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Preparation method for muscle accellular material capable of slow-releasing IGF-1 growth factors

An IGF-1, growth factor technology, applied in tissue regeneration, medical science, prosthesis, etc., can solve the lack of microenvironment required for cell growth and development, maintain it between 10-24 hours, and cannot achieve muscle repair and regeneration, etc. problem, to achieve the effect of low immune rejection, promoting muscle regeneration, and promoting regeneration

Inactive Publication Date: 2018-09-11
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The results of applying IGF-1 alone or adding IGF-1 to artificial synthetic materials to repair large-scale muscle defects are not ideal, because the proliferation and differentiation of muscle satellite cells need to be several days later (3-7 days), and the current reported The maximum potency of IGF-1 release is only maintained between 10-24 hours
Moreover, compared with natural muscle ECM, artificial synthetic materials lack the microenvironment required for natural cell growth and development, and cannot achieve good muscle repair and regeneration.

Method used

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  • Preparation method for muscle accellular material capable of slow-releasing IGF-1 growth factors
  • Preparation method for muscle accellular material capable of slow-releasing IGF-1 growth factors
  • Preparation method for muscle accellular material capable of slow-releasing IGF-1 growth factors

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1: Preparation and regenerative repair of decellularized muscle material with spontaneous and sustained release of IGF-1 growth factor

[0042] 1. Preparation of novel decellularized muscle materials

[0043] (1) Materials: Cut the erector spinae of pigs into lengths, widths and thicknesses of 2 cm according to the direction of the muscle fibers. Rinse them with normal saline containing 10 KIU / ml protease inhibitors for 3 times, each time for 20 minutes, and then fully rinse them with sterile PBS to remove them. blood and other impurities.

[0044] (2) The decellularization steps are as follows:

[0045] ①. In the PBS buffer solution containing 10K IU / ml protease inhibitor, shake at 100 rpm on a low temperature (4°C) shaker for 48 hours, and then freeze and thaw with liquid nitrogen for 3 cycles (-80°C / 37°C);

[0046] ②. In the PBS buffer containing 0.1M KCL, add penicillin and streptomycin (20KIU / ml, 20g / ml) mixed antibacterial solution 1:1, and shake at 100r...

Embodiment 2

[0062] Example 2: Preparation and regenerative repair of decellularized muscle material with spontaneous and sustained release of IGF-1 growth factor

[0063] 1. Preparation of novel decellularized muscle materials

[0064] (1) Materials: Cut the erector spinae of pigs into length, width and thickness of 4 cm according to the direction of the muscle fibers, rinse them with normal saline containing 10 KIU / ml protease inhibitors for 3 times, each time for 20 min, and then fully rinse with sterile PBS to remove blood and other impurities.

[0065] (2) The decellularization steps are as follows:

[0066] ①. In the PBS buffer solution containing 30K IU / ml protease inhibitor, shake at 100 rpm for 36 hours on a low-temperature (4°C) shaker, and then freeze and thaw with liquid nitrogen for 3 cycles (-80°C / 37°C);

[0067] ②. In the PBS buffer containing 0.6M KCL, add penicillin and streptomycin (20K IU / ml, 20g / ml) mixed antibacterial solution 1:1, shake at 100rpm on a low temperatur...

Embodiment 3

[0072] Example 3: Preparation and regenerative repair of decellularized muscle material with spontaneous and sustained release of IGF-1 growth factor

[0073] 1. Preparation of novel decellularized muscle materials

[0074] (1) Materials: The erector spinae of sheep were cut into 4 cm in length, width and thickness according to the direction of muscle fiber, rinsed with normal saline containing 10 KIU / ml protease inhibitor for 3 times, each time for 20 min, fully rinsed with sterile PBS to remove blood and other impurities.

[0075] (2) The decellularization steps are as follows:

[0076] ①. In the PBS buffer solution containing 50K IU / ml protease inhibitor, shake at 100 rpm for 24 hours on a low-temperature (4°C) shaker, and then freeze and thaw with liquid nitrogen for 3 cycles (-80°C / 37°C);

[0077] ②. In the PBS buffer containing 1.2M KCL, add penicillin and streptomycin (20K IU / ml, 20g / ml) mixed antibacterial solution 1:1, shake at 100rpm on a low temperature (4°C) shak...

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Abstract

The invention provides a preparation method for a muscle accellular material capable of slow-releasing IGF-1 growth factors. The method comprises the following steps: enabling a skeletal muscle tissueof a mammal in arbitrary sizes to be processed by PBS containing a protease inhibitor, PBS buffer solution containing KCl, PBS buffer solution containing Triton X, PBS buffer solution containing SDS,and PBS buffer solution containing DNA enzyme, and acquiring the muscle accellular material capable of spontaneously and continuously releasing the IGF-1 growth factors. The method is capable of simultaneously performing complete decellularization treatment on cells in the muscle tissue, and gentle in condition without ECM damage, rapid and stable. The obtained muscle accellular material is capable of spontaneously and continuously releasing the IGF-1 growth factors, has the advantages of good biocompatibility, strong plasticity, high biomechanical strength and the like, and can be used for repairing muscle injury and regression related diseases caused by various pathogenesis clinically.

Description

technical field [0001] The present invention mainly relates to the medical field for repairing and regenerating skeletal muscle tissue of the whole body, and specifically relates to the preparation of a novel muscle decellularized material. The biological scaffold can spontaneously and continuously release IGF-1-1 growth factor, and preferably Retaining the extracellular matrix components of natural muscle tissue, the material can promote the repair and regeneration of large area muscle defects. Background technique [0002] Although skeletal muscle has a certain self-repair ability after injury, acute muscle damage of more than 20% of the area often leads to irreparable defects and is often replaced by a large amount of scar fibrous tissue, which we call large area muscle defects. Currently, there is no effective treatment that can promote the repair and regeneration of large-scale muscle defects. [0003] During skeletal muscle extracellular matrix (ECM) remodeling and mu...

Claims

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Application Information

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IPC IPC(8): A61L27/36A61L27/54
CPCA61L27/3604A61L27/367A61L27/3687A61L27/54A61L2300/414A61L2430/30
Inventor 林贤丰范顺武王刚良王艺芸陈家鑫王晟毓
Owner ZHEJIANG UNIV
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