A preparation method of tendon-bonded decellularized material derived from natural tissue
A decellularization and tissue technology, applied in the field of preparation of tendon and bone materials, can solve the problems of unsatisfactory mechanical properties of materials, severe immune response of materials, incomplete removal of cells, etc., and achieve a good extracellular microenvironment and good material extraction. , good biocompatibility
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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 PBS buffer solution containing 4% Triton X-100, add penicillin and streptomycin mixed antibacterial solution 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 concentrati...
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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