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Preparation method and application of bionic silicified collagen material

A technology of silicified gel and raw materials, applied in the direction of prosthesis, medical science, etc., can solve the problems of limiting the industrial application of biomimetic silicified collagen materials, dose-dependent toxicity, etc.

Active Publication Date: 2021-07-27
FOURTH MILITARY MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] The existing biomimetic silicified collagen materials were prepared in the past by using polypropylene ammonium chloride as a collagen pretreatment agent, but polypropylene ammonium chloride has dose-dependent toxicity, which limits the industrial application of biomimetic silicified collagen materials

Method used

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  • Preparation method and application of bionic silicified collagen material
  • Preparation method and application of bionic silicified collagen material
  • Preparation method and application of bionic silicified collagen material

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1: Preparation of biomimetic siliconized collagen material

[0032] Mix Silbond 40, absolute ethanol, deionized water and 37% hydrochloric acid (pH regulator) at room temperature according to the molar ratio of 1.875:396.79:12.03:0.0218 (mass ratio is 15:182.8:2.167:0.008) to obtain the volume percentage 3% orthosilicic acid solution;

[0033] Mix the above-mentioned 3% orthosilicic acid solution with 36mM (choline chloride, molecular weight: 139.62; Sigma-Aldrich, St.Louis, MO, USA) in equal volumes to prepare a pretreatment solution, and determine the pH value of the pretreatment solution Adjusted to 5.5, the concentration of orthosilicic acid adjusted to 1.5% by volume;

[0034] Trim the three-dimensionally restructured type I collagen sponge (ACE collagen, Ace Surgical Supply Co., Inc, MA, USA) into a collagen block with a diameter of 0.3 cm, rinse it with Milli-Q deionized water three times and set aside;

[0035] The four schemes shown in Table 1 were us...

Embodiment 2

[0061] Example 2: Effects of Silicified Collagen Scaffold Extraction on the Morphology and Neuropeptide Expression of Dorsal Root Ganglion Cells

[0062] 2.1 Preparation of biomimetic siliconized collagen scaffold extract

[0063] According to the method 1.4 in Example 1, soak the dried biomimetic siliconized collagen scaffold (100mg) in 10ml Tris hydrochloric acid buffer solution (PH=7.4) at 37°C, take out the material after 24 hours, and collect the buffer solution;

[0064] The above-mentioned buffer solution was mixed with Neurobasal complete medium containing 20ng / mL nerve growth factor (R&D Systems, Minneapolis, MN, USA) according to a ratio of 1:2, and used as an extract medium for the cultivation of dorsal root ganglion cells ( extract group), and the dorsal root ganglion cells of the control group were cultured with Neurobasal complete medium containing 20ng / mL nerve growth factor (control group). The Neurobasal-A complete medium configuration method is to add 0.5mM / ...

Embodiment 3

[0097] Example 3: Effects of Silicic Acid on Morphology and Neuropeptide Expression of Dorsal Root Ganglion Cells

[0098] 3.1 Isolation and culture of dorsal root ganglion cells

[0099] With embodiment 2.

[0100] 3.2 Immunofluorescence staining experiment

[0101] In order to investigate the effect of silicic acid stimulation on DRG cells, dilute an appropriate amount of sodium orthosilicate in the complete medium to prepare fresh silicon-containing medium, and the final silicon concentration is between 5 μM and 40 μM; the isolated DRG Ganglion cells were inoculated on cell slides in a 24-well plate, cultured in different concentrations of silicon-containing medium for 3 days, and immunofluorescent staining experiments were performed; the specific method was the same as in Example 2;

[0102] 3.3 Determination of the effect of silicic acid on the morphology of dorsal root ganglion cells

[0103] After the dorsal root ganglion cells were cultured in 5, 10, 20, and 40 μM s...

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Abstract

The invention discloses a preparation method and application of a bionic silicified collagen material. The preparation method disclosed by the invention comprises the step of soaking ACE collagen in a pretreatment solution to obtain a bionic silicified collagen material, wherein the pretreatment solution is composed of choline chloride and an orthosilicic acid solution. The bionic silicified collagen material constructed by the invention has good physical and chemical properties, and can promote the growth of peripheral sensory nerve axons and activate a sensory nerve mTOR signal channel to secrete Semaphorins 3A.

Description

technical field [0001] The invention relates to bionic collagen material technology, in particular to a preparation method and application of bionic silicified collagen material. Background technique [0002] The existing preparation method of biomimetic silicified collagen materials is to use polypropylene ammonium chloride as a collagen pretreatment agent, but polypropylene ammonium chloride has dose-dependent toxicity, which limits the industrial application of biomimetic silicified collagen materials. Contents of the invention [0003] Aiming at the defects or deficiencies of the prior art, the present invention provides a method for preparing a biomimetic silicified collagen material. [0004] Therefore, the preparation method provided by the present invention comprises soaking ACE collagen in a pretreatment solution to obtain a biomimetic siliconized collagen material, and the pretreatment solution is composed of choline chloride and orthosilicic acid solution. [0...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08J9/40C08L89/00A61L27/24A61L27/54A61L27/56
CPCC08J9/40A61L27/24A61L27/54A61L27/56C08J2389/00A61L2300/10A61L2300/216A61L2300/412
Inventor 焦凯牛丽娜马雨轩万美辰万千千
Owner FOURTH MILITARY MEDICAL UNIVERSITY
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