A kind of nerve growth factor injectable in situ hydrogel, preparation and application
A technology of nerve growth factor and injection water, applied in the field of nerve tissue engineering and regenerative medicine, can solve problems such as short half-life, achieve good tissue compatibility, reduce toxic side effects, and strong practicability.
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Embodiment 1
[0050] Preparation of HA-CS-NGF injectable in situ hydrogel
[0051] (1) Add 0.1g of HA to 10ml of deionized water in an ice bath at 4°C, stir magnetically for 50 minutes to fully dissolve, then gradually add 0.1g CS hydrochloride, stir magnetically for 1 hour to mix evenly Adjust the pH to 4.7, then weigh a certain mass of EDC and NHS into the above reaction solution, so that the molar masses of EDC and NHS are 30mM and 30mM, respectively, and react for 2 hours to obtain the HA-CS sol.
[0052] (2) Mix the HA-CS sol prepared in (1) with 0.5ml of 0.5μg / ml NGF solution at 4°C, and stir for 30min to make it evenly mixed.
[0053] Structure and morphology characterization of HA-CS-NGF injectable in situ hydrogel
[0054] Adjust the pH of the above-prepared system to 7.4 and place it in a constant temperature water bath at 37°C (simulating the physiological environment of the human body) until the sol in the bottle cannot flow completely, then vacuum freeze-dry for 12-48 hours be...
Embodiment 2
[0061] Preparation of HA-CS-NGF injectable in situ hydrogel
[0062] (1) Add 0.2g of HA into 10ml of deionized water in an ice bath at 4°C, stir magnetically for 50min to fully dissolve, then gradually add 0.1gCS hydrochloride, stir magnetically for 1h to mix evenly Adjust the pH to 5.4, then weigh a certain mass of EDC and NHS into the above reaction solution, so that the molar masses of EDC and NHS are 70mM and 30mM, respectively, and react for 1 hour to obtain the HA-CS sol.
[0063] (2) Mix the HA-CS sol prepared in (1) with 0.5ml of 0.75μg / ml NGF solution at 4°C in a certain ratio, and stir for 30min to make it evenly mixed.
[0064] Structure and morphology characterization of HA-CS-NGF injectable in situ hydrogel
[0065] Adjust the pH of the above-prepared system to 7.4 and place it in a constant temperature water bath at 37°C (simulating the physiological environment of the human body) until the sol in the bottle cannot flow completely, then vacuum freeze-dry for 12-...
Embodiment 3
[0074] Preparation of HA-CS-NGF injectable in situ hydrogel
[0075] (1) Add 0.2g of HA to 10ml of deionized water in an ice bath at 4°C, stir magnetically for 50min to fully dissolve, then gradually add 0.3gCS hydrochloride, stir magnetically for 1h to mix evenly Adjust the pH to 6, then weigh a certain mass of EDC and NHS into the above reaction solution, so that the molar masses of EDC and NHS are 10mM and 10mM, respectively, and react for 2 hours to obtain the HA-CS sol.
[0076] (2) Mix the HA-CS sol prepared in (1) with 0.5ml of 1μg / ml NGF solution at 4°C in a certain ratio, and stir for 30min to make it evenly mixed.
[0077] Structure and morphology characterization of HA-CS-NGF injectable in situ hydrogel
[0078] Adjust the pH of the above-prepared system to 7.4 and place it in a constant temperature water bath at 37°C (simulating the physiological environment of the human body) until the sol in the bottle cannot flow completely, then vacuum freeze-dry for 12-48 hou...
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