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Calcium phosphate material marked by fluorescein isothiocyanate and preparation method thereof

A technology of fluorescein isothiocyanate and fluorescein, which is used in luminescent materials, biochemical equipment and methods, chemical instruments and methods, etc. problem, to achieve the effect of high fluorescence intensity and clear imaging

Inactive Publication Date: 2010-02-10
HUAZHONG UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the biocompatibility of quantum dots needs to be further studied; while the physical adsorption is reversible, the binding force between the adsorbed hydroxyapatite material and fluorescein isothiocyanate is small, and it is easy to delabel, which is difficult to apply to complex In vivo environment is also not conducive to the preservation of labeled calcium and phosphorus materials

Method used

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  • Calcium phosphate material marked by fluorescein isothiocyanate and preparation method thereof
  • Calcium phosphate material marked by fluorescein isothiocyanate and preparation method thereof
  • Calcium phosphate material marked by fluorescein isothiocyanate and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Select needle-shaped hydroxyapatite particles with a length of 200nm and a width of 40nm, and the process of covalently binding fluorescein isothiocyanate is as follows:

[0034] (1) Disperse 1.0 g of hydroxyapatite powder in 50 ml of toluene under magnetic stirring, and stir at a constant speed for 30 minutes;

[0035] (2) Add 0.02 g of γ-aminopropyltriethoxysilane (21 μl) to the toluene suspension in step (1) above, and keep stirring for 12 hours;

[0036] (3) centrifugal separation, centrifugal washing with absolute ethanol 3 times, and drying overnight at 50°C;

[0037] (4) Disperse 0.2 g of the γ-aminopropyltriethoxyaminosilane-modified hydroxyapatite powder obtained in the above step (3) in 20 ml of methanol, and stir magnetically for 30 min;

[0038] (5) Prepare 5-6 mg of fluorescein isothiocyanate into a 2 mg / ml methanol solution, take 1 ml and slowly add the suspension obtained in step (4) above, and continue stirring at room temperature for 3 hours;

[0039]...

Embodiment 2

[0043] Select nano-β-TCP particles with a particle size of 300nm, and the process of covalently binding fluorescein isothiocyanate is:

[0044] (1) Disperse 1.0g of β-TCP powder in 50ml of toluene under magnetic stirring, and stir at a constant speed for 30 minutes;

[0045] (2) Add 0.02 g of γ-aminopropyltriethoxysilane (21 μl) to the toluene suspension in step (1) above, and keep stirring for 12 hours;

[0046] (3) centrifugal separation, centrifugal washing with absolute ethanol 3 times, and drying overnight at 50°C;

[0047] (4) Disperse 0.2 g of the γ-aminopropyltriethoxysilane-modified β-TCP powder obtained in step (3) above in 20 ml of methanol, and stir magnetically for 30 minutes;

[0048] (5) Prepare 5-6 mg of fluorescein isothiocyanate into a 2 mg / ml methanol solution, take 1 ml and slowly add to the suspension obtained in step (4) above, and continue stirring at room temperature for 3 hours;

[0049] (6) Centrifugal separation, centrifugal washing with absolute e...

Embodiment 3

[0053] Acicular zinc-containing hydroxyapatite (ZnCa 9 (PO 4 ) 6 (OH) 2 ) particles, the process of covalently binding fluorescein isothiocyanate is:

[0054] (1) Disperse 1.0 g of hydroxyapatite powder in 50 ml of xylene under magnetic stirring, and stir at a constant speed for 30 minutes;

[0055] (2) Add 0.02 g of γ-aminopropyltrimethoxysilane (21 μl) to the xylene suspension in the above step (1), and keep stirring for 12 hours;

[0056] (3) centrifugal separation, centrifugal washing with absolute ethanol 3 times, and drying overnight at 50°C;

[0057] (4) Disperse 0.2 g of the γ-aminopropyltrimethoxysilane-modified hydroxyapatite powder obtained in step (3) above in 20 ml of ethanol, and stir magnetically for 30 min;

[0058] (5) Prepare 5-6 mg of fluorescein isothiocyanate into a 2 mg / ml ethanol solution, take 1 ml and slowly add the suspension obtained in step (4) above, and continue stirring at room temperature for 3 hours;

[0059] (6) Centrifugal separation, c...

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Abstract

The invention discloses a calcium phosphate material marked by fluorescein isothiocyanate and a preparation method thereof. In the invention, an amino-silane coupling agent is firstly adopted to carry out surface modification on the calcium phosphate material, silicon-oxygen-phosphorus (Si-O-P) covalent bond is formed on the surface of the calcium phosphate material, an amino group in the coupling agent is then utilized to covalently combine with iso-thiocyanate in fluorescein, and the stable covalence-type calcium phosphate material can be marked by fluorescein. The compounded material has high fluorescence intensity, stable chemical property and good biocompatibility, and is suitable for tracing the calcium phosphate material in cells in complex internal and external body environment. The calcium phosphate material marked by fluorescein isothiocyanate has important application in the aspects of material biocompatibility evaluation and material-cell interaction, and the like.

Description

technical field [0001] The invention belongs to the field of biological materials, in particular to a calcium phosphorus material covalently labeled with fluorescein isothiocyanate and a preparation method thereof. Background technique [0002] Artificial bones and dental calcium-phosphorus materials, such as β-tricalcium phosphate, hydroxyapatite, bioactive glass, etc., because their main components are similar to the inorganic components of natural bones and teeth, they can act as scaffolds after being implanted in organisms , and can induce new bone growth. At present, calcium phosphorus materials have been widely used in orthopedics and dentistry. [0003] Artificially synthesized nano-hydroxyapatite is an excellent hard tissue replacement material. The composite bioactive bone repair material prepared by its nano-crystals, collagen, and bone growth factors has both osteoconductive and osteoinductive effects, which can improve bone grafting. Osteogenic capacity of the ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C09K11/06C12Q1/02A61K49/00
Inventor 张胜民仇志烨刘健王深琪周磊
Owner HUAZHONG UNIV OF SCI & TECH
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