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Preparation method of tendon conjunction bone decellularization material of natural tissue source

A decellularized and tissue-based technology, which is applied in the field of preparation of tendon and bone materials, can solve the problems of unsatisfactory mechanical properties of materials, serious immune reactions of materials, and incomplete cell removal, etc., and achieve a good extracellular microenvironment and wide range of materials , good biocompatibility effect

Active Publication Date: 2016-08-24
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the decellularized materials for tendon and bone are in the initial stage of research. The methods for preparing tendon and bone materials in the existing reports are not thorough in cell removal, and the immune reaction of the material is serious, and the damage to the ECM is large, and the mechanical properties of the material are poor. Neither is ideal

Method used

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  • Preparation method of tendon conjunction bone decellularization material of natural tissue source
  • Preparation method of tendon conjunction bone decellularization material of natural tissue source
  • Preparation method of tendon conjunction bone decellularization material of natural tissue source

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Example 1 Preparation and Research of Decellularized Tendon and Bone Material

[0039] 1. Preparation of tendon-bonded acellular matrix

[0040] (1) Material collection: Take the Achilles tendon and bone of a mature domestic pig, mechanically cut off the fat attached to the Achilles tendon, and then cut the Achilles tendon into 20mm (length) X 5mm (width) X 3mm (thickness), the size of the calcaneus is 3mm (thickness) )X7mm(diameter)

[0041] (2) Rinse 3 times in normal saline containing 10KIU / ml protease inhibitor, 20min each time;

[0042] (3) In the PBS buffer solution containing 10KIU / ml protease inhibitor, shake at 150rpm on a low-temperature (4°C) shaker for 24 hours;

[0043] (4) In the 4% PBS buffer solution containing Triton X-100, add penicillin and streptomycin mixed antibacterial solution at 1:1, the concentration is 20KIU / ml, shake at 150rpm on a low temperature (4°C) shaker for 24 hours ;

[0044] (5) In the PBS buffer solution containing SDS at a conc...

Embodiment 2

[0055] Example 2 Preparation and Research of Decellularized Tendon and Bone Material

[0056] (1) Material collection: take the Achilles tendon and bone of mature domestic pigs, mechanically cut off the fat attached to the Achilles tendon, and then cut the Achilles tendon into 40mm (length) X20mm (width) X5mm (thickness), the size of the calcaneus is 5mm (thickness) )X25mm (diameter)

[0057] (2) Rinse 3 times in normal saline containing 30KIU / ml protease inhibitor, 20min each time;

[0058] (3) In the PBS buffer solution containing 30KIU / ml protease inhibitor, shake at 150rpm on a low-temperature (4°C) shaker for 48 hours;

[0059] (4) Add penicillin and streptomycin (20KIU / ml, 20g / ml) mixed antibacterial solution 1:1 in PBS buffer solution containing Triton X-100 at a concentration of 8%, and shake at low temperature (4°C) at 150rpm Shock for 30 hours;

[0060] (5) In the PBS buffer solution containing SDS with a concentration of 8%, shake at 150 rpm on a low-temperature ...

Embodiment 3

[0065] Example 3 Preparation and Research of Decellularized Tendon and Bone Material

[0066] (1) Material collection: take the Achilles tendon and bone of a mature domestic pig, mechanically cut off the fat attached to the Achilles tendon, and then cut the Achilles tendon into 40mm (length) X20mm (width) X3mm (thickness), the size of the calcaneus is 4mm (thickness) )X15mm (diameter)

[0067] (2) Rinse 3 times in normal saline containing 50KIU / ml protease inhibitor, 20min each time;

[0068] (3) In the PBS buffer solution containing 50KIU / ml protease inhibitor, shake at 150rpm on a low temperature (4°C) shaker for 36 hours;

[0069] (4) Add penicillin and streptomycin (20KIU / ml, 20g / ml) mixed antibacterial solution 1:1 in PBS buffer solution containing Triton X-100 at a concentration of 4%, and shake at low temperature (4°C) at 150rpm Shock for 48 hours;

[0070] (5) In the PBS buffer solution containing SDS at a concentration of 0.5%, shake at 150 rpm on a low-temperature...

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Abstract

The invention discloses a preparation method of a tendon conjunction bone decellularization material of a natural tissue source. The heel tendon conjunction bone tissue of any size of a mammal is treated by normal saline containing protease inhibitor, PBS containing protease inhibitor, a PBS buffer solution containing Triton X, a PBS buffer solution containing SDS and a PBS buffer solution containing DNA enzyme, and the tendon conjunction bone decellularization material is obtained. Complete decellularization treatment can be carried out on tendon cells and bone cells on tendon conjunction bone tissue and cartilage cells on the interface at the same time, conditions are mold, no ECM damage is caused, the process is rapid and stable, and the obtained tendon conjunction bone material has the advantages of being good in biocompatibility, strong in plasticity, high in biomechanical strength and the like, and can be used for repairing tendon degenerative diseases caused by various etiological factors clinically, tendon severe tear and tendon conjunction bone regeneration and repair obstacles.

Description

technical field [0001] The invention mainly relates to the biological field for tissue or organ repair and regeneration, in particular to a method for preparing a tendon joint bone material derived from natural tissue. Background technique [0002] Tendon tears are a common condition that often occurs during high-intensity sports. Most patients are young people, especially among athletes. It will cause tendon damage, resulting in a decrease in exercise capacity, and even loss of exercise function in severe cases, which will bring great inconvenience to life. Bone and tendon tears are more serious. The existing surgical treatment has no effective solution for this kind of tear. The fibrocartilage layer between the tendon and bone often degenerates in the natural body repair after conventional suturing, and the scar tissue with poor mechanical properties is replaced. The probability of re-cutting after surgery is high, and the normal activities of patients are greatly restr...

Claims

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Application Information

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IPC IPC(8): A61L27/36A61L27/50
CPCA61L27/3604A61L27/3608A61L27/3662A61L27/3687A61L27/50A61L2430/10A61L2430/40
Inventor 林贤丰王晟毓宋立阳范顺武
Owner ZHEJIANG UNIV
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