Preparation method of microcarrier-cell compound with induction activity and application of compound

A technology for inducing activity and complexes, applied in the field of biomedical materials, it can solve the problem of limited bone defect repair effect, low specific surface area, low effective adhesion area cell culture-related parameters, and lack of combined use of osteoblasts and fibroblasts. It can improve the content of matrix components and the degree of calcification, improve the utilization efficiency of seed cells, and improve the effect of bone defect repair.

Inactive Publication Date: 2014-08-20
FOURTH MILITARY MEDICAL UNIVERSITY
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Problems solved by technology

[0002] At present, in the process of tissue engineering bone construction, during the use of seed cells, many steps such as digestion and collection of seed cells are required to destroy the extracellular matrix when the seed cells are combined with the scaffold material, which will undoubtedly lead to the activity of the seed cells used. In addition, a single type of seed cell-osteoblast is mostly used, because it is the main and direct cell component of osteogenesis
[0003] The reason for this problem is that the scaffold materials used for tissue engineering bone construction are mostly bulky due to the need to provide sufficient bone function replacement, so their specific surface area, effective adhesion area and other cell culture-related parameters are relatively low. , which is not conducive to the in vitro cultivation of seed cells; and the scaffolds currently used for in vitro cultivation of seed cells, such as microcarriers, cannot be used for in vivo transplantation due to various reasons such as poor degradability, poor biocompatibility, and high cost, which in turn leads to the loss of seed cells. During the construction of tissue engineered bone, repeated digestion and separation steps are required
In addition, in the construction of bone tissue engineering, the current selection of seed cells...

Method used

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  • Preparation method of microcarrier-cell compound with induction activity and application of compound

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preparation example Construction

[0025] see figure 1 , the present invention has the preparation method of inducing active microcarrier-cell complex comprising the following steps:

[0026] 1) Preparation of decalcified bone matrix microparticles: Store frozen bovine cortical bone at -80°C for 24-36 hours, then freeze and pulverize; screen out particles with a particle size of less than 200um; degrease and decalcify the particles to obtain decalcified Calcium bone matrix microparticles; the specific conditions for degreasing and decalcifying treatment are that the particles are put into the degreasing mixed solution and stirred for 2-6 hours to obtain the degreased particles; wherein the mixed solution is methanol and chloroform by volume ratio (1- 3): obtained by mixing 1, and adding 1g of granules to every 3mL of the mixed solution; adding the degreased granules to 0.5mol / L hydrochloric acid solution, stirring for 48-72h, to obtain decalcified bone matrix microparticles, wherein, Add 1 g of defatted partic...

Embodiment 1

[0033]1) Preparation of decalcified bone matrix microparticles: Store frozen bovine cortical bone at -80°C for 24 hours, then freeze and pulverize; screen out particles with a particle size of less than 200um; degrease and decalcify the particles to obtain decalcified bone Substrate microparticles; the specific conditions for degreasing and decalcifying treatment are as follows: put the particles into the degreasing mixture and stir for 6 hours to obtain degreased particles; wherein the mixture is obtained by mixing methanol and chloroform at a volume ratio of 1:1, And every 3mL of the mixed solution was added with 1g of particles; the degreased particles were added to 0.5mol / L hydrochloric acid solution, and stirred for 60 hours to obtain decalcified bone matrix microparticles. Add 1 g of defatted granules to hydrochloric acid.

[0034] 2) Preparation of acellular dermal matrix microparticles: take the full-thickness skin, first carry out sterilization treatment, soak in 1mol...

Embodiment 2

[0039] 1) Preparation of decalcified bone matrix microparticles: Store frozen bovine cortical bone at -80°C for 36 hours, then freeze and pulverize; screen out particles with a particle size of less than 200um; degrease and decalcify the particles to obtain decalcified bone Substrate microparticles; the specific conditions for degreasing and decalcifying treatment are as follows: put the particles into the degreasing mixture and stir for 2 hours to obtain degreased particles; wherein the mixture is obtained by mixing methanol and chloroform at a volume ratio of 2:1, And every 3mL of the mixed solution was added with 1g of granules; the degreased granules were added to 0.5mol / L hydrochloric acid solution, stirred for 72h to obtain decalcified bone matrix microparticles. Add 1 g of defatted granules to hydrochloric acid.

[0040] 2) Preparation of acellular dermal matrix microparticles: take the full-thickness skin, first sterilize it, soak it in 1mol / L NaCl solution for 12 hour...

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Abstract

The invention discloses a preparation method of a microcarrier-cell compound with induction activity and an application of the compound. The preparation method comprises the following steps: respectively using decalcified bone matrix particles and acellular dermal matrix micro particles to culture bone marrow mesenchymal stem cells for 7-21 days, differentiating into osteoblasts and fibroblasts to obtain an osteoblast-particle compound and a fibroblast-particle compound, and then, mixing evenly. The activity of the seed cells of the prepared product are higher, the product can be used for preparing repair material for treating lacunar bone defect and segmental bone defect, the in vivo repair effect is better, and the in vivo repair effect of tissue-engineered bone for more than two times; supporting cell components are added in the construction process of the tissue-engineered bone to promote the bone formation effect of bone parenchymal cells in bone formation in vivo, the new bone strength, the matrix component content and the calcification extent can be improved for 1.5-2 times, the utilization rate of the seed cells is obviously improved, the type of the bone repair seed cells is enriched, and the bone defect repair effect is greatly improved.

Description

technical field [0001] The invention belongs to the field of biomedical materials, and specifically relates to a preparation method and application of a microcarrier-cell complex with inducible activity. Background technique [0002] At present, in the process of tissue engineering bone construction, during the use of seed cells, many steps such as digestion and collection of seed cells are required to destroy the extracellular matrix when the seed cells are combined with the scaffold material, which will undoubtedly lead to the activity of the seed cells used. In addition, a single type of seed cell-osteoblast is mostly used, because it is the main and direct cell component of osteogenesis. [0003] The reason for this problem is that the scaffold materials used for tissue engineering bone construction are mostly bulky due to the need to provide sufficient bone function replacement, so their specific surface area, effective adhesion area and other cell culture-related param...

Claims

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Application Information

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IPC IPC(8): A61L27/44A61L27/38A61L27/36A61L27/54
Inventor 邹继伟张智勇裴国献
Owner FOURTH MILITARY MEDICAL UNIVERSITY
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