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Preparation method of decellularized heterogeneous small blood vessels

A decellularization and blood vessel technology, applied in the field of biomaterials, can solve the problems of damaged mechanical properties of blood vessels, large damage, and destruction of the three-dimensional structure of heterogeneous blood vessels, and achieve satisfactory mechanical properties, favorable infiltration, and good biomechanical properties and structure. Effect

Active Publication Date: 2020-09-11
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 heterogeneous small blood vessels
  • Preparation method of decellularized heterogeneous small blood vessels
  • Preparation method of decellularized heterogeneous small blood vessels

Examples

Experimental program
Comparison scheme
Effect test

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 decellularized heterogeneous small blood vessels. By adopting a method for combining ultrasound, perfusion, detergent and enzyme, cells in a biological material matrix are broken through a cavitation effect of high-frequency ultrasonic waves, then the detergent is pressed into a materialin a perfusion mode to further wash cell components and part of immunogenic substances; after the primary action of ultrasound and the detergent, the cells are basically broken, most of cell components are washed away, most of nucleic acid substances are further removed under the action of DNase-I and RNase-A, and the residual quantity is less than 100 ng / mg; and finally, a good decellularizationeffect is achieved through repeated sterile PBS solution lavage.

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 Applications(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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