Schiff base hydrogel material with side chain grafted with phenylboronic acid and active oxygen eliminating function and preparation method of Schiff base hydrogel material
A technology of phenylboronic acid and active oxygen, which is applied to wave energy or particle radiation treatment materials, medical preparations with non-active ingredients, medical preparations containing active ingredients, etc., can solve problems such as high reaction sensitivity and promote removal response, reducing the outbreak of inflammation, and inhibiting the further deterioration of myocardial infarction
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Embodiment 1
[0036] Step 1: Dissolve HA (1 g, 0.01 mmol) in 2-fold concentration PBS (adjusted with 1 M HCl) at pH = 6, then add ADH (13.05 g, 75 mmol) to adjust the pH to keep it constant. After complete dissolution, EDC (3.9g) and NHS (1.15g) were added in sequence, while the pH was kept at 6. The reaction was carried out in a water bath at 37°C for 4h, and then reacted overnight. After the reaction, the pH of the reaction solution was adjusted to neutral. The reaction mixture was dropped into 800 mL ice ethanol for precipitation. The precipitate was collected by centrifugation, dissolved in water, dialyzed (MWCO=14kDa) for 5 days, and freeze-dried to obtain the product HA-ADH.
[0037] Step 2: Dissolve HA-ADH (400 mg) and PBA (105.8 mg) in 8 mL of a mixed solvent (volume ratio water:DMSO=3:1), and add 1 drop of 10% HCl solution as a catalyst. Drop the reaction mixture into 80mL of ice ethanol for precipitation, centrifuge to collect the precipitate, redissolve the precipitate in water...
Embodiment 2
[0042] Step 1: Dissolve HA (1 g, 0.01 mmol) in 2-fold concentration PBS (adjusted with 1 M HCl) at pH = 6, then add ADH (13.05 g, 75 mmol) to adjust the pH to keep it constant. After complete dissolution, EDC (3.9g) and NHS (1.15g) were added in sequence, while the pH was kept at 6. The reaction was carried out in a water bath at 37°C for 4h, and then reacted overnight. After the reaction, the pH of the reaction solution was adjusted to neutral. The reaction mixture was dropped into 800 mL ice ethanol for precipitation. The precipitate was collected by centrifugation, dissolved in water, dialyzed (MWCO=14kDa) for 5 days, and freeze-dried to obtain the product (HA-ADH).
[0043] Step 2: Dissolve HA-ADH (400 mg) and PBA (105.8 mg) in 8 mL of a mixed solvent (volume ratio water:DMSO=3:1), and add 2 drops of 10% HCl solution as a catalyst. The reaction mixture was dropped into 80mL of ice ethanol for precipitation, centrifuged to collect the precipitate, and the precipitate was ...
Embodiment 3
[0047] To detect the protective effect of hydrogel material R gel on the oxidative stress of cardiomyocytes stimulated by hydrogen peroxide. H9C2 cells were inoculated in DMEM containing 10% fetal bovine serum at a density of 3×10 5 . After 24 h, use fresh medium, medium containing 500 μM H2O2 or medium containing 500 μM H 2 o 2 Replace the culture medium with the culture medium of the hydrogel. After 1 h of treatment, DCFH-DA (1:2000 v / v in DMEM) was added. After 30 min, rinse 3 times with PBS to remove excess probe. Fluorescence quantification was performed using a fluorescence microscope (Olympus, IX81, Japan) and a BD FACSCalibur flow cytometer (BD Biosciences, USA).
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