II type collagen sponge bracket and uses thereof

A collagen sponge and collagen technology, which is applied in the field of biomedicine, can solve the problems of difficulty in material scaffold forming research and animal experiments, difficult to use as a cell culture scaffold material, and complicated extraction method of type II collagen, etc., achieves good repair effect, and increases three-dimensional stability. The effect of improving the elasticity and elastic modulus

Inactive Publication Date: 2013-01-16
广州市红十字会医院
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, due to the complexity of the extraction method of type II collagen, the research on the formation of material scaffolds and animal experiments are relatively difficult. At present, there are many literature reports on the extraction methods of type II collagen, but most of the extractions are mixtures or crude extracts. Chinese patent application No. 200810045418 " A preparation method of medical grade type Ⅱ collagen" and Ye Chunting, Li Siming, etc. "The development of type Ⅱ collagen and its application in chondrocyte culture" in "Chinese Journal of Orthopedic Surgery", Volume 8, No. 1, January 2001 Application" and "Preparation and Detection of High Purity Porcine Cartilage Type II Collagen" in "Journal of Biomedical Engineering" disclose the current ideal extraction method of type II collagen, which can obtain high purity type II collagen
Based on this, the extracted type II collagen is freeze-dried, and the obtained collagen sponge is easily soluble in water, which is difficult to be used as a cell culture scaffold material; there are also reports in the literature that gel type II collagen is used as a tissue engineering graft material, but it lacks Mechanical strength, quickly dissolved and absorbed by the body after transplanted into the body, because the type II collagen scaffold decomposes and absorbs too fast, the cartilage tissue has not yet formed, and it is difficult to achieve the purpose of cartilage repair

Method used

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  • II type collagen sponge bracket and uses thereof
  • II type collagen sponge bracket and uses thereof
  • II type collagen sponge bracket and uses thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] 1. Preparation of high-purity type Ⅱ collagen [Reference: Li Siming, Ye Chunting, etc., Preparation and detection of high-purity porcine cartilage type Ⅱ collagen, Journal of Biomedical Engineering, 2001, 18 (4) 592-594]:

[0032] Material:

[0033] Fresh pig articular cartilage: obtained from the limb articular cartilage of freshly slaughtered pigs in the slaughterhouse

[0034] Guanidine hydrochloride: Farco 500g / bottle

[0035] Pepsin: Sigma 100g / bottle

[0036] The rest are domestic analytical reagents.

[0037] method:

[0038] Cut the hyaline cartilage from freshly slaughtered pig limb joints, cut into thin slices, degrease, mash, and homogenate, then stir with 10 times the volume of 4M (pH7.5) guanidine hydrochloride for 24 hours, and centrifuge; the sediment diameter is fully washed , Weigh the precipitated cartilage particles with a wet weight of about 260g, hydrolyze with pepsin at a mass ratio of 1 / 50 under acidic conditions for 24 to 48 hours, centrifuge...

experiment example 1

[0044] Experimental example 1. Pore diameter detection of type II collagen sponge scaffold material

[0045] The type II collagen sponge material was prepared according to the above method, and divided into stock solution non-crosslinked group, stock solution crosslinked group, concentrated non-crosslinked group, and concentrated crosslinked group.

[0046] Collagen autofluorescence layered scanning of collagen sponges with various pore diameters was carried out by laser confocal microscope. The excitation wavelength was 488nm, and the emission wavelength was 520nm. The autofluorescence signal intensity was measured to determine the pore size of the sponge material; using CLSM image The analysis system is used for operation and analysis, and then the SPSS13.0 software package is used to make statistics on the data.

[0047] As shown in Figure 2, the laser confocal microscopy results show that the collagen sponges of each group have obvious autofluorescence, indicating that the...

experiment example 2

[0051] Experimental Example 2: Solubility Detection of Type II Collagen Scaffold Materials

[0052] The type II collagen sponge material was prepared according to the above method, which was divided into a cross-linked group and an uncross-linked group.

[0053] Cut the type II collagen sponge material into 1×1cm 2 Put it into a 6-well culture plate, add 3ml of DMEM medium, and observe the condition of the sponge at 37°C.

[0054] The results showed that the non-cross-linked type II collagen sponge material swelled after being added to the culture medium, and partially dissolved, and the material became soft and translucent; after 7 days, the material was completely dissolved. The appearance of the cross-linked sponge material remains unchanged after being added to the culture medium, and it can still maintain a complete state up to 50 days after being planted into the cells.

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Abstract

The present invention belongs to biomedicine field, relating to a II type collagen sponge bracket used in renovating cartilage of tissue engineering. The II type collagen sponge bracket of the invention is made from the extracted high-purity II type collagen solution, using polyethyleneglycol concentration method to concentrate the II type collagen solution, then using EDC and NHS dual cross linking agent to cross linkage the concentrated II type collagen to obtain. The II type collagen sponge bracket of the invention possess uniform communication aperture suitable for cell growth and preferable mechnical intensity and three-dimensional stability of aperature structure, being able to be used as bracket materials to culture together with seeds cells for a long time in water-solubility culture medium. The II type collagen sponge bracket of the invention can be used as tissue engineering implant, bracket materials of cell culture or drug delivery for application of preventing and treatingof renovating cartilage damnification.

Description

technical field [0001] The invention belongs to the field of biomedicine and relates to a type II collagen sponge support for cartilage repair in tissue engineering. Background technique [0002] There is no clinically effective treatment for cartilage defects. Tissue engineered cartilage transplantation is currently one of the latest and most promising treatment techniques for cartilage defects, that is, seed cells cultured in vitro are planted in natural or synthetic materials that can be degraded and degraded in the body. Absorbable material scaffolds, and then implant the composite into the defect in animals or humans to form tissues with cartilage-like structure and function, thereby completing the repair and reconstruction of the defect. [0003] Tissue engineering has higher requirements on scaffold materials, which determine the quality of tissue engineered cartilage. In order to find ideal cartilage graft materials, scholars at home and abroad have done a lot of re...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61L27/24A61L27/56
Inventor 陈鸿辉杨小红秦胜男
Owner 广州市红十字会医院
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