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Method for removing cell components and/or nucleic acid substances of homogeneous or heterogeneous biological materials

A biomaterial and cell component technology, applied in medical science, prosthesis, etc., can solve the problems of three-dimensional structure damage of extracellular matrix, long processing time, influence of mechanical properties of acellular tissue, etc., and achieve low residual immunogenicity, The effect of short processing time and low damage

Active Publication Date: 2021-12-21
THE SECOND XIANGYA HOSPITAL OF CENT SOUTH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0007]Although there are many decellularization methods at home and abroad, there are still some problems: (1) The corresponding chemical reagents can cause the destruction of the three-dimensional structure of the extracellular matrix, which is harmful to the The mechanical properties of decellularized tissue are negatively affected; (2) The corresponding treatment time lasts for a long time, especially the single or only combination of the two methods, and the treatment process is more than one week

Method used

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  • Method for removing cell components and/or nucleic acid substances of homogeneous or heterogeneous biological materials
  • Method for removing cell components and/or nucleic acid substances of homogeneous or heterogeneous biological materials
  • Method for removing cell components and/or nucleic acid substances of homogeneous or heterogeneous biological materials

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preparation example Construction

[0048] In the preparation method, the key point is the combination of detergent and enzyme. After the initial action of the detergent, the cell membrane ruptures, the cells are basically broken, and most of the cell components are also washed away, but there are still chromosomes of the nucleus, Small substances such as DNA, RNA, and broken phospholipids stay in the biological material, and further DNase-I and RNase-A actions are required to remove most of the nucleic acid substances; finally, the residuals are removed by repeated rinsing with sterile PBS solution Base fragments to achieve satisfactory immunogenicity removal effect.

[0049] In the described preparation method, the key point is soaking with bromogeramine. In the present invention, the soaking of bromogeramine can have better bactericidal effect and prevent bacterial pollution in the subsequent treatment and preservation process.

[0050] In the preparation method, the key points are the concentration and acti...

Embodiment 1

[0054] Embodiment 1 Early stage treatment

[0055] The bovine pericardium was provided by the local slaughterhouse, and the specimens were collected in the slaughterhouse. After the samples were taken out, they were stored in pre-cooled PBS solution and delivered to the laboratory within 4 hours. Wash with sterile PBS solution containing antibiotics (penicillin and streptomycin, the concentrations are 100U / ml, 100ug / ml respectively), immerse in 0.1% bromogeramine for 30min, wash pericardium with sterile PBS containing antibiotics for 10min×3 times, store at 4°C .

Embodiment 2

[0056] Embodiment 2 sample pruning

[0057] Use a magnifying glass to observe whether there are obvious knife wounds, cut off all areas that do not meet the requirements, carefully remove the fatty tissue on the material matrix with scissors and tweezers, and hold the tweezers parallel to the material to avoid damage to the material. This process should be as gentle as possible. To maximize the protection of tissue fibers from being damaged, so as not to affect its mechanical properties.

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Abstract

The invention relates to the field of medical materials, and particularly relates to a method for removing cell components and / or nucleic acid substances of homogeneous or heterogeneous biological materials. The method adopts a method of combining a detergent and an enzyme, fully utilizes the membrane rupture and decontamination capabilities of the novel detergent, and simultaneously utilizes the specific decomposition effect of the ribozyme to effectively remove the cell components. The method has the advantages of short time and good decellularization effect.

Description

technical field [0001] The present invention relates to the field of medical materials, in particular to a method for removing cellular components and / or nucleic acid substances of the same or heterogeneous biological materials. Background technique [0002] Heart valve disease is a disease with high morbidity and high fatality rate. The rate of heart valve disease in the world is increasing year by year with the increasing aging. It has become one of the major life-threatening medical care problems in the world. one. Advanced valvular disease results in loss of self-repair function, and valve replacement becomes the only surgical option. Nearly 300,000 valve replacement surgeries are performed each year worldwide, and this number is expected to triple by 2050. At present, artificial heart valves are mainly composed of two types: mechanical heart valves and bioprosthetic heart valves, and have been widely used for decades. Both types of artificial valves have their own ad...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/36
CPCA61L27/3604A61L27/3687
Inventor 宋明哲姜振林吴忠仕唐贞洁齐晓科刘育宏解鑫隆吴其应
Owner THE SECOND XIANGYA HOSPITAL OF CENT SOUTH UNIV
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