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Preparation method of novel decellularized tendon or ligament bracket

A decellularization and tendon technology, which is applied in the field of preparation of bioengineered medical materials, can solve the problems of long decellularization period, large damage to extracellular matrix, and difficulty in removing cell components.

Active Publication Date: 2016-09-21
拜欧迪赛尔成都生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Because ligaments and tendons are dense connective tissues, the fiber bundles are surrounded by intima and adventitia, making it difficult to remove cell components, so the decellularization cycle is long, and the decellularization process greatly damages the extracellular matrix
At present, when preparing tendon / ligament for in vivo repair, the methods used are difficult to effectively decellularize and remove xenoantigens while retaining the structural shape, biomechanical properties and biological properties of tendon / ligament tissue

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0013] Get fresh (obtained within 3 hours after slaughter) pig or beef thigh tendon or ligament tissue, remove the outer membrane and synovial tissue on the surface, and keep a 1X2cm tendon / ligament tissue piece for future use.

[0014] 1. Protect the tendon / ligament tissue with protective solution throughout the process. The formula of the protective solution is: RPMI-1640 medium, add chondroitin sulfate 20g / L, dextran 15g / L, erythromycin 5mg / L, adjust the pH to 7.2 , the osmotic pressure is 400mOsm;

[0015] 2. Seal the tendon / ligament tissue piece in a plastic bag filled with protective solution;

[0016] 3. Under the condition of 500MPa high static pressure, treat 8 times, each time is 5 minutes;

[0017] 4. After taking out the tendon / ligament, place it in a protective solution containing 5% Triton X-100+2500U / ml DNase at a temperature of 30°C, set the shaker speed to 100 rpm, and treat for 5 hours;

[0018] 5. After the detergent and enzyme treatment, take out the tend...

Embodiment 2

[0020] Get fresh (obtained within 3 hours after slaughter) pig or beef thigh tendon or ligament tissue, remove the outer membrane and synovial tissue on the surface, and keep a 1X2cm tendon / ligament tissue piece for future use.

[0021] 1. Protect the tendon / ligament tissue with protective solution throughout the process. The formula of the protective solution is: RPMI-1640 medium, add 50g / L of chondroitin sulfate, 20g / L of hyaluronic acid, 6mg / L of erythromycin, and adjust the pH value To 7.2, the osmotic pressure is 450mOsm;

[0022] 2. Seal the tendon / ligament tissue piece in a plastic bag filled with protective solution;

[0023] 3. Under the condition of 600MPa high static pressure, treat 6 times, each time is 6 minutes;

[0024] 4. After taking out the tendons / ligaments, place them in a protective solution containing 5% sodium dodecylsulfonate + 4000U / ml DNase at a temperature of 25°C, set the shaker speed to 100 rpm, and treat for 5 Hour;

[0025] 5. After the deterg...

Embodiment 3

[0027] Get fresh (obtained within 3 hours after slaughter) pig or beef thigh tendon or ligament tissue, remove the outer membrane and synovial tissue on the surface, and keep a 1X2cm tendon / ligament tissue piece for future use.

[0028] 1. Protect the tendon / ligament tissue with protective solution throughout the process. The formula of the protective solution is: RPMI-1640 medium, add chondroitin sulfate 30g / L, hyaluronic acid 50g / L, neomycin 5mg / L, adjust the pH value To 7.2, the osmotic pressure is 450mOsm;

[0029] 2. Seal the tendon / ligament tissue piece in a plastic bag filled with protective solution;

[0030] 3. Under the condition of 800MPa high static pressure, treat 4 times, each time is 5 minutes;

[0031] 4. After taking out the tendons / ligaments, place them in a protective solution containing 2% sodium alginate + 5000U / ml DNase at a temperature of 30°C, set the shaker speed to 100 rpm, and treat for 8 hours;

[0032] 5. After the detergent and enzyme treatment,...

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Abstract

The invention relates to a preparation method of a decellularized tendon or ligament bracket, and aims at providing a decellularized tendon or ligament collagen fiber material with good biomechanical characteristic and good biocompatibility. The preparation method comprises the following steps: acquiring animal tendon or ligament with the adventitia or synovial tissue removed, wherein during the whole decellularization process, a mixed protecting liquid containing a ligament or tendon support structure protective agent is adopted; pre-separating cells in the ligament or tendon tissue by adopting a high hydrostatic pressure technology; digesting the residual cell nuclei by adopting a compound nuclease method; removing loose broken cell sheets by adopting a detergent; and carrying out rinsing by adopting the mixed protecting liquid. Through the efficient physical means, the adoption of small amount of enzyme and the detergent removing means, the cellular constituent in the ligament or tendon can be sufficiently removed, and the protecting liquid is adopted for the whole-process protection, so that on the basis of guaranteeing the complete removal of the cell nuclei, while the immunogenicity of the tendon or ligament tissue is reduced to the maximum, the three-dimensional structure and arrangement polarity of the normal collagenous fibers of the tendon or ligament tissue are reserved.

Description

technical field [0001] The invention relates to the field of preparation of bioengineering medical materials, in particular to a preparation method of decellularized tendon / ligament scaffold. Background technique [0002] Poor healing ability after repair of ligament and tendon injury is a common clinical problem, and the use of substitutes for ligament and tendon repair and reconstruction is urgently needed clinically. Currently tendon and ligament substitutes mainly include autografts, allografts or xenografts, and synthetic material grafts, but all have their unavoidable shortcomings. Although autologous ligament transplantation is the most effective method of repair and reconstruction, it cannot be widely used clinically due to the limited range and quantity of materials obtained. Allogeneic or heterogeneous tendon has strong antigenicity, and the inevitable postoperative immune rejection is its biggest shortcoming. Artificial synthetic materials such as carbon fiber, ...

Claims

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Application Information

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IPC IPC(8): A61L27/36
CPCA61L27/3604A61L27/3662A61L27/3687A61L27/3691A61L2430/10
Inventor 史真史伟云
Owner 拜欧迪赛尔成都生物科技有限公司
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