Hydrogel coated HIF-1 alpha overexpression lentiviral vector compound for repairing brachial plexus avulsion and preparation method

A technology of HIF-1 and lentiviral vector, which is applied in the fields of botany equipment and methods, biochemical equipment and methods, nervous system diseases, etc., can solve the problems such as undiscovered brachial plexus avulsion injury, and achieve the promotion of angiogenesis and movement Functional recovery, simple process method, effect of promoting nerve regeneration and functional recovery

Inactive Publication Date: 2019-06-07
GUANGDONG MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The HIF-1α gene has been deeply studied by researchers in ischemia-hypoxic injury, but there is no relevant report on its application in the repair of brachial plexus avulsion injury

Method used

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  • Hydrogel coated HIF-1 alpha overexpression lentiviral vector compound for repairing brachial plexus avulsion and preparation method
  • Hydrogel coated HIF-1 alpha overexpression lentiviral vector compound for repairing brachial plexus avulsion and preparation method
  • Hydrogel coated HIF-1 alpha overexpression lentiviral vector compound for repairing brachial plexus avulsion and preparation method

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Experimental program
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Embodiment 1

[0030] A method for preparing a hydrogel-wrapped HIF-1α overexpression lentiviral vector complex for repairing brachial plexus avulsion nerve injury, comprising the following steps:

[0031] Step 1, synthesis of HIF-1α overexpression lentiviral vector: pHBLV-CMVIE-IRES-Puro synthesized by Hanheng Biotechnology Co., Ltd. (Shanghai, China) was selected as the lentiviral vector for the overexpression of HIF-1α, wherein, HIF-1α primer sequences are as follows: ggatctatttttggggtgaattcgccaccATGGA GGGCGCCGGCG (forward primer); tagaactagtctcgaggaattcTC AGTTAACTTGATCCAAA (reverse primer);

[0032] Step 2, preparation of hydrogel stock solution: PF-127 sterile powder and water are configured into a suspension with a certain mass-to-volume ratio, and then the suspension is shaken until the hydrogel is fully dissolved to obtain a mixed solution, Then, after the mixed solution is filtered and sterilized through a filter membrane, a sterile PF-127 hydrogel stock solution is obtained;

[00...

Embodiment 2

[0035] A method for preparing a hydrogel-wrapped HIF-1α overexpression lentiviral vector complex for repairing brachial plexus avulsion nerve injury, comprising the following steps:

[0036] Step 1, synthesis of HIF-1α overexpression lentiviral vector: pHBLV-CMVIE-IRES-Puro synthesized by Hanheng Biotechnology Co., Ltd. (Shanghai, China) was selected as the lentiviral vector for the overexpression of HIF-1α, wherein, The HIF-1α primer sequence is as follows: ggatctatttttggggtgaattcgccaccATGGA GGGCGCCGGCG (forward primer); tagaactagtctcgaggaattcTC AGTTAACTTGATCCAAA (reverse primer); in this embodiment, the titer of the lentiviral vector is 2 × 10 8 TU / mL.

[0037] Step 2, preparation of hydrogel stock solution: PF-127 sterile powder and deionized water are configured into a suspension with a mass volume ratio of 20%, and then the suspension is placed on a horizontal shaker at 2°C to Shake at a speed of 65 rpm for 18 hours until the hydrogel is fully dissolved to obtain a mixed...

Embodiment 3

[0042] A method for preparing a hydrogel-wrapped HIF-1α overexpression lentiviral vector complex for repairing brachial plexus avulsion nerve injury, comprising the following steps:

[0043] Step 1, synthesis of HIF-1α overexpression lentiviral vector: pHBLV-CMVIE-IRES-Puro synthesized by Hanheng Biotechnology Co., Ltd. (Shanghai, China) was selected as the lentiviral vector for the overexpression of HIF-1α, wherein, The HIF-1α primer sequence is as follows: ggatctatttttggggtgaattcgccaccATGGA GGGCGCCGGCG (forward primer); tagaactagtctcgaggaattcTC AGTTAACTTGATCCAAA (reverse primer); in this embodiment, the titer of the lentiviral vector is 2 × 10 8 TU / mL.

[0044] Step 2, preparation of hydrogel stock solution: PF-127 sterile powder and deionized water are configured into a suspension with a mass volume ratio of 15%, and then the suspension is placed on a horizontal shaker at 0° C. Shake at a speed of 60 rpm for 24 hours until the hydrogel is fully dissolved to obtain a mixed ...

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Abstract

The invention relates to the technical field of biology, in particular to a hydrogel coated HIF-1 alpha overexpression lentiviral vector compound for repairing brachial plexus avulsion and a preparation method. The preparation method of the hydrogel coated HIF-1 alpha overexpression lentiviral vector compound for repairing brachial plexus avulsion comprises the steps of step I, synthetizing an HIF-1 alpha overexpression lentiviral vector; step II, preparing a hydrogel stock solution; and step III, preparing the hydrogel and HIF-1 alpha overexpression lentiviral vector compound. According to the hydrogel coated HIF-1 alpha overexpression lentiviral vector compound prepared by the preparation method, local delivery and slow release effects of the HIF-1 alpha overexpression lentiviral vectorcan be realized, advantageous influence is provided for regenerative repair and revascularization of BPA posterior nerve, the overexpression of local HIF-1 alpha genes of BPA damage is realized, BPA damage repair is effectively promoted, and angiogenesis and movement function restoration are promoted.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a hydrogel-wrapped HIF-1α overexpression lentiviral vector complex for repairing brachial plexus avulsion nerve injury and a preparation method thereof. Background technique [0002] Brachial plexus avulsion (BPA) is a serious central-peripheral nervous system junctional injury. With the development of modern transportation, the number of patients with brachial plexus avulsion caused by accidents is increasing day by day. After the nerve root is severed from the spinal cord, it is often accompanied by a large number of motor neurons (motor neurons, MNs) loss and death, resulting in upper limb motor sensory dysfunction, which greatly affects the quality of life of patients (Wang et al., 2015). Due to the complex condition of BPA and its special anatomical location, the current treatment of such patients focuses on the treatment of nerve injury. Nerve root transplantation is often use...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K48/00A61K38/17A61K9/06A61K47/10A61P25/00A61P25/02C12N15/867
Inventor 吴洪福曾丽妮王裕辉王涛丁璐张晓敏马锴璇林婉冯宇泽
Owner GUANGDONG MEDICAL UNIV
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