Method for constructing simulated outer cell membrane structure coating on material surface by RAFT (reversible addition fragmentation chain transfer) polymerization technology
A technology imitating the outer membrane and coating of cells, which is applied in the field of surface science and biomedical polymer materials, can solve the problems of wide molecular weight distribution of PMPC, unsatisfactory modification effect, and harsh coating conditions, so as to improve biocompatibility Anti-platelet adhesion, good anti-platelet effect, simple operation
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Embodiment 1
[0015] The preparation of 4-cyanopentanoic acid dithiobenzoic acid RAFT chain transfer agent is completed in the following three steps:
[0016] The first step is to synthesize sodium dithiobenzoate aqueous solution, which is to add 23g of sodium metal to 500mL of methanol to obtain a methanol solution of sodium methoxide, then add 32g of sublimed sulfur, and then pass N 2 , Add 57 mL of benzyl chloride dropwise, and the dropwise addition is completed in one hour. Reflux reaction at 67°C for 10 hours, then rapidly cool the reaction solution to 7°C, remove the methanol by rotary evaporation after suction filtration, add 500 mL of distilled water and then filter again, then wash with 200 mL ether three times, add 200 mL ether and 1 mol / L of hydrochloric acid aqueous solution 500 mL. The dithiobenzoic acid generated after protonation is extracted into the ether phase, the water phase is separated, and 300 mL of water is added, and then 600 mL of 1 mol / L NaOH solution is added, ...
Embodiment 2
[0020] Weigh 4-cyanovaleric acid dithiobenzoic acid RAFT chain transfer agent, dicyclohexylcarbodiimide (DCC), 4-dimethylaminopyridine (DMAP) in a ratio of 10:10:1 Each substance was dissolved in dichloromethane, and the concentration of the chain transfer agent was 0.001-0.002 mmol / mL. The cover glass with primary amino groups on the surface was immersed in this solution, protected from light, and reacted at room temperature for 48 hours. After rinsing the surface with dichloromethane, tetrahydrofuran, and distilled water, the surface was vacuum-dried at 30°C for 5 hours for later use.
[0021] Dissolve MPC and 4,4’-azobis(cyanovaleric acid) (V-501) (the ratio of the two substances is 10:1) in distilled water, and the concentration of MPC is 5g / 100mL. Re-immerse the coverslip with surface-bound RAFT chain transfer agent, pass N 2 After reacting at 70°C for 12 hours, the surface of the substrate was rinsed with distilled water, and dried in vacuum at 30°C for 5 hours. By us...
Embodiment 3
[0024] The platelet adhesion experiment refers to the literature method (Chen Y. M. et al. Biomaterials 2007, 28(10), 1752-1760), add 3.8wt% sodium citrate solution to healthy human blood at a volume ratio of 9:1, mix well, centrifuge at 1000 rp for 10 min, and take the upper layer of light yellow Platelet-rich plasma (PRP) was used for experiments. Soak the sample in PBS buffer for 2 h, then take out the sample, rinse with PBS buffer three times and wash off excess liquid from the edge of the sample with filter paper. Pipette 20 muL PRP was added dropwise to the middle part of the sample surface, placed in a petri dish, and incubated at 37 °C for 2 h in a carbon dioxide incubator filled with distilled water. Then rinse the sample with PBS buffer to wash away the platelets that are not firmly adhered. Soak the sample in the previous step into 2.5% glutaraldehyde solution for fixation for 1 h, and then rinse the surface of the sample with PBS buffer and distilled water re...
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