Method for preparing tendon composite muscle decellularized material derived from natural tissue

A decellularization and tendon technology, used in tissue regeneration, medical science, prosthesis, etc., can solve the problems of rupture, complex junction, poor mechanical properties of muscle scaffolds, etc., to achieve strong biomechanical properties, low immune rejection, good The effect of biocompatibility

Inactive Publication Date: 2018-08-31
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The widespread application of tissue engineering has made some progress in overcoming this disease. Current research is mainly focused on preparing a simple muscle scaffold to induce muscle fiber regeneration, but the effect is not ideal.
First: Under physiological conditions, muscle fibers are connected to tendons or myofascia, and it is through the connection of muscles and tendons that a network structure is formed, and a single muscle fiber forms a sum of contractions to produce efficient movement, while a simple muscle scaffold lacks this structure
Second: The mechanical properties of simple muscle scaffolds are poor (maximum stress is 0.1-5Mpa), and they are easy to rupture under high tension of large abdominal defects or limb m

Method used

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  • Method for preparing tendon composite muscle decellularized material derived from natural tissue
  • Method for preparing tendon composite muscle decellularized material derived from natural tissue
  • Method for preparing tendon composite muscle decellularized material derived from natural tissue

Examples

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Comparison scheme
Effect test

Embodiment 1

[0040] Example 1: Preparation method and regenerative repair research of tendon composite muscle decellularized material derived from natural tissue

[0041] 1. Preparation of Tendon Composite Muscle Decellularized Material

[0042] (1) Materials: Take the tendon compound muscle material from the Achilles tendon of pigs, cut the tendon into length, width and thickness of 2*1.0*0.2cm according to the fiber direction, and the length, width and height of the muscle are 0.3cm. IU / ml protease inhibitor normal saline rinsed 3 times, 30min each time, fully rinsed with sterile PBS to remove blood and other impurities.

[0043] (2) The decellularization steps are as follows:

[0044] 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);

[0045] ②. In the PBS buffer containing 0.1M KCL, add penicillin and streptomycin (20KIU / ml, 20...

Embodiment 2

[0061] Example 2: Preparation and regenerative repair of decellularized tendon and muscle materials

[0062] 1. Preparation of Tendon Composite Muscle Decellularized Material

[0063] (1) Materials: Take the tendon compound muscle material from the Achilles tendon of pigs, cut the tendon into length, width and thickness of 4.0*2.0*0.4cm according to the fiber direction, and the length, width and height of the muscle are 0.6cm. IU / ml protease inhibitor normal saline rinsed 3 times, 30min each time, fully rinsed with sterile PBS to remove blood and other impurities.

[0064] (2) The decellularization steps are as follows:

[0065] ①. In the PBS buffer solution containing 30K IU / ml protease inhibitor, shake at 100 rpm for 48 hours on a low-temperature (4°C) shaker, and then freeze and thaw with liquid nitrogen for 3 cycles (-80°C / 37°C);

[0066] ②. In the PBS buffer containing 0.5M KCL, add penicillin and streptomycin (20KIU / ml, 20g / ml) mixed antibacterial solution 1:1, and sha...

Embodiment 3

[0071] Example 3: Preparation method and regenerative repair research of tendon composite muscle decellularized material derived from natural tissue

[0072] 1. Preparation of Tendon Composite Muscle Decellularized Material

[0073] (1) Materials: The tendon compound muscle material was taken from the Achilles tendon of sheep, and the tendon was cut into length, width and thickness according to the fiber direction of 4.0*2.0*0.6cm, and the length, width and height of the muscle were 0.6cm. IU / ml protease inhibitor normal saline rinsed 3 times, 30min each time, fully rinsed with sterile PBS to remove blood and other impurities.

[0074] (2) The decellularization steps are as follows:

[0075] 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);

[0076] ②. In the PBS buffer containing 0.5M KCL, add penicillin and streptomy...

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Abstract

The invention discloses a method for preparing a tendon composite muscle decellularized material derived from natural tissue. The method comprises the following step: treating tendon composite muscletissue of any size with a PBS (Phosphate Buffer Solution) with a protease inhibitor, a PBS with KCl, a PBS with KI, a PBS with Triton X, a PBS with SDS (Sodium Dodecyl Sulfate) and a PBS with DNA (Deoxyribonucleic Acid) enzymes, thereby obtaining the tendon composite muscle decellularized material derived from natural tissue. By adopting the method, cells on tissue can be subjected to complete decellularization treatment which is gentle in condition, free of ECM (Extracellular Matrix) damage and rapid and stable. The obtained material not only is capable of simulating layered structures of natural tendon composite muscle to the maximum extent and relatively well retaining extracellular matrix components, but also has the advantages of very low immunogenicity, relatively good mechanical properties, and the like, can be applied to repairing clinical muscle damage and regression related diseases caused by various disease reasons.

Description

technical field [0001] The present invention mainly relates to the medical field for repairing and regenerating skeletal muscle tissue of the whole body, in particular to the preparation of a tendon composite muscle decellularized material derived from natural tissue. It retains the layered structure of the natural tendon compound muscle, extracellular matrix components, has strong mechanical properties, and the material can promote the repair and regeneration of muscle defects. Background technique [0002] Muscle injuries caused by trauma, unreasonable exercise, degenerative diseases, etc. are very common in clinical practice and seriously affect the quality of life of patients. The widespread application of tissue engineering has made some progress in overcoming this disease. Current research is mainly focused on preparing a simple muscle scaffold to induce muscle fiber regeneration, but the effect is not ideal. First: Under physiological conditions, muscle fibers are co...

Claims

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Application Information

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IPC IPC(8): A61L27/36
CPCA61L27/3633A61L27/367A61L27/3687A61L27/3691A61L2430/30
Inventor 林贤丰王晟毓范顺武
Owner ZHEJIANG UNIV
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