CN104649283B - A kind of preparation method of nanometer β-calcium silicates/selenium bioactive composite material - Google Patents
A kind of preparation method of nanometer β-calcium silicates/selenium bioactive composite material Download PDFInfo
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- CN104649283B CN104649283B CN201510030287.7A CN201510030287A CN104649283B CN 104649283 B CN104649283 B CN 104649283B CN 201510030287 A CN201510030287 A CN 201510030287A CN 104649283 B CN104649283 B CN 104649283B
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Abstract
The invention discloses the preparation method of the bioactive composite material of a kind of nanometer β calcium silicates/selenium, it is characterized in that, (1) under room temperature, enuatrol and calcium chloride are joined containing deionized water, dehydrated alcohol, normal hexane there-necked flask in, after heating in water bath backflow certain time, prepare calcium oleate through post processing.(2) prepared calcium oleate is joined containing ethanol, oil-dissolving solvent there-necked flask in, make it fully dissolve at 45 DEG C, be subsequently added the aqueous solution of sodium silicate, heating in water bath backflow certain time.(3) reaction system being cooled to 0 DEG C, be subsequently adding the aqueous solution of sodium selenite, after being sufficiently stirred for, add Vitamin C aqueous acid, under room temperature, reaction is completely.(4) by prepared sample respectively with the alternately washing of deionized water, ethanol, after drying, sample is calcined in Muffle furnace the composite i.e. obtaining nanometer β calcium silicates/selenium.Antibacterial experiment shows that this composite is respectively 92 ± 4% and 95 ± 4% to the bacteriostasis rate of escherichia coli and staphylococcus aureus.Nanometer β calcium silicates/selenium composite of the present invention has a wide range of applications as antibiotic property bioactive materials.
Description
Technical field
The invention belongs to the preparation field of nano material, particularly to a kind of nanometer β-calcium silicates with antibiotic property
The preparation method of the bioactive composite material of/selenium.
Background technology
In recent years, calcium silicates is studied widely in field of biology.Experiment in vivo and vitro proves, calcium silicates has
Good induction osteoid apatite Forming ability and Bioactivity, and in simulated body fluid, bone-like apatite
The speed that stone is formed on calcium silicates surface is more than the speed formed at other biological glass and glass-ceramic surface.
In vitro cell experiment shows, calcium silicates no cytotoxicity and have good biological activity, the calcium of dissolution, silicon from
Son has the effect promoting Oesteoblast growth, and further research also indicates that calcium-silica-base material compares calcium-phosphorio
Material has more preferably osteogenic activity, and is applied on osseous tissue and joint as filling and repair materials.
Additionally, it is inevitable complication in Implant that antibacterial infects, although through strict sterilization and aseptic
Operation, the infection rate still suffering from 0.5-3% exists.Selenium is trace element necessary to human body, and nanometer selenium is also
There is the strongest antibacterial functions, research show nanometer selenium can effectively suppress the growth of staphylococcus aureus with
And a series of infection (Tran, P.A.Webster, the T.J.Int.J. that treatment causes due to staphylococcus aureus
Nanomed.2001,6,1553-1558)；All demonstrating the probiotic bacteria rich in selenium by internal and experiment in vitro can
The colibacillary growth caused a disease with suppression, experiment in vitro confirms after 96h, and the probiotic bacteria rich in selenium is (every
Gram probiotic bacteria is containing 0.509 microgram selenium) just can suppress colibacillary growth；By inoculated colibacillary always
Mus has injected a certain amount of probiotic bacteria rich in selenium, after 28 days, has been injected the old of probiotic bacteria containing selenium
The survival rate of Mus is apparently higher than (Yang J, Huang K, Qin S, Zhao Z, the Chen F.Dig not injected
Dis Sci.2009；54 (2): 246-254).If designing the phase interaction being i.e. conducive to regulation tissue with storeroom
With, there are again double effect bioactive materials of antibiotic property, make treatment reparation and prevention infection be integrated, will
There is important medical application be worth.
Summary of the invention
The technical problem to be solved is to provide a kind of bioactive composite material with antibiotic property
Preparation method.
In order to solve the problems referred to above, the invention provides a kind of nanometer β-calcium silicates/selenium bioactive composite material
Preparation method, it is characterised in that comprise the following steps:
Step 1): under room temperature, enuatrol, calcium chloride are added separately to containing dehydrated alcohol, normal hexane with
And in the there-necked flask of deionized water, after heating in water bath backflow, prepare calcium oleate through post processing；
Step 2): prepared calcium oleate is joined containing ethanol, oil-dissolving solvent there-necked flask in, 45
Making it fully dissolve at DEG C, be subsequently added the aqueous solution of sodium silicate, heating in water bath refluxes；
Step 3): reaction system is cooled to 0 DEG C, is subsequently adding the aqueous solution of sodium selenite, after being sufficiently stirred for,
Adding Vitamin C aqueous acid, under room temperature, reaction is completely；
Step 4): by prepared sample respectively with the alternately washing of deionized water, ethanol, after drying, by sample
Muffle furnace is calcined and i.e. obtains nanometer β-calcium silicates/selenium composite.
Preferably, described step 1) in the temperature of water-bath be 70 DEG C～75 DEG C, return time is 2-4h.
Preferably, described step 1) in the concentration of enuatrol be 3.0mol/L, its solution temperature is 45 DEG C.
Preferably, described step 1) in the molar concentration of calcium chloride be 3.0mol/L.
Preferably, described step 2) in the molar concentration of calcium oleate be 0.25mol/L.
Preferably, described step 2) oil-soluble solvent is hexamethylene, normal hexane or toluene.
Preferably, described step 2) in the concentration of sodium silicate solution be 0.25mol/L.
Preferably, described step 2) in the temperature of water-bath be 70 DEG C～75 DEG C, return time is 2-4h.
Preferably, described step 3) in the molar concentration of sodium selenite be 0.3-0.5mol/L.
Preferably, described step 3) in the molar concentration of ascorbic acid be 0.6-1.0mol/L.
Preferably, described step 3) response time be 2-4h.
Preferably, described step 4) in alternately washing particularly as follows: first with absolute ethanol washing, then 8000
Rpm centrifugation 10min removes supernatant；It is being washed with deionized, then centrifugal point of 8000rpm
Remove supernatant from 15min, so circulate dehydrated alcohol-deionized water wash 3 times；Finally use dehydrated alcohol
Washing, under 8000rpm, centrifugation 10min is precipitated.
Preferably, described step 4) in be dried temperature be 60 DEG C.
Preferably, described step 4) in the calcining heat of Muffle furnace be 460-800 DEG C, calcination time is 2h.
Sign and the evaluation of anti-microbial property to the material that benzene invention provides
One, In Vitro Bacteriostasis performance test
The test of nanometer β-calcium silicates/selenium composite anti-microbial property is adopted with the following method: experimental strain uses big
Enterobacteria and staphylococcus aureus.Take a certain amount of strain to be inoculated in the LB fluid medium of 50mL,
On the constant-temperature table of 37 DEG C, 160rpm cultivates 18～24h, in case antibacterial test.In laboratory sample and test
Equipment used is through 121 DEG C of high temperature sterilize 20min；Bacterium solution after former cultivation is diluted 102 times；Take dilution
After bacterium solution 1mL be inoculated in equipped with nanometer β-calcium silicates, the vial of nanometer β-calcium silicates/selenium composite
In；After sample and bacterium solution act on 18～24h in the constant incubator of 37 DEG C, it is separately added into the nothing of 10mL
Bacterium normal saline, covers tightly bottle cap, with hand rolling 30s, is eluted by antibacterial；Bacterium solution physiology after eluting
Take 100 μ L after the saline suitable multiple of dilution on LB solid medium, be coated with flat board, parallel painting three,
Count after the constant incubator of 37 DEG C cultivates 18～24h and average, then carry out conversion and obtain vial
In bacteria living number.It is calculated as follows the bacteriostasis rate of antimicrobial sample:
In formula:
CtThe meansigma methods of the bacterial population that-control sample records after inoculating and cultivating 18～24h；
TtThe meansigma methods of the bacterial population that-experiment sample records after inoculating and cultivating 18～24h.
The present invention has a following prominent beneficial effect:
1) concise in technology of the present invention, simple to operate；
2) antimicrobial efficiency of nanometer β prepared by the present invention-calcium silicates/selenium compound is high, to escherichia coli and gold
Staphylococcus aureus bacteriostasis rate be respectively 92 ± 4% and 95 ± 4%.
The present invention uses oil-water interfaces method to synthesize the bioactive composite material of nanometer β-calcium silicates/selenium.Profit
Interface method, refers to that chemical reaction occurs at biphase interface, and usually two mutual exclusive oil-water interfaces occur
Reaction.Utilize two phases in reaction, reactant can be dispersed in respectively in upper and lower two liquid, make product
Thing synthesizes in biphase interface.Therefore the present invention mainly synthesizes nanometer β-calcium silicates by oil-water interfaces and receives
The composite of rice selenium combines the excellent biological activity of calcium silicates and the effective antibiotic property of nanometer selenium, plays it each
In advantage biologically, will have more preferable application prospect.
Accompanying drawing explanation
Fig. 1 is the process chart of the present invention；
Fig. 2 is the complex X ray diffracting spectrum of nanometer β obtained by the present invention-calcium silicates/selenium；
Fig. 3 is the complex EDS collection of illustrative plates of nanometer β obtained by the present invention-calcium silicates/selenium；
Fig. 4 is the complex TEM figure of nanometer β obtained by the present invention-calcium silicates/selenium；
Fig. 5 is the antibacterial experiment design sketch of the complex of nanometer β obtained by the present invention-calcium silicates/selenium；
Fig. 6 is that the complex of nanometer β obtained by the present invention-calcium silicates/selenium is to escherichia coli and golden yellow Fructus Vitis viniferae
The experimental result cartogram of coccus.
Detailed description of the invention
For making the present invention become apparent, hereby with preferred embodiment, and accompanying drawing is coordinated to be described in detail below.
Embodiment 1
(1) weighing 9.1g enuatrol, 45 DEG C are dissolved in the deionized water of 10.0mL.Treat that enuatrol is the most molten
Xie Hou, adds the calcium chloride water of 3.0mol/L.35.0mL normal hexane, and 20.0 are added in system
ML dehydrated alcohol.Heating in water bath is to backflow, magnetic agitation 2h.
(2) after question response completes, being cooled to 0 DEG C, have the solid of white to separate out, vacuum filters, and is dried preparation oil
Acid calcium.
(3) 2.3g calcium oleate, 45 DEG C of dehydrated alcohol being dissolved in the hexamethylene of 15.0mL, 10.0mL are weighed
In.After calcium oleate is completely dissolved, add the sodium silicate aqueous solution of 0.25mol/L.Heating in water bath extremely backflow,
Magnetic agitation 2h.
(4) after above-mentioned reaction completes, it is cooled to 0 DEG C, adds the sodium selenite aqueous solution of 0.3mol/L, fill
After dividing stirring, add the aqueous ascorbic acid of 0.6mol/L, react 2h.
(5) after reaction terminates, absolute ethanol washing, then remove supernatant at 8000rpm centrifugation 10min
Liquid；Deionized water wash, removes supernatant at 8000rpm centrifugation 15min, so circulates anhydrous second
Alcohol-deionized water wash 3 times；Finally with absolute ethanol washing, under 8000rpm, centrifugation 10min obtains
To precipitation.In air dry oven, 60 DEG C are dried, and are finally placed in 460 DEG C of calcining 2h in Muffle furnace and i.e. obtain nanometer
The composite of β-calcium silicates/selenium；When calcining heat is 800 DEG C, it is thus achieved that product be β-calcium silicates.
(6) nanometer β-calcium silicates/selenium composite is used for the antibacterial reality of escherichia coli and staphylococcus aureus
Test.
Embodiment 2
(1) weighing 9.1g enuatrol, 45 DEG C are dissolved in the deionized water of 10.0mL.Treat that enuatrol is the most molten
Xie Hou, adds the calcium chloride water of 3.0mol/L.35.0mL normal hexane, and 20.0 are added in system
ML dehydrated alcohol.Heating in water bath is to backflow, magnetic agitation 3h.
(2) after question response completes, being cooled to 0 DEG C, have the solid of white to separate out, vacuum filters, and is dried preparation oil
Acid calcium.
(3) 2.3g calcium oleate, 45 DEG C of dehydrated alcohol being dissolved in the normal hexane of 15.0mL, 10.0mL are weighed
In.After calcium oleate is completely dissolved, add the sodium silicate aqueous solution of 0.25mol/L.Heating in water bath extremely backflow,
Magnetic agitation 3h.
(4) after above-mentioned reaction completes, it is cooled to 0 DEG C, adds the sodium selenite aqueous solution of 0.4mol/L, fill
After dividing stirring, add the aqueous ascorbic acid of 0.8mol/L, react 3h.
(5) after reaction terminates, with absolute ethanol washing, then in 8000rpm centrifugation 10min removal
Clear liquid；It is washed with deionized, then removes supernatant at 8000rpm centrifugation 15min, so follow
Ring dehydrated alcohol-deionized water wash 3 times；Finally with absolute ethanol washing, centrifugation under 8000rpm
10min is precipitated.In air dry oven, 60 DEG C are dried, and are finally placed in 460 DEG C of calcining 2h in Muffle furnace and are
Obtain the composite of nanometer β-calcium silicates/selenium.
Embodiment 3
(1) weighing 9.1g enuatrol, 45 DEG C are dissolved in the deionized water of 10.0mL.Treat that enuatrol is the most molten
Xie Hou, adds the calcium chloride water of 3.0mol/L.35.0mL normal hexane, and 20.0 are added in system
ML dehydrated alcohol.Heating in water bath is to backflow, magnetic agitation 4h.
(2) after question response completes, being cooled to 0 DEG C, have the solid of white to separate out, vacuum filters, and is dried preparation oil
Acid calcium.
(3) weighing 2.3g calcium oleate, 45 DEG C are dissolved in the dehydrated alcohol of the toluene of 15.0mL, 10.0mL.
After calcium oleate is completely dissolved, add the sodium silicate aqueous solution of 0.25mol/L.Heating in water bath is to backflow, magnetic force
Stirring 4h.
(4) after above-mentioned reaction completes, it is cooled to 0 DEG C, adds the sodium selenite aqueous solution of 0.5mol/L, fill
After dividing stirring, add the aqueous ascorbic acid of 1.0mol/L, react 4h.
(5) after reaction terminates, first with absolute ethanol washing, then remove at 8000rpm centrifugation 10min
Supernatant；It is washed with deionized, then removes supernatant at 8000rpm centrifugation 15min, so
Circulation dehydrated alcohol-deionized water wash 3 times；Finally with absolute ethanol washing, it is centrifuged under 8000rpm and divides
It is precipitated from 10min.In air dry oven, 60 DEG C are dried, and are finally placed in 460 DEG C of calcining 2h in Muffle furnace
I.e. obtain the composite of nanometer β-calcium silicates/selenium.
As it is shown in figure 1, be the complex reacting flow chart of nanometer β-calcium silicates/selenium.
Fig. 2 is the complex XRD figure spectrum of nanometer β-calcium silicates/selenium that embodiment 1-3 prepares, wherein, (a)
It is under 460 DEG C of calcination conditions；B () is under 800 DEG C of calcination conditions.
Fig. 3 is the EDS collection of illustrative plates of the complex of nanometer β-calcium silicates/selenium that embodiment 1-3 prepares.
Fig. 4 is the TEM figure of the composite of nanometer β-calcium silicates/selenium that embodiment 1 prepares.
Fig. 5 is the antibacterial effect figure of nanometer β-calcium silicates/selenium composite that embodiment 1 prepares, and is respectively big
Enterobacteria blank experiment (a1), β-calcium silicates (b1), β-calcium silicates/selenium (c1)；Staphylococcus aureus
Blank experiment (a2), β-calcium silicates (b2), β-calcium silicates/selenium (c2).
Fig. 6 is the escherichia coli that prepare of embodiment 1 and staphylococcus aureus cultivates 24h with two groups of sample contacts
After surface adhesion clump count statistics (* represents sample β-CaSiO3/ and blank between variant) (average
Value ± standard deviation, n=5, * * P ＜ 0.01).
Claims (14)
1. the preparation method of nanometer β-calcium silicates/selenium bioactive composite material, it is characterised in that comprise the following steps:
Step 1): under room temperature, is added sequentially to enuatrol, calcium chloride in the there-necked flask containing dehydrated alcohol, normal hexane and deionized water, after heating in water bath backflow, prepares calcium oleate through post processing；
Step 2): prepared calcium oleate is joined containing ethanol, oil-dissolving solvent there-necked flask in, make it fully dissolve at 45 DEG C, be subsequently added the aqueous solution of sodium silicate, heating in water bath refluxes；
Step 3): reaction system is cooled to 0 DEG C, is subsequently adding the aqueous solution of sodium selenite, after being sufficiently stirred for, adds Vitamin C aqueous acid, and under room temperature, reaction is completely；
Step 4): by the alternately washing of prepared sample deionized water, ethanol, after drying, sample is calcined in Muffle furnace and i.e. obtains nanometer β-calcium silicates/selenium composite.
2. the preparation method of nanometer β-calcium silicates/selenium bioactive composite material as claimed in claim 1, it is characterised in that in described step 1), the temperature of water-bath is 70 DEG C ~ 75 DEG C, and return time is 2-4 h.
3. the preparation method of nanometer β-calcium silicates/selenium bioactive composite material as claimed in claim 1, it is characterised in that in described step 1), the concentration of enuatrol is 3.0 mol/L, and its solution temperature is 45 DEG C.
4. the preparation method of nanometer β-calcium silicates/selenium bioactive composite material as claimed in claim 1, it is characterised in that in described step 1), the molar concentration of calcium chloride is 3.0 mol/L.
5. the preparation method of nanometer β-calcium silicates/selenium bioactive composite material as claimed in claim 1, it is characterised in that described step 2) in the molar concentration of calcium oleate be 0.25 mol/L.
6. the preparation method of nanometer β-calcium silicates/selenium bioactive composite material as claimed in claim 1, it is characterised in that described step 2) oil-soluble solvent is hexamethylene, normal hexane or toluene.
7. the preparation method of nanometer β-calcium silicates/selenium bioactive composite material as claimed in claim 1, it is characterised in that described step 2) in the concentration of sodium silicate solution be 0.25 mol/L.
8. the preparation method of nanometer β-calcium silicates/selenium bioactive composite material as claimed in claim 1, it is characterised in that described step 2) in the temperature of water-bath be 70 DEG C ~ 75 DEG C, return time is 2-4 h.
9. the preparation method of nanometer β-calcium silicates/selenium bioactive composite material as claimed in claim 1, it is characterised in that in described step 3), the molar concentration of sodium selenite is 0.3-0.5 mol/L.
10. the preparation method of nanometer β-calcium silicates/selenium bioactive composite material as claimed in claim 1, it is characterised in that in described step 3), the molar concentration of ascorbic acid is 0.6-1.0 mol/L.
The preparation method of 11. nanometer β-calcium silicates/selenium bioactive composite materials as claimed in claim 1, it is characterised in that the response time of described step 3) is 2-4 h.
The preparation method of 12. nanometer β-calcium silicates/selenium bioactive composite materials as claimed in claim 1, it is characterized in that, alternately washing in described step 4) is particularly as follows: first use absolute ethanol washing, and then under 8000 rpm, centrifugation 10 min removes supernatant；Being washed with deionized, then under 8000 rpm, centrifugation 15 min removes supernatant, so circulates dehydrated alcohol-deionized water wash 3 times again；Finally with absolute ethanol washing, under 8000 rpm, centrifugation 10 min is precipitated.
The preparation method of 13. nanometer β-calcium silicates/selenium bioactive composite materials as claimed in claim 1, it is characterised in that the temperature being dried in described step 4) is 60 DEG C.
The preparation method of 14. β-calcium silicates/selenium bioactive composite materials as claimed in claim 1, it is characterised in that in described step 4), the calcining heat of Muffle furnace is 460-800 DEG C, and calcination time is 2 h.
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| CN105256354A (en) * | 2015-10-29 | 2016-01-20 | 东华大学 | Preparing method of 316 L stainless steel with surface containing nanometer Se particles |
| CN105217681A (en) * | 2015-10-29 | 2016-01-06 | 东华大学 | A kind of method by regulating temperature of reaction to prepare different crystal forms nanometer titanium dioxide hafnium particle |
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| CN202280385U (en) * | 2011-01-18 | 2012-06-20 | 东莞市东上板业有限公司 | Antibiotic calcium silicate plate |
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| JPS58151323A (en) * | 1982-03-04 | 1983-09-08 | Tokuyama Soda Co Ltd | Modified calcium silicate |
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| JP2005279456A (en) * | 2004-03-30 | 2005-10-13 | Denki Kagaku Kogyo Kk | Water treatment material and water treatment method |
| CN202280385U (en) * | 2011-01-18 | 2012-06-20 | 东莞市东上板业有限公司 | Antibiotic calcium silicate plate |
| CN103387262A (en) * | 2013-07-19 | 2013-11-13 | 东华大学 | A preparation method for oil-soluble nano zirconium dioxide particles in a tetragonal crystal form by adopting an oil-water interface method |
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