Rapid preparation of accellular porcine intestinal submucosa

A submucosa and decellularization technology, applied in medical science, tissue regeneration, prostheses, etc., can solve the problems of reducing the mechanical properties of the final ECM, incompleteness, cumbersome complexity, etc., to maintain chemical composition and mechanical properties, and ensure compatibility Sexual, non-toxic promoting effect

Active Publication Date: 2018-11-13
承功(厦门)生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, the commonly used decellularization methods are cumbersome and complicated, mainly including: physical methods, chemical methods and digestive enzymatic methods, and the obtained products are either incompletely decellularized, residual antigenicity, or over-treated, which reduces the mechanical properties of the final ECM. performance

Method used

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  • Rapid preparation of accellular porcine intestinal submucosa
  • Rapid preparation of accellular porcine intestinal submucosa
  • Rapid preparation of accellular porcine intestinal submucosa

Examples

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preparation example Construction

[0032] A rapid preparation method of acellular porcine small intestinal submucosa includes the following steps:

[0033] S1: Material pretreatment: Cut the fresh pig jejunum, clean it repeatedly, and then cut it to an appropriate length as needed, generally 10-20cm long; scrape off the serosal layer, mucosal layer and muscle layer of the jejunum with a blunt spatula. Rinse it with deionized water, then place it in the newly prepared pretreatment solution, soak it for 30-60 minutes, and then set it aside.

[0034] S2: One-time decellularization treatment: Rinse the pretreated submucosa of the pig small intestine with deionized water, drain it, transfer it to the cell lysate, sonicate for 10-30 minutes or soak for 30-120 minutes with shaking, and then use it for another 30 minutes. Rinse with deionized water at -45℃ for 10-60 minutes;

[0035] S3: Degreasing treatment: the submucosa of the pig small intestine after a decellularization treatment is rinsed with deionized water, drained,...

Embodiment 1

[0065] 1. Preparation of pretreatment liquid

[0066] Weigh sodium silicate (Na2SiO 3 ) 100g, 60g hydrogen peroxide, add 700ml deionized water and mix well; measure 20-60nm colloidal silver 40mg, then slowly add colloidal silver while stirring the hydrogen peroxide solution, adjust the pH to 2.0 with 1N HCl after addition, Finally, add deionized water to 1 liter to obtain the pretreatment solution for rapid preparation of decellularized porcine intestinal submucosa (p-SIS) of this example.

[0067] 2. Preparation of cell lysate

[0068] Weigh 1g of borax and 10.3g of sodium carbonate, add 700ml of deionized water and mix well; add 200ml of pre-prepared 5M NaCl mother liquor and 40ml of 0.5M EDTA mother liquor, then slowly add 10ml of liquid NP-40(1g / ml), after the addition, adjust the pH to 11 with 1N NaOH, and finally add deionized water to 1 liter to obtain the cell lysate for rapid preparation of decellularized pig small intestinal submucosa (p-SIS) of this example. Each mother...

Embodiment 2

[0086] This example is basically the same as Example 1, except that the rapid preparation method of decellularized porcine small intestinal submucosa (p-SIS) partially omitted step 5) (that is, the step of refolding treatment).

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Abstract

The invention provides a rapid preparation of accellular porcine intestinal submucosa. The method sequentially comprises the steps of decellularization treatment performed before degreasing treatment,the degreasing treatment, and secondary decellularization treatment and renaturation treatment which are performed after the degreasing treatment. The rapid preparation of the accellular porcine intestinal submucosa not only can quickly and effectively remove all relevant antigen components of all cells and the like in tissues, but also maintains the good chemical composition and mechanical properties of extracellular matrix (ECM).

Description

Technical field [0001] The invention belongs to the technical field of preparation of medical tissue repair materials, and in particular relates to a rapid preparation method of decellularized porcine small intestinal submucosa (p-SIS) using porcine small intestine as a material. Background technique [0002] Natural biological tissue repair materials are based on the principles of regenerative medicine, molecular biology and immunology, and are extracted from natural biological tissues through a series of techniques except antigens. It is a kind of ultramicro tissue scaffold--" "Cell scaffold", after it is implanted in the human body, it can play a very good role as a scaffold, induce tissue cells to migrate to the scaffold, start the body's own repair function, and make cells gradually grow new tissues at this position to fill the damaged The missing tissue at the site completes organ tissue regeneration. [0003] The porcine small intestinal submucosa (p-SIS) tissue repair mate...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/36
CPCA61L27/3629A61L27/3687A61L27/3691A61L2430/40
Inventor 邹潮徐爱华朱征
Owner 承功(厦门)生物科技有限公司
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