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Preparation method of cartilage regeneration carrier based on collagen

A technology of collagen and procollagen, applied in the field of preparation of cartilage regeneration carriers, can solve the problems of loss of cartilage function, difficult operation, hardening, etc., and achieve the effect of reducing operation difficulty and economic cost, and excellent clinical practicability

Inactive Publication Date: 2019-10-08
杭州蓝朗生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] At present, a variety of materials and schemes have been used clinically for the repair of cartilage injuries, but there are varying degrees of application defects. For example, the prototype of microfracture surgery is subchondral bone drilling, which is simple and effective, and the short-term effect is good, but the newly formed The repair tissue is not real articular cartilage, but more fibrocartilage components, which will harden and lose cartilage function over time
For example, matrix-induced chondrocyte transplantation technology, as the most widely used cartilage tissue engineering technology in clinical practice, can indeed provide a good choice for cartilage repair, but this technology requires a one-stage operation to obtain autologous cells for isolation, culture and induction of differentiation , and then perform the second-stage surgical transplantation, the operation is difficult, time-consuming and expensive

Method used

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Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0017] A preparation method of a collagen-based cartilage regeneration carrier, comprising the steps of extracting a collagen stock solution, preparing a concentration, homogenizing, and defoaming.

[0018] The extraction steps of collagen stock solution are as follows:

[0019] ①Thaw the beef Achilles tendon, remove blood, foreign matter and fascia, stir in ethanol solution for 2-8min, and mix sodium hypochlorite and EDTA-Na 2 Stir in the mixed solution of TritionX-114 for 2-10min, and stir for 10-30min in the TritionX-114 solution. The concentration of ethanol solution is 75%, the mixed solution is 0.05-0.35% sodium hypochlorite and 0.05-0.2% EDTA-Na 2 For the mixed solution, the concentration of the TritionX-114 solution is 0.1-2%.

[0020] ②The bovine Achilles tendon defat film treated in step 1 is soaked in ethanol solution for 2-8 minutes, soaked in sodium hypochlorite solution for 2-10 minutes, and soaked in TritionX-114 solution for 10-30 minutes. The concentration o...

Embodiment 1

[0027] 1. Prepare a collagen solution with a concentration of 8mg / ml.

[0028] a. According to the filling volume, 900mL collagen solution needs to be prepared. The final concentration of the acid solution is 50mM, and the collagen concentration of the stock solution is 30mg / ml. It is necessary to weigh 240g of the collagen stock solution, add 330mL of 68mM acetic acid, and then disperse with a high-speed disperser for 10 minutes until no Obvious collagen particles, then add 330mL acetic acid, and disperse for 3 minutes to make the protein solution basically uniform.

[0029] b. Use a mixer to stir the above protein solution for 60 minutes (the stirring speed is such that the stirring paddle can stir the protein), so that the prepared collagen solution is uniform and free of particles.

[0030] c. Use 4% NaOH solution to adjust the pH of the protein solution to 5.0.

[0031] 2. Homogeneous collagen solution.

[0032] Homogenize the collagen solution obtained in step 1 using ...

Embodiment 2

[0036] 1. Prepare a collagen solution with a concentration of 10mg / ml.

[0037] a. According to the filling volume, 900mL collagen solution needs to be prepared. The final concentration of the acid solution is 20mM, and the collagen concentration of the stock solution is 30mg / ml. It is necessary to weigh 300g of the collagen stock solution, add 300mL of 30mM acetic acid, and then disperse with a high-speed disperser for 7 minutes until no Obvious collagen particles, then add 300mL acetic acid, disperse for 5 minutes to make the protein solution basically uniform.

[0038] b. Use an electric mixer to stir the above protein solution for 120min (stirring speed is the state that the stirring paddle can stir the protein), so that the prepared collagen solution is uniform and free of particles.

[0039] c. Use 2% NaOH solution to adjust the pH of the protein solution to 6.5.

[0040] 2. Homogeneous collagen solution.

[0041] Homogenize the collagen solution obtained in step 1 usi...

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PUM

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Abstract

The invention discloses a preparation method of a cartilage regeneration carrier based on collagen. The preparation method includes the steps that a collagen crude extract is prepared by washing and processing bovine achilles tendons, enzymolysis and supernate taking through centrifugation; collagen precipitation is obtained by salting out and centrifugation of a neutral salt, and a phosphate buffer solution is added to the collagen precipitation to obtain a collagen stock solution; a certain volume of acid is added to the collagen stock solution, and the mixture is dispersed for 2-10 min through a disperser; and a certain volume of acid is added again, the mixture is dispersed for 1-10 min through the disperser and mixed for 10-180 min through a mixer, after homogenizing the protein solution with a homogenizer and the disperser or the mixer, and the collagen solution is subpackaged into centrifugal cups for balancing, and centrifuged for 5-35 min at 1000-2500 r / min.

Description

technical field [0001] The invention relates to a preparation method of a cartilage regeneration carrier, in particular to a preparation method of a collagen-based cartilage regeneration carrier. Background technique [0002] At present, a variety of materials and schemes have been used clinically for the repair of cartilage injuries, but there are varying degrees of application defects. For example, the prototype of microfracture surgery is subchondral bone drilling, which is simple and effective, and the short-term effect is good, but the newly formed The repair tissue is not real articular cartilage, but more fibrocartilage components, which will harden and lose cartilage function over time. For example, matrix-induced chondrocyte transplantation technology, as the most widely used cartilage tissue engineering technology in clinical practice, can indeed provide a good choice for cartilage repair, but this technology requires a one-stage operation to obtain autologous cell...

Claims

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Application Information

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IPC IPC(8): A61L27/24A61L27/50A61L27/54C12P21/06C07K1/30
CPCA61L27/24A61L27/50A61L27/54A61L2300/252A61L2300/412A61L2430/06C07K14/78C12P21/06
Inventor 郭文弘何拥军李巧美刘苏汶郭文静夏伟轩秀玲
Owner 杭州蓝朗生物技术有限公司