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
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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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