Preparation method and use of decellularized tendon material

A decellularized tendon technology, applied in the field of preparation of decellularized tendon materials, can solve the problems of no significant change in DNA residue and high immunogenicity, and achieve the effect of simple preparation method, low cost and good application prospects

Active Publication Date: 2014-12-17
WEST CHINA HOSPITAL SICHUAN UNIV
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

In addition, Jiang Yanlin et al. also found that on the basis of repeated freeze-thaw and nuclease treatment of calf Achilles tendon, and then treated with α-galactosidase for 24 hours, the content of α-galactosyl antigen (α-gal) decreased significantly

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  • Preparation method and use of decellularized tendon material
  • Preparation method and use of decellularized tendon material
  • Preparation method and use of decellularized tendon material

Examples

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Embodiment 1

[0040]Embodiment 1 Preparation of calf tendon decellularized material of the present invention

[0041] (1) Take the Achilles tendon of the hindlimb of a newborn calf (birth day 1) and wash it three times with PBS or normal saline;

[0042] (2) Use a vernier caliper to measure the thickness of each tendon, which is about 4-6mm, and compress it on a uniaxial compression testing machine, and the compression rate is 80% (such as figure 1 shown). The thickness of the long ribbon tendon after compression processing is about 0.8-1.2mm, and the length is 8cm;

[0043] (3) Take out the compressed long ribbon-shaped Achilles tendon for repeated freeze-thaw treatment: freeze in liquid nitrogen at -196°C for 1 minute, then rewarm in 37°C normal saline for 5 minutes, and repeat this operation 5 times;

[0044] (4) Nuclease treatment: Wash the Achilles tendon after repeated freezing and thawing three times with PBS, place it in a solution containing DNase and RNase (DNase concentration i...

Embodiment 2

[0045] Embodiment 2 Preparation of calf tendon decellularized material of the present invention

[0046] (1) Take the Achilles tendon of the hindlimb of a newborn calf (birth day 1) and wash it three times with PBS or normal saline;

[0047] (2) Use a vernier caliper to measure the thickness of each tendon, which is about 4-6mm, and compress it on a uniaxial compression testing machine, and the compression rate is 80% (such as figure 1 shown). The thickness of the long ribbon tendon after compression processing is about 0.8-1.2mm, and the length is 8cm;

[0048] (3) Repeated freezing and thawing of the compressed long ribbon-shaped Achilles tendon: freezing in liquid nitrogen at -196°C for 1 minute, then rewarming in saline at 37°C for 5 minutes, and repeating this operation 5 times;

[0049] (4) Nuclease treatment: wash the Achilles tendon after repeated freezing and thawing with PBS three times, place it in a container containing DNase and RNase (DNase concentration is 150...

Embodiment 3

[0051] Example 3 Preparation of calf tendon decellularized material of the present invention

[0052] (1) Take the Achilles tendon of the calf (30 days after birth) and wash it twice with PBS or normal saline;

[0053] (2) Use a vernier caliper to measure the thickness of each tendon, which is about 4-6mm, and compress it on a uniaxial compression testing machine, and the compression rate is 70% (such as figure 1 shown).

[0054] (3) Take the compressed long ribbon-shaped Achilles tendon for repeated freeze-thaw treatment: freeze in liquid nitrogen at -196°C for 3 minutes, then rewarm in normal saline at 25°C for 10 minutes, and repeat this operation 4 times;

[0055] (4) Nuclease treatment: Wash Achilles tendon twice with PBS after repeated freezing and thawing, place in a container containing DNase and RNase (DNase concentration is 120IU / ml, RNase concentration is 80μg / ml), seal and store at 37°C Treated on a constant temperature shaker for 24 hours;

[0056] (5) Wash wit...

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Abstract

The invention discloses a preparation method of a decellularized tendon material. The preparation method comprises the following steps of 1, taking fresh tendon and carrying out cleaning, 2, compressing the tendon along the thickness direction at a compression ratio of 60-90%, 3, carrying out repeated freezing-thawing on the compressed tendon, and 4, treating the tendon subjected to the step 3 by nuclease and carrying out cleaning. The invention also discloses the decellularized tendon material obtained by the preparation method and a use thereof. The decellularized tendon material realizes thorough decellularization and has low immunogenicity, good biomechanical properties and good biocompatibility. The preparation method has simple processes, a low cost and a good application prospect.

Description

technical field [0001] The invention relates to a preparation method and application of a decellularized tendon material, belonging to the field of biological materials. Background technique [0002] Tendon is a kind of dense connective tissue with less blood supply and less cell specific gravity, mainly composed of highly axially arranged collagen fibers, so it can provide maximum strain resistance when bearing load, and is a good tendon / ligament injury or defective prosthetic stents. However, the tendon tissue of allogeneic or xenogeneic origin has various immunogenic substances on its cells, such as the DNA in the nucleus will cause immune rejection in all animals, and the α-galactosyl antigen (α-gal) on the cell membrane will It causes immune rejection in primates, and the effect of in vivo repair is unsatisfactory, and there are safety hazards at the same time. Therefore, when preparing tendon for in vivo repair, various decellularization methods are usually used to r...

Claims

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Application Information

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IPC IPC(8): A61L27/36
Inventor 秦廷武江燕林宁良菊
Owner WEST CHINA HOSPITAL SICHUAN UNIV
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