Preparation and differentiation methods of micro-nano topological structure for promoting mesenchymal stem cell differentiation

A topological structure and qualitative stem cell technology, applied in the field of micro-nano biomaterials and its application, can solve the problems of poor induction and differentiation effect, cumbersome operation process, large time consumption and workload, etc., to enhance the induction ability of osteotropic differentiation, The effect of firm binding and high proliferation rate

Inactive Publication Date: 2017-05-31
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this operation process is cumbersome and the effect of inducing differentiation is not good.
Especially when the combination of cells and bone repair scaffold materials is implanted into the body, the mesenchymal stem cells must be fully induced into osteoblasts in the early stage before they can be implanted into the body, which will consume a lot of time and workload

Method used

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  • Preparation and differentiation methods of micro-nano topological structure for promoting mesenchymal stem cell differentiation
  • Preparation and differentiation methods of micro-nano topological structure for promoting mesenchymal stem cell differentiation
  • Preparation and differentiation methods of micro-nano topological structure for promoting mesenchymal stem cell differentiation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] (1) Prepare a water-soluble silk protein solution, adjust the concentration of the silk protein solution to 1 mg / mL, and perform ultrasonic treatment for 10 min.

[0042] (2) CaCl 2 The particles were added to 100 mL of silk protein solution in step (1) for premineralization for 1 h, and the CaCl 2 completely dissolved, making the CaCl 2 The final concentration was 20mM.

[0043] (3) 100 mL of Na with a molar concentration of 12 mM 2 HPO 4The solution was slowly dripped into the solution in step (2) through a constant flow pump device for mineralization, the dropping rate was controlled at 10mL / min, and a magnetic stirrer was used to stir during the period. The mineralization temperature was controlled between 4° C., and the pH of the system was kept constant at 9.5 by dropping the solution of 0.1 M NaOH.

[0044] After the dropwise addition, the stirring was continued at a relatively fast speed for 2 hours, so that the apatite crystals were layered on the fibroin ...

Embodiment 2

[0051] (1) Prepare a water-soluble silk protein solution, adjust the silk protein solution to 1 mg / mL, and perform ultrasonic treatment for 10 min.

[0052] (2) CaCl 2 The particles were added to 100 mL of silk protein solution in step (1) for premineralization for 1 h, and the CaCl 2 completely dissolved, making the CaCl 2 The final concentration was 20 mM.

[0053] (3) 100 mL of Na with a molar concentration of 12 mM 2 HPO 4 The solution was slowly dripped into the solution in step (2) through a constant flow pump device for mineralization, the dropping rate was controlled at 10mL / min, and a magnetic stirrer was used to stir during the period. The mineralization temperature was controlled between 40° C., and the pH of the system was kept constant at 9.5 by dropping the solution of 0.1 M NaOH. Continue to stir at a faster speed for 24 hours after the dropwise addition, so that the apatite crystals are laminated on the AS template.

[0054] (4) Take out the solution in s...

Embodiment 3

[0060] (1) Prepare a water-soluble silk protein solution, adjust the silk protein solution to 1 mg / mL, and perform ultrasonic treatment for 10 min.

[0061] (2) CaCl 2 The particles were added to 100 mL of silk protein solution in step (1) for premineralization for 1 h, and the CaCl 2 completely dissolved, making the CaCl 2 The final concentration was 20 mM.

[0062] (3) 100 mL of Na with a molar concentration of 12 mM 2 HPO 4 The solution was slowly dripped into the solution in step (2) through a constant flow pump device for mineralization, the dropping rate was controlled at 10mL / min, and a magnetic stirrer was used to stir during the period. The mineralization temperature was controlled between 40° C., and the pH of the system was kept constant at 9.5 by dropping the solution of 0.1 M NaOH. After the dropwise addition, the stirring was continued at a relatively fast speed for 24 hours, so that the apatite crystals were laminated on the fibroin template.

[0063] (4) ...

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Abstract

The invention discloses preparation and differentiation methods of a micro-nano topological structure for promoting mesenchymal stem cell differentiation. A water-soluble fibroin solution is prepared, CaCl2 is added into the water-soluble fibroin solution for pre-mineralization, stirring is continuously carried out, and CaCl2 is dissolved; a Na2HPO4 solution is slowly dropped into the obtained solution through a constant flow pump device for mineralization; stirring continues to be carried out with a magnetic stirrer, so that apatite crystals are laminated on the water-soluble fibroin solution; the solution is centrifuged, supernate is poured away, sediment mineralizer is collected, deionized water is added, and the materials are mixed to be uniform; the steps are repeated, the obtained solution is dropwise added into a plastic plate or a mold, curing molding is carried out, and fibroin and apatite are compounded into a film with a micro-nano structure characteristic. The material is high in mechanical strength, micro-nano topological morphology is controllable, excellent differentiation inducibility to bone cells is shown, the material can be applied to repairing and replacing of bone defects, and the methods are used for biological material surface modification and modification and have wide application prospects.

Description

technical field [0001] The invention relates to a micro-nano biological material and its application, in particular to a preparation and differentiation method of a micro-nano topological structure for promoting the differentiation of mesenchymal stem cells, and the obtained material is applied to the repair and replacement of bone defects. [0002] technical background [0003] It is an important method to treat bone defects by simulating the main components of natural bones and using bionic methods to prepare bio-scaffold materials for the repair and replacement of bone defects. The surface properties of bone repair and bone replacement materials are of great significance to the integration of cells on the material, which can directly affect the cell behavior of cells on the material. Many studies have shown that compared with smooth or non-periodic surfaces, the nanotopological structure of the surface of materials (generally metals and synthetic polymers), such as nano-gr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/077
CPCC12N5/0654C12N2500/05C12N2501/998C12N2506/1346
Inventor 杨明英帅亚俊
Owner ZHEJIANG UNIV
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