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A kind of articular cartilage vitrification cryoprotectant solution and cartilage preservation method

A technology of vitrification and articular cartilage, which is 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 effect of improving storage effect, prolonging storage time, and expanding application

Active Publication Date: 2018-02-02
TAISHAN MEDICAL UNIV
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  • Abstract
  • Description
  • Claims
  • 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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  • A kind of articular cartilage vitrification cryoprotectant solution and cartilage preservation method
  • A kind of articular cartilage vitrification cryoprotectant solution 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 vitrified cryoprotective solution for articular cartilage and a cartilage preservation method. Each 1000ml vitrified cryoprotectant solution contains: 99.9% dimethyl sulfoxide 150-250ml, acetamide 110-200g, 1,2-propylene glycol 70 ~120ml, polyethylene glycol 10~80g, trehalose 20~100g, the rest is deionized water. Soak the osteochondral pieces in gradient vitrification protection solution at 4°C for 10 minutes, seal them in refrigerated tubes, and put them in liquid nitrogen for storage; take them out before transplantation for rewarming, rewarm in a water bath at 37°C for 10 seconds, and then Put them into 10% and 5% sucrose solutions at 37°C successively and stay for 5 minutes each, and finally rinse with phosphate buffer solution for 3 times before transplantation. The invention enables long-term preservation of articular cartilage tissue, maintains a relatively high survival rate of cartilage cells, improves the preservation effect of osteochondral grafts, and is used for cartilage tissue bank and 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 Patents(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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