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Method for preparing re-cellularized biological valve material

A bioprosthetic valve and cellularization technology, applied in the field of biomedical engineering, can solve unseen problems

Active Publication Date: 2013-09-18
江苏先进无机材料研究院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there have been no reports and related patents on the preparation of bioprosthetic valve materials using NDGA cross-linked decellularized animal heart valves.

Method used

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  • Method for preparing re-cellularized biological valve material
  • Method for preparing re-cellularized biological valve material
  • Method for preparing re-cellularized biological valve material

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] Use the balanced salt buffer D-Hanks solution at 1-6°C as the storage and transportation solution, take fresh porcine aortic heart valves from the slaughterhouse within 2 hours, cut the valve leaflets at 4°C within 4 hours, and use Wash with D-Hanks solution. The prepared decellularized porcine heart aortic valve was washed clean to obtain the following: figure 1 Structure and shape of acellular bioprosthetic valve materials. Under the scanning electron microscope, the extracellular matrix fibers of the valve material are loosely arranged in bundles, without fiber swelling and fracture. The valve material is porous, the extracellular matrix fibers are naturally arranged, and there is no cell debris remaining on the surface. The valve material was soaked in a D-Hanks special solution with 2.5 mg / ml nordihydroguaiaretic acid added at a rate of 2 ml per piece, and cross-linked at 37° C. for 48 hours while shaking at 120 rpm. Then add 2ml of D-Hanks solution to each piece...

Embodiment 2

[0059] Prepare the porcine decellularized tissue engineering valve natural material according to the aforementioned method and clean it up, add 1.5 mg / ml D-Hanks special solution of nordihydroguaiaretic acid at a ratio of 2 ml to each valve material Example 1 Method handover The resulting cross-linked biological valve material can also maintain the original shape of the valve, soft, non-shrinking, porous, and natural arrangement of extracellular matrix fibers. image 3 The maximum tensile strength of the acellular valve material cross-linked by 1.5 mg / ml nordihydroguaiaretic acid was 13.18 MPa. Figure 5 It was shown that only about 10% of the 1.5 mg / ml nordihydroguaiaretic acid cross-linked decellularized valve material was degraded after 24 hours of enzymatic hydrolysis. It shows that this concentration of nordihydroguaiaretic acid also has a cross-linking effect on the decellularized tissue engineering valve material, and has good mechanical properties and stability against...

Embodiment 3

[0061] Get fresh porcine aortic heart valve from slaughterhouse by the method of embodiment 1 and clean up. After washing three times with D-Hanks solution, valve cells were decellularized with decellularization solution at 4°C for 96 hours. Use D-Hanks solution to shake the decellularized heart valve material at 24°C (normal temperature) at 120rpm to wash valve cell residues, debris, free protein, nucleic acid and other macromolecules 5 times, each time for more than 10 hours. Add 0.1mg / ml, 0.5mg / ml, 1.5mg / ml and 5mg / ml nordihydroguaiaretic acid D-Hanks solution at a ratio of 10ml per valve, at 24°C, at a shaking speed of 240rpm , cross-linked for 2 hours ( Figure 9 3, 4, 5 and 6 in), 72 hours ( Figure 9 11, 12, 13 and 14) and 240 hours ( Figure 9 7, 8, 9 and 10 in ). Add 20ml of D-Hanks solution to each valve to soak the valve, and wash at 120rpm at room temperature. The bioprosthetic valve material of the present invention prepared in this way was subjected to the a...

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Abstract

The invention relates to a method for preparing a cellularized biological valve material, which is characterized by comprising the following steps: performing cell removal treatment for a heart valve of a pig aorta, then using nordihydroguaiaretic acid solution to cross link, and performing post treatment after the cross linking. The concentration of the used cross linking agent is 0.1 to 10 mg / ml. The cross linking conditions comprise that the shake cross linking is carried out for 2 to 240 hours at 60 to 360rpm at the temperature of between 1 and 40 DEG C, then the heart valve is taken out and soaked into D-Hanks solution, and the heart valve is post-treated for 3 to 40 days at the temperature of between 1 and 6 DEG C so as to obtain the cellularized biological valve material. By adjusting the concentrations of the cross linking agent the nordihydroguaiaretic acid solution, the cross linking time, the cleaning time and the like, the adjustment and control in the aspects of cross linking mechanical strength, enzymolysis resistance, cross linking agent release, tissue regeneration capacity promotion and the like are realized.

Description

technical field [0001] The present invention relates to a cross-linking preparation method of a biological valve material, more precisely, the present invention relates to a method for preparing a recellularized biological valve material by cross-linking nordihydroguaiaretic acid as a cross-linking agent , belonging to the field of biomedical engineering. Background technique [0002] Heart valve disease is an important disease that endangers human health. The mainstay of treatment is heart valve replacement. Since Harken and Starr first replaced the aortic valve and mitral valve with artificial cage-type mechanical valves in 1960, artificial heart valves have received great attention and attention from the medical community, and thus have made significant progress. [0003] At present, there are two types of artificial heart valves: mechanical valves and biological valves. The use of isotropic low-temperature pyrolytic carbon (or titanium oxide) as a material (or modifie...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61L27/36A61L27/56
Inventor 常江翟万银
Owner 江苏先进无机材料研究院
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