CN103738932A - Nano-hydroxyapatite and preparation method thereof - Google Patents

Abstract

The invention discloses nano-hydroxyapatite and a preparation method thereof. The particular preparation method is that a regenerated silk fibroin membrane prepared by directly dissolving fibroin fibers in formic acid/calcium salt or hexafluoroisopropanol/calcium salt solutions is used as a template, and spherical nano-hydroxyapatite is prepared by aging, filtration, water washing, alcohol washing and drying at normal temperature in a phosphate solution. The preparation method and equipment of the invention is simple and low in cost, can realize continuous operation, and thus effectively shortens the preparation period; the hydroxyapatite material prepared by the method is excellent in quality, stable in performance, and uniform in particle size, has main morphology of particles, and has a particle size of 10-500 nm.

Description

A kind of nano-hydroxyapatite and preparation method thereof

technical field

The invention relates to an inorganic nano material and a preparation method thereof, in particular to a nano-hydroxyapatite and a preparation method thereof.

Background technique

Hydroxyapatite is an important inorganic component that constitutes the hard tissue of the human body. It has good biocompatibility, bone conduction performance, high chemical stability, and does not produce harmful substances after degradation. It can promote the growth of new bone when implanted in the body. An ideal implant carrier material. The preparation methods of hydroxyapatite can be divided into dry preparation and wet preparation. Wet preparation mainly adopts chemical precipitation method, hydrothermal synthesis method and sol-gel method, etc.

With the development of nanomaterial preparation technology, the preparation of nano-hydroxyapatite has ushered in new opportunities for development. Using technologies such as biomimetic mineralization and template induction, nano-sized hydroxyapatite can be obtained at room temperature, normal pressure, and aqueous solution. stone products, thereby effectively improving product quality and reducing energy consumption.

The prior art discloses a variety of methods for preparing hydroxyapatite. For example, Chinese patent application CN102491298A discloses a method for obtaining nano-hydroxyapatite by mixing silk fibroin nanofibers with calcium sources and phosphate sources; Application CN102491299A discloses a method for preparing nano-hydroxyapatite using silk fibroin nanosphere gel as a template. This method requires silk fibroin aqueous solution to be treated under an electrostatic field. First, silk fibroin nanosphere gel is obtained, and then Mix with calcium source and phosphate source, and react to obtain nano-hydroxyapatite; Chinese patent CN101880034B discloses the preparation of a porous hydroxyapatite, which uses organic small molecular acid to form a porous structure through self-assembly, and prepares The width of the hydroxyapatite nanosheets is 50-200 nm, and the pore size of the material is 0.5-5 μm; Chinese patent application CN102070131A discloses a method for preparing hydroxyapatite by using eggshells as raw materials and using a hydrothermal synthesis process. Synthetic hydroxyapatite powder has structural characteristics similar to natural bone HA; Chinese patent application CN102167300A discloses a preparation method of hexagonal columnar micropowder hydroxyapatite material, which uses calcium carbonate and calcium hydrogen phosphate as the initial reaction According to the principle of biomineralization, hexagonal columnar micropowder hydroxyapatite powder with a particle size of 1-3 μm was synthesized by wet pulverization and hydrothermal synthesis; in addition, Neyvis Almora-Barrios et al. used collagen as a template, alkaline Calcium source and phosphate source are mixed under conditions to prepare hydroxyapatite (see: Neyvis Almora-Barrios, Nora H. De Leeuw. Cryst. Molecular Dynamics Simulation of the Early Stages of Nucleation of Hydroxyapatite at a Collagen Template Growth Des. 12( 2012) 756-763). The above methods can obtain hydroxyapatite nanomaterials, but the preparation process is cumbersome and the required raw materials are expensive, which is not suitable for large-scale production.

Contents of the invention

The purpose of the present invention is to provide a preparation method of nano-hydroxyapatite.

In order to achieve the above object, the technical scheme adopted in the present invention is: a kind of preparation method of nanometer hydroxyapatite, comprises the steps:

(a) Preparation of silk fibroin film: degumming and drying the silk to obtain silk fibroin fibers, dissolving the silk fibroin fibers to obtain a silk fibroin solution, then drying, washing with water, and drying again to prepare a regenerated silk fibroin film;

(b) Preparation of hydroxyapatite: the phosphate is formulated into a phosphate aqueous solution with a concentration of 0.05mol/L to saturation, and the pH value of the phosphate aqueous solution is adjusted to 4 to 11, and then the silk fibroin prepared in step (a) is The protein film is placed in a phosphate aqueous solution, and after standing and aging, filtering, washing with water, washing with alcohol, and drying, nano-hydroxyapatite is obtained.

In the above technical scheme, the dissolving method of the silk fibroin fiber in the step (a) is to dissolve the silk fibroin fiber with formic acid/calcium salt mixed solution or hexafluoroisopropanol/calcium salt mixed solution; the silk fibroin fiber and calcium salt The mass ratio of the salt is 99:1-10:90; wherein the calcium salt is selected from one or more of CaCl ₂ , Ca(OH) ₂ , Ca(NO ₃ ) ₂ , and Ca(OC ₂ H ₅ ) ₂ .

In the above technical solution, the mass concentration of the silk fibroin solution in the step (a) is 0.1%-60%.

In the above technical scheme, in the step (b), the phosphate is selected from one or more of (NH ₄ ) ₂ HPO ₄ , H ₃ PO ₄ , Na ₃ PO ₄ , K ₂ HPO ₄ ; adjusting the phosphate solution The reagent for the pH value is ammonia water or hydrochloric acid; the static aging reaction time is 0.5-30 days, the drying temperature is 20-120° C., and the drying time is 1-72 hours.

In the above technical solution, the particle size of the hydroxyapatite prepared in the step (b) is 10-500 nm.

The present invention also claims to protect the nano-hydroxyapatite prepared by the above method.

Due to the use of the above-mentioned technical solutions, the present invention has the following advantages compared with the prior art:

(1) The present invention has developed a new method for preparing nano-hydroxyapatite, first preparing a regenerated silk fibroin film; then using the silk fibroin film as a template to induce the formation of nano-hydroxyapatite in the solution, The raw materials used are conventional products, cheap and easy to get;

(2) The preparation method provided by the present invention is simple, the conditions are mild and controllable, and nano-hydroxyapatite with controllable size and shape can be obtained, which has positive practical significance.

Description of drawings

FIG. 1 is a SEM image of nano-hydroxyapatite in Example 1.

Detailed ways

The present invention will be further described below in conjunction with embodiment:

Embodiment one

A preparation method of nano-hydroxyapatite, comprising the steps of:

(1) Natural mulberry silk was degummed by boiling 0.05% sodium carbonate solution for 30 minutes, and the pure silk fibroin fiber was obtained after repeated 3 times; the silk fibroin fiber was dissolved in the mixed solution of formic acid/calcium chloride, Wherein the mass ratio of silk fibroin fiber and calcium chloride is 1:0.5, magnetically stirred at room temperature for 3 hours to obtain a regenerated silk fibroin solution with a mass fraction of 5%, the regenerated silk fibroin solution is formed into a film, dried, washed with water, and then dried , to obtain a regenerated silk fibroin film;

(2) Prepare (NH ₄ ) ₂ HPO ₄ into 0.5mol/L (NH ₄ ) ₂ HPO ₄ aqueous solution, and adjust the pH value of the solution to 11 with ammonia water; put the regenerated silk fibroin film into 0.5mol/L ( NH ₄ ) ₂ HPO ₄ aqueous solution, aged for 3 hours at room temperature, filtered, washed with deionized water and absolute ethanol for 10 minutes, and dried at 60°C for 72 hours to obtain nano-hydroxyapatite materials.

The scanning electron microscope image of the above-prepared nano-hydroxyapatite is shown in Figure 1; it can be seen from the figure that the hydroxyapatite is granular and has a size of 20-100 nm.

Embodiment two

(1) Natural mulberry silk was degummed by boiling with 0.05% sodium carbonate solution for 30 minutes, and the pure silk fibroin fiber was obtained after repeated 3 times; the silk fibroin fiber was dissolved in a mixed solution of formic acid/calcium nitrate, wherein The mass ratio of silk fibroin fiber and calcium nitrate is 1:1, and magnetically stirred at room temperature for 3 hours to obtain a regenerated silk fibroin solution with a mass fraction of 20%. The regenerated silk fibroin solution is formed into a film, dried, washed with water, and then dried to obtain regenerated silk fibroin film;

(2) Prepare H ₃ PO ₄ into 0.1mol/L H ₃ PO ₄ aqueous solution, and adjust the pH value of the solution to 7 with ammonia water; put the regenerated silk fibroin film into 0.1mol/L H ₃ PO ₄ aqueous solution, Stand at room temperature for aging reaction for 120 hours, filter, wash with deionized water and absolute ethanol for 10 minutes, and then dry at 120° C. for 1 hour to obtain nano-hydroxyapatite materials.

Embodiment three

(1) Natural tussah silk was degummed by boiling with 0.5% sodium carbonate solution for 30 minutes, and obtained pure silk fibroin fiber after repeating 3 times; the silk fibroin fiber solution was dissolved in hexafluoroisopropanol/calcium chloride In the mixed solution, wherein the mass ratio of silk fibroin fiber and calcium chloride is 20:1, magnetically stirred at room temperature for 6h to obtain a regenerated silk fibroin solution with a mass fraction of 0.2%, the regenerated silk fibroin solution is formed into a film, and dried , washed with water, and then dried to obtain a regenerated silk fibroin film;

(2) Prepare (NH ₄ ) ₂ HPO ₄ into 0.2mol/L (NH ₄ ) ₂ HPO ₄ aqueous solution, and adjust the pH value of the solution to 6 with hydrochloric acid; place the regenerated silk fibroin film in 0.2mol/L ( In NH ₄ ) ₂ HPO ₄ aqueous solution, stand at room temperature for aging reaction for 24 hours, filter, wash with deionized water and absolute ethanol for 10 minutes, and then dry at 90°C for 20 hours to obtain hydroxyapatite material.

Embodiment four

(1) Natural mulberry silk was degummed by boiling with 0.5% sodium carbonate solution for 30 minutes, and obtained pure silk fibroin fiber after repeating 3 times; the silk fibroin fiber solution was dissolved in hexafluoroisopropanol/calcium hydroxide In the mixed solution, wherein the mass ratio of silk fibroin fiber and calcium hydroxide is 1:2, magnetically stirred at room temperature for 6h to obtain a regenerated silk fibroin solution with a mass fraction of 20.0%, the regenerated silk fibroin solution is formed into a film, and dried , washed with water, and then dried to obtain a regenerated silk fibroin film;

(2) Prepare Na ₃ PO ₄ into saturated Na ₃ PO ₄ aqueous solution, and adjust the pH value of the solution to 8 with hydrochloric acid; put the regenerated silk fibroin film into saturated Na ₃ PO ₄ aqueous solution, and let it stand at room temperature for aging reaction After 72 hours, filter, wash with deionized water and absolute ethanol for 10 minutes, and then dry at 50° C. for 40 hours to obtain hydroxyapatite material.

Embodiment five

(1) Natural mulberry silk was degummed by boiling in 0.5% sodium carbonate solution for 30 min, and the pure silk fibroin fiber was obtained after repeated 3 times; the silk fibroin fiber solution was dissolved in hexafluoroisopropanol/Ca(OC ₂ H ₅ ) ₂ in the mixed solution, wherein the mass ratio of silk fibroin fiber and Ca(OC ₂ H ₅ ) ₂ is 1:9, magnetically stirred at room temperature for 6h, to obtain a regenerated silk fibroin solution with a mass fraction of 60.0%, The regenerated silk fibroin solution is formed into a film, dried, washed with water, and then dried to obtain a regenerated silk fibroin film;

(2) Prepare K ₂ HPO ₄ into 0.05mol/L K ₂ HPO ₄ aqueous solution, and adjust the pH of the solution to 10 with hydrochloric acid; put the regenerated silk fibroin film into 0.05mol/L K ₂ HPO ₄ aqueous solution, Stand at room temperature for aging reaction for 10 days, filter, wash with deionized water and absolute ethanol for 10 minutes, and then dry at 100°C for 10 hours to obtain hydroxyapatite material.

Claims (9)

1. a preparation method of nanometer hydroxyapatite, is characterized in that, comprises the steps: (a) Preparation of silk fibroin film: degumming and drying the silk to obtain silk fibroin fibers, dissolving the silk fibroin fibers to obtain a silk fibroin solution, then drying, washing with water, and drying again to prepare a regenerated silk fibroin film; (b) Preparation of hydroxyapatite: prepare phosphate into a phosphate aqueous solution with a concentration of 0.05mol/L~saturated, and adjust the pH value of the phosphate aqueous solution to 4~11, and then the regenerated silk prepared in step (a) The plain protein film is put into phosphate aqueous solution, and after standing and aging, filtering, washing with water, washing with alcohol, and drying, nano-hydroxyapatite is obtained. 2. the preparation method of nano-hydroxyapatite according to claim 1 is characterized in that: the dissolving mode of silk fibroin fiber in the described step (a) is to use formic acid/calcium salt mixed solution or hexafluoroisopropanol The silk fibroin fiber/calcium salt mixed solution is dissolved; the mass ratio of the silk fibroin fiber to the calcium salt is 99:1-10:90. 3. The preparation method of nano-hydroxyapatite according to claim 2, characterized in that: the calcium salt is selected from CaCl ₂ , Ca(OH) ₂ , Ca(NO ₃ ) ₂ , Ca(OC ₂ H ₅ ) One or more of ₂ . 4. The method for preparing nano-hydroxyapatite according to claim 1, characterized in that: the mass concentration of the silk fibroin solution in the step (a) is 0.1% to 60%. 5. The preparation method of nano-hydroxyapatite according to claim 1, characterized in that: in the step (b), the phosphate is selected from (NH ₄ ) ₂ HPO ₄ , H ₃ PO ₄ , Na ₃ PO ₄ , NaH ₂ PO ₄ , K ₂ HPO ₄ or one or more. 6. The preparation method of nano-hydroxyapatite according to claim 1, characterized in that: the reagent for adjusting the pH value of the phosphate aqueous solution in the step (b) is ammonia water or hydrochloric acid. 7. The preparation method of nano-hydroxyapatite according to claim 1, characterized in that: the static aging time in the step (b) is 0.5 to 30 days, the drying temperature is 20 to 120 ° C, and the drying time For 1 ~ 72h. 8. The method for preparing nano-hydroxyapatite according to claim 1, characterized in that: the particle size of the hydroxyapatite prepared in the step (b) is 10-500 nm. 9. The nano-hydroxyapatite prepared by the method for preparing nano-hydroxyapatite according to claim 1.

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