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Collagen membrane with highly oriented and crystalline collagen fiber structure and preparation method thereof

A technology of collagen fibers and collagen membranes, applied in medical science, prostheses, tissue regeneration, etc., can solve the problems of inconvenient material collection, limited orientation, mechanical inability to approach natural tendon, etc., and achieve fast and effective preparation and easy operation Effect

Active Publication Date: 2022-06-14
江苏博创生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, one of the shortcomings of the reported technology is that because the isoelectric point region is usually located at a certain position in the electrolyte, it is not convenient to obtain materials, and it is impossible to use the electrode shape to shape the collagen material (such as obtaining tubular, irregular shapes, etc.) Collagen material), the second is that its orientation is completely dependent on the electric field, and the degree of orientation is limited, so the mechanics cannot be close to the natural tendon
The third is that the distance between the two electrodes is very small, generally 1-2 mm, which is inconvenient to operate (An electrochemical fabrication process for the assembly of anisotropically oriented collagen bundles, Biomaterials 29 (2008) 3278–3288; Tenogenic Induction of Human MSCs by Anisotropically Aligned Collagen Biotextiles , Adv. Funct. Mater. 2014, 24, 5762–5770)

Method used

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  • Collagen membrane with highly oriented and crystalline collagen fiber structure and preparation method thereof
  • Collagen membrane with highly oriented and crystalline collagen fiber structure and preparation method thereof
  • Collagen membrane with highly oriented and crystalline collagen fiber structure and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] Example 1: Assembly of short-range oriented amorphous collagen materials by EDP technology

[0052] (1) Collagen solution configuration: Accurately weigh 400 mg of type I collagen into 40 mL of ultrapure water, dropwise add glacial acetic acid and stir well to promote complete dissolution of the collagen, and adjust the pH of the final solution to 3.5. Put it in a dialysis bag (M Wcut off =7.0kDa) and put it into a beaker containing 1000ml of water and 15ml of glacial acetic acid, and dialyzed at 4°C for 72h to remove small molecular impurities. A collagen viscous fluid of 10 mg / ml was obtained after dialysis. By changing the quality of the collagen raw material added, the final concentration of collagen solution can be adjusted in the range of 1-20 mg / ml. When the concentration exceeds 20 mg / ml, the solution loses fluidity.

[0053] (2) Add 100 μl / ml of hydrogen peroxide to the collagen solution described in step (1), stir evenly, centrifuge at 8000 rpm / min at 4°C to...

Embodiment 2： and preparation example I

[0060] (1) Mechanical stretching

[0061] The collagen membrane E-Col was prepared according to the method of Example 1, and it was cut into a rectangular spline with a length of 30 mm and a width of 10 mm. Immerse several rectangular splines of E-Col in ultrapure water for 5 minutes, and then use Electro-Force 3200 biodynamic tester to stretch the length of the collagen membrane to a strain of 200%, so that the internal The microfibrils are further oriented along the force direction, forming a long-range oriented collagen material. Finally, the stretched E-Col was soaked in ethanol to temporarily fix the orientation structure.

[0062] (2) Ion incubation

[0063] Carry out ion incubation of the long-range oriented collagen material in step (1): fix both ends of the strip-shaped collagen membrane material in a petri dish with tape to keep the collagen membrane subject to continuous external force without shrinking, and then place the strip on the petri dish. 0.1M PBS buffer...

Embodiment 3

[0067] Example 3: Preparation Example II of Collagen Film with Highly Oriented and Crystalline Collagen Fiber Structure

[0068] (1) Mechanical stretching

[0069] The collagen membrane E-Col was prepared according to the method of Example 1, and it was cut into a rectangular spline with a length of 20 mm and a width of 20 mm. Immerse several rectangular splines of E-Col in ultrapure water for 10 minutes, and then use Electro-Force 3200 biodynamic tester to stretch the length of the collagen membrane to a strain of 50%, so that the internal collagen membrane is stretched to 50% strain. The microfibrils are further oriented along the force direction, forming a long-range oriented collagen material. Then, the stretched E-Col was soaked in ethanol to temporarily fix the orientation structure.

[0070] (2) Ion incubation

[0071] Incubate the highly oriented collagen material in step (1): fix both ends of the strip-shaped collagen membrane material in the petri dish with tape, ke...

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Abstract

The invention belongs to the field of biomacromolecule assembly, and relates to a collagen membrane preparation method which comprises the following steps: firstly, taking a short-range oriented collagen microfiber membrane, and stretching along the length direction of the collagen membrane, so that microfibers in the collagen membrane are further oriented along the stress direction to form a highly oriented collagen material; secondly, carrying out ion incubation on the highly-oriented collagen material to induce rearrangement of an internal microfiber structure so as to form large-diameter collagen fibers in a crystalline state with D-band characteristics; and finally, carrying out chemical crosslinking through photo-crosslinking, glutaraldehyde crosslinking or genipin or polyphenol. According to the method, the long-range oriented collagen material can be obtained, and the characteristics such as appearance, microstructure, Young modulus and crystal structure are highly similar to those of natural tendons. The preparation method disclosed by the invention is simple and convenient to operate, does not need complex instruments and equipment, and can be used for quickly and effectively preparing the collagen membrane material highly similar to natural tendons or ligaments.

Description

technical field [0001] The invention belongs to the field of biological macromolecular assembly, and relates to a collagen film preparation technology with highly oriented and crystalline collagen fiber structure, the obtained collagen film and applications. Background technique [0002] Collagen is one of the most abundant proteins in vertebrates. Due to its low immunogenicity, high biocompatibility, and its ability to promote cell proliferation and wound healing, it has been widely used in various biomedical materials. Collagen molecule is a triple helix structure. Under the guidance of some endogenous signals in the body, it can be assembled in a hierarchical order, that is, starting from the triple helix structure, going through different hierarchical structures such as collagen microfibrils, collagen fibrils, and collagen fibers. The specific hierarchy of collagen molecules determines the unique properties of each specific tissue. For example, in tendons and ligaments...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/24A61L27/50
CPCA61L27/24A61L27/50A61L2430/10
Inventor 屈雪万浩然雷淼刘昌胜
Owner 江苏博创生物科技有限公司
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