Preparation method of decellularized xenogeneic small blood vessels

A decellularized and xenogeneic technology, applied in the field of the preparation of decellularized xenogeneic small blood vessels, can solve the problems of destroying the three-dimensional structure of xenogeneic blood vessels, large damage, and damage to the mechanical properties of blood vessels.

Active Publication Date: 2022-05-13
THE SECOND XIANGYA HOSPITAL OF CENT SOUTH UNIV
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Problems solved by technology

However, trypsin has great damage to the structure of extracellular matrix, which can destroy the three-dimensional structure of xenografts and damage the mechanical properties of blood vessels.

Method used

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  • Preparation method of decellularized xenogeneic small blood vessels
  • Preparation method of decellularized xenogeneic small blood vessels
  • Preparation method of decellularized xenogeneic small blood vessels

Examples

Experimental program
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Embodiment 1

[0063] 1. Preparation of decellularized xenogeneic small vessels

[0064] 1.) Pre-treatment: Take out the bilateral internal mammary arteries from the cattle (about 400-500Kg in body weight) in the slaughterhouse, separate the fat and adventitia within 30 minutes of warm ischemia time, and use sterile PBS solution containing antibiotics (penicillin) and streptomycin (100U / ml, 100ug / ml respectively), washed several times, soaked in 0.1% bromogeramine for 30min, rinsed the inside and outside of the lumen with sterile PBS containing antibiotics for 10min×3 times, and stored at low temperature at 4°C.

[0065] 2.) Decellularization treatment: three-step method

[0066] ①High-frequency ultrasonic wave + perfusion + detergent: cut the blood vessel into a length of 10 cm, connect one end of the perfusion tube and leave the other end unconnected, and place the perfusion rate at 90ml / min in a specially designed container (see figure 1 ); Use sterile PBS containing antibiotics to prepa...

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Abstract

The invention relates to the technical field of biological materials, in particular to a preparation method of acellular xenogeneic small blood vessels. The method adopts the combination of ultrasound, perfusion, detergent and enzyme, uses the cavitation effect of high-frequency ultrasound to break the cells in the biological material matrix, and then uses the perfusion method to press the detergent into the material to further wash the cell components and some immunogenic substances; after the initial action of ultrasound and detergent, the cells were basically broken, and most of the cell components were also washed away, and further through the action of DNase-I and RNase-A, most of the nucleic acid substances were removed, and the residual The amount is less than 100ng / mg; finally, through repeated lavage with sterile PBS solution, a good decellularization effect is achieved.

Description

technical field [0001] The invention relates to the technical field of biomaterials, in particular to a method for preparing decellularized heterogeneous small blood vessels. Background technique [0002] Coronary atherosclerotic heart disease (coronary heart disease) mortality rate ranks first in various diseases, and is on the rise. Coronary artery bypass grafting (CABG) is one of the main treatments for coronary heart disease. The United States alone needs more than 600,000 small-diameter graft vessels (inner diameter 3-5mm) for CABG every year, and my country needs about 150,000 per year, and it is growing rapidly. At present, such transplanted blood vessels mainly use autologous arteries or great saphenous veins, but 30% of patients cannot obtain ideal autologous blood vessels due to problems such as underlying diseases, amputation, and secondary surgery. At the same time, the acquisition of autologous blood vessels will inevitably bring trauma to the body. Unfortunat...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61L27/36A61L27/50
CPCA61L27/507A61L27/3691A61L27/3687A61L27/50A61L27/3625A61L2430/40
Inventor 刘育宏陈春阳吴忠仕唐贞洁卢婷袁浩泳刘四喜解鑫隆钱涛
Owner THE SECOND XIANGYA HOSPITAL OF CENT SOUTH UNIV
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