Composite cross-linked bioprosthetic valve and preparation method thereof
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A bioprosthetic valve and composite technology, applied in tissue regeneration, medical science, prosthesis, etc., can solve the problems of inability to endothelialize, toxicity, etc., achieve good anti-collagen enzymolysis and elastolysis ability, thin thickness, and wide application range effect
Pending Publication Date: 2021-09-24
THE SECOND XIANGYA HOSPITAL OF CENT SOUTH UNIV
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And due to the toxicity of its residual aldehyde groups, degradation of elastin, inability to endothelialize, and inflammatory reactions, the average lifespan of biological valves is only 12-15 years
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Embodiment 1
[0043] Example 1 Decellularization of fresh bovine pericardium
[0044]Soak fresh bovine pericardium with 0.1% bromogeramine, disinfect for 30 minutes, and stir once every 10 minutes;
[0045] Place the sterilized fresh bovine pericardium in a solution with a mass percent concentration of 0.25% Trixon-100, shake it at 37°C for 48 hours, and change the solution every 12 hours;
[0046] Use 3U / mL of DNase, 0.03mg / mL of RNase, 2.5mmol / L of MgCl 2 , 0.1mmol / L of CaCl 2 Prepare a mixed solution, place the bovine pericardium treated with Trixon-100 in the mixed solution, shake at 37°C for 24 hours, and change the solution every 12 hours;
[0047] Then use distilled water to shake and rinse for 48 hours, and change the solution every 8 hours;
[0048] Then use PBS buffer to rinse for 24 hours, change the medium, and then use new PBS buffer to preserve the decellularized bovine pericardium.
[0049] Among them, the PBS buffer formula is: per 10XPBS 1L: NACL 80g, KCL 2g, KH 2 PO ...
Embodiment 2
[0050] Example 2 Cross-linking decellularized bovine pericardium
[0051] 1) The decellularized bovine pericardium obtained in Example 1 was placed in a 1% tannic acid solution (using 70% ethanol-water solution as a solvent, using hydrochloric acid to adjust pH=5), at room temperature (25 ±5°C) shaking treatment for 48h;
[0052] 2) Put tannic acid-decellularized bovine pericardium in 1mmol / L neomycin solution (use 0.05mml / L morpholineethanesulfonic acid solution as solvent, use hydrochloric acid to adjust pH=6), normal temperature (25± 5°C) for 1 h with shaking.
[0053] 3) Put tannic acid-neomycin-decellularized bovine pericardium in 10 mmol / L riboflavin 5-monophosphate solution (using PBS buffer as a solvent), and irradiate for 12 hours under ultraviolet light with a wavelength of 360 nm;
[0054] 4) Rinse with PBS buffer, and then store with new PBS buffer.
[0055] Among them, the PBS buffer formula is: per 10XPBS 1L: NACL 80g, KCL 2g, KH 2 PO 4 2g, NA 2 HPO 4 *12...
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Abstract
The application discloses a preparation method of a composite cross-linked bioprosthetic valve. The preparation method comprises the following steps: 1, treating a decellularized bioprosthetic valve with tannic acid; 2, treating the tannic acid treated bioprosthetic valve with neomycin; 3, treating the neomycin treated bioprosthetic valve with 5-riboflavin monophosphate under the condition of ultraviolet irradiation; and 4, rinsing to obtain the composite cross-linked bioprosthetic valve. The application further provides a composite cross-linked bioprosthetic valve. According to the preparation method of the composite cross-linked bioprosthetic valve and the prepared composite cross-linked bioprosthetic valve, which are provided by the application, photooxidation of riboflavin is adopted and synergic with tannic acid and neomycin to carry out cross-linking on decellularized bovine pericardium, so that the aim of cross-linking three main components of an extracellular matrix is fulfilled, and the bioprosthetic valve which is of a good extracellular matrix structure, has an excellent mechanical effect and heat shrinkage temperature, is thin in thickness, can have a wider application range and has good collagen enzymolysis resistance and elastic enzymolysis resistance can be obtained.
Description
technical field [0001] The present application relates to the technical field of biological materials, in particular to a composite cross-linked biological valve and a preparation method thereof. Background technique [0002] The human heart has four heart valves, which open and close normally, cooperate with the contraction of the myocardium to make blood circulate throughout the body and maintain normal life activities. However, due to congenital malformations, acquired factors such as rheumatic heart disease and heart degeneration, heart valve abnormalities will be found, which seriously threatens human life. Valve replacement is the mainstay of treatment for advanced valvular disease. Worldwide, more than 250,000 valve replacement surgeries are performed each year, and it is estimated that by 2050, there will be more than 500,000 valve replacement surgeries per year. [0003] At present, there are two main options for artificial valves: mechanical valves and biological...
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