Preparation method for novel cartilage acellular matrix ink
A technology of acellular matrix and cartilage, applied in medical science, tissue regeneration, prosthesis, etc., can solve the problems of difficult processing and poor solubility, and achieve improved solubility, good fluidity, improved processing performance and enzyme solubility Effect
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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 10 mg / ml, the concentration of pepsin is 1 mg / ml, and the concentration of hydrochloric acid is 0.01M), stir at room temperature for 12 hours, dissolve and then add 0.1 M NaOH to adjust the pH to 7.0-8.0, then add methacrylic anhydride dropwise (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...
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