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Preparation method of a novel cartilage decellularized matrix ink

A kind of acellular matrix, cartilage technology, applied in medical science, tissue regeneration, prosthesis and other directions, can solve the problems of difficult processing, poor solubility, etc., to achieve the effect of improving solubility, improving processing performance, and shortening dissolution time

Active Publication Date: 2021-05-11
DONGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The technical problem to be solved by the present invention is to provide a method for preparing a novel cartilage decellularized matrix ink, so as to overcome the defects of difficult processing and poor solubility caused by the dense texture of the decellularized cartilage matrix in the prior art

Method used

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  • Preparation method of a novel cartilage decellularized matrix ink
  • Preparation method of a novel cartilage decellularized matrix ink
  • Preparation method of a novel cartilage decellularized matrix ink

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] The cartilage decellularized matrix was prepared by the following two methods, wherein the first method was used for the heating method, and the second method was used for the grinding method. The first method is to cut the cartilage into 0.5cm×0.5cm×2cm size, put it in a watch glass, soak it in 1mol / L NaCl aqueous solution at 37℃ for 24h, use 4% NaOH solution to ablate it, and clean it with B-type ultrasonic cleaner intermittently shaking for 16h , replace the ablation solution once every 4 hours, then soak in distilled water for 24 hours, replace the distilled water every 4 hours until the solution becomes clear and the pH value is neutral, and finally soak the prepared cartilage decellularized matrix in physiological saline (drops Add penicillin and gentamicin solution) and store in a 4°C refrigerator for later use. The second method is to cut the cartilage into thin slices (thickness 1mm), freeze at -80°C, and then grind it into coarse powder, stir the powder in 1% ...

Embodiment 2

[0043] The cartilage decellularized matrix was cut into 1 cm thick slices, placed in 100°C water, heated continuously for 12 hours, cooled to room temperature to prepare the cartilage decellularized matrix solution, and the cartilage decellularized matrix solution was prepared by grinding for 2 weeks as a control, such as figure 2 with image 3As shown, it can be seen that the solution obtained by the grinding method is milky white, the solution is opaque, and contains some insoluble particles; the solution obtained by the heating method is a clear and transparent colorless solution, indicating that the heating method promotes the dissolution of the cartilage decellularized matrix.

Embodiment 3

[0045] (1) The cartilage decellularized matrix was cut into 1 cm thick slices, placed in 100°C water, heated continuously for 7 hours, cooled to room temperature, then frozen at -80°C, and vacuum-dried.

[0046] (2) Add the cartilage acellular matrix obtained by freeze-drying into the hydrochloric acid solution of pepsin (the concentration of the acellular matrix is ​​10mg / ml, the concentration of pepsin is 1mg / ml, and the concentration of hydrochloric acid is 0.01M), stir at room temperature for 12h, and then add 0.1 M NaOH to adjust the pH to 7.0-8.0, then dropwise add methacrylic anhydride (the material ratio of methacrylic anhydride to cartilage decellularized matrix is ​​1ml / g), react at 20°C for 24h, and then use a dialysis bag with a molecular weight cut-off of 14000 for dialysis, Freeze at -80°C and dry in vacuum to obtain a methacrylic acid-modified acellular matrix.

[0047] (3) Dissolve the acellular matrix modified with methacrylic anhydride in step (2) in hydrochl...

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Abstract

The invention relates to a preparation method of a novel cartilage decellularized matrix ink. The method comprises: preparing cartilage decellularized matrix solution, preparing acrylamide decellularized cartilage matrix, and preparing cartilage decellularized matrix ink. The method is simple, convenient and feasible. It uses natural acellular matrix and methacrylic anhydride as raw materials, and has a wide range of sources. The dissolution time of cartilage acellular matrix is ​​greatly shortened, and the dissolution performance is greatly improved. It is expected to become an ideal tissue engineering scaffold. It has a broader application prospect in cartilage repair; the prepared acellular matrix bioink has good fluidity and has the characteristics of photocrosslinking.

Description

technical field [0001] The invention belongs to the field of bio-ink preparation, in particular to a preparation method of novel cartilage decellularized matrix ink. Background technique [0002] Cartilage defects such as joint pain, deformity, and meniscus damage are becoming a common phenomenon due to factors such as sports trauma, disease, and age-induced degeneration. [0003] Existing clinical treatment methods include joint replacement surgery and non-surgical treatment. The results of joint replacement surgery are good for a while, but the risks and complications of surgery can easily lead to more painful conditions for patients. Non-surgical treatment can relieve pain and improve joint function in some patients, but the treatment effect of most patients is not good. [0004] Cartilage acellular matrix is ​​a cell-free extracellular matrix component and structure derived from autologous or allogeneic tissue, and its chemical components include collagen, glucosamine,...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61L27/36A61L27/38
CPCA61L27/3612A61L27/3687A61L27/3691A61L27/3817A61L27/3834A61L2430/06
Inventor 莫秀梅余凡李亚强李军王晓宇徐小青刘明月吴晶磊
Owner DONGHUA UNIV