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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.

Active Publication Date: 2017-11-17
武汉渝联生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Afterwards, the environment-sensitive properties of the hydrogel can be used to realize the slow release of NGF in the body, effectively solving the problems of NGF activity reduction and burst release caused by short half-life, diffusion or rapid degradation, and maintaining good biological activity of NGF , promote nerve axon extension and myelination, so as to achieve the purpose of repairing nerves

Method used

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  • A kind of nerve growth factor injectable in situ hydrogel, preparation and application
  • A kind of nerve growth factor injectable in situ hydrogel, preparation and application
  • A kind of nerve growth factor injectable in situ hydrogel, preparation and application

Examples

Experimental program
Comparison scheme
Effect test

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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Abstract

The invention relates to NGF (Nerve Growth Factor) injectable in-situ hydrogel as well as preparation and application thereof. The NGF injectable in-situ hydrogel is obtained by adding EDC(1-ethyl amide crosslinking agent-3-(3-dimethyl amino propyl) carbide imine) and NHS (N-hydroxysuccinimide) into an aqueous solution system of hyaluronic acid and chitosan hydrochloride, and adding an NGF after HA-NHS-active ester intermediate by the hyaluronic acid under the action of the EDC / NHS under an acidic condition. The hydrogel has a pH sensitive feature; the conversion process of solution-gel can be completed under physiological conditions of a human body; the NGF injectable in-situ hydrogel can be subjected to in-situ curing under the action of pH in an acceptor at a nerve damage part after the NGF injectable in-situ hydrogel is injected into nerve tissue; the traumatic property of a surgery is avoided. The NGF can be released slowly in vitro by utilizing the environmentally sensitive feature of the hydrogel; the problems of low activity, burst release and the like caused by the facts that a half-life period is short and the diffusion and the degradation are too quick of the NGF are effectively solved; high bioactivity of the NGF is kept; the extension and the myelination of a nerve axon are promoted; therefore, the purpose of repairing nerves is achieved.

Description

technical field [0001] The invention belongs to the fields of nerve tissue engineering and regenerative medicine, and in particular relates to an injectable in situ hydrogel of nerve growth factor, its preparation and its application in nerve repair. Background technique [0002] Peripheral nerve injury is a common clinical disease. At present, nerve anastomosis, autologous nerve transplantation, allogeneic nerve transplantation, autologous non-nerve tissue transplantation, biomaterial replacement and other methods are generally used in clinical nerve repair. Among them, the processing of biomaterials into tubular structures of nerve conduits is the focus of current research. However, when encountering long-distance and thick nerve defects, such catheters made of biological materials lack the support of microenvironmental substances such as bioactive substances, and the repair effect is very limited. [0003] Nerve growth factor (NGF) is a nerve cell growth regulator with ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61L27/20A61L27/56A61L27/54
Inventor 徐海星张凌溪许沛虎郑芙蓉金鸣李安召汪志辉
Owner 武汉渝联生物科技有限公司
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