A self-assembled porous polymer membrane for glomerular filtration membrane repair and its preparation method and application
A porous polymer, self-assembly technology, used in medical science, prostheses, antibodies, etc., to achieve the effect of reducing toxic effects, good effects, and good repair effects
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Embodiment 1
[0031] To prepare a macroporous self-assembled porous polymer membrane, the steps are as follows:
[0032] 1) Add 1×10 to the flask -4 Mole of ethylene glycol and polyethylene glycol (the molar ratio of ethylene glycol and polyethylene glycol is 1:1), add 0.1 mole of methyl ester and butyl acetate (the molar ratio of methyl ester and butyl acetate is 1:1), And use nitrogen to remove the air in the reaction bottle and then seal it, and stir and react at 110°C for 15 hours;
[0033] 2) Dissolve the above obtained material with ethyl acetate and precipitate with ethanol, and repeat this process 4 times;
[0034] 3) The obtained product was vacuum-dried at 70 °C for 10 hours to obtain neutral polymer particles;
[0035] 4) Dissolve 1 mol of polymer particles in chloroform, add 1 mol of acetic anhydride, stir and react at 80 °C for 20 hours to obtain negatively charged polymer particles;
[0036] 5) Dissolve 1 g of negatively charged polymer particles in 30 ml of chloroform and ...
Embodiment 2
[0040] Prepare a kind of self-assembled porous polymer film of small size hole, the steps are as follows:
[0041] 1) Dissolve 1 g of the negatively charged polymer particles obtained in Example 1 in 25 ml of chloroform and butyl acetate, add 100 ml of polyethylene glycol and methanol under stirring at 40 °C, and then quench with liquid nitrogen For 3 min, pour 1000 ml of distilled water to exchange the solvent to obtain a negatively charged self-assembled porous polymer film;
[0042]2) Disperse the negatively charged self-assembled porous polymer membrane in PB buffer solution with pH=5.5-6, stir for 15 minutes, add EDAC and Sulfo-NHS, stir for 15 minutes, adjust the pH to 7-7.5, add anti-nephrin (the nephrin antibody on the glomerulus, purchased from Santa Crus, USA), stirred at room temperature for 4 hours, filtered and washed 3 times, and obtained a self-assembled porous polymer membrane for glomerular filtration membrane repair; the negative The molar ratio of charged s...
Embodiment 3
[0045] Podocytes grown to 80% cells were digested with trypsin to make a concentration of 5×10 4 / ml of cell suspension, inoculate 100 μL of cell suspension into a 96-well plate, and store at 37 °C, 5% CO 2 Cultured for 24 h, replaced with serum-free medium and cultured for 6 h to keep the cells in a stationary phase, and added 10 μL of different concentrations of self-assembled porous polymer membranes (0.01 mg / ml, 0.1 mg / ml, 1 mg / ml, 10 mg / ml) respectively. ml), and a blank control with only culture medium and a normal control with cells but no self-assembled porous polymer membrane were set up. Three replicate wells were set up for each concentration, and the culture plate was incubated in the incubator for 12 h. After incubation, 10 μL of CCK-8 solution was added to each well, and the absorbance at 450 nm wavelength was measured with a microplate reader 2 h later. Repeat 3 times. see results image 3 , when the material concentration is as high as 10 mg / ml, there is al...
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