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Method for preparing SIS tissue repair material

A tissue repair and solution technology, applied in the field of preparation of SIS tissue repair materials, can solve the problems that do not involve material degreasing and decellularization, insufficient removal of organic solvents, degreasing and deproteinization disinfection, etc. Rejection and allergic reactions, safety-enhancing effects

Active Publication Date: 2009-05-20
BEIJING DATSING BIO TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] At present, in the field of biomaterials, there are also many reports on the preparation of SIS as tissue repair materials and related animal experiments, but the process is simple, and most of them are limited to decellularization and descaling alone. The problem of insufficient disinfection leads to certain immunity and antigenicity of the material, and some methods have some problems of insufficient removal of organic solvents, which leads to a certain degree of antigenicity of the material to the receptor
There are also related reports on the preparation and use of injectable and powdered SIS abroad, but the preparation method is relatively complicated, and it does not involve the steps of degreasing and decellularizing the material, so the material will also have certain antigenicity, and in There are also certain restrictions on the use of

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032]Fresh pig small intestines within 4 hours of slaughter were taken, the serosa layer, mucous membrane layer and muscular layer of the small intestine were removed and the submucosa was retained, rinsed with pure water, and rinsed after disinfection with iodine. Soak the prepared SIS in a mixed solution of 0.5% peracetic acid-10% ethanol for disinfection treatment, the mass volume ratio of SIS to the solution is 1:5, soak for 1 hour; after soaking, use pure water to ultrasonically clean it twice , 20 minutes each time and change the water; take it out and rinse it, put it into 2% SDS solution for descaling treatment, the mass volume ratio of SIS and solution is 1:5, soak for 18 hours; ultrasonic cleaning 2 times, each time 20 minutes and change the water; rinse until the solution is clear; soak for 16 hours with 1mmol / L NaCl phosphate buffered saline solution (PBS), the mass volume ratio of SIS to solution is 1:10. Wash with pure water until the pH value is between 5.8; th...

Embodiment 2

[0034] The fresh sheep small intestine within 4 hours of slaughter was taken, the serosa layer, mucous membrane layer and muscular layer of the small intestine were removed and the submucosa layer was retained, rinsed with pure water, and rinsed after disinfection with iodine. Soak the prepared SIS in the mixed solution of 1.5% peracetic acid-10% ethanol by immersing the SIS prepared by mechanical method, the mass volume ratio of SIS and solution is 1:5, soak for 40 minutes; after soaking, use Ultrasonic cleaning with pure water 2 times, 20 minutes each time and changing the water; after rinsing, soak in a mixed solution of 1mmol / L HCl and 1mmol / LNaCl (pH value between 0 and 1) for 6 hours for decellularization , the mass volume ratio of SIS and solution is 1:10; ultrasonic cleaning with pure water 2 times for 20 minutes each time, after rinsing clean, soak for 16 hours with 1mmol / L NaCl phosphate buffered saline solution (PBS), SIS and solution The mass to volume ratio is 1:1...

Embodiment 3

[0036] Fresh dog small intestines within 4 hours of slaughter were taken, the serosa layer, mucous membrane layer and muscular layer of the small intestine were removed and the submucosa was retained, rinsed with pure water, and rinsed after disinfection with iodine. Soak the prepared SIS in 0.05% sodium hydroxide for 1 hour, and soak in 1.5% SDS solution for 10 hours to carry out decellularization and descaling treatment. The SIS and solution The mass-volume ratio is 1:5, and a fresh solution is changed every 5 hours; after soaking, ultrasonically clean it twice with pure water, and change the water for 20 minutes each time; after rinsing, put in polyacrylic acid with a mass-volume ratio of 0.1% Soak in the solution for 4 hours to decellularize again, the mass volume ratio of SIS to the solution is 1:5, ultrasonically clean with pure water to a pH value of 7.0; soak the material in a solution containing 0.8% peracetic acid and 12% alcohol Disinfect for 5 hours, the mass volum...

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Abstract

The invention provides a method for preparing an SIS tissue restoring material. The method comprises the following steps: carrying out sterilizing treatment, degreasing treatment, cell free treatment and absterging treatment according to an arbitrary sequence at the lower layer of small intestinal mucosa; and finally preparing the material through freezing and drying. The SIS tissue restoring material can be used as an independent and common support vector for a growth factor, a cell, a drug and the like, and becomes a tissue engineering material through in-vitro culture. The material is a natural and nontoxic biological material without antigenicity and immunity. The SIS tissue restoring material can be used for restoring hernia, defect of skin, urinary system tissue defect, mucosa defect and other periphery and internal parenchyma defects.

Description

technical field [0001] The present invention relates to a preparation method of a tissue repair material, in particular to a preparation method of an SIS (small intestinal submucosa, small intestinal submucosa) tissue repair material. Background technique [0002] The mammalian small intestine is mainly composed of 4 layers, which are the mucosal layer, submucosa layer, muscular layer and serosa layer from inside to outside. The submucosa is composed of a layer of dense connective tissue, mainly composed of type I and type III collagen fibers, containing a small number of cells and some large blood vessels, lymphatic vessels and nerves. Small intestinal submucosa (hereinafter referred to as SIS) is a biologically derived material of natural extracellular matrix. Because SIS has non-immunogenicity, antimicrobial activity, can promote tissue regeneration, etc., and can be metabolized through biodegradation in vivo, it has been widely studied. [0003] SIS is generally prepar...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/36A61L31/14
Inventor 李次会
Owner BEIJING DATSING BIO TECH
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