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Preparation method of gradient mineralized bone extracellular matrix material

A bone extracellular matrix, gradient technology, applied in medical science, prosthesis, tissue regeneration, etc., can solve the irreversible damage of natural bone ECM scaffold materials, loss of minerals and growth active factors, affecting the biological regeneration and repair activity of scaffolds, etc. problem, to achieve the effect of good biomechanical properties, rich bioactive components, and low immunogenicity

Active Publication Date: 2019-09-13
ZHEJIANG DISAI BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, since the process of bone repair needs to go through the stages of hematoma organization, callus formation, and callus shaping, and the application of biomaterials is often before and after the hematoma organization stage, which results in mature, dense, fully mineralized bone. Bone ECM material cannot be well integrated with new bone to promote regeneration
It has also been reported in the literature that completely decalcified bone matrix materials are used for the repair and regeneration of bone tissue. In the traditional treatment process, strong acid and long-term EDTA-2Na soaking decalcification [10] often cause irreversible damage to the natural bone ECM scaffold material. , and lead to the decline of biomechanical properties, the loss of minerals and growth active factors, and the change of three-dimensional microstructure, which seriously affects the bioregeneration and repair activity of scaffolds.

Method used

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  • Preparation method of gradient mineralized bone extracellular matrix material
  • Preparation method of gradient mineralized bone extracellular matrix material
  • Preparation method of gradient mineralized bone extracellular matrix material

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] Example 1 Preparation of porcine scapula specific part demineralized cancellous bone ECM material

[0052] 1) Take fresh pork scapula, wash it 4 times with sterile normal saline, take out the cancellous bone with a 6mm drill, and cut it into cylindrical bone pieces with a height of about 2mm with a scalpel;

[0053] 2) Rinse with sterile saline for 2 hours after collecting the materials, send them to the irradiation center, and sterilize them with 25w of radiation;

[0054] 3) Rinse with deionized water containing 20KIU / ml protease inhibitor for 3 times, each time for 10 minutes, to remove blood, adipose tissue and other impurities;

[0055] 4) Prepare high-temperature sterilized 1L glass bottles containing 500ml deionized water and 20 embedding boxes. Put 3 sterilized bone fragments into embedding boxes on a sterile operating table, put them into embedding boxes, put 10ml of deionized aqueous solution containing 15% acetone, and shake at a constant temperature of 10°C...

Embodiment 2

[0065] Example 2 Detection of bone ECM material with 90% and 60% mineralization degree of porcine scapula

[0066] 1) Take fresh pork scapula, wash it 4 times with sterile normal saline, take out the cancellous bone with a 6mm drill, and cut it into cylindrical bone pieces with a height of about 2mm with a scalpel;

[0067] 2) Rinse with sterile saline for 2 hours after collecting the materials, send them to the irradiation center, and sterilize them with 40w of radiation;

[0068] 3) Rinse with deionized water containing 10KIU / ml protease inhibitor for 3 times, each time for 20 minutes, to remove blood, adipose tissue and other impurities;

[0069] 4) Prepare high-temperature sterilized 1L glass bottles containing 500ml deionized water and 20 embedding boxes. Put 3 sterilized bone fragments into embedding boxes on a sterile operating table, put them into embedding boxes, put 10ml of deionized aqueous solution containing 15% acetone, and shake at a constant temperature of 10°...

Embodiment 3

[0083] Example 3 Characterization of bone ECM material with 90% and 60% mineralization degree of porcine scapula

[0084] 1) Take fresh pork scapula, wash it 4 times with sterile normal saline, take out the cancellous bone with a 6mm drill, and cut it into cylindrical bone pieces with a height of about 2mm with a scalpel;

[0085]2) Rinse with sterile saline for 2 hours after collecting the materials, send them to the irradiation center, and sterilize them with 5w of radiation;

[0086] 3) Rinse twice with deionized water containing 50KIU / ml protease inhibitor, each time for 10 minutes, to remove blood, adipose tissue and other impurities;

[0087] 4) Prepare high-temperature sterilized 1L glass bottles containing 500ml deionized water and 20 embedding boxes. Put 3 sterilized bone fragments into embedding boxes on a sterile operating table, put them into embedding boxes, put them into 10ml of deionized aqueous solution containing 10% acetone, and shake at a constant temperatu...

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Abstract

The invention discloses a gradient mineralized bone extracellular matrix material and a preparation method thereof. The method comprises the following steps of: carrying out immunogenicity removal treatment, namely decellularization on bone tissues derived from natural tissues, and carrying out gradient demineralization treatment on the obtained decellularized bone so as to obtain the gradient mineralized bone extracellular matrix material. The method enlarges the porosity of the bone matrix material and the exposure degree of collagen on the surface of the bone matrix material, effectively releases growth factors, well improves the adhesion degree of the material to cells, and regulates and controls the up-regulation of genes and proteins related to cell regeneration. The method not onlywell retains the biomechanical characteristics and the three-dimensional microstructure of a natural bone ECM scaffold, but also plays a positive role in osteogenesis, angiogenesis and collagen mineralization in the early stage of fracture, thereby increasing the colonization adhesion of cells and promoting the differentiation induction of the cells. The gradient mineralized bone extracellular matrix material derived from natural tissues exhibit greater potential in promoting mesenchymal stem cell differentiation and osteogenesis than non-demineralized or fully-demineralized bone matrix materials.

Description

technical field [0001] The invention belongs to the technical field of bone tissue repair and regeneration, and in particular relates to a gradient mineralized bone extracellular matrix material derived from natural tissue and a preparation method thereof. Background technique [0002] Bone regeneration and repair is a long process, especially for severe bone defects that are difficult to repair by themselves. At present, bone graft material has been considered as an alternative treatment with broad application prospects. Natural bone tissue, especially natural cancellous bone matrix (CBM), is rich in collagen, growth factors and other substances that promote cell growth and bone tissue repair. Not only that, but it also contains a lot of minerals (including Ca 2+ 、PO4 -3 , Mg 2+ Plasma). In recent years, more and more studies have shown that mineral ions (Ca 2+ 、PO4 -3 , Mg 2+ etc.) play an extremely important role in the regulation and promotion of new bone tissue ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/36A61L27/50
CPCA61L27/3633A61L27/3608A61L27/365A61L27/3687A61L27/3691A61L27/50A61L2300/412A61L2430/02A61L2430/40A61L27/3683A61L27/54
Inventor 林贤丰柳世杰王艺芸芦雅智范顺武
Owner ZHEJIANG DISAI BIOTECHNOLOGY CO LTD
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