Growth factor controllable slow-releasing system composite multilayer membrane promoting ossification and preparation method thereof

A technology of composite multi-layer film and growth factor, applied in the direction of prosthesis, medical science, etc., can solve the problems of lack of biological activity and cell histocompatibility, lack of biological regulation function, lack of cell-specific recognition sites, etc. Achieving good attracting and receiving ability, improving cell affinity and improving biocompatibility

Inactive Publication Date: 2015-07-08
SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, most synthetic polymer materials such as polylactic acid (PLA), polycaprolactone (PCL) and polyurethane (PU) are biologically inert on the surface, do not have biological activity and cell tiss

Method used

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  • Growth factor controllable slow-releasing system composite multilayer membrane promoting ossification and preparation method thereof
  • Growth factor controllable slow-releasing system composite multilayer membrane promoting ossification and preparation method thereof
  • Growth factor controllable slow-releasing system composite multilayer membrane promoting ossification and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] Example 1 Layer-by-layer self-assembly method to construct a composite multi-layer film of growth factor controllable slow-release system that promotes ossification

[0058] 1. Preparation

[0059] S1. Using hexamethylenediamine to treat the surface of the substrate for amination treatment;

[0060] S2. Place the surface-aminated substrate in the heparin / casein solution (negatively charged macromolecule assembly solution) prepared with HEPES, and soak for 15 minutes to make the PU surface adsorb a layer of negatively charged casein and heparin; then move to Rinse 3 times in HEPES solution with a pH value of 7;

[0061] The concentration of casein in the heparin / casein solution is 0.2 mg / mL and the concentration of heparin is 4 mg / mL;

[0062] S3. Place the assembled polyanion electrolyte substrate obtained in S2 in aFGF / collagen solution (positively charged macromolecule assembly solution) and soak for 15 minutes to absorb monolayer collagen and aFGF; then move to H...

Embodiment 2

[0066] Example 2 Regulation of the content of heparin in the composite multilayer film by the number of assembled layers

[0067] The composite multilayer film was prepared according to the method in Example 1, and the content of heparin in the layer-by-layer self-assembly process was determined by the toluidine blue method.

[0068] 1. Determination of the content of heparin loaded on the composite multilayer membrane

[0069](1) Assemble according to Implementation 1. After each layer of heparin is assembled, soak the composite multilayer membrane in 500 μL of toluidine blue buffer, and shake for 30 minutes; the formula of the toluidine blue buffer is: Dissolve 0.2g NaCl, 100uL 36% concentrated hydrochloric acid and 5mg toluidine blue in 1000mL deionized water.

[0070] (2) Add 1mL of n-hexane, shaker and mix well for 20min;

[0071] (3) Remove the organic phase, remove the lower aqueous phase, add to a 96-well plate, and detect the absorbance with a UV spectrophotometer...

Embodiment 3

[0075] Example 3 Regulating the Content of Growth Factors in Multilayer Films by Adjusting the Number of Assembled Layers

[0076] The composite multilayer film was prepared according to the method of Example 1, and the successful construction of the casein / growth factor composite multilayer film was determined by fluorescence tracking after rhodamine-labeled acidic fibroblast growth factor.

[0077] 1. Use rhodamine (RD) to label acidic fibroblast growth factor (aFGF) to obtain rhodamine-labeled acidic fibroblast growth factor (RD-aFGF).

[0078] 2. RD-aFGF was substituted for unlabeled aFGF, and assembled according to Example 1. After each layer of aFGF was assembled, its fluorescence intensity was detected with a microplate reader.

[0079] 3. Repeat step 2 to complete the construction of the casein / growth factor composite multilayer film, and analyze the relationship between the fluorescence intensity and the number of assembled layers. The results are attached figure...

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Abstract

The invention discloses a surface modification method of a bio-material promoting ossification and a growth factor controllable slow-releasing system composite multilayer membrane promoting the ossification. In particular, the invention relates to a biological multilayer membrane which is constructed on the surface of the bio-material in a layer-by-layer electrostatic self-assembly manner through a layer-by-layer self-assembly technology and a growth factor slow-controllable-releasing technology with casein and heparin as a negative electric macro-molecular layer and collagen and a growth factor as a positive electric macro-molecular layer, wherein the obtain the multilayer membrane is in a structure of a circulation of a casein layer/heparin layer and a collagen/growth factor layer. In the invention, the four biological macro-molecules having excellent physico-chemical properties are deposited onto a same substrate material alternatingly, so that not only is the biological activity of the growth factor protected, but also the load amount and slow releasing of the growth factor can be controlled and adjusted. In addition, the multilayer membrane is excellent in biological mineralization promoting and ossification promoting capabilities and meanwhile can improve adhesion, growth and proliferation of mesenchymal stem cells. The multilayer membrane can be widely used in tissue repair engineering of cartilage, bone, blood vessel, nerve, cardiac valve and the like.

Description

Technical field [0001] The invention is the field of organizational engineering medical materials.More specifically, a composite multi -layer film and its preparation method involving a cricket -to -be -mixed -release system that contributes to ossification systems. Background technique [0002] Polymer biomass materials are widely used in artificial organs, tissue engineering, shape plastic surgery, and drug control due to good physical mechanical performance and chemical stability.However, most of the artificial synthetic polymer materials such as polystumin (PLA), polystone (PCL), and polyurethane (PU), etc., are biological inertia, do not have biological activity and cell tissue compatibility, lack of lack ofCell -specific identification sites also lack efficient biological regulatory functions. Therefore, biological materials that have more interaction with matrix are increasingly valued.The surface modification technology of artificially synthesized polymer biomass can not ...

Claims

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Application Information

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IPC IPC(8): A61L27/40A61L27/18A61L27/22A61L27/54
Inventor 龚逸鸿庄良婷李燕曹智楠李裕民蒋庆
Owner SUN YAT SEN UNIV
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