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Method for preparing heterogenic bone with cell-removing matrix

A technology of acellular matrix and production method, which is applied in the field of production of acellular matrix heterogeneous bone, can solve the problems of unacceptable by patients, limited sources, difficulty in autologous tissue modeling, etc., and achieve the effect of reducing antigenicity and enriching foam

Inactive Publication Date: 2007-01-31
芮钢
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Autologous bone transplantation is the most ideal bone transplantation method, but due to the limited bone source and bone donor site, and the difficulty of autologous tissue modeling, its clinical application is limited; allogeneic bone transplantation has the risk of spreading human diseases , there is a certain immune rejection, which makes people worry about the application, and at the same time, the source is also limited
At present, the artificial bone in clinical application is expensive and expensive, and it is not easy for patients to accept it.
Acellular matrix heterogeneous bone, which is taken from animals, such as pigs and cattle, has abundant sources and is easy to obtain, process and store in large quantities. These bone tissues retain a natural dense pore structure and have a good bone growth scaffolding effect. The proliferation, differentiation, and osteogenesis of cells provide a natural three-dimensional space structure, which has good osteoinductive activity and a degradation rate that matches the growth rate of new bone. However, since xenogeneic bone antigens mainly exist in bone cells, decellularization must be performed. However, the existing decellularized treatment methods make these xenograft bones expensive, which affects the popularization and application of these bone graft materials

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] Embodiment 1, a method for making an acellular matrix xenograft bone of the present invention, which is made by the following steps:

[0018] The first step, decontamination: use ordinary detergent to wash the oil and other impurities on the bone surface of the xenograft bone, then rinse it with distilled water, and dry it to obtain a complete bone matrix.

[0019] The second step, protein removal: put the bone matrix obtained after the above decontamination step into 18% hydrogen peroxide H at room temperature or in an oven at 37°C. 2 o 2 Soak in water for 35 hours, and then rinse with double distilled water until it is clean and free of foam; the hydrogen peroxide H used 2 o 2 The concentration should not be too thick, if it exceeds 20%, it is easy to damage the strength of bones.

[0020] The third step, deesterification: at room temperature such as 25 ° C, put the bone matrix obtained after the above deproteinization step into a 2:1 chloroform:methanol mixture an...

Embodiment 2

[0026] Embodiment 2, a method for making an acellular matrix xenograft bone of the present invention, which is made by the following steps:

[0027] The first step, decontamination: use ordinary detergent to wash the oil and other impurities on the bone surface of the xenograft bone, then rinse it with distilled water, and dry it to obtain a complete bone matrix.

[0028] The second step, protein removal: put the bone matrix obtained after the above decontamination step into 15% hydrogen peroxide H at room temperature or in an oven at 37°C. 2 o 2 Soak in water for 48 hours, and then rinse with double distilled water until it is clean and free of foam.

[0029] The third step, deesterification: at room temperature such as 25 ° C, put the bone matrix obtained after the above deproteinization step into a 1:1 chloroform:methanol mixture and soak for 48 hours in a continuous shaking manner, and then use Rinse with double distilled water until there is no smell of chloroform and m...

Embodiment 3

[0033] Embodiment 3, a method for making an acellular matrix xenograft bone of the present invention, which is made by the following steps:

[0034]The first step, decontamination: use ordinary detergent to wash the oil and other impurities on the bone surface of the xenograft bone, then rinse it with distilled water, and dry it to obtain a complete bone matrix.

[0035] The second step, protein removal: put the bone matrix obtained after the above decontamination step into 20% hydrogen peroxide H at room temperature or in an oven at 37°C. 2 o 2 Soak in water for 24 hours, and then rinse with double distilled water until it is clean and free of foam.

[0036] The third step, deesterification: at room temperature such as 25°C, put the bone matrix obtained after the above deproteinization step into a 3:1 chloroform:methanol mixture and soak for 24 hours in a continuous shaking manner, and then use Rinse with double distilled water until there is no smell of chloroform and meth...

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Abstract

A process for preparing the foreign bone without cell matrix and biologic ejection includes such steps as washing with ordinary detergent, flushing with distilled water, removing protein by H2O2, flushing with distilled water, removing fat by the mixture of chloroform and methanol, flush with distilled water, removing cells by sodium lauryl sulfate, proteinase depressant and trypsase, flushing by distilled water, and sterilizing by epoxy ethane or Co-60.

Description

technical field [0001] The invention relates to a decellularized bone, in particular to a method for making a decellularized matrix heterogeneous bone. Background technique [0002] Acellular tissue matrix is ​​a new type of biological material that uses physical or chemical methods to decellularize allogeneic or heterogeneous tissues, thereby removing related antigens that cause rejection during tissue transplantation, and is used to repair damaged tissues. With the rapid development of society and the continuous improvement of transportation and mechanization, high-speed, high-energy trauma can cause serious and complex fractures, often causing bone defects and fracture nonunions. Autologous bone transplantation is the most ideal bone transplantation method, but due to the limited bone source and bone donor site, and the difficulty of autologous tissue modeling, its clinical application is limited; allogeneic bone transplantation has the risk of spreading human diseases ,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/00A61F2/28
Inventor 芮钢王者生高景恒
Owner 芮钢
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