serum DMEM medium containing 2mmol / L glutamine, 100U / ml penicillin and streptomycin. Cells were placed at 37°C, 5% CO 2 Cultivate in an incubator, change the medium every 2 to 3 days, and pass the cells to about 80% in 3 to 5 days.
[0061] (2) Prepare quercetin-containing chitosan nanoparticles according to the method in Example 2, and then use the CCK-8 method to study its inhibitory effect on colon cancer cells. The specific operation method is: take Caco-2 cells in the logarithmic growth phase to make a single cell suspension, and press 1*10 per well 4 Cells were seeded in a 96-well plate (100 μL per well) at 37°C, 5% CO 2 Cultivate in a cell incubator with saturated humidity for 24 hours. When the cells adhere to the wall and grow to 80% fusion, add different concentrations of chitosan nanoparticles containing quercetin, and set 5 duplicate holes for each concentration. The cells were used as neg...
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