Method for inducing oriented differentiation bone marrow mesenchymal stem cells into osteocytes

A bone marrow mesenchymal, directed differentiation technology, applied in the field of biomedical engineering, to achieve the effect of simple operation and accelerated interaction

Active Publication Date: 2017-09-05
SICHUAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The differentiation of MSCs is closely related to the microenvironment in which they grow, but the

Method used

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  • Method for inducing oriented differentiation bone marrow mesenchymal stem cells into osteocytes
  • Method for inducing oriented differentiation bone marrow mesenchymal stem cells into osteocytes
  • Method for inducing oriented differentiation bone marrow mesenchymal stem cells into osteocytes

Examples

Experimental program
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Effect test

Embodiment 1

[0060] Example 1 Using the method of the present invention to induce bone marrow mesenchymal stem cells

[0061] Press Ca 10 (PO 4 ) 6 (OH) 2 The molar ratio of calcium to phosphorus (1.67), under constant stirring, the 2.8mol / L diammonium hydrogen phosphate aqueous solution is added dropwise in the 2.4mol / L calcium nitrate aqueous solution, and the reaction solution is kept at 70°C, and the concentration is 25% by mass percentage. Ammonia water was used to keep the pH at 10.5. After the reaction, it was allowed to stand for 10 minutes, and then aged for 24 hours to obtain a hydroxyapatite suspension with a solid content of 34%. Then add the pore-forming agent deionized water to the hydroxyapatite suspension, and dilute it into a hydroxyapatite suspension with a solid content of 17% for later use. Using plasma spraying equipment, the nozzle adopts atomizing nozzle, and uses liquid plasma spraying method to form a coating with a thickness of 120 microns, a porosity of 50%, ...

Embodiment 2

[0062] Example 2 Using the method of the present invention to induce bone marrow mesenchymal stem cells

[0063] Press Ca 10 (PO 4 ) 6 (OH) 2 The molar ratio of calcium to phosphorus (1.67), under constant stirring, the diammonium hydrogen phosphate aqueous solution with a molar concentration of 2.8mol / L is added dropwise to the calcium nitrate aqueous solution with a molar concentration of 2.4mol / L, and the reaction solution is kept at 70°C. Ammonia water with a mass percentage concentration of 25% was used to control the pH at 10.5, and after the reaction, it was allowed to stand for 10 minutes, and then aged for 24 hours to obtain a hydroxyapatite suspension with a solid content of 34%. The pore-forming agent deionized water was added to the hydroxyapatite suspension, and it was diluted to form a hydroxyapatite suspension with a solid content of 17% for later use. Using plasma spraying equipment, the nozzle adopts an atomizing nozzle, and uses a liquid plasma spraying m...

Embodiment 3

[0064] Example 3 Using the method of the present invention to induce bone marrow mesenchymal stem cells

[0065] Press Ca 10 (PO 4 ) 6 (OH) 2 The molar ratio of calcium to phosphorus (1.67), under constant stirring, the diammonium hydrogen phosphate aqueous solution with a molar concentration of 2.8mol / L is added dropwise to the calcium nitrate aqueous solution with a molar concentration of 2.4mol / L, and the reaction solution is kept at 70°C. Ammonia water with a mass percentage concentration of 25% was used to control the pH at 10.5, and after the reaction, it was allowed to stand for 10 minutes, and then aged for 24 hours to obtain a hydroxyapatite suspension with a solid content of 34%. The hydroxyapatite suspension was dried at 70-100°C for 24h. Atmospheric plasma spraying is used to finally form a dense hydroxyapatite coating with a thickness of 120 microns on the biomedical metal substrate Ti-6Al-4V. Dissolve collagen in acetic acid solution with a concentration of 7m...

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Abstract

The invention belongs to the field of biomedical engineering, and concretely relates to a method for inducing oriented differentiation bone marrow mesenchymal stem cells into osteocytes. A simple, ecologic, safe and effective method for inducing oriented differentiation bone marrow mesenchymal stem cells into osteocytes is lacked in the prior art. The method for inducing oriented differentiation bone marrow mesenchymal stem cells into osteocytes, provided by the invention, comprises the following steps: 1, constructing a porous calcium phosphate scaffold; 2, compounding collagen to the scaffold obtained in step 1 to form a fiber network structure; and 3, inoculating the fiber network with bone marrow mesenchymal stem cells, pre-culturing the bone marrow mesenchymal stem cells for 4-48 h, and culturing the bone marrow mesenchymal stem cells in an ossification induction medium. The calcium phosphate-collagen scaffold constructed in the invention provides a suitable microenvironment, and the collagen is compounded to the calcium phosphate scaffold to realize the cascade amplification of a signal among mediated cells; and the bone marrow mesenchymal stem cells are cultured in the ossification induction medium to enhance the expression of ossification associated transcription factors and promote the differentiation of the bone marrow mesenchymal stem cells into the osteocytes.

Description

technical field [0001] The invention belongs to the field of biomedical engineering, and in particular relates to a method for inducing directional differentiation of bone marrow mesenchymal stem cells into osteoblasts. Background technique [0002] Bone marrow mesenchymal stem cells (BMSCs) are a kind of adult stem cells that can self-renew and have multi-directional differentiation potential. Under appropriate induction conditions, they can differentiate into cartilage, bone, muscle, fat, nerve, etc. Various tissue cells. MSCs have become a research hotspot in the field of basic medicine and clinical tissue and organ damage repair and regeneration, but there are still some technical problems, such as low differentiation efficiency or non-directional differentiation to form heterogeneous cells or even abnormal differentiation into tumor cells. In the application of bone tissue engineering, directional induction of MSCs to differentiate into osteoblasts is a key step in the...

Claims

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Application Information

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IPC IPC(8): C12N5/077
CPCC12N5/0654C12N2533/18C12N2533/54
Inventor 何静吴方姚瑞娟郭江龙吴尧
Owner SICHUAN UNIV
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