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Preparation method for decellularized periosteal matrix gel material sourced from natural tissue

An acellular bone and gel material technology, applied in tissue regeneration, prosthesis, medical science, etc., can solve the problems of inability to obtain sufficient specific surface area, low retention of bioactive substances, and no periosteal ECM gel. Biological activity, promoting in situ regeneration, increasing the effect of repair function

Active Publication Date: 2019-09-17
ZHEJIANG DISAI BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, the previous preparation of acellular matrix gel materials cannot obtain sufficient specific surface area by shredding, which is not conducive to full contact with digestive juice, and the retention of biologically active substances is low.
In addition, there is currently no standardized preparation of periosteal ECM gel

Method used

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  • Preparation method for decellularized periosteal matrix gel material sourced from natural tissue
  • Preparation method for decellularized periosteal matrix gel material sourced from natural tissue
  • Preparation method for decellularized periosteal matrix gel material sourced from natural tissue

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] The preparation of the femoral periosteum ECM gel of embodiment 1 natural tissue source

[0049] 1. Remove the muscle, fascia and fat on the adult pig femur with a blade, and cut the femoral periosteum as a raw material;

[0050] 2. Rinse the material obtained in step 1 with sterile water for 3 times, 20 minutes each time, to remove residual muscle, fascia, fat and other tissues, and irradiate with 24kGy gamma rays for 24 hours to kill possible bacteria and viruses ;

[0051] 3. Treat the material obtained in step 2 with a volume ratio of tributyl phosphate to deionized water of 2% for 24 hours;

[0052] 4. Soak the material obtained in step 3 in a solution with a concentration of 1% TritonX-100, shake on a constant temperature 23°C shaker at 200rpm for 12 hours, rinse with sterile water for 3 times, 20 minutes each time, to remove residual TritonX-100 solution;

[0053] 5. Soak the material obtained in step 4 in a 1% SDS solution, shake at a constant temperature of 2...

Embodiment 2

[0061] The preparation of the scapula periosteum ECM gel of embodiment 2 natural tissue sources

[0062] 1. Use a blade to remove the muscles, fascia and fat on the scapula of an adult pig, and cut the periosteum of the scapula as a raw material;

[0063] 2. The decellularization treatment steps are the same as steps 2-7 in the above-mentioned embodiment 1; wherein the volume ratio of tributyl phosphate and deionized water in step 3 is 1%; the treatment time is 12 hours;

[0064] 3. The material obtained in step 7 is irradiated with gamma rays at a dose of 40 kGy for 10 hours to kill bacteria and viruses that may exist;

[0065] 4. Pre-cool the sterilized decellularized periosteum material obtained in step 8 in a refrigerator at -80°C, then put it into a freeze dryer to dry for 24 hours, keep the vacuum at 1.5mtorr, the temperature is -50°C, and the pumping rate of the vacuum pump is 195L / min , 60Hz;

[0066] 5. Transfer the freeze-dried material obtained in step 9 to a grin...

Embodiment 3

[0069] The preparation of the femoral periosteum ECM gel of embodiment 3 natural tissue sources

[0070] 1. Remove the muscle, fascia and fat on the adult pig femur with a blade, and cut the femoral periosteum as a raw material;

[0071] 8. The decellularization treatment steps are the same as steps 2-7 in the above-mentioned embodiment 1; wherein the volume ratio of tributyl phosphate and deionized water in step 3 is 5%; the treatment time is 16 hours;

[0072] 2. The material obtained in step 7 is irradiated with gamma rays at a dose of 5 kGy for 48 hours to kill bacteria and viruses that may exist;

[0073] 3. Pre-cool the sterilized decellularized periosteum material obtained in step 8 in a refrigerator at -80°C, then put it into a freeze dryer to dry for 24 hours, keep the vacuum at 1.5mtorr, the temperature is -50°C, and the pumping rate of the vacuum pump is 195L / min , 60Hz;

[0074] 4. Transfer the freeze-dried material obtained in step 9 to a grinder, the frequency ...

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Abstract

The invention discloses a preparation method for a decellularized periosteal matrix gel material sourced from natural tissue. By conducting freeze drying on decellularized periosteal matrix sourced from mammal, bioactive molecules are kept to the maximum degree; the decellularized periosteal matrix gel material sourced from natural tissue is ground into powder, the grain size of the powder is controlled, the contact area of a solution is increased, in this way, digestion is promoted, and finally the standard and uniform decellularized periosteal matrix gel having a good osteogenic activity potential is obtained. In addition, EDC crosslinking growth factors have a good promoting effect when regenerated bone tissue is regenerated excellently and grows into with vascularization. The decellularized periosteal matrix gel material can be used for repairing bone defects, bone disunion and other kinds of bone regeneration obstacles caused by all kinds of clinical pathogeneses.

Description

technical field [0001] The invention belongs to the technical field of bone tissue repair and regeneration, and in particular relates to a preparation method of an acellular periosteum matrix gel material derived from natural tissue. Background technique [0002] Bone defect is a common clinical disease, which often causes bone nonunion, delayed union or nonunion, and local dysfunction, which causes pain to patients, restricts their mobility, and seriously affects their quality of life. Although traditional synthetic materials can provide sufficient mechanical support, they have poor biological activity, insufficient osteoinductivity, are not easy to degrade, and even cause foreign body reactions. At present, autologous bone grafting and allogeneic bone grafting are the main methods for treating bone defects and osteonecrosis. There is an urgent need to develop new bone tissue engineering materials with good bone healing and regenerative repair effects. [0003] Extracellu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/22A61L27/36A61L27/52A61L27/54
CPCA61L27/227A61L27/3608A61L27/365A61L27/3687A61L27/3691A61L27/52A61L27/54A61L2300/414A61L2430/02A61L2430/40C08L89/00
Inventor 林贤丰邱朋程范顺武
Owner ZHEJIANG DISAI BIOTECHNOLOGY CO LTD
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