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Method for preparing collagen sponge under acid condition

A technology of collagen sponge and acidic conditions, applied in the field of biomedical materials, can solve the problems of high cost, cumbersome and incomplete operation process, and achieve the effects of reducing preparation cost, low cost and simplifying the production process

Inactive Publication Date: 2017-02-01
HANGZHOU S EVANS KETUO STEM CELL TECH RES CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the existing extraction steps of collagen, methods such as freeze-thaw, hypertonicity, liquid high pressure, ultrasound, and protease digestion are often used to achieve the purpose of decellularization, and the operation process is cumbersome and incomplete
Some even directly buy commercially available collagen powder to prepare collagen sponge, which is expensive

Method used

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  • Method for preparing collagen sponge under acid condition
  • Method for preparing collagen sponge under acid condition
  • Method for preparing collagen sponge under acid condition

Examples

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Effect test

preparation example Construction

[0033] A preparation method of collagen sponge under acidic conditions, comprising the steps of:

[0034] Step 1: Obtain the skin of a fresh mammal, shave and degrease, obtain the dermis with an electric dermatome, wash with clear water, and dry.

[0035] Step 2. Mix the dermis obtained in step 1, quicklime and water, and shake at room temperature for 1-2 hours; wherein, the mass-volume ratio of quicklime and water is 1-5g:100ml, and immerse the dermis in the solution. The functions of quicklime include killing pathogens, removing DNA, removing grease, lysing cells, and disinfecting. Quicklime treatment of mammalian skin raw materials is beneficial to obtain high-purity collagen at low cost.

[0036] Step 3: Take out the dermis layer from the solution in Step 2, wash and immerse in the eluent, shake at room temperature for 24-48 hours, change the solution every 12 hours; the eluent includes 5% Tween-80, 5% Tween-20 , 1% Triton X-100 or 1% SDS. The role of the eluent is to e...

Embodiment 1

[0045] A preparation method of collagen sponge under acidic conditions, comprising the steps of:

[0046] Step 1. Obtain the skin of fresh mammals (such as pigs and cattle), shave and degrease, obtain the dermis with an electric dermatome, wash with clear water, and dry.

[0047] Step 2. Mix the dermis obtained in step 1, quicklime, and water, and shake at room temperature for 1.5 hours; wherein, the mass-volume ratio of quicklime to water is 3g:100ml, and the dermis is immersed in the solution.

[0048] Step 3: Take out the dermis layer from the solution in Step 2, wash and immerse in the eluent 5% Tween-80, shake at room temperature for 36 hours, and change the solution every 12 hours.

[0049] Step 4, taking out the dermis layer from the eluate in Step 3, washing and freeze-drying, and breaking into granules.

[0050]Step 5. Mix the granules prepared in step 4 with 0.5 mol / L acetic acid solution containing 0.5 mg / ml pepsin, stir at 37°C for 36 hours, centrifuge at 12000 rp...

Embodiment 2

[0057] The difference between this embodiment and embodiment 1 is that the pH is adjusted to 5 in step 8, and the others are the same.

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Abstract

The invention discloses a method for preparing a collagen sponge under an acid condition. The method comprises the following steps: mixing a corium layer of mammal skin, quicklime and water, and conducting oscillating at room temperature; conducting eluting by virtue of an eluent; conducting freeze-drying and crushing the freeze-dried material into particles; mixing the particles with an acetic acid solution containing pepsase, conducting stirring and centrifuging, and taking the supernatant; conducting salting out so as to precipitate collagen and purifying the collagen; and then, dissolving the collagen in an acetic acid solution, regulating pH to 1-5, injecting the obtained material into a mould, conducting freeze-drying and cross-linking treatment, and conducting packaging and sterilizing, so that the finished collagen sponge product is obtained. According to the preparation method provided by the invention, a high-purity raw material can be directly extracted by virtue of a two-step method, and in a process of preparing the collagen sponge, rather than a rigid requirement on neutral pH of a collagen solution, the ideal collagen sponge is prepared under a low-pH condition, so that a production process is effectively simplified and preparation cost is reduced.

Description

technical field [0001] The invention relates to the technical field of biomedical materials, in particular to a method for preparing a collagen sponge under acidic conditions. Background technique [0002] Collagen is the most abundant protein in mammals, accounting for 25%-33% of the total protein in the body. Collagen is very rich in bone, cartilage, muscle bonds and skin. The structure of collagen is characterized by a triple helical structure. According to the composition and sequence of amino acids in the polypeptide chain, collagen can be subdivided into 27 types such as type I, type II, and type III. As one of the most useful biomaterials, collagen has broad application prospects in the field of biomedical materials. [0003] Collagen sponge is a new type of biomedical material made of collagen with high purity and low immunogenicity. The collagen sponge has an appropriate pore size and three-dimensional structure, allowing cells to migrate freely between the pores ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P21/06C07K14/78C07K1/14
CPCC12P21/06C07K14/78
Inventor 董伟仁李锋潘若浪戴玲华王金福
Owner HANGZHOU S EVANS KETUO STEM CELL TECH RES CO LTD
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