Preparation method of a novel anticoagulant stent coating for capturing endothelial progenitor cells EPCs
An endothelial progenitor cell and anti-coagulation technology, which is applied in the field of biomedical engineering materials, can solve the problems of poor anti-coagulation performance and blood compatibility, and achieve the effect of excellent anti-coagulation ability, strong application potential, and small raw material input
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Embodiment approach 2
[0033] 1. Prepare a sample grafted with CD133 using a gold sheet;
[0034] 2. Then wash the QCM-D channel with SDS and UP water in sequence;
[0035] 3. Place the sample, then open the software, set the temperature to 37°C, and test the sensitivity of the instrument;
[0036] 4. Start to run the air to check whether the QCM-D gold sheet is intact; then pass into the PBS buffer
[0037] 5. After the baseline runs flat, inject αMEM (containing 10% bovine serum albumin) medium solution for about 30 minutes to obtain a relatively flat baseline;
[0038] 6. Pass through the EPCs cell solution containing bovine serum albumin (the cell density is about 10^4 / ml); finally wait for about 8 hours, and then observe the changes in frequency and dissipation. After the experiment is over, analyze the data.
Embodiment 1
[0040] 1. The titanium oxide stent was ultrasonically cleaned three times with SDS, alcohol (ethanol), and RO water (deionized water) for 10 minutes each time, and then dried;
[0041] 2. Hydroxylation treatment of slides: Put the slides in concentrated sulfuric acid and hydrogen peroxide according to the volume ratio of 7:3 (V%:V%=7:3) and soak in 100°C for 2 hours; then wash with RO water for 3 hours times, 5 minutes each time;
[0042] 3. Then immediately put the above sample into Tris-base (10mM, pH8.5) buffer solution, put it in a shaker and shake it tightly for about 1 hour, and then shake it open for about 11 hours. Then use RO water to ultrasonically clean 3 times, and dry with cold air.
[0043] 4. Then configure 10ml of EDC / NHS / MES (1omM / 20mM / 50mM) PBS solution with pH 5.4, then dilute the concentration of CD133 solution with EDC to 0.8-3.2ug / ml and the concentration of Fucoidan to 50-400ug / ml; 50ul each of the CD133 solution and Fucoidan were dropped onto the surf...
Embodiment 2
[0045] 1. The titanium oxide stent was ultrasonically cleaned three times with SDS, alcohol (ethanol), and RO water (deionized water) for 10 minutes each time, and then dried;
[0046] 2. Hydroxylation treatment of slides: Put the slides in concentrated sulfuric acid and hydrogen peroxide according to the volume ratio of 7:3 (V%:V%=7:3) and soak in 100°C for 2 hours; then wash with RO water for 3 hours times, 5 minutes each time;
[0047] 3. Immediately put the above sample into Tris-base (10mM, pH8.5) buffer solution, soak for 8h, then ultrasonically clean it with RO water for 3 times, each time for 5min, and then continue to soak for 3 times in a row. The glass slides of the dopamine film are sealed and preserved;
[0048] 4. Then configure 10ml of EDC / NHS / MES (5mM / 10mM / 50mM) PBS solution with pH 7.4, then dilute the concentration of CD133 solution with EDC to 0.8-3.2ug / ml and the concentration of Fucoidan to 50-400ug / ml; 50ul of CD133 solution and Fucoidan were dropped on...
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