CN109401157A - A kind of amorphous calcium phosphate-polyacrylic acid hybrid nano-material and its preparation method and application - Google Patents
A kind of amorphous calcium phosphate-polyacrylic acid hybrid nano-material and its preparation method and application Download PDFInfo
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Abstract
The invention discloses a kind of amorphous calcium phosphate-polyacrylic acid hybrid nano-materials and its preparation method and application, belong to biomedical materials field.Under polyacrylic acid molecule and magnesium ion synergistic stability effect, amorphous calcium phosphate-polyacrylic acid hybrid nano-material is prepared by wet chemistry method.The polyacrylic acid molecule of doping can hinder crystal to generate jointly with complexing calcium ions with magnesium ion.Compared with existing calcium phosphate crystal material, amorphous calcium phosphate-polyacrylic acid hybrid nano-material degradation faster, can be used for the coated carrier of the functional moleculars such as gene, drug.Such as fluorescent molecule (such as fluorescein isothiocynate, indocyanine-green) is coated in hybrid material, fluorography effect may be implemented.Compared with traditional fluorography preparation, amorphous calcium phosphate-polyacrylic acid hybrid nano-material coats fluorescent molecule, has the function of the quick release fluorescent molecule under weakly acidic condition, is a kind of fluorescence carrier that biocompatibility is splendid.
Description
Technical field
The present invention relates to biomedical material technologies, and in particular to a kind of amorphous calcium phosphate-polyacrylic acid hydridization is received
Rice material and its preparation method and application.
Background technique
The function that bio-medical hybrid material combines organic molecule and inorganic material respectively has, in bio-medical material
In highlighted its researching value to become more and more important.Currently, nano-hybrid material be widely used in genophore, pharmaceutical carrier and
[document 1:Wu, Y.et al., the Devising new lipid-coated calcium such as Dental Erosion and regeneration
phosphate/carbonate hybrid nanoparticles to control release in endosome for
Efficient gene delivery.J.Mater.Chem.B, 2017,5 (34): p.7194-7203. document 2:Sailor,
M.et al.,Hybrid Nanoparticles for Detection and Treatment of
Cancer.Adv.Mater., 2012,24 (28): p.3779-3802. document 3:Elsharkawy, S.et al., Protein
disorder–order interplay to guide the growth of hierarchical mineralized
structures.Nat.Commun.,2018,9(2145):p.1-12]。
Fluorescent nano particle is that fluorescent reagent is passed through embedding, the covalently means such as key connection introducing organic or inorganic nanometer material
In material, and nano particle is allowed to undertake the functions such as the detection of small organic molecule fluorescent dye, label.Traditional fluorescent reagent carrier packet
Silica, carbon ball etc. are included, is difficult to [document 4:Veeranarayanan, S.et al., the FITC labeled that degrades in vivo
silica nanoparticles as efficient cell tags:Uptake and photostability Study
In endothelial cells.J.Fluoresc., 2012,22 (2): p.537-548. document 5:Li, C, et al.,
Mesoporous carbon nanospheres featured fluorescent aptasensor for multiple
diagnosis of cancer in vitro and in vivo.ACS Nano,2015,9(12):p.12096-12103.]。
And main component of the calcium phosphate as human body inner bone tissues, there is good biocompatibility and degradability, catabolite can quilt
Human body utilizes [document 6:Rezwan, K.et al., Biodegradable and bioactive porous again
polymer/inorganic composite scaffolds for bone tissue engineering,
Biomaterials, 2006,27 (18): being a kind of fluorescence carrier that biocompatibility is splendid p.3413-3431.].
Summary of the invention
Object of the present invention is to can not degrade, mention in vivo for the problem that existing fluorescent reagent carrier organism poor compatibility
For a kind of amorphous calcium phosphate-polyacrylic acid hybrid nano-material and its preparation method and application, which has good biological phase
It is capacitive, degradable, it can be used as fluorescent molecule carrier.
To achieve the above object, technical scheme is as follows:
A kind of preparation method of amorphous calcium phosphate-polyacrylic acid hybrid nano-material, the material use wet-chemical legal system
It is standby, specifically comprise the following steps:
(1) mixed solution of calcium nitrate and magnesium nitrate is prepared；
(2) mixed solution of the calcium nitrate of step (1) preparation and magnesium nitrate is added dropwise to diammonium hydrogen phosphate and polyacrylic acid
Mixed solution in, gained mixed material is stirred at room temperature generates emulsion after a certain period of time；
(3) it by emulsion obtained by step (2) successively after eccentric cleaning and dialysis cleaning, then is freeze-dried, that is, is obtained
Obtain the amorphous calcium phosphate-polyacrylic acid hybrid nano-material.
The process of above-mentioned steps (1) specifically: first by Ca (NO3)2·4H2O and Mg (NO3)2·6H2O is dissolved separately in
In deionized water, calcium nitrate solution and magnesium nitrate solution is obtained, then calcium nitrate solution is mixed with magnesium nitrate solution, obtain nitric acid
The mixed solution of calcium and magnesium nitrate；Wherein: the concentration of calcium nitrate solution is 1-50g/L, and the concentration of magnesium nitrate solution is 0-
8.14g/L, and molar percentage (Mg/ (the Ca+Mg)) range for meeting Mg element in solution is 0-15%.
In above-mentioned steps (2), the mixed solution of the diammonium hydrogen phosphate and polyacrylic acid is by diammonium hydrogen phosphate and poly- third
Olefin(e) acid is dissolved in deionized water, and is obtained after adjusting pH by ammonium hydroxide；Wherein: the concentration of diammonium hydrogen phosphate is 0.5-25g/L,
The concentration of polyacrylic acid is 0.05-5g/L；The pH of the mixed solution of diammonium hydrogen phosphate and polyacrylic acid is 7-10.
In above-mentioned steps (2), mixing time 30min-2h.
The process of above-mentioned steps (3) specifically: supernatant is removed into the centrifugation of emulsion obtained by step (2), and be added go from
Sub- water ultrasonic disperse sediment；Then unreacted polyacrylic acid reagent is removed using dialysis membrane dialysis；Finally freeze-drying obtains
Obtain amorphous calcium phosphate-polyacrylic acid hybrid nano-material powder.
The amorphous calcium phosphate of the preparation-polyacrylic acid hybrid nano-material particle size range is 50-500nm.
The amorphous calcium phosphate-polyacrylic acid hybrid nano-material particle, can be phagocytized by cells, and then by lysosome
Cladding dissolution, has good biocompatibility.The material is coated applied to fluorescent molecule, and the fluorescent reagent includes different sulphur cyanogen
Sour fluorescein (FITC) and indocyanine-green (ICG).
Application process are as follows: when preparing the hybrid nano-material, fluorescent reagent is added to the mixing of calcium nitrate and magnesium nitrate
In solution, make to contain fluorescent reagent in prepared hybrid nano-material；The fluorescent reagent used accounts for the hybrid nano-material
Mass percent be 0.2-5%.The hybrid nano-material containing fluorescent reagent of preparation can be in (pH=under weakly acidic condition
It 5.0-6.5) dissolves, and discharges fluorescent molecule, to realize enhancing developing function.
Compared with prior art, the beneficial effects of the present invention are embodied in:
1. the present invention prepares amorphous phosphorus by wet chemistry method under polyacrylic acid molecule and magnesium ion synergistic stability effect
Sour calcium-polyacrylic acid hybrid nano-material.Using Magnesium-doped and polyacrylic acid hydridization, hybrid material can effectively improve
Stability.The polyacrylic acid molecule of doping can hinder crystal to generate jointly with complexing calcium ions with magnesium ion.
2, compared with existing calcium phosphate crystal material, amorphous calcium phosphate-polyacrylic acid hybrid nano-material degradation is more
Fastly, it can be used for the coated carrier of the functional moleculars such as gene, drug.Such as fluorescent molecule (such as different sulphur cyanogen is coated in hybrid material
Sour fluorescein, indocyanine-green) fluorography effect may be implemented.
3. the amorphous calcium phosphate with fluorography function-polyacrylic acid hybrid nano-material of the method for the present invention preparation,
Compared with existing fluorescent reagent carrier, amorphous calcium phosphate-polyacrylic acid hybrid particulates can dissolve and fast under weakly acidic condition
Quick-release amplification quantity fluorescent molecule to realize enhancing developing function, such as realizes enhancing development in tumor locus, is a kind of biology
The splendid fluorescence carrier of compatibility.
Detailed description of the invention
Fig. 1 is amorphous calcium phosphate-polyacrylic acid hybrid nano-material XRD result.
Fig. 2 is amorphous calcium phosphate-polyacrylic acid hybrid nano-material particle diameter distribution.
Fig. 3 is amorphous calcium phosphate-polyacrylic acid hybrid nano-material TEM shape appearance figure.
Fig. 4 is amorphous calcium phosphate-polyacrylic acid hybrid nano-material of various concentration and the cell morphology that cell co-cultures
Figure；Wherein: (A) 70 μ g/mL, (B) 35 μ g/mL, (C) 17.5 μ g/mL, (D) 8.8 μ g/mL, (E) 4.4 μ g/mL, (F) blank pair
According to group；Scale size is 100 microns.
Fig. 5 is the Synchrotron Radiation Soft X ray imaging that amorphous calcium phosphate-polyacrylic acid hybrid nano-material enters cell.
Fig. 6 is that cell dead dye liquor living contaminates in amorphous calcium phosphate-polyacrylic acid hybrid nano-material and cell co-culture experiments
Chromatic graph；Wherein: (A) 70 μ g/mL, (B) 35 μ g/mL, (C) 17.5 μ g/mL, (D) 8.8 μ g/mL, (E) 4.4 μ g/mL, (F) blank
Control group；Figure Green is living cells, and red is dead cell (scale is 100 microns).
Fig. 7 is that amorphous calcium phosphate-polyacrylic acid hybrid nano-material of package FITC fluorescent molecule enters the fluorescence of cell
Dye image (green is the hybrid material of package fluorescence, and blue portion is nucleus).
Fig. 8 is the amorphous calcium phosphate-polyacrylic acid hybrid nano-material infared spectrum for wrapping up ICG fluorescent molecule.
Specific embodiment
Of the invention is described further below with reference to embodiment:
The present invention is that wet chemistry method preparation has nanoscale amorphous calcium phosphate-polyacrylic acid hybrid material method, the party
Method mixes calcium nitrate with a certain amount of magnesium nitrate (fluorescent reagent), is dissolved in the mixing after water to diammonium hydrogen phosphate and polyacrylic acid
It is added dropwise in solution.It is stirred at room temperature and generates emulsion after a certain period of time, eccentric cleaning, further dialysis cleaning, then freezes
It is dry to obtain amorphous calcium phosphate-polyacrylic acid hybrid nano-material.
Specifically comprise the following steps:
(1) by Ca (NO3)2·4H2O、Mg(NO3)2·6H2O is dissolved in deionized water by certain molar ratio, is obtained molten
Liquid A.
(2) it weighs diammonium hydrogen phosphate and polyacrylic acid is dissolved in deionized water, ammonium hydroxide is added dropwise dropwise and adjusts pH=7-10,
Obtain solution B.
(3) solution A is added dropwise in solution B, is stirred 1 hour at room temperature.
(4) supernatant is removed in centrifugation after reacting, and deionized water is added, and ultrasonic disperse sediment obtains nano particle dispersion
Liquid is dialysed using dialysis membrane and removes unreacted polyacrylic acid in dispersion liquid.
(5) freeze-drying, which obtains, has amorphous calcium phosphate-polyacrylic acid hybrid nano-material powder.
(6) preparation has amorphous calcium phosphate-polyacrylic acid hybrid nano-material of fluorography, needs in above-mentioned steps
(1) a certain amount of fluorescent molecule is added in the solution A mentioned in.Continue above-mentioned steps (2)-(5), can prepare with fluorescence
The amorphous calcium phosphate of developing function-polyacrylic acid hybrid nano-material.
Embodiment 1
Solution is prepared: weighing the Ca (NO of 240mg3)2·4H2Mg (the NO of O and 36mg3)2·6H2O is dissolved in 50mL deionization
In water, it is uniformly mixing to obtain mixed solution A.Weigh (the NH of 110mg4)2HPO4200mL deionized water is dissolved in the PAA of 40mg
In, it is uniformly mixing to obtain mixed solution B, ammonium hydroxide is added dropwise dropwise, adjusts pH to 9.5.
Wet-chemical precipitating: by the prepared mixed solution A of previous step, mixed solution B is added dropwise to the speed of 1mL/min
In.With the revolving speed magnetic agitation of 500rpm, the time is 1 hour.
Cleaning: after reaction, centrifugal treating, revolving speed 4000rpm.Supernatant is removed after centrifugation, collects sediment, and
50mL deionized water is added, ultrasonic disperse obtains dispersion liquid.
Dialysis: by the dialysis membrane of dispersion liquid injection 8000-14000Da, dialysis cleaning 24 hours changes water one in every 6 hours
It is secondary.Purpose is to remove the polyacrylic acid molecule not wrapped up.
Dry: the dispersion liquid after dialysis is put into -20 DEG C of refrigerators and freezes 12 hours, and it is cold to be then placed in freeze drier progress
It is lyophilized dry, obtains amorphous calcium phosphate-polyacrylic acid hybrid nano-material the molar percentage Mg/ (Ca+Mg) of doping (wherein magnesium)
=10%).XRD testing result shows that the material is amorphous state (Fig. 1), and average grain diameter is 244nm (Fig. 2).Fig. 3 is nano material
Microscopic appearance figure.
Show that cell growth state is good (Fig. 4) with cell co-culture experiments.Display amorphous phosphoric acid is imaged using grenz ray
Calcium-polyacrylic acid hybrid nanomaterial can be phagocytized by cells (Fig. 5), and then coated and dissolved by lysosome.Dead decoration method card living
Clear-cells existing state is good (Fig. 6).The above results prove amorphous calcium phosphate-polyacrylic acid hybrid nano-material acellular poison
Property, there is good biocompatibility.
Embodiment 2
Solution is prepared: weighing 267mg Ca (NO3)2·4H2O is dissolved in 50mL deionized water, and it is molten to be uniformly mixing to obtain mixing
Liquid A.Weigh 110mg (NH4)2HPO4It is dissolved in 200mL deionized water with 40mg PAA, is uniformly mixing to obtain mixed solution B, by
Ammonium hydroxide is added dropwise in drop, adjusts pH to 9.5.
Wet-chemical precipitating: by the prepared mixed solution A of previous step, mixed solution B is added dropwise to the speed of 1mL/min.
With the revolving speed magnetic agitation of 500rpm, the time is 1 hour.
Cleaning: after reaction, centrifugal treating, revolving speed 4000rpm.Supernatant is removed after centrifugation, collects sediment, and
50mL deionized water is added, ultrasonic disperse obtains dispersion liquid.
Dialysis: by the dialysis membrane of dispersion liquid injection 8000-14000Da, dialysis cleaning 24 hours changes water one in every 6 hours
It is secondary.Purpose is to remove the polyacrylic acid molecule not wrapped up.
Dry: the dispersion liquid after dialysis is put into -20 DEG C of refrigerators and freezes 12 hours, and it is cold to be then placed in freeze drier progress
It is lyophilized dry, obtains amorphous calcium phosphate-polyacrylic acid hybrid nano-material.
Embodiment 3
Solution is prepared: weighing 227mg Ca (NO3)2·4H2O and 54mg Mg (NO3)2·6H2O is dissolved in 50mL deionized water
In, it is uniformly mixing to obtain mixed solution A.Weigh 110mg (NH4)2HPO4It is dissolved in 200mL deionized water, stirs with 40mg PAA
Mixed solution B uniformly is obtained, ammonium hydroxide is added dropwise dropwise, adjusts pH to 9.5.
Wet-chemical precipitating: by the prepared mixed solution A of previous step, mixed solution B is added dropwise to the speed of 1mL/min.
With the revolving speed magnetic agitation of 500rpm, the time is 1 hour.
Cleaning: after reaction, centrifugal treating, revolving speed 4000rpm.Supernatant is removed after centrifugation, collects sediment, and
50mL deionized water is added, ultrasonic disperse obtains dispersion liquid.
Dialysis: by the dialysis membrane of dispersion liquid injection 8000-14000Da, dialysis cleaning 24 hours changes water one in every 6 hours
It is secondary.Purpose is to remove the polyacrylic acid molecule not wrapped up.
Dry: the dispersion liquid after dialysis is put into -20 DEG C of refrigerators and freezes 12 hours, and it is cold to be then placed in freeze drier progress
It is lyophilized dry, obtains amorphous calcium phosphate-polyacrylic acid hybrid nano-material the molar percentage Mg/ (Ca+Mg) of doping (wherein magnesium)
=15%).
Embodiment 4
Solution is prepared: weighing 240mg Ca (NO3)2·4H2O, 36mg Mg (NO3)2·6H2O and 8mg FITC is dissolved in 50mL
In deionized water, it is uniformly mixing to obtain mixed solution A.Weigh 110mg (NH4)2HPO4200mL deionization is dissolved in 40mg PAA
In water, it is uniformly mixing to obtain mixed solution B, ammonium hydroxide is added dropwise dropwise, adjusts pH to 9.5.
Wet-chemical precipitating: by the prepared mixed solution A of previous step, mixed solution B is added dropwise to the speed of 1mL/min.
With the revolving speed magnetic agitation of 500rpm, the time is 1 hour.
Cleaning: after reaction, centrifugal treating, revolving speed 4000rpm.Supernatant is removed after centrifugation, collects sediment, and
50mL deionized water is added, ultrasonic disperse obtains dispersion liquid.
Dialysis: by the dialysis membrane of dispersion liquid injection 8000-14000Da, dialysis cleaning 24 hours changes water one in every 6 hours
It is secondary.Purpose is to remove the fluorescent molecule that do not wrap up and polyacrylic acid molecule.
Dry: the dispersion liquid after dialysis is put into -20 DEG C of refrigerators and freezes 12 hours, and it is cold to be then placed in freeze drier progress
It is lyophilized dry, obtains the amorphous calcium phosphate with fluorography-polyacrylic acid hybrid nano-material (wherein moles hundred of magnesium doping
Divide than Mg/ (Ca+Mg))=10%, 3%) fluorescent molecule weight percent is about.After being co-cultured with cell, fluorescent staining image
(Fig. 7) shows that amorphous calcium phosphate-polyacrylic acid hybrid nano-material enters cell, has preferable fluorescence intensity.
Embodiment 5
Solution is prepared: weighing 267mg Ca (NO3)2·4H2O and 8mg FITC is dissolved in 50mL deionized water, is stirred evenly
Obtain mixed solution A.Weigh 110mg (NH4)2HPO4It is dissolved in 200mL deionized water, is uniformly mixing to obtain mixed with 40mg PAA
Solution B is closed, ammonium hydroxide is added dropwise dropwise, adjusts pH to 9.5.
Wet-chemical precipitating: by the prepared mixed solution A of previous step, mixed solution B is added dropwise to the speed of 1mL/min.
With the revolving speed magnetic agitation of 500rpm, the time is 1 hour.
Cleaning: after reaction, centrifugal treating, revolving speed 4000rpm.Supernatant is removed after centrifugation, collects sediment, and
50mL deionized water is added, ultrasonic disperse obtains dispersion liquid.
Dialysis: by the dialysis membrane of dispersion liquid injection 8000-14000Da, dialysis cleaning 24 hours changes water one in every 6 hours
It is secondary.Purpose is to remove the fluorescent molecule that do not wrap up and polyacrylic acid molecule.
Dry: the dispersion liquid after dialysis is put into -20 DEG C of refrigerators and freezes 12 hours, and it is cold to be then placed in freeze drier progress
It is lyophilized dry, obtains the amorphous calcium phosphate with fluorography-polyacrylic acid hybrid nano-material (wherein fluorescent molecule weight hundred
Divide than for 3%).
Embodiment 6
Solution is prepared: weighing 240mg Ca (NO3)2·4H2O, 36mg Mg (NO3)2·6H2O and 8mg ICG is dissolved in 50mL
In deionized water, it is uniformly mixing to obtain mixed solution A.Weigh 110mg (NH4)2HPO4200mL deionization is dissolved in 40mg PAA
In water, it is uniformly mixing to obtain mixed solution B, ammonium hydroxide is added dropwise dropwise, adjusts pH to 9.5.
Wet-chemical precipitating: by the prepared mixed solution A of previous step, mixed solution B is added dropwise to the speed of 1mL/min.
With the revolving speed magnetic agitation of 500rpm, the time is 1 hour.
Cleaning: after reaction, centrifugal treating, revolving speed 4000rpm.Supernatant is removed after centrifugation, collects sediment, and
50mL deionized water is added, ultrasonic disperse obtains dispersion liquid.
Dialysis: by the dialysis membrane of dispersion liquid injection 8000-14000Da, dialysis cleaning 24 hours changes water one in every 6 hours
It is secondary.Purpose is to remove the fluorescent molecule that do not wrap up and polyacrylic acid molecule.
Dry: the dispersion liquid after dialysis is put into -20 DEG C of refrigerators and freezes 12 hours, and it is cold to be then placed in freeze drier progress
It is lyophilized dry, obtains the nano amorphous calcium phosphate with fluorography-polyacrylic acid hybrid material (wherein moles hundred of magnesium doping
Divide than Mg/ (Ca+Mg))=10%, 3%) fluorescent molecule weight percent is.Infrared results prove amorphous calcium phosphate-polypropylene
Sour hybrid nano-material successfully wraps up ICG fluorescent reagent (Fig. 8).
Embodiment 7
Solution is prepared: weighing 267mg Ca (NO3)2·4H2O and 8mg ICG is dissolved in 50mL deionized water, is stirred evenly
Obtain mixed solution A.Weigh 110mg (NH4)2HPO4It is dissolved in 200mL deionized water, is uniformly mixing to obtain mixed with 40mg PAA
Solution B is closed, ammonium hydroxide is added dropwise dropwise, adjusts pH to 9.5.
Wet-chemical precipitating: by the prepared mixed solution A of previous step, mixed solution B is added dropwise to the speed of 1mL/min.
With the revolving speed magnetic agitation of 500rpm, the time is 1 hour.
Cleaning: after reaction, centrifugal treating, revolving speed 4000rpm.Supernatant is removed after centrifugation, collects sediment, and
50mL deionized water is added, ultrasonic disperse obtains dispersion liquid.
Dialysis: by the dialysis membrane of dispersion liquid injection 8000-14000Da, dialysis cleaning 24 hours changes water one in every 6 hours
It is secondary.Purpose is to remove the fluorescent molecule that do not wrap up and polyacrylic acid molecule.
Dry: the dispersion liquid after dialysis is put into -20 DEG C of refrigerators and freezes 12 hours, and it is cold to be then placed in freeze drier progress
It is lyophilized dry, obtains the amorphous calcium phosphate with fluorography-polyacrylic acid hybrid nano-material (wherein fluorescent molecule weight hundred
Divide than for 3%).
Claims (10)
1. a kind of amorphous calcium phosphate-polyacrylic acid hybrid nano-material preparation method, it is characterised in that: the material is using wet
Chemical method preparation, specifically comprises the following steps:
(1) mixed solution of calcium nitrate and magnesium nitrate is prepared；
(2) mixed solution of the calcium nitrate of step (1) preparation and magnesium nitrate is added dropwise to the mixed of diammonium hydrogen phosphate and polyacrylic acid
It closes in solution, gained mixed material is stirred at room temperature generates emulsion after a certain period of time；
(3) it by emulsion obtained by step (2) successively after eccentric cleaning and dialysis cleaning, then is freeze-dried, i.e. acquisition institute
State amorphous calcium phosphate-polyacrylic acid hybrid nano-material.
2. amorphous calcium phosphate according to claim 1-polyacrylic acid hybrid nano-material preparation method, feature exist
In: the process of step (1) specifically: first by Ca (NO3)2·4H2O and Mg (NO3)2·6H2O is dissolved separately in deionized water
In, calcium nitrate solution and magnesium nitrate solution are obtained, then calcium nitrate solution is mixed with magnesium nitrate solution, obtains calcium nitrate and nitric acid
The mixed solution of magnesium；Wherein: the concentration of calcium nitrate solution is 1-50g/L, and the concentration of magnesium nitrate solution is 0-8.14g/L, and full
Molar percentage (Mg/ (the Ca+Mg)) range of Mg element is 0-15% in sufficient mixed solution.
3. amorphous calcium phosphate according to claim 1-polyacrylic acid hybrid nano-material preparation method, feature exist
In: in step (2), the mixed solution of the diammonium hydrogen phosphate and polyacrylic acid is to be dissolved in diammonium hydrogen phosphate and polyacrylic acid
In deionized water, and obtained after adjusting pH by ammonium hydroxide；Wherein: the concentration of diammonium hydrogen phosphate is 0.5-25g/L, polyacrylic acid
Concentration is 0.05-5g/L；The pH of the mixed solution of diammonium hydrogen phosphate and polyacrylic acid is 7-10.
4. amorphous calcium phosphate according to claim 1-polyacrylic acid hybrid nano-material preparation method, feature exist
In: in step (2), mixing time 30min-2h.
5. amorphous calcium phosphate according to claim 1-polyacrylic acid hybrid nano-material preparation method, feature exist
In: the process of step (3) specifically: supernatant is removed into the centrifugation of emulsion obtained by step (2), and deionized water ultrasound point is added
Dissipate sediment；Then using the dialysis membrane dialysis unreacted polyacrylic acid reagent in place to go；Finally freeze-drying obtains amorphous phosphoric acid
Calcium-polyacrylic acid hybrid nano-material powder.
6. a kind of amorphous calcium phosphate-polyacrylic acid hybrid nano-material prepared using claim 1 the method, feature
Be: the particle size range of the material is 50-500nm.
7. amorphous calcium phosphate according to claim 6-polyacrylic acid hybrid nano-material, it is characterised in that: the amorphous
Calcium phosphate-polyacrylic acid hybrid nano-material particle, can be phagocytized by cells, and then is coated and dissolve by lysosome, have good
Biocompatibility.
8. the application of amorphous calcium phosphate according to claim 6-polyacrylic acid hybrid nano-material, it is characterised in that: should
Material is coated applied to fluorescent molecule, and the fluorescent reagent includes fluorescein isothiocynate (FITC) and indocyanine-green (ICG).
9. the application of amorphous calcium phosphate according to claim 8-polyacrylic acid hybrid nano-material, it is characterised in that: answer
With process are as follows: when preparing the hybrid nano-material, fluorescent reagent is added in the mixed solution of calcium nitrate and magnesium nitrate, makes institute
Contain fluorescent reagent in the hybrid nano-material of preparation；The fluorescent reagent used accounts for the mass percent of the hybrid nano-material
For 0.2-5%.
10. the application of amorphous calcium phosphate according to claim 9-polyacrylic acid hybrid nano-material, it is characterised in that:
The hybrid nano-material containing fluorescent reagent of preparation (pH=5.0-6.5) can dissolve under weakly acidic condition, and discharge fluorescence
Molecule, to realize enhancing developing function.
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