A method for oxidizing oligosaccharides to cross-link collagen
A technology for oxidizing oligosaccharides and collagen, which is applied in the field of biomedical materials and leather industry, can solve the problems of oxidized polysaccharides with large molecular weight, low cross-linking efficiency, and large reaction steric hindrance, and achieve active chemical properties, small molecular weight, and cross-linking conditions easy effect
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Embodiment 1
[0032] (1) Weigh 5 grams (dry weight) of acellular porcine dermal matrix into an Erlenmeyer flask, and add 80ml of sodium carbonate-sodium bicarbonate buffer solution with a pH of 9.4;
[0033] (2) Weigh 1 g of oxidized seaweed oligosaccharide with an oxidation degree of 45%, add it to the above-mentioned Erlenmeyer flask, shake well, and close the mouth of the bottle;
[0034] (3) Place the above-mentioned Erlenmeyer flask in a water bath with a temperature-controllable cradle, adjust the reaction temperature to 37°C, and adjust the rotation speed to 150r / min;
[0035] (4) React for 24 hours, take out the Erlenmeyer flask, discard the reaction solution, add 50ml of 0.1mol / L ammonium chloride solution and shake for 0.5h, then discard the solution, add 100ml of water for injection to wash for 0.5 hours, discard and wash solution, repeated washing three times;
[0036] (5) Take out the cross-linked acellular porcine dermal matrix, and freeze-dry it in a freeze dryer.
Embodiment 2
[0038] (1) Weigh 3 grams of collagen film;
[0039] (2) Weigh 0.5 g of oxidized isomaltooligosaccharide with a degree of oxidation of 100%, place it in a beaker filled with 50 ml of phosphate buffer solution of pH 7.4, and shake it to completely dissolve it;
[0040] (3) Put the collagen film into the above beaker and close the mouth of the beaker;
[0041] (3) Put a magnetic stir bar into the above beaker, place the beaker on the magnetic stirrer, adjust the reaction temperature to 25°C, and the stirring speed to 1000r / min;
[0042] (4) After 40 hours, discard the reaction solution, add 50ml of distilled water to wash for 0.5 hours, and repeat the washing three times;
[0043] (5) Take out the cross-linked collagen membrane and air-dry it on an ultra-clean bench.
Embodiment 3
[0045] (1) Weigh 100 grams of collagen sponge, put it in a 30-liter reaction kettle, add 19 liters of 0.5mol / L acetic acid solution, and stir thoroughly at 4°C until the collagen sponge is completely dissolved;
[0046] (2) Weigh 10 grams of oxidized chitooligosaccharide with a degree of oxidation of 10%, and dissolve it in 1 liter of 0.5mol / L acetic acid solution at a temperature of 4°C at 4°C;
[0047] (3) Slowly add the oxidized chitosan oligosaccharide solution into the collagen solution, and keep stirring at 4°C to carry out the cross-linking reaction;
[0048] (4) After reacting for 16 hours, use sodium hydroxide solution to adjust the pH value to be weakly alkaline (pH value is about 7.50); slowly add solid ammonium sulfate to make the concentration about 1.5 mol / L, stir continuously until it is completely dissolved, and then let stand Overnight (10-12h); the next day, centrifuge (8000 r / min, 15 min), discard the supernatant, add 2 liters of 0.5mol / L acetic acid solut...
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