A chitosan-based drug-loaded thermosensitive hydrogel and its application in the preparation of animal vaccines and sustained-release drugs
A temperature-sensitive hydrogel, chitosan technology, which is applied in the directions of non-active ingredients medical preparations, active ingredients-containing medical preparations, pharmaceutical formulas, etc. Poor and other problems, to achieve the effect of enhancing mechanical strength and good epidemic prevention effect
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Embodiment 1
[0028] Embodiment 1: Chitosan-based thermosensitive hydrogel and preparation method thereof
[0029] The preparation method of the chitosan-based thermosensitive hydrogel comprises the following steps (10ml system):
[0030] (1) Dissolve 0.2g chitosan in 5ml 0.2M acetic acid (mass volume ratio concentration is 4%), dissolve 0.1g polyvinyl alcohol in 4ml ultrapure water (mass volume ratio concentration is 2.5%), heat and dissolve in 1ml ultrapure water 0.78g beta-sodium glycerophosphate (mass volume ratio concentration is 78%); The mass unit in the described mass volume ratio is g, and the unit of volume is ml;
[0031] (2) Slowly drop the β-sodium glycerophosphate (GP) solution into the polyvinyl alcohol (PGA) solution under stirring at room temperature; stir evenly to obtain the polyvinyl alcohol / β-sodium glycerophosphate mixed solution (PGA / GP solution);
[0032] (3) Mixing the polyvinyl alcohol / sodium β-glycerophosphate mixed solution with the chitosan solution to obtain t...
Embodiment 2
[0035] Embodiment 2: Cytocompatibility experiment of chitosan-based thermosensitive hydrogel
[0036] Firstly, the chitosan-based thermosensitive hydrogel glue is applied to the film. Then the chitosan-based thermosensitive hydrogel membrane was cut into circular membranes with a diameter of 1.1 cm. Soak the circular diaphragm in 25%, 50%, and 75% alcohol in sequence, for 2-3 hours each time. Soak with 75% sterile alcohol for 1-2d for the last time. Then transfer the diaphragm to the sterile D-Hanks buffer for soaking, about 10 hours each time, and change the buffer at least 4-5 times. Finally, put it in D-Hanks buffer and soak it for later use.
[0037] 100 microliters of DMEM medium containing 10% fetal bovine serum (FBS) was added to a 48-well cell culture plate, and then the corresponding membrane was placed in a predetermined culture well, and each group had 5 parallel samples at each time point. After the diaphragm is placed, take the cell culture flask, wash, change...
Embodiment 3
[0039] Embodiment 3: Cytotoxicity experiment of chitosan-based thermosensitive hydrogel
[0040] Take the mouse fibroblast L929 in the logarithmic growth phase, digest with 0.25% trypsin and adjust the cell concentration to 1×10 4 / ml, the cell suspension was seeded in a 96-well cell culture plate, and 200 μL of the cell suspension was added to each well. in CO 2 Incubator (37°C, 5% CO 2 ) after pre-cultivation for 24 hours, discard the original culture solution, and replace it with the DMEM medium that has extracted the chitosan-based thermosensitive hydrogel membrane for more than 24 hours, add normal medium to the control group, and set up 12 parallel samples in each group. Place the culture plate at 37 °C, 5% CO2 Continue to grow in the incubator. The culture plate was taken out on the 3rd and 5th day respectively, and the state of the cells was observed under a microscope. 20 μL of 5 mg / ml MTT was added to each well and incubated for 4 hours routinely. Discard the cul...
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