Arthrodial cartilage proto micro-carrier

A technology of articular cartilage and microcarriers, which is applied in the field of biomedical tissue engineering, can solve the problems of incomplete separation of cells and microcarriers, affecting the quality of engineered cartilage, and damage to chondrocytes, achieving cheap reagents, avoiding loss, and growing state Good results

Inactive Publication Date: 2006-11-29
GENERAL HOSPITAL OF PLA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The currently commercially available microcarriers are not only expensive, but also need to digest the chondrocytes from the microcarriers during use, and there is bound to be some damage or loss of chondrocytes
In addition, there is also the possibility that the cell-microcarriers cannot be completely separated, and the microcarriers are also implanted in the patient's body, which not only affects the quality of the regenerated engineered cartilage, but may also cause harm to the patient's body

Method used

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  • Arthrodial cartilage proto micro-carrier
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  • Arthrodial cartilage proto micro-carrier

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] 1. Take 1kg of fresh pig articular cartilage, freeze-dry it, pulverize it with a pulverizer, and sieve to obtain particles with a diameter of 200μm for use;

[0025] 2. Add 10 times the volume of 0.25% trypsin to the granules, keep shaking at 37°C for 48h and then wash with distilled water 3 times, use 1% Triton X-100 at 4°C to shake for 72h, then rinse with distilled water for 48h;

[0026] 3. Centrifuge at 1000 rpm for 10 minutes, take the precipitate, mix the precipitate with distilled water in a volume of 1:3, and put it in a freeze dryer for 12 hours to freeze-dry it. After 25kGy cobalt 60 Sterilized by irradiation, digested with 10 times the volume of 0.25% trypsin for 15 minutes, stirred, then pulverized into granules, and washed with distilled water 3 times to obtain.

Embodiment 2

[0028] 1. Take 4kg of fresh bovine joint cartilage, freeze-dried, crush it with a pulverizer, and sieve to obtain particles with a diameter of 150μm for use;

[0029] 2. Add 10 times the volume of 0.25% trypsin (purchased from Sigma) to the granules, continue shaking at 37°C for 48h, and then wash 3 times with distilled water, then use 1% Triton X-100 at 4°C to continue shaking for 72h, then Rinse with distilled water for 48h;

[0030] 3. Centrifuge at 1000 rpm for 10 minutes, take the precipitate, mix the precipitate with distilled water in a volume of 1:3, and put it in a freeze dryer for 12 hours to freeze-dry it. After 25kGy cobalt 60 Sterilized by irradiation, digested with 10 times the volume of 0.25% trypsin (purchased from Sigma) for 15 minutes, stirred, pulverized into granules, and washed with distilled water 3 times to obtain.

Embodiment 3

[0032] 1. Take 0.5kg of fresh human articular cartilage, freeze-dried, crush it with a pulverizer, and sieve to obtain particles with a diameter of 180μm for use;

[0033] 2. Add 10 times the volume of 0.25% trypsin (purchased from Sigma) to the granules, continue shaking at 37°C for 48h, and then wash 3 times with distilled water, then use 1% Triton X-100 at 4°C to continue shaking for 72h, then Rinse with distilled water for 48h;

[0034] 3. Centrifuge at 1000 rpm for 10 minutes, take the precipitate, mix the precipitate with distilled water in a volume of 1:3, and put it in a freeze dryer for 12 hours to freeze-dry it. After 25kGy cobalt 60 Sterilized by irradiation, digested with 0.25% trypsin for 15 minutes, stirred, then pulverized into granules, and washed with distilled water 3 times to obtain.

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Abstract

A proto-microcarrier for arthrodial cartilage is prepared from fresh joint cartilage through freeze drying, pulverizing, sieving adding trypsin to the particles, oscillating at 37 deg.C for 24-48 hr, washing with distilled water three times, adding TritonX-100, oscillating at 4 deg.C for 48-72 hr, flushing with distilled water for 48 hr, centrifugal separation of deposit, mixing with distilled water, freeze drying, UV irradiating, Co60 irradiation, digestion by trypsin, stirring, pulverizing and washing with distilled water three times.

Description

Technical field [0001] The invention relates to a microcarrier, in particular to a preparation method of articular cartilage genic microcarrier and a product prepared by the method, and belongs to the field of biomedical tissue engineering. Background technique [0002] Tissue engineering technology is the application of biotechnology to prepare tissues identical to the human body in vitro to repair human diseased or defective tissues, also known as regenerative medicine. [0003] The repair of cartilage defects mainly relies on engineered cartilage tissue. An important condition for the construction of cartilage tissue engineering is the large-scale expansion of chondrocytes in vitro. The use of bioreactor and microcarrier technology to cultivate chondrocytes is a way to efficiently expand chondrocytes. The currently commercially available microcarriers are not only expensive, but also need to digest chondrocytes from the microcarriers during use, which is bound to cause damage ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61F2/30A61L27/36
Inventor 卢世璧张建党袁玫黄靖香孙明学
Owner GENERAL HOSPITAL OF PLA
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