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Protein-modified PLGA microsphere and tissue-engineered nerve constructed by protein-modified PLGA microsphere

A protein modification and tissue engineering technology, applied in the field of biomedicine, can solve the problems of immunogenicity, difficulty in the source of seed cells, clinical application limitations, etc., and achieve the effect of promoting peripheral nerve regeneration, promoting neuronal process growth, and regular morphology.

Active Publication Date: 2020-08-18
NANTONG UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the source of seed cells is often difficult, or the allogeneic immunogenicity and other problems, the clinical application is limited; when adding growth factors to the scaffold material, it is necessary to consider maintaining stability and efficiency according to their respective physical and chemical properties

Method used

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  • Protein-modified PLGA microsphere and tissue-engineered nerve constructed by protein-modified PLGA microsphere
  • Protein-modified PLGA microsphere and tissue-engineered nerve constructed by protein-modified PLGA microsphere
  • Protein-modified PLGA microsphere and tissue-engineered nerve constructed by protein-modified PLGA microsphere

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] The preparation of embodiment 1 lipid-soluble extract of Brucea javanica

[0037] The lipid-soluble extract of Brucea javanica was prepared according to the method disclosed in CN2020103158417.

[0038] Bructus javanica was purchased from Nantong Hospital of Traditional Chinese Medicine, washed with deionized water, dried in an oven at 60-65°C for 2 days, and then ground into powder (300g). Accurately weigh 50 g of Brucea javanica dry powder, put it into an extraction vessel, and soak it in mixed solutions of absolute ethanol and ethyl acetate of different concentrations for 48 hours at room temperature (22°C). Grouping: 95% absolute ethanol + 5% ethyl acetate; 85% absolute ethanol + 15% ethyl acetate; 75% absolute ethanol + 25% ethyl acetate; 65% absolute ethanol + 35% ethyl acetate; 55% absolute ethanol + 45% ethyl acetate; 45% absolute ethanol + 55% ethyl acetate; 35% absolute ethanol + 65% ethyl acetate; 25% absolute ethanol + 75% ethyl acetate; 15% Absolute ethan...

Embodiment 2

[0039] The extraction effect of the dehydrated alcohol and ethyl acetate mixed solution of embodiment 2 different concentrations

[0040] In order to detect the effect of the fat-soluble mixture of Brucea javanica extracted with different concentrations of absolute ethanol and ethyl acetate on the growth of nerve cells, the experiment took the nerve cell line PC12 cells as the observation model.

[0041] PC12 cells were cultured in DMEM complete medium (containing 10% horse serum, 5% fetal bovine serum, 100 U / ml penicillin, 100 μg / ml streptomycin) in a petri dish, placed in 5% CO 2 , Cultivated in a 37°C incubator. The medium was changed every two days, and the cells were passaged when they grew to 80% confluence. In the experiment, PC12 cells in the logarithmic growth phase were taken, and 5×10 4 / ml Seed cells in 24-well culture plate, 400μl per well. After 24 hours, it was replaced with DMEM medium containing 1% horse serum and 1% fetal bovine serum. The experiments wer...

Embodiment 3

[0045] Example 3 Cytotoxicity Detection of Brucea Brucea Fat-Soluble Extract

[0046] The fat-soluble extract of Brucea javanica obtained in Example 1 was freeze-dried overnight (EYELA FDU-1200, Tokyo), and the obtained powder was put into a 15ml centrifuge tube and stored at 4°C. Prepare a 10μg / mL solution with sterile deionized water, centrifuge at 8000g for 10min, and then prepare the solution to the concentration required for the experiment, and then use a 0.2mm nylon syringe filter (Millipore, USA) to filter and sterilize the cultured PC12 Cytotoxicity tests were performed in cell lines. The CCK8 kit was purchased from Japan Dojin Chemical Research Institute.

[0047] Take PC12 cells in the logarithmic growth phase, digest, count and resuspend, and adjust the cell density to 5×10 5 / mL, inoculate 100 μL per well in a 96-well plate; after the cells adhere to the wall, discard the medium and wash with 0.01 MPBS for 5 min×2 times. The negative control group was 1% horse s...

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Abstract

The invention discloses a protein modified PLGA microsphere and a tissue engineered nerve constructed by the same. According to the invention, microspheres are loaded with active substances for treating peripheral nerve injury and are combined with tissue-engineered nerves, and researches show that the prepared tissue-engineered nerves can effectively promote nerve regeneration after peripheral nerve injury.

Description

technical field [0001] The invention belongs to the field of biomedicine, and in particular relates to a protein-modified PLGA microsphere and a tissue-engineered nerve constructed therefrom, which can be used for repairing peripheral nerve defects. Background technique: [0002] Peripheral nerve injury is a common clinical disease, which usually leads to paralysis, paralysis and even loss of autonomous control of corresponding body parts, seriously affecting the quality of life of patients. At present, the effect of surgical repair by simply suturing stumped nerves is not satisfactory, and autologous nerve transplantation, which is the gold standard for repairing peripheral nerve defects, is not suitable for clinical application due to the difficulty in the source of autologous nerve donors, secondary damage caused by transplantation, and many other factors. restricted. Therefore, constructing suitable tissue-engineered nerves to replace autologous nerves to repair periphe...

Claims

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Application Information

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IPC IPC(8): A61L27/18A61L27/22A61L27/24A61L27/50A61L27/54
CPCA61L27/24A61L27/227A61L27/18A61L27/54A61L27/50A61L2300/30A61L2300/412A61L2300/414A61L2300/602A61L2430/32C08L67/04
Inventor 汤欣周友浪顾晓松
Owner NANTONG UNIVERSITY