Cartilago articularis vitrification cryoprotectant and cartilage preservation method

A technology for vitrification and freezing of articular cartilage, applied in the field of biomedicine, can solve the problems of low survival rate of chondrocytes and short storage time in vitro, and achieve the effects of improving the storage effect, prolonging the storage time, and expanding the promotion and application.

Active Publication Date: 2015-09-30
TAISHAN MEDICAL UNIV
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  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The object of the present invention is to provide a vitrified cryoprotective solution for articular cartilage grafts and its preservation method, aiming to solve the existing long-term continuous maintenance of the activity of articular cartilage grafts and the low survival rate of chondrocytes and short storage time in vitro The problem

Method used

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  • Cartilago articularis vitrification cryoprotectant and cartilage preservation method

Examples

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Embodiment 1

[0029] Example 1: Each 1000ml vitrification solution includes: 99.9% dimethyl sulfoxide 160ml, acetamide 125g, 1,2-propanediol 80ml, polyethylene glycol (8000) 30g, trehalose 40g, the rest is deionized water , prepare the raw materials into a solution, stir evenly, and adjust the pH value to 7.30-7.50; wash the fresh allogeneic osteochondral graft with phosphate buffer for 3-5 times; vitrify the osteochondral block at 4°C Solution: 1 / 3 concentration of vitrification protection solution, 2 / 3 concentration of vitrification protection solution, 3 / 3 concentration of vitrification protection solution, etc., soaked for 10 minutes in sequence, sealed in a refrigerated tube, marked, and then directly at the fastest speed Put into liquid nitrogen for storage. Rewarm the cartilage blocks at the 8th week: After taking out the frozen cartilage blocks, immediately rewarm them in a 37°C water bath for 10 seconds, then put them into 37°C 10% and 5% sucrose solutions for 5 minutes each, and f...

Embodiment 2

[0030] Example 2: Each 1000ml vitrification solution includes: 99.9% dimethyl sulfoxide 205ml, acetamide 155g, 1,2-propanediol 100ml, polyethylene glycol (8000) 60g, trehalose 60g, the rest is deionized water , prepare the raw materials into a solution, stir evenly, and adjust the pH value to 7.30-7.50; wash the fresh allogeneic osteochondral graft with phosphate buffer for 3-5 times; vitrify the osteochondral block at 4°C Solution: 1 / 3 concentration vitrification protection solution, 2 / 3 concentration vitrification protection solution, 3 / 3 vitrification protection solution, soaked step by step for 10 minutes, packaged in a refrigerated tube, marked well, and then put directly into it at the fastest speed Store in liquid nitrogen. Rewarm the cartilage blocks at the 8th week: After taking out the frozen cartilage blocks, immediately rewarm them in a 37°C water bath for 10 seconds, then put them into 37°C 10% and 5% sucrose solutions for 5 minutes each, and finally phosphate buf...

Embodiment 3

[0031] Example 3: Each 1000ml vitrification solution includes: 225ml of 99.9% dimethyl sulfoxide, 190g of acetamide, 115ml of 1,2-propanediol, 70g of polyethylene glycol (8000), 90g of trehalose, and the rest is deionized water , prepare the raw materials into a solution, stir evenly, and adjust the pH value to 7.30-7.50; wash the fresh allogeneic osteochondral graft with phosphate buffer for 3-5 times; vitrify the osteochondral block at 4°C Solution: 1 / 3 concentration of vitrification protection solution, 2 / 3 concentration of vitrification protection solution, 3 / 3 concentration of vitrification protection solution, etc., soaked for 10 minutes in sequence, sealed in a refrigerated tube, marked, and then directly at the fastest speed Put into liquid nitrogen for storage. Rewarm the cartilage blocks at the 8th week: After taking out the frozen cartilage blocks, immediately rewarm them in a 37°C water bath for 10 seconds, then put them into 37°C 10% and 5% sucrose solutions for 5...

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Abstract

The invention discloses a cartilago articularis vitrification cryoprotectant and a cartilage preservation method. Each 1000ml of a vitrification cryoprotectant comprises 150-250ml of 99.9% dimethyl sulfoxide, 110-200g of acetamide, 70-120ml of 1,2-propylene glycol, 10-80g of polyethylene glycol, 20-100g of trehalose and the balance of deionized water. The cartilage preservation method comprises the following steps: soaking cartilage bone blocks at 4 DEG C by a gradient vitrification cryoprotectant sequentially for 10 minutes, packaging by a refrigerating pipe, putting into liquid nitrogen and storing; taking out before transplanting, rewarming, and carrying out water bath rewarming at 37 DEG C for 10 seconds; then putting cartilage bone blocks in a 10% saccharose solution and a 5% saccharose solution at 37 DEG C respectively, and remaining for 5 minutes; and finally washing by a phosphate buffer for 3 times so as to transplant and apply. According to the invention, the cartilago articularis tissues can be stored for a long time, the relatively high cell survival rate of cartilage is maintained, and the osteochondral graft preservation effect is enhanced; the cartilago articularis vitrification cryoprotectant and the cartilage preservation method are applied to the cartilage tissue library and the clinical cartilage transplantation.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a vitrification cryoprotectant solution for articular cartilage and a cartilage preservation method. Background technique [0002] In recent years, with the improvement of people's living standards in our country and the comprehensive popularization of sports, the number of patients with articular cartilage injuries has increased significantly. A series of encouraging progress has been made in the repair of articular cartilage defects by allogeneic osteochondral transplantation. Cartilage transplantation is one of the very promising repair methods for the treatment of cartilage lesions. Because the allogeneic cartilage graft material is relatively easy to obtain and can be prefabricated into any shape and size, it is convenient to make the normal functional articular cartilage into various shapes and implant it in the bone defect to restore the shape of the carti...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02
Inventor 亓建洪谢地李昊宋洪强陈彬董军
Owner TAISHAN MEDICAL UNIV
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