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Animal bladder decellularizing matrix as well as preparation method and application thereof

A decellularized matrix and decellularized technology, applied in the field of biomedical materials, can solve the problems of large amount of chemical reagents and incomplete antigen removal, and achieve the effect of low concentration of chemical reagents, reducing the risk of transmission, and mild nature.

Active Publication Date: 2017-11-28
BEIJING NATON TECH GRP CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Most of the existing porcine bladder acellular matrix is ​​decellularized by chemical reagents, which has defects such as incomplete removal of antigens and large amount of chemical reagents

Method used

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  • Animal bladder decellularizing matrix as well as preparation method and application thereof
  • Animal bladder decellularizing matrix as well as preparation method and application thereof
  • Animal bladder decellularizing matrix as well as preparation method and application thereof

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preparation example Construction

[0023] In one embodiment of the preparation method according to the present invention, the animal bladder is preferably a common mammalian bladder, such as a common pig, cow, etc., more preferably a low-cost pig bladder.

[0024] In one embodiment of the preparation method according to the present invention, the submucosa of the animal bladder can be obtained by any existing method. For example, after rewarming the deep-frozen bladder tissue, inject purified water, fasten the end and gradually increase the water pressure until the mucosal layer cracks, and then bluntly scrape off the mucosal layer, and only keep the submucosa.

[0025] In one embodiment of the preparation method according to the present invention, in the decellularization step of the decellularized solution, the decellularized solution contains trypsin with a mass concentration of 0.01% to 0.2%, pancreatin with a mass concentration of 0.1% to 1.0%. Ammonium sulfate and a degreasing agent with a mass concentrat...

Embodiment 1

[0070] Purchasing pig bladders with qualified indicators, sorting, cleaning and storing them in a deep low temperature environment from -20°C to -196°C. Bluntly remove excess fat and other tissues, wash with purified water, and place in a -80°C ultra-low temperature refrigerator for deep freezing for 15 hours. The porcine bladder was taken out from the deep-low temperature refrigerator and fixed in a purified water pressurized flushing device, pressurized and flushed until cracks appeared in the mucosal layer of the bladder, and then the mucosal layer was bluntly scraped off, leaving only the submucosa. Put the bladder submucosa in a -80°C refrigerator for 6 hours, take out the bladder submucosa from the deep-low temperature refrigerator, and soak it in a container filled with purified water at room temperature until it is completely thawed. At this point, a freeze-thaw process is completed. So repeated freezing and thawing 3 times. A virus inactivation solution is prepared, ...

Embodiment 2

[0072] Purchasing pig bladders with qualified indicators, sorting, cleaning and storing them in a deep low temperature environment from -20°C to -196°C. Bluntly remove excess fat and other tissues, wash with purified water, and place in a -80°C ultra-low temperature refrigerator for deep freezing for 15 hours. The porcine bladder was taken out from the deep-low temperature refrigerator and fixed in a purified water pressurized flushing device, pressurized and flushed until cracks appeared in the mucosal layer of the bladder, and then the mucosal layer was bluntly scraped off, leaving only the submucosa. Put the bladder submucosa in a -80°C refrigerator for 7 hours, take out the bladder submucosa from the deep-low temperature refrigerator, and soak it in a container filled with purified water at room temperature until it is completely thawed. At this time, a freeze-thaw process is completed. So repeated freezing and thawing 5 times. A virus inactivation solution is prepared, s...

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Abstract

The invention provides a preparation method of an animal bladder decellularizing matrix. The preparation method comprises the steps of extracting a strata submucosum of an animal bladder, and carrying out virus inactivation, degreasing and decellularization, wherein the decellularization step sequentially comprises a repeated freeze thawing step and a decellularization solution decellularization step, the freezing temperature in the freeze thawing step is (-70)-(-90) DEG C, and the freezing time is 5-15 hours. The invention further provides the animal bladder decellularizing matrix prepared by virtue of the preparation method and the application of the animal bladder decellularizing matrix. According to the preparation method, the bladder decellularizing matrix is prepared by combining the repeated freeze thawing with the decellularization solution decellularization, elastic fibers and a collagen net structure of an extracellular matrix are furthest preserved, and meanwhile, antigens are thoroughly removed. The animal bladder decellularizing matrix can be applied to the repairing fields of bladders, urethrae and ureters.

Description

technical field [0001] The invention relates to the field of biomedical materials, in particular to a preparation method of animal bladder decellularized matrix, the obtained decellularized matrix and its application. Background technique [0002] For defects of the bladder, urethra, and ureter, three types of tissue replacement materials are currently used clinically: synthetic polymer materials, allogeneic acellular matrix, and heterogeneous acellular matrix. The use of non-degradable synthetic polymer materials such as polysilicone and polytetrafluoroethylene is prone to tissue necrosis, fistula, stenosis, extravasation, and stone formation, while the use of degradable synthetic polymer materials such as polylactic acid and polyglycolic acid Molecular materials are used for bladder, urethra, and ureteral defects. Although the material is easy to obtain and convenient to prepare, the inflammatory response is strong during the degradation process, which affects the differen...

Claims

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Application Information

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IPC IPC(8): A61L27/36
CPCA61L27/3633A61L27/3679A61L27/3683A61L27/3687A61L2430/22A61L2430/40
Inventor 张国强柳轶男杨新广董骧
Owner BEIJING NATON TECH GRP CO LTD
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