CN102491298A - Preparation method for linear nanometre hydroxylapatite - Google Patents
Preparation method for linear nanometre hydroxylapatite Download PDFInfo
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- CN102491298A CN102491298A CN2011103584909A CN201110358490A CN102491298A CN 102491298 A CN102491298 A CN 102491298A CN 2011103584909 A CN2011103584909 A CN 2011103584909A CN 201110358490 A CN201110358490 A CN 201110358490A CN 102491298 A CN102491298 A CN 102491298A
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Abstract
The invention provides a preparation method for linear nanometre hydroxylapatite. The preparation method comprises the following steps of: concentrating silk fibroin aqueous solution of a first mass concentration to a second mass concentration, and then performing sealed cultivation to acquire the silk fibroin nanofibre aqueous solution; mixing the silk fibroin nanofiber aqueous solution with a phosphate radical source and a calcium source for reaction to acquire the linear nanometre hydroxylapatite. The invention further provides a preparation method for the nanometre hydroxylapatite. The method comprises the following steps of: firstly, concentrating silk fibroin aqueous solution of a first mass concentration to a second mass concentration, and then performing a sealed cultivation to the mixture to acquire the silk fibroin aqueous solution, wherein in the process, the silk fibroin is self-organized to be silk fibroin nanofiber, so that silk fibroin nanofiber aqueous solution is acquired ; and using the silk fibroin nanofiber aqueous solution as a template to synthetize nanometre hydroxylapatite on the surface of the silk fibroin nanofiber. The silk fibroin interacts with the hydroxylapatite, so that the union of the hydroxylapatite cannot be restrained, but also, the linear nanometre hydroxylapatite can be acquired.
Description
Technical field
The invention belongs to technical field of nano material, relate in particular to a kind of preparation method of linear nano Win 40350.
Background technology
Silk fibroin is the structural protein that a kind of occurring in nature extensively exists; Can from the domestic silkworm silk fiber, extract; Not only the source is sufficient, process for extracting is simple; And have adhesion, amplification and the Differentiation of excellent biological compatibility, biodegradability, good mechanical performance and good pair cell, be the good organic substrate material of a kind of development prospect.Silk fibroin can be prepared into various ways such as solution, film, porous support, gel, has a good application prospect at biomedical sector.
Win 40350 (Hydroxyapatite, HA, Ca
10(PO
4)
6(OH)
2) be the weakly alkaline calcium phosphate salt that are slightly soluble in water, belong to hexagonal system, be one type of inorganic materials with good biological activity, biocompatibility and absorption property.In vivo; Win 40350 since with organic substrate generation special combination such as a small amount of collagen, chitosan; Formed better, the different inorganic/organic composite material of inducibility with medium property; Biological intravital Win 40350 not only has special mechanical property, optical property and complicated pattern, and in the biomineralization process, and biological physical efficiency is controlled the mineralisation process of Win 40350 on from nanometer to micron-sized yardstick accurately.In recent years; The research of relevant biomimetic mineralization is very noticeable; Its major cause is that not only this field is in the joint of Life Science and Inorganic Chemistry, biophysics and Materials science; Characteristics with tangible subject crossing and infiltration, it provides new thinking for crystalline material and biological intelligence material that synthetic has unique exquisite pattern what is more important, and energy consumption is lower in the building-up process; Meeting the Materials science requirements of green environmental protection, is one of focus of present materialogy research.
The Win 40350 of line style has important purposes substituting of tooth and long bone; Particularly with the nature bone linear nano Win 40350 similar with the nanostructure in the dental tissue; Having better promotion bone and dental tissue regenerated ability than common Win 40350, is the ideal material that bone and tooth are repaired.At present, prepare the method that the linear nano Win 40350 mainly adopts magnetic field, electric field and template, wherein; Adopt the preparation method of magnetic field, electric field technology to have energy consumption height, the harsh problem of preparation condition; The development of bionic technology has brought new opportunity to develop for the preparation of hydroxy apatite powder, utilizes the bionic technology, can in normal temperature, normal pressure, water solution system, obtain the Win 40350 product of size pattern homogeneous; Thereby effectively improve the quality of products, cut down the consumption of energy.
Prior art discloses the multiple report that utilizes bionics techniques preparation orientation Win 40350, is published in document " the Bacteriophage Bundles with Prealigned Ca of " CHEMISTRY OF MATERIALS " in 2010 like investigators such as FukeWang
2+Initiatethe Oriented Nucleationand Growth of Hydroxylapatite " in point out, utilize phage bundle to do template, can form the Win 40350 of orientation, but phage is not suitable for directly being used for tissue repair; Investigators such as Anton Ficai were published in the document " Self-assembled collagen/hydroxyapatite composite materials " of " Chemical Engineering Journal " and point out in 2010; On the collagen of orientations, carry out mineralising and handle the Win 40350 that can form orientation; But it utilizes collagen as template; Cost an arm and a leg, and be not suitable for large-scale production.
Summary of the invention
In view of this, the technical problem that the present invention will solve is to provide a kind of preparation method of linear nano Win 40350, and preparation method provided by the invention is simple, mild condition, can obtain dispersiveness linear nano Win 40350 preferably.
The invention provides a kind of preparation method of linear nano Win 40350, may further comprise the steps:
Seal cultivation after the silk fibroin water solution of first mass concentration is concentrated into second mass concentration, obtain the silk fibroin nano-fiber aqueous solution;
The said silk fibroin nano-fiber aqueous solution is mixed with phosphoric acid root and calcium source, obtain the linear nano Win 40350 after the reaction.
Preferably, said first mass concentration is 0.1%～10%, and said second mass concentration is 10%～50%.
Preferably, said spissated temperature is 0 ℃～90 ℃.
Preferably, the temperature that said sealing is cultivated is 0 ℃～70 ℃, and the time that said sealing is cultivated is 1 day～20 days.
Preferably, said phosphoric acid root is phosphoric acid, sodium phosphate or calcium phosphate.
Preferably, said calcium source is calcium hydroxide or nitrocalcite.
Preferably, the temperature of said reaction is 50 ℃～80 ℃.
The present invention also provides a kind of preparation method of nanometer hydroxyapatite, may further comprise the steps:
Silk fibroin water solution is dried to fibroin protein film, said fibroin protein film is soluble in water, repeat drying-dissolution process, obtain silk fibroin nano-fiber solution;
Said silk fibroin nano-fiber solution is mixed with phosphoric acid root and calcium source, obtain the linear nano Win 40350 after the reaction.
Preferably, said exsiccant temperature is 0 ℃～60 ℃.
Preferably, the number of times of said repetition drying-dissolution process is 1 time～5 times.
Compared with prior art; The present invention seals cultivation after at first the silk fibroin water solution of first mass concentration being concentrated into second mass concentration; In this process, silk fibroin is self-assembled into silk fibroin nano-fiber, obtains the silk fibroin nano-fiber aqueous solution; Be template with this silk fibroin nano-fiber aqueous solution then; The synthesis of nano Win 40350 on the silk fibroin nano-fiber surface; Silk fibroin and Win 40350 interact, and not only can suppress the reunion of Win 40350, and can access the linear nano Win 40350.Preparing method provided by the invention need not to use noxious solvent, the preparation method is simple, mild condition is controlled; Can not destroy the secondary structure of silk fibroin; Can obtain the silk fibroin nano-fiber of controllable size, thereby obtain the linear nano Win 40350 of controllable size; Silk fibroin and nanometer hydroxyapatite interact, and can realize the hydroxyapatite crystal growth and assemble the control of behavior, the linear nano Win 40350 emulsion that acquisition can stable existence; The linear nano hydroapatite particles that obtains after said emulsion is centrifugal, washing, the drying can be scattered in the water once more, can not form reunion, helps the further utilization of linear nano Win 40350.
Description of drawings
The AFM analysis of spectra of the silk fibroin nano-fiber solution that Fig. 1 provides for the embodiment of the invention;
The infrared spectrogram of the silk fibroin nano-fiber solution that Fig. 2 provides for the embodiment of the invention;
The stereoscan photograph of the linear nano Win 40350 that Fig. 3 provides for the embodiment of the invention;
The infrared spectrum of the linear nano Win 40350 that Fig. 4 provides for the embodiment of the invention.
Embodiment
The invention provides a kind of preparation method of linear nano Win 40350, may further comprise the steps:
Seal cultivation after the silk fibroin water solution of first mass concentration is concentrated into second mass concentration, obtain the silk fibroin nano-fiber aqueous solution;
The said silk fibroin nano-fiber aqueous solution is mixed with phosphoric acid root and calcium source, obtain the linear nano Win 40350 after the reaction.
The present invention seals cultivation after at first the silk fibroin water solution of first mass concentration being concentrated into second mass concentration, and in this process, silk fibroin is self-assembled into silk fibroin nano-fiber, obtains the silk fibroin nano-fiber aqueous solution; Be template with this silk fibroin nano-fiber aqueous solution then; The synthesis of nano Win 40350 on the silk fibroin nano-fiber surface; Silk fibroin and Win 40350 interact, and not only can suppress the reunion of Win 40350, and can access the linear nano Win 40350.
The present invention seals cultivation after at first the silk fibroin water solution of first mass concentration being concentrated into second mass concentration, obtains the silk fibroin nano-fiber aqueous solution.Said silk fibroin water solution is preferably according to following method preparation:
Silk is handled in sodium carbonate solution, obtained silk fibroin;
Said silk fibroin is dissolved in the solvent, in the aqueous solution, dialyses then, obtain silk fibroin water solution.
Silk is handled in sodium carbonate solution, and the sericin that silk is outside is removed, and obtains silk fibroin.The mass concentration of said sodium carbonate solution is preferably 0.1%～1%, and more preferably 0.3%～0.8%; Said treatment temperature is preferably 80 ℃～120 ℃, more preferably 90 ℃～110 ℃; The time of said processing is preferably 40min～80min, more preferably 50min～70min; Said processing is specially boiling, and in boiling process, outside sericin and the yellow soda ash of silk reacts, thereby separates with silk fibroin, obtains silk fibroin.
After disposing, use deionized water to wash the product that obtains repeatedly, obtain silk fibroin 60 ℃ of following dryings then.After obtaining silk fibroin, said silk fibroin is dissolved in the solvent, is contained in the dialysis tubing then and in deionized water, dialyses, obtain silk fibroin water solution.The solvent that said solvent is preferably lithium-bromide solution or is made up of calcium chloride, water and ethanol.Dialysis time is preferably 3 days～and 4 days, whenever between dialysis period change primary water at a distance from 2h, obtain silk fibroin water solution.
In the present invention, first mass concentration of said silk fibroin water solution is preferably 0.1%～10%, and more preferably 0.5%～8%, most preferably be 1%～5%; Said second mass concentration is preferably 10%～50%, and more preferably 15%～45%, most preferably be 20%～40%.The present invention does not have particular restriction to said spissated method, is preferably to adopt the method for evaporating solvent to concentrate, and said spissated temperature is preferably 0 ℃～90 ℃, more preferably 4 ℃～85 ℃, most preferably is 5 ℃～80 ℃.In concentration process, silk fibroin is orientated, and forms linear structure.
After said silk fibroin water solution is concentrated into second concentration; It is sealed cultivation, and when sealing was cultivated, silk fibroin was self-assembled into and is nanofiber; Obtain the silk fibroin nano-fiber aqueous solution; The temperature that said sealing is cultivated is preferably 0 ℃～70 ℃, more preferably 5 ℃～65 ℃, most preferably is 10 ℃～60 ℃; The time that said sealing is cultivated is preferably 1 day～and 20 days, more preferably 2 days～18 days, most preferably be 3 days～15 days.
After obtaining the silk fibroin nano-fiber aqueous solution; Synthesize the line style nanometer hydroxyapatite with this aqueous solution as template solution; Be about to the said silk fibroin nano-fiber aqueous solution and mix, obtain nanometer hydroxyapatite after the reaction, specifically may further comprise the steps with phosphoric acid root and calcium source:
The said silk fibroin nano-fiber aqueous solution is scattered in the phosphoric acid root aqueous solution, obtains mixing solutions, said mixing solutions is joined in the aqueous solution of calcium source, obtain the Win 40350 emulsion after the reaction;
With obtaining the linear nano Win 40350 after said Win 40350 separation of emulsions, washing, the drying.
The said silk fibroin nano-fiber aqueous solution is scattered in the phosphoric acid root aqueous solution, obtains mixing solutions; Said phosphoric acid root is preferably phosphoric acid, sodium phosphate or calcium phosphate, more preferably phosphoric acid; The volumetric molar concentration of phosphorus is preferably 0.01mol/L～0.1mol/L in the said phosphoric acid root, more preferably 0.02mol/L～0.08mol/L.
With said mixing solutions and calcium source aqueous solution, said phosphoric acid root and said calcium source react, and form hydroxyapatite crystal at the silk fibroin nano-fiber particle surface, thereby obtain line style Win 40350 emulsion; Said calcium source is preferably calcium hydroxide or nitrocalcite, more preferably calcium hydroxide; The temperature of said reaction is preferably 50 ℃～80 ℃, more preferably 60 ℃～70 ℃.
In the present invention, said silk fibroin is preferably 3: 7 with the mass ratio of the line style Win 40350 that obtains～and 9: 1, more preferably 4: 6～8: 2, most preferably be 4: 5～7: 3.
In said Win 40350 emulsion; Silk fibroin can interact with its surperficial hydroxyapatite crystal; Win 40350 can be dispersed in the silk fibroin macromolecule network, thereby suppresses the reunion of Win 40350, make said Win 40350 emulsion can be in water stable existence; Help the homodisperse of line style Win 40350 in other matrix, obtain stay-in-grade, as to be used as tooth or bone renovating material matrix material.
After obtaining the Win 40350 emulsion, with obtaining the linear nano Win 40350 after said separation of emulsions, washing, the drying.Said separation is preferably spinning, and the rotating speed during spinning is preferably more than the 10000rpm, more preferably more than the 11000rpm; Said washing is preferably water and washs, and washing times is preferably more than 3 times; Said drying is preferably vacuum-drying.
In the present invention, also can may further comprise the steps as the synthetic line style nanometer hydroxyapatite of template solution with the silk fibroin nano-fiber aqueous solution:
The said silk fibroin nano-fiber aqueous solution is scattered in the aqueous solution of calcium source, obtains mixing solutions, the phosphoric acid root aqueous solution is joined in the said mixing solutions, obtain the Win 40350 emulsion after the reaction;
With obtaining the linear nano Win 40350 after said Win 40350 separation of emulsions, washing, the drying.
Except the addition sequence of raw material, this method and aforesaid method are basic identical.
Obtain the linear nano Win 40350, said nanometer hydroxyapatite is carried out electron-microscope scanning, the result shows that it has linear structure.
After obtaining the linear nano Win 40350, it is scattered in the water once more, good dispersion property can not form reunion.
The linear nano Win 40350 that the present invention obtains has with the nature bone structure similar with the inorganic components in the tooth, can be used as tissue renovation material or pharmaceutical carrier etc. and is used for biomedical sector.
The present invention seals cultivation after at first the silk fibroin water solution of first mass concentration being concentrated into second mass concentration, and in this process, silk fibroin is self-assembled into silk fibroin nano-fiber, obtains the silk fibroin nano-fiber aqueous solution; Be template with this silk fibroin nano-fiber aqueous solution then; The synthesis of nano Win 40350 on the silk fibroin nano-fiber surface; Silk fibroin and Win 40350 interact, and not only can suppress the reunion of Win 40350, and can access the linear nano Win 40350.Preparing method provided by the invention need not to use noxious solvent, the preparation method is simple, mild condition is controlled; Can not destroy the secondary structure of silk fibroin; Can obtain the silk fibroin nano-fiber of controllable size, thereby obtain the linear nano Win 40350 of controllable size; Silk fibroin and nanometer hydroxyapatite interact, and can realize the hydroxyapatite crystal growth and assemble the control of behavior, the linear nano Win 40350 emulsion that acquisition can stable existence; The linear nano hydroapatite particles that obtains after said emulsion is centrifugal, washing, the drying can be scattered in the water once more, can not form reunion, helps the further utilization of linear nano Win 40350.
The present invention also provides a kind of preparation method of nanometer hydroxyapatite, may further comprise the steps:
Silk fibroin water solution is dried to fibroin protein film, said fibroin protein film is soluble in water, repeat drying-dissolution process, obtain silk fibroin nano-fiber solution;
Said silk fibroin nano-fiber solution is mixed with phosphoric acid root and calcium source, obtain the linear nano Win 40350 after the reaction.
In this technical scheme; Make silk fibroin water solution at first be dried to fibroin protein film, then that the fibroin protein film that obtains is soluble in water, repeat drying-dissolution process; In this process; The silk fibroin self-assembly obtains silk fibroin nano-fiber solution, is the linear nano Win 40350 that template prepares good dispersion property with this silk fibroin nano-fiber.In this technical scheme, obtain the silk fibroin nano-fiber gel except silk fibroin water solution being repeated drying-dissolution process, other, like the concentration of silk fibroin water solution, phosphoric acid root, calcium source etc., all identical with technique scheme.
The present invention repeats drying and forming-film-dissolved operation with silk fibroin water solution, and multiplicity is preferably 1 time～5 times, more preferably 2 times～3 times.The present invention does not have particular restriction to said drying and forming-film and dissolution process, and said exsiccant temperature is preferably 0 ℃～60 ℃, more preferably 4 ℃～55 ℃, most preferably is 5 ℃～50 ℃.After repeating drying-dissolved process, obtain silk fibroin nano-fiber at last.
The present invention at first repeats drying-dissolution process with silk fibroin water solution, makes the silk fibroin self-assembly, obtains the silk fibroin nano-fiber aqueous solution; Be template with this silk fibroin nano-fiber aqueous solution then; The synthesis of nano Win 40350 on the silk fibroin nano-fiber surface; Silk fibroin and Win 40350 interact, and not only can suppress the reunion of Win 40350, and can access the linear nano Win 40350.Preparing method provided by the invention need not to use noxious solvent, the preparation method is simple, mild condition is controlled; Can not destroy the secondary structure of silk fibroin; Can obtain the silk fibroin nano-fiber of controllable size, thereby obtain the linear nano Win 40350 of controllable size; Silk fibroin and nanometer hydroxyapatite interact, and can realize the hydroxyapatite crystal growth and assemble the control of behavior, the linear nano Win 40350 emulsion that acquisition can stable existence; The linear nano hydroapatite particles that obtains after said emulsion is centrifugal, washing, the drying can be scattered in the water once more, can not form reunion, helps the further utilization of linear nano Win 40350.
In order to further specify the present invention, the preparation method of nanometer hydroxyapatite provided by the invention is described in detail below in conjunction with embodiment.
's 0.5% Na with silk in mass concentration
2CO
3100 ℃ are boiled 60min in the solution, remove the outside sericin of silk; The product that uses deionized water rinsing to obtain repeats 3 times, obtains silk fibroin after 60 ℃ of dryings;
27 gram silk fibroins are dissolved in the 100mL lithium-bromide solution, the silk fibroin protein solution that obtains are contained in dialysis tubing are immersed in the deionized water dialysis 3 days, during every two hours change primary water, obtain purified silk fibroin protein solution, its mass concentration is 5.9%;
Said silk fibroin protein solution room temperature under stink cupboard is slowly dry, silk fibroin concentration is brought up to 25%, be placed into 4 ℃ then and cultivated 2 days, obtain silk fibroin nano-fiber solution;
Said silk fibroin nano-fiber solution is carried out AFM analysis and IR spectroscopy; The result is referring to Fig. 1 and Fig. 2; The AFM analysis of spectra of the silk fibroin nano-fiber solution that Fig. 1 provides for the embodiment of the invention; The infrared spectrogram of the silk fibroin nano-fiber solution that Fig. 2 provides for the embodiment of the invention can know that by Fig. 1 and Fig. 2 the present invention has obtained silk fibroin nano-fiber.
Said silk fibroin nano-fiber solution is dispersed in the phosphate aqueous solution that phosphorus acid ion concentration is 0.06mol/L obtains mixing solutions, and the adjustment silk fibroin with synthetic after the Win 40350 mass ratio be 6: 4; Measure the said mixing solutions of 20mL and be contained in the beaker, said mixing solutions is added drop-wise in the calcium hydroxide aqueous solution, obtain the Win 40350 emulsion with the speed of 180mL/60min; Said Win 40350 emulsion is at room temperature placed, but its stable existence; After said emulsion is centrifugal, washing, the drying, obtain nano-hydroapatite particles.
Said hydroapatite particles is carried out electron-microscope scanning and IR spectroscopy; The result is referring to Fig. 3 and Fig. 4; The stereoscan photograph of the nanometer hydroxyapatite that Fig. 3 provides for the embodiment of the invention; The infrared spectrum of the nanometer hydroxyapatite that Fig. 4 provides for the embodiment of the invention can know that by Fig. 3 and Fig. 4 the present invention has prepared the linear nano Win 40350.
Said nano-hydroapatite particles is scattered in the water once more, and good dispersion property can not form reunion.
Embodiment 2
's 0.5% Na with silk in mass concentration
2CO
3100 ℃ are boiled 60min in the solution, remove the outside sericin of silk; The product that uses deionized water rinsing to obtain repeats 3 times, obtains silk fibroin after 60 ℃ of dryings;
27 gram silk fibroins are dissolved in the 100mL lithium-bromide solution, the silk fibroin protein solution that obtains are contained in dialysis tubing are immersed in the deionized water dialysis 3 days, during every two hours change primary water, obtain purified silk fibroin protein solution, its mass concentration is 5.9%;
Said silk fibroin protein solution room temperature under stink cupboard is slowly dry, silk fibroin concentration is brought up to 40%, be placed into 4 ℃ then and cultivated 18 days, obtain silk fibroin nano-fiber solution;
Said silk fibroin nano-fiber solution is carried out AFM analysis and IR spectroscopy, and the result shows that the present invention has obtained silk fibroin nano-fiber.
Said silk fibroin nano-fiber solution is dispersed in the phosphate aqueous solution that phosphorus acid ion concentration is 0.06mol/L obtains mixing solutions, and the adjustment silk fibroin with synthetic after the Win 40350 mass ratio be 3: 7; Measure the said mixing solutions of 20mL and be contained in the beaker, said mixing solutions is added drop-wise in the calcium hydroxide aqueous solution, obtain the Win 40350 emulsion with the speed of 180mL/60min; Said Win 40350 emulsion is at room temperature placed, but its stable existence; After said emulsion is centrifugal, washing, the drying, obtain nano-hydroapatite particles.
Said hydroapatite particles is carried out electron-microscope scanning and IR spectroscopy, and the result shows that the present invention has prepared the linear nano Win 40350.
Said nano-hydroapatite particles is scattered in the water once more, and good dispersion property can not form reunion.
Embodiment 3
's 0.5% Na with silk in mass concentration
2CO
3100 ℃ are boiled 60min in the solution, remove the outside sericin of silk; The product that uses deionized water rinsing to obtain repeats 3 times, obtains silk fibroin after 60 ℃ of dryings;
27 gram silk fibroins are dissolved in the 100mL lithium-bromide solution, the silk fibroin protein solution that obtains are contained in dialysis tubing are immersed in the deionized water dialysis 3 days, during every two hours change primary water, obtain purified silk fibroin protein solution, its mass concentration is 5.9%;
Said silk fibroin protein solution 60 ℃ of slow down dryings, is brought up to 15% with silk fibroin concentration, be placed into 70 ℃ then and cultivated 18 days, obtain silk fibroin nano-fiber solution;
Said silk fibroin nano-fiber solution is carried out AFM analysis and IR spectroscopy, and the result shows that the present invention has obtained silk fibroin nano-fiber.
Said silk fibroin nano-fiber solution is dispersed in the phosphate aqueous solution that phosphorus acid ion concentration is 0.06mol/L obtains mixing solutions, and the adjustment silk fibroin with synthetic after the Win 40350 mass ratio be 8: 2; Measure the said mixing solutions of 20mL and be contained in the beaker, said mixing solutions is added drop-wise in the calcium hydroxide aqueous solution, obtain the Win 40350 emulsion with the speed of 180mL/60min; Said Win 40350 emulsion is at room temperature placed, but its stable existence; After said emulsion is centrifugal, washing, the drying, obtain nano-hydroapatite particles.
Said hydroapatite particles is carried out electron-microscope scanning and IR spectroscopy, and the result shows that the present invention has prepared the linear nano Win 40350.
Said nano-hydroapatite particles is scattered in the water once more, and good dispersion property can not form reunion.
Embodiment 4
's 0.5% Na with silk in mass concentration
2CO
3100 ℃ are boiled 60min in the solution, remove the outside sericin of silk; The product that uses deionized water rinsing to obtain repeats 3 times, obtains silk fibroin after 60 ℃ of dryings;
27 gram silk fibroins are dissolved in the 100mL lithium-bromide solution, the silk fibroin protein solution that obtains are contained in dialysis tubing are immersed in the deionized water dialysis 3 days, during every two hours change primary water, obtain purified silk fibroin protein solution, its mass concentration is 5.9%;
Said silk fibroin protein solution room temperature under stink cupboard is slowly dry, obtain the silk fibroin film; Said silk fibroin Film Fractionation in water, behind the drying and forming-film again, redissolve-drying three times, is dissolved in the water the film that obtains again, obtains silk fibroin nano-fiber solution;
Said silk fibroin nano-fiber solution is carried out AFM analysis and IR spectroscopy, and the result shows that the present invention has obtained silk fibroin nano-fiber.
Said silk fibroin nano-fiber solution is dispersed in the phosphate aqueous solution that phosphorus acid ion concentration is 0.06mol/L obtains mixing solutions, and the adjustment silk fibroin with synthetic after the Win 40350 mass ratio be 6: 4; Measure the said mixing solutions of 20mL and be contained in the beaker, said mixing solutions is added drop-wise in the calcium hydroxide aqueous solution, obtain the Win 40350 emulsion with the speed of 180mL/60min; Said Win 40350 emulsion is at room temperature placed, but its stable existence; After said emulsion is centrifugal, washing, the drying, obtain nano-hydroapatite particles.
Said hydroapatite particles is carried out electron-microscope scanning and IR spectroscopy, and the result shows that the present invention has prepared the linear nano Win 40350.
Said nano-hydroapatite particles is scattered in the water once more, and good dispersion property can not form reunion.
Embodiment 5
's 0.5% Na with silk in mass concentration
2CO
3100 ℃ are boiled 60min in the solution, remove the outside sericin of silk; The product that uses deionized water rinsing to obtain repeats 4 times, obtains silk fibroin after 60 ℃ of dryings;
27 gram silk fibroins are dissolved in the 100mL lithium-bromide solution, the silk fibroin protein solution that obtains are contained in dialysis tubing are immersed in the deionized water dialysis 3 days, during every two hours change primary water, obtain purified silk fibroin protein solution, its mass concentration is 5.9%;
Said silk fibroin protein solution is dry under 50 ℃, obtain the silk fibroin film; With said silk fibroin Film Fractionation in water, repeat drying-dissolving 2 times after, obtain silk fibroin nano-fiber solution;
Said silk fibroin nano-fiber solution is carried out AFM analysis and IR spectroscopy, and the result shows that the present invention has obtained silk fibroin nano-fiber.
Said silk fibroin nano-fiber solution is dispersed in the phosphate aqueous solution that phosphorus acid ion concentration is 0.06mol/L obtains mixing solutions, and the adjustment silk fibroin with synthetic after the Win 40350 mass ratio be 7: 3; Measure the said mixing solutions of 20mL and be contained in the beaker, said mixing solutions is added drop-wise in the calcium hydroxide aqueous solution, obtain the Win 40350 emulsion with the speed of 180mL/60min; Said Win 40350 emulsion is at room temperature placed, but its stable existence; After said emulsion is centrifugal, washing, the drying, obtain nano-hydroapatite particles.
Said hydroapatite particles is carried out electron-microscope scanning and IR spectroscopy, and the result shows that the present invention has prepared the linear nano Win 40350.
Said nano-hydroapatite particles is scattered in the water once more, and good dispersion property can not form reunion.
Embodiment 6
's 0.5% Na with silk in mass concentration
2CO
3100 ℃ are boiled 60min in the solution, remove the outside sericin of silk; The product that uses deionized water rinsing to obtain repeats 3 times, obtains silk fibroin after 60 ℃ of dryings;
27 gram silk fibroins are dissolved in the 100mL lithium-bromide solution, the silk fibroin protein solution that obtains are contained in dialysis tubing are immersed in the deionized water dialysis 3 days, during every two hours change primary water, obtain purified silk fibroin protein solution, its mass concentration is 5.9%;
Said silk fibroin protein solution is concentrated into 30% under 70 ℃, is placed into 4 ℃ of sealings then and cultivated 18 days, obtain silk fibroin nano-fiber solution;
Said silk fibroin nano-fiber solution is carried out AFM analysis and IR spectroscopy, and the result shows that the present invention has obtained silk fibroin nano-fiber.
Said silk fibroin nano-fiber solution is dispersed in the calcium hydroxide aqueous solution that calcium ion concn is 0.02mol/L obtains mixing solutions, and the adjustment silk fibroin with synthetic after the Win 40350 mass ratio be 3: 7; Measure the said mixing solutions of 20mL and be contained in the beaker, phosphoric acid is added drop-wise in the said mixing solutions, obtain the Win 40350 emulsion with the speed of 180mL/60min; Said Win 40350 emulsion is at room temperature placed, but its stable existence; After said emulsion is centrifugal, washing, the drying, obtain nano-hydroapatite particles.
Said hydroapatite particles is carried out electron-microscope scanning and IR spectroscopy, and the result shows that the present invention has prepared the linear nano Win 40350.
Said nano-hydroapatite particles is scattered in the water once more, and good dispersion property can not form reunion.
The above only is a preferred implementation of the present invention; Should be pointed out that for those skilled in the art, under the prerequisite that does not break away from the principle of the invention; Can also make some improvement and retouching, these improvement and retouching also should be regarded as protection scope of the present invention.
Claims (10)
1. the preparation method of a linear nano Win 40350 may further comprise the steps:
Seal cultivation after the silk fibroin water solution of first mass concentration is concentrated into second mass concentration, obtain the silk fibroin nano-fiber aqueous solution;
The said silk fibroin nano-fiber aqueous solution is mixed with phosphoric acid root and calcium source, obtain the linear nano Win 40350 after the reaction.
2. preparation method according to claim 1 is characterized in that, said first mass concentration is 0.1%～10%, and said second mass concentration is 10%～50%.
3. preparation method according to claim 1 is characterized in that, said spissated temperature is 0 ℃～90 ℃.
4. preparation method according to claim 1 is characterized in that, the temperature that said sealing is cultivated is 0 ℃～70 ℃, and the time that said sealing is cultivated is 1 day～20 days.
5. preparation method according to claim 1 is characterized in that, said phosphoric acid root is phosphoric acid, sodium phosphate or calcium phosphate.
6. preparation method according to claim 1 is characterized in that, said calcium source is calcium hydroxide or nitrocalcite.
7. preparation method according to claim 1 is characterized in that, the temperature of said reaction is 50 ℃～80 ℃.
8. the preparation method of a nanometer hydroxyapatite may further comprise the steps:
Silk fibroin water solution is dried to fibroin protein film, said fibroin protein film is soluble in water, repeat drying-dissolution process, obtain silk fibroin nano-fiber solution;
Said silk fibroin nano-fiber solution is mixed with phosphoric acid root and calcium source, obtain the linear nano Win 40350 after the reaction.
9. preparation method according to claim 8 is characterized in that, said exsiccant temperature is 0 ℃～60 ℃.
10. preparation method according to claim 8 is characterized in that, the number of times of said repetition drying-dissolution process is 1 time～5 times.
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Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103738932A (en) * | 2013-12-13 | 2014-04-23 | 苏州大学 | Nano-hydroxyapatite and preparation method thereof |
| CN104211037A (en) * | 2014-08-07 | 2014-12-17 | 华中农业大学 | Method for catalytically synthesizing biomimetic material hydroxyapatite from phosvitin and polypeptide thereof |
| CN106567151A (en) * | 2016-11-03 | 2017-04-19 | 武汉理工大学 | Calcium fluoride collagen micro-fiber composite and preparation method thereof |
| CN106672933A (en) * | 2016-12-02 | 2017-05-17 | 中国科学院上海硅酸盐研究所 | Hydroxyapatite super-long nanowire-based macro-scale ordered structure material and preparation method thereof |
| CN109569347A (en) * | 2018-12-14 | 2019-04-05 | 苏州大学 | A kind of method of hydrophobic material water phase transfer |
| CN110078049A (en) * | 2019-05-27 | 2019-08-02 | 杭州电子科技大学 | A kind of hollow Nano transition metal oxide/carbon composite and preparation method thereof |
| CN110305202A (en) * | 2019-07-03 | 2019-10-08 | 浙江大学 | A method of improving fibroin albumen biomineralisation capabilities |
| CN114249982A (en) * | 2022-01-25 | 2022-03-29 | 武汉纺织大学 | Preparation method and application of high-strength high-modulus silk material |
| CN115581801A (en) * | 2022-09-29 | 2023-01-10 | 苏州大学 | Calcium phosphate mineralized silk micro-nano fiber membrane and preparation method thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101474429A (en) * | 2009-01-22 | 2009-07-08 | 浙江理工大学 | Method for preparing hydroxylapatite-silk fibroin compound stent material using two-step method |
| CN101844789A (en) * | 2010-06-18 | 2010-09-29 | 苏州大学 | Method for preparing calcium carbonate microspheres |
-
2011
- 2011-11-14 CN CN2011103584909A patent/CN102491298B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101474429A (en) * | 2009-01-22 | 2009-07-08 | 浙江理工大学 | Method for preparing hydroxylapatite-silk fibroin compound stent material using two-step method |
| CN101844789A (en) * | 2010-06-18 | 2010-09-29 | 苏州大学 | Method for preparing calcium carbonate microspheres |
Non-Patent Citations (2)
| Title |
|---|
| 刘琳等: "预处理对丝素蛋白膜调控羟基磷灰石晶体生长的影响", 《高等学校化学学报》 * |
| 崔彪: "丝素调控纳米羟基磷灰石复合材料制备工艺研究", 《中国优秀硕士学位论文全文数据库 医药卫生科技辑》 * |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103738932A (en) * | 2013-12-13 | 2014-04-23 | 苏州大学 | Nano-hydroxyapatite and preparation method thereof |
| CN103738932B (en) * | 2013-12-13 | 2016-04-20 | 苏州大学 | A kind of nanometer hydroxyapatite and preparation method thereof |
| CN104211037A (en) * | 2014-08-07 | 2014-12-17 | 华中农业大学 | Method for catalytically synthesizing biomimetic material hydroxyapatite from phosvitin and polypeptide thereof |
| CN106567151A (en) * | 2016-11-03 | 2017-04-19 | 武汉理工大学 | Calcium fluoride collagen micro-fiber composite and preparation method thereof |
| CN106672933A (en) * | 2016-12-02 | 2017-05-17 | 中国科学院上海硅酸盐研究所 | Hydroxyapatite super-long nanowire-based macro-scale ordered structure material and preparation method thereof |
| CN106672933B (en) * | 2016-12-02 | 2019-01-08 | 中国科学院上海硅酸盐研究所 | The preparation method of hydroxyapatite overlong nanowire macro-scale ordered structural material |
| CN109569347A (en) * | 2018-12-14 | 2019-04-05 | 苏州大学 | A kind of method of hydrophobic material water phase transfer |
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