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Freezing protection solution for articular cartilage and freezing preservation method for articular cartilage

A technology of cryoprotective solution and articular cartilage, which is applied in the preservation, application, and animal husbandry of human or animal bodies, and can solve the problems of difficult cleaning of DMSO, low survival rate of articular chondrocytes, and short storage time

Inactive Publication Date: 2019-04-19
徐艳红
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] (1) The cell survival rate of articular cartilage is still low, and the storage time is short;
[0007] (2) Dimethyl sulfoxide is used as the main cryoprotective component, but DMSO has cytotoxicity and osmotic damage. After thawing, it needs to be fully washed to remove DMSO, but it is difficult to completely wash away the DMSO that has entered the cell

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Embodiment 1 A kind of cryoprotective liquid for articular cartilage

[0025] Described cryoprotective liquid, by weight, comprises the following components:

[0026] 20 parts of carboxymethyl starch, 3 parts of rhamnose, 2 parts of cetene, 3 parts of microcrystalline cellulose, 5 parts of galactomannose, 5 parts of muramyl dipeptide, 4 parts of tripalmitoyl pentapeptide, α - 1 part of arbutin, 70 parts of deionized water.

[0027] The preparation method of described cryoprotective liquid is:

[0028] After mixing and stirring the components evenly, first carry out homogenization treatment at 3MPa for 3 minutes, and then use ultrasonic treatment, the ultrasonic frequency is 18HZ, the time is 10 minutes, and the temperature is 40°C. After ultrasonic treatment, adjust the pH to 7 to obtain Stable cryoprotectant.

Embodiment 2

[0029] Embodiment 2 The method for cryopreserving articular cartilage using the above-mentioned cryoprotectant solution

[0030] Include the following steps:

[0031] (1) rinse

[0032] After the fresh articular cartilage grafts were washed once with phosphate buffered saline, they were rinsed with cryoprotectant solution, which was pre-cooled to 3°C in advance;

[0033] The rinsing flow rate of the cryoprotectant solution is 2ml / min, the rinsing time is 5 minutes, and the temperature of the cryoprotectant solution is 3°C.

[0034] (2) Add part of the cryoprotectant solution dropwise

[0035] Part of the cryoprotectant solution was added dropwise to the articular cartilage graft under nitrogen protection for 5 minutes; the amount of the added cryoprotectant solution accounted for 50% of the total amount; the temperature of the cryoprotectant solution was 3°C.

[0036] (3) Add the remaining cryoprotectant

[0037] Add the remaining protective solution and soak for 10 minute...

Embodiment 3

[0043] Embodiment 3 The method for cryopreserving articular cartilage using the above-mentioned cryoprotectant solution

[0044] Include the following steps:

[0045] (1) rinse

[0046] Fresh articular cartilage grafts were washed once with phosphate buffered saline, and rinsed with cryoprotective solution, which was pre-cooled to 4°C in advance;

[0047] The rinsing flow rate of the cryoprotectant solution is 3ml / min, the rinsing time is 4 minutes, and the temperature of the cryoprotectant solution is 4°C.

[0048] (2) Add part of the cryoprotectant solution dropwise

[0049] Part of the cryoprotectant solution was added dropwise to the articular cartilage graft under nitrogen protection for 7 minutes; the amount of cryoprotectant solution added accounted for 55% of the total amount; the temperature of the cryoprotectant solution was 4°C.

[0050] (3) Add the remaining cryoprotectant

[0051] Add the remaining protective solution and soak for 10 minutes at 4°C; transfer t...

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Abstract

The invention provides a freezing protection solution for the articular cartilage. The freezing protection solution is prepared from, by weight, 18-22 parts of carboxymethyl starch, 2-3 parts of rhamnose, 1-2 parts of cetene, 2-4 parts of microcrystalline cellulose, 4-6 parts of galactomannan, 4-6 parts of muramyl dipeptide, 3-5 parts of palmitoyl pentapeptide-3, 0.5-1.5 parts of alpha-arbutin and70 parts of deionized water. The invention further provides a freezing preservation method for the articular cartilage by adopting the freezing protection solution for the articular cartilage. The method comprises the steps of leaching, dripping of the partial freezing protection solution, addition of a remaining freezing protection solution and program cooling. According to the freezing protection solution, a freezing protection agent, harmful to cells, such as dimethyl sulfoxide is not used; according to the freezing preservation method for the articular cartilage, in the eighth week afterthe articular cartilage is frozen for storage, the articular cartilage is taken, and after temperature recovery, the cell survival rate of the articular cartilage is 82-85%, and the cell survival rateof the articular cartilage is 78-80% in the tenth week.

Description

technical field [0001] The invention relates to a cryoprotective liquid for articular cartilage and a cartilage cryopreservation method, belonging to the technical field of biological materials. Background technique [0002] With age, articular cartilage is prone to damage and wear. Cartilage transplantation is one of the most important methods for the treatment of cartilage damage. The cryopreserved allogeneic soft transplantation has achieved good therapeutic effects in the treatment of joint soft defects. However, it is difficult to achieve long-term preservation of biological materials after they are separated from the donor. With the continuous development of cryopreservation technology, studies have shown that the metabolism of isolated biological materials in deep low temperature environments will gradually slow down or even stop metabolism. [0003] CN104938478A discloses a vitrification cryoprotectant solution for articular cartilage and a cartilage preservation met...

Claims

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Application Information

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IPC IPC(8): A01N1/02
CPCA01N1/0226
Inventor 徐艳红
Owner 徐艳红
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