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Method for preparing soft tissue decellular matrix through supercritical fluid technology

A technology for decellularized matrix and soft tissue, applied in the fields of biomedicine, regenerative medicine, and clinical medicine, can solve problems such as immune rejection, cytotoxicity, and time-consuming removal, and achieve reduced immunogenicity, high decellularization efficiency, and The effect of shortening the time period

Inactive Publication Date: 2017-09-15
GUANGZHOU SUN SHING BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The existing soft tissue decellularized matrix preparation process mostly uses solutions such as protease and sodium dodecyl sulfate (SDS) to remove cells in the tissue, but these reagents have a severe effect on the tissue, severely damage the matrix components, and easily lead to the degradation of the tissue matrix
In addition, these reagents are not easy to remove, may cause cytotoxicity if left in the tissue, and have poor biocompatibility
[0005] The traditional soft tissue decellularized matrix preparation process is easy to cause the matrix components to be easily destroyed, and the reagent residue is serious, which may cause severe immune rejection when implanted in the tissue
[0006] The traditional soft tissue decellularized matrix preparation process may lead to matrix degradation and decrease in mechanical properties
[0007] The traditional soft tissue decellularized matrix preparation process takes a lot of time to remove residual cells and residual reagents in the tissue

Method used

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  • Method for preparing soft tissue decellular matrix through supercritical fluid technology
  • Method for preparing soft tissue decellular matrix through supercritical fluid technology

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] Decellularization of soft tissue using supercritical fluid technology, taking the submucosa of pig small intestine as an example, includes the following steps:

[0052] Pretreatment: Take the jejunum section of pig small intestine, scrape off the serosa layer, muscular layer and mucosa layer under low temperature environment, rinse with clean water until clarified, and obtain the submucosa of pig small intestine;

[0053] Disinfection: Soak with 0.02wt% peracetic acid solution and 4% ethanol solution for 2 hours, rinse with water until the pH is neutral;

[0054] Decellularization: with supercritical CO 2 The fluid was treated for 2 hours, the pressure was 20MPa, the temperature was 37°C, and absolute ethanol was added as an entrainer. The volume of absolute ethanol was 20 times the volume of the porcine small intestinal submucosa, and the pressure release rate was 0.5MPa / min;

[0055] Rinse: with supercritical CO 2 The fluid was treated for 20 hours, the pressure was...

Embodiment 2

[0058] Decellularization of soft tissue using supercritical fluid technology, taking the submucosa of pig small intestine as an example, includes the following steps:

[0059] Pretreatment: Take the jejunum section of pig small intestine, scrape off the serosa layer, muscular layer and mucosal layer in a low temperature environment, rinse with water until clear, and obtain the small intestine submucosa;

[0060] Disinfection: Soak with a peracetic acid solution with a concentration of 0.08wt% or a solution of bromogeramine with a concentration of 0.1wt% for 0.5h, rinse with water until the pH is neutral;

[0061] Decellularization: with supercritical CO 2 The fluid was treated for 0.5h, the pressure was 30MPa, the temperature was 37°C, and a mixture of absolute ethanol and acetone was added as an entrainer. The volume of the mixture of absolute ethanol and acetone was 30 times the volume of the porcine small intestinal submucosa, and the pressure release rate was 0.2 MPa / min;...

Embodiment 3

[0065] Decellularization of porcine small intestinal submucosa using supercritical fluid technology includes the following steps:

[0066] Pretreatment: Take the jejunum section of pig small intestine, scrape off the serosa layer, muscular layer and mucosa layer under low temperature environment, rinse with clean water until clarified, and obtain the submucosa of pig small intestine;

[0067] Disinfection: Soak with a peracetic acid solution with a concentration of 0.04wt% and an ethanol solution with a concentration of 5% for 1 hour, rinse with water until the pH is neutral;

[0068] Decellularization: with supercritical CO 2 The fluid was treated for 3.5 hours, the pressure was 10MPa, the temperature was 37°C, and a mixture of absolute ethanol and acetone was added as an entrainer. The volume of the mixture of absolute ethanol and acetone was 10 times the volume of the porcine small intestinal submucosa, and the pressure release rate was 1MPa / min;

[0069] Rinse: with sup...

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Abstract

The present invention discloses a preparation method of a soft tissue decellular matrix, and relates to the technical field of clinical medicine, biomedical medicine and regenerative medicine. According to the present invention, the natural soft tissue decellular matrix is prepared by using a supercritical fluid technology, wherein the natural soft tissue comprises small intestine tissue, blood vessel tissue, muscle tissue, heart tissue, valve tissue, lung tissue, spleen tissue, kidney tissue, liver tissue, stomach tissue, gallbladder tissue, fat tissue, cartilage tissue, trachea tissue, esophagus tissue, bladder tissue, ureter tissue, oviduct tissue, or uterus tissue. According to the present invention, with the preparation method, the damage on the soft tissue matrix component is low, and the obtained soft tissue decellular matrix has advantages of low immunogenicity and good biocompatibility.

Description

technical field [0001] The invention relates to the technical fields of clinical medicine, biomedicine and regenerative medicine, in particular to a method for preparing soft tissue decellularized matrix by supercritical fluid technology. Background technique [0002] The decellularized matrix is ​​to decellularize the tissue / organ with an appropriate method, remove the cellular components in the tissue that may cause rejection, and maintain the three-dimensional structure of the tissue / organ. Compared with non-animal-derived materials such as metals and plastics, the composition and structure of decellularized matrix are closer to human tissue, and its biocompatibility is better, which makes it the most widely used material in tissue engineering repair. The process of decellularization is to remove cells in tissues / organs through physical methods, chemical methods, and biological enzymes to minimize the content of immunogenic substances such as cellular components. The eff...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/00
CPCC12N5/0081
Inventor 曾晨光梁丽金杨习锋全大萍
Owner GUANGZHOU SUN SHING BIOTECH CO LTD
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