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A kind of preparation method of natural tissue-derived cartilage combined bone decellularization material

A technology of decellularization and cartilage, which is applied in the field of preparation of cartilage combined with bone decellularization materials, achieving the effect of good extracellular microenvironment, high efficiency, convenience and uniformity

Active Publication Date: 2019-11-05
THE AFFILIATED SIR RUN RUN SHAW HOSPITAL OF SCHOOL OF MEDICINE ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, no one has reported the use of bone-cartilage decellularized materials to repair cartilage or cartilage-linked bone lesions.

Method used

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  • A kind of preparation method of natural tissue-derived cartilage combined bone decellularization material
  • A kind of preparation method of natural tissue-derived cartilage combined bone decellularization material
  • A kind of preparation method of natural tissue-derived cartilage combined bone decellularization material

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Example 1 Preparation and research of decellularized pine cartilage combined with bone material

[0039] (1) Take the porcine femoral head cartilage joint bone, transfer it to an environment at 4°C, and rinse it with sterile saline for 3 times, 20 minutes each time, to remove blood, residual muscle tissue, hair, ligament and other tissues; the size of the sample depends on the actual clinical operation It depends on the needs, usually the size is 1cm*0.5cm*1cm.

[0040] (2) In 1000ml of normal saline buffer solution containing 1% protease inhibitor (the content of protease inhibitor is 10KIU / ml) at a molar concentration of 1000ml, shake at 150rpm on a shaker at a constant temperature of 45°C for 24 hours, and rinse with normal saline for 5 hours;

[0041] (3) In 1000ml of organic solvent solution (chloroform and methanol solution with equal volume ratio), degrease with constant temperature 45°C shaker at 150rpm for 12 hours, and rinse with normal saline for 5 hours;

...

Embodiment 2

[0048] Example 2 Preparation and Research of Decellularized Cartilage Combined with Bone Material

[0049] Get pig femoral head cartilage joint bone, step (2) in 1000ml containing protease inhibitor physiological saline buffer solution (concentration is 5%, protease inhibitor content is 10KIU / ml), constant temperature 45 ℃ of shaker 150rpm shakes 8 hours; Carry out with reference to the method of Example 1 to obtain the decellularized cartilage combined with bone material.

Embodiment 3

[0050] Example 3 Preparation and Research of Acellular Cartilage Combined with Bone Material

[0051] Get the porcine femoral head cartilage joint bone, step (2) in 1000ml of the physiological saline buffer solution containing protease inhibitor (concentration is 3%, protease inhibitor content is 10KIU / ml), constant temperature 45 DEG C shaker 150rpm shakes 12 hours; Carry out with reference to the method of Example 1 to obtain the decellularized cartilage combined with bone material.

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Abstract

The invention discloses a method for preparing synchondrosis bone acellular materials from natural tissue origins.The method particularly includes treating optional synchondrosis bone tissues of mammals by the aid of normal saline buffer solution with protease inhibitors, organic solvent solution, PBS (phosphate buffer solution) with Triton X, PBS with SDS (sodium dodecyl sulfate) and PBS with DNA (deoxyribonucleic acid) enzymes to obtain the synchondrosis bone acellular materials.The method has the advantages that the optional synchondrosis bone tissues of the whole bodies of the mammals are subjected to thorough decellularization treatment, so that the synchondrosis bone acellular materials with high homogeneity can be efficiently, conveniently and quickly obtained; the integrity of the original ECM (extra cellular matrixes) can be kept while allogenic or xenogeneic cells with immunogenicity are removed, and the synchondrosis bone acellular materials can be applied to orthopedic diseases at optional positions of the whole bodies of the mammals.

Description

technical field [0001] The invention belongs to the technical field of osteochondral tissue repair and regeneration, and in particular relates to a method for preparing a cartilage-bonded decellularized material derived from natural tissue. Background technique [0002] At present, diseases such as osteoarthritis and cartilage defect are serious threats to human health, and articular cartilage repair is still a major clinical problem. For this reason, artificial biomaterials are being gradually applied to patients' cartilage tissue repair. However, due to deficiencies in biocompatibility, degradability, and difficulty in regeneration of hyaline cartilage, there is still no suitable material that can truly replace healthy cartilage tissue and realize its original physiological function requirements. In addition, cartilage damage often causes subchondral bone damage and degeneration. Osteochondral lesions are very common in clinical practice, which restricts joint activities ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61L27/36A61L27/50
CPCA61L27/3608A61L27/3612A61L27/365A61L27/3687A61L27/50A61L2430/02
Inventor 林贤丰陈家鑫张琪陈蕴缤王晟毓
Owner THE AFFILIATED SIR RUN RUN SHAW HOSPITAL OF SCHOOL OF MEDICINE ZHEJIANG UNIV
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