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Preparation method for cellfree intestinum tenue submucosa biological material

A technology of small intestinal submucosa and biomaterials, applied in medical science, prostheses, etc., can solve problems such as SIS growth factors can not play a role, product protein activity is not considered, and the decellularization method is not perfect, so as to facilitate clinical suture fixation, Unrestricted sources of raw materials, the effect of protecting protein activity

Active Publication Date: 2009-02-18
SHAANXI RUISHENG BIOTECH
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  • Summary
  • Abstract
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AI Technical Summary

Problems solved by technology

The problem with these products is that the decellularization method is not perfect. For example, the treatment method of Surgisis only uses low-concentration peracetic acid treatment, so there are many residual cells and the decellularization effect is not good. It will cause the body’s immune rejection during use; Restore is ten layers SIS is superimposed, which makes the above problems more prominent, and its DNA residue is high after testing; and the natural antigenic components (such as α-ga1) existing in the extracellular matrix are not considered to be removed
In the end, the current acellular SIS products still have a large immune rejection reaction, and they only become temporary substitutes after implantation, and it is difficult to survive in the body for a long time; in addition, the current technical methods do not fully consider how to effectively remove the cells carried by SIS. Viruses, there is a potential risk of virus infection; the protection of protein activity in the product is not considered in the freeze-drying process, so that the growth factors contained in the SIS cannot play a role

Method used

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Examples

Experimental program
Comparison scheme
Effect test

example 1

[0017] 1). Pre-treatment: Remove the intestinal contents from the porcine jejunum, rinse it with distilled water, cut it into required pieces, put it in an aqueous solution containing 0.2% peracetic acid and 10% ethanol for 1 hour, and use Rinse with PBS; mechanically scrape off the small intestinal mucosa, muscle layer and serosa, rinse with PBS, and obtain the small intestinal submucosa;

[0018] 2). Decellularization treatment: put the pre-treated small intestinal submucosa into 0.5M NaOH solution, soak for 20 minutes at 6±2°C, and rinse with PBS until neutral;

[0019] 3). Enzyme treatment: put the decellularized submucosa into a mixed solution containing 40U / ml DNase and 10U / ml α-galactosidase, soak at 35°C for 1 hour, and rinse with PBS;

[0020] 4). Preparation of membrane-like product of acellular small intestinal submucosa: Soak the enzyme-treated submucosa in cryoprotectant solution for 30 minutes. The cryoprotectant solution is Hanks balanced salt solution with 1% d...

example 2

[0023] 1). Pre-treatment: Remove the intestinal contents from the bovine jejunum, rinse it with distilled water, cut it into required pieces, put it in an aqueous solution containing 0.4% peracetic acid and 5% ethanol for 2 hours, and use Rinse with PBS; spread the submucosa fragments on the platform to mechanically scrape the small intestinal mucosa, muscular layer and serosa, and rinse with PBS to obtain the small intestinal submucosa;

[0024] 2). Decellularization treatment: put the pretreated small intestinal submucosa into 1M NaOH solution, soak for 30 minutes at 4±2°C, and rinse with PBS until neutral;

[0025] 3). Enzyme treatment: put the decellularized small intestinal submucosa into a mixed solution containing 50U / ml DNase and 20U / ml α-galactosidase, soak at 37°C for 30 minutes, and rinse with PBS;

[0026] 4). Preparation of microparticles of acellular small intestinal submucosa: Soak the enzyme-treated submucosa in cryoprotectant solution for 30 minutes. The cryop...

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Abstract

The invention relates to a method for preparing a biomaterial of acellular small intestinal submucosa, which comprises the steps of preposition treatment, acellular treatment, enzyme treatment, preparations of membranous products and particle products. Compared with the prior art, the biomaterial has higher bioactivity and biocompatibility, no obvious immune rejection and no toxic effect on cells; besides, the biomaterial has a certain mechanical strength and toughness, and variable shape, size and thickness, thereby being convenient for clinical suturing and fixing. At the same time, the preparation method has the advantages of unlimited raw material source, cell-free residues, no ethical issues and being capable of effectively inactivating virus. The prepared products are applicable to the biomedical engineering fields, such as repairing defections of body tissue, serving as tissue filling materials, repairing facial depression deformity, serving as tissue reinforcements to replace fascia, repairing membranous defections and malnourished and infected surfaces of wound, serving as materials for biodegradable stents, and serving as injectable filling materials.

Description

technical field [0001] The invention belongs to the technical field of tissue engineering biomaterials, and in particular relates to a preparation method of decellularized small intestinal submucosa biomaterials. Background technique [0002] Extracellular matrix (Extracellular matrix, ECM) is the main tissue component of various tissues and organs. It constitutes the three-dimensional structure of various tissues and organs and the storage of nutrients. Plays an important role in organ repair. The small intestinal submucosa (SIS) is derived from the small intestine and is a natural extracellular matrix biomaterial, mainly composed of type I and type III fibrous collagen, containing a small amount of type IV and V collagen, and also contains amino groups. Dextran and glycoproteins, etc., the amino dextran contained mainly include hyaluronic acid, heparin, heparan acetate sulfate, chondroitin sulfate A and dermatan sulfate. Since SIS is a complex component structure of natu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/26
Inventor 王爱军
Owner SHAANXI RUISHENG BIOTECH
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