CN103147111B - A kind of pure titanium differential arc oxidation coating and application thereof - Google Patents
A kind of pure titanium differential arc oxidation coating and application thereof Download PDFInfo
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- CN103147111B CN103147111B CN201310096457.2A CN201310096457A CN103147111B CN 103147111 B CN103147111 B CN 103147111B CN 201310096457 A CN201310096457 A CN 201310096457A CN 103147111 B CN103147111 B CN 103147111B
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Abstract
The invention discloses a kind of pure titanium differential arc oxidation coating and application thereof.Pure titanium differential arc oxidation coating provided by the present invention is by matrix, and the oxide film of the porous surface be combined with described matrix forms; Described matrix is titanium; Described oxide film is the anatase titanium dioxide containing element silicon and calcium constituent; The mass percentage of described element silicon in described oxide film is 2.2%; The mass percentage of described calcium constituent in described oxide film is 1.5%.Experiment proves, compared with pure Titanium base, pure titanium differential arc oxidation coating provided by the present invention has stronger cell compatibility, when cultivating 7 days, the proliferation rate of pure Titanium base superficial cell is in contrast only 0.713, and the proliferation rate of pure titanium differential arc oxidation coating superficial cell provided by the present invention can up to 1.172.
Description
Technical field
The present invention relates to a kind of pure titanium differential arc oxidation coating and application thereof.
Background technology
Titanium and titanium alloys is most widely used osseous tissue equivalent material, because it has good intensity, toughness and biocompatibility, but its biological activity and self-bone grafting poor-performing.Can improve the biological activity on its surface to titanium alloy surface modification, conventional method of modifying has plasma spraying, electrochemical deposition, collosol and gel etc.
Differential arc oxidation (Microarcoxidation, MAO) also known as micro-plasma oxidation (Microplasmaoxidation, MPO), the combination that to be a kind of be by electrolytic solution and corresponding electrical parameter, at the TRANSIENT HIGH TEMPERATURE High Pressure that aluminium, magnesium, titanium and titanium alloys surface rely on arc discharge to produce, grow the new technology of the ceramic film based on substrate metal oxide, because its preparation method is simple, film base conjugation is high, cost is low, on material with complex, evenly can prepare coating, can introduce advantages such as the useful trace elements of biological activity extensively concerned simultaneously.
Calcium is mineral substance the abundantest in human body, participates in the whole vital process of human body, is human life basis.Playing an important role to growing of bone, is the foundation stone of bone health.Calcium constituent, in human physiological metabolism, is responsible for the important task of courier, almost participates in all cells activity.Calcium constituent directly regulates the excitability of N&M.It plays the various effects self can supported in human body down.
Silicon is absorbed by the body in metasilicic acid form in water, is mainly distributed among human body skin and reticular tissue, in Skeleton process, has physiological effect, promotes skeleton development growth.Silicon also participates in the metabolism of polysaccharide, is the main component forming some Glucoamino polyvalent carboxylics.Silicon is relevant with cardiovascular diseases, and people, as scarce silicon, can cause the cardiovascular diseasess such as sacroiliitis, arteriosclerosis, coronary heart disease.
Not yet there is the report introducing Si, Ca element in differential arc oxidation coating at present, more this coating is not used for the report of cytocompatibility Journal of Sex Research.
Summary of the invention
The object of this invention is to provide a kind of pure titanium differential arc oxidation coating and application thereof.
Pure titanium differential arc oxidation coating provided by the present invention, by matrix, and the dense oxidation film of the porous surface of combining closely with described matrix forms;
Described matrix is titanium; Described oxide film is the anatase titanium dioxide containing element silicon and calcium constituent;
The mass percentage of described element silicon in described oxide film is 2.2%; The mass percentage of described calcium constituent in described oxide film is 1.5%.
The diameter in described hole is specially 2 μm.
Another object of the present invention is to provide a kind of method preparing described pure titanium differential arc oxidation coating.
The method of the described pure peptide differential arc oxidation coating of preparation provided by the present invention, titanium is specifically comprised the steps: to be placed in electrolytic solution, with described titanium for anode, take stainless steel as negative electrode, voltage be 200V, under frequency is 600Hz, dutycycle is the condition of 8%, oxidation 5min, obtains described pure titanium differential arc oxidation coating;
The solute of described electrolytic solution is water glass, sodium hydroxide, lime acetate, disodium ethylene diamine tetraacetate, and solvent is water; The proportioning of described water glass, described sodium hydroxide, described lime acetate, described disodium ethylene diamine tetraacetate and described water is 14.2g:20g:8.8g:15g:9L; The pH of described electrolytic solution is 7.2.
In the above-mentioned methods, the titanium being placed in described electrolytic solution is the titanium through following pre-treatment: by titanium sheet successively after the sand papering of 300# or 800# or 1500#, with acetone, dehydrated alcohol, deionized water successively ultrasonic cleaning, 50 DEG C of oven dry, obtain the titanium after processing.
Described pure titanium differential arc oxidation coating is also belonging to protection scope of the present invention as the application in animal body hard tissue alternate material or implant.
Described animal body sclerous tissues can be joint, bone, tooth etc.Described implant can be joint prosthesis, artificial bone, spine correcting rod, intramedullary pin, dental implant or skull bone etc.
Described pure titanium differential arc oxidation coating is also belonging to protection scope of the present invention as the application in cell cultures base material.
In preparation, described pure titanium differential arc oxidation coating promotes that the application in the product of cell proliferation also belongs to protection scope of the present invention.
Pure titanium differential arc oxidation coating main component provided by the present invention is anatase titanium dioxide, and successfully introduces element silicon and calcium constituent at coatingsurface.Experiment proves, compared with pure Titanium base, pure titanium differential arc oxidation coating provided by the present invention has stronger cell compatibility.When cultivating 7 days, the proliferation rate of pure Titanium base superficial cell is in contrast only 1.8, and the proliferation rate of pure titanium differential arc oxidation coating superficial cell provided by the present invention can up to 4.3.
Accompanying drawing explanation
Fig. 1 is the X ray diffracting spectrum of pure titanium differential arc oxidation coating and titanium sheet.
Fig. 2 is scanning electron microscope diagram spectrum and the X-ray energy spectrum thereof of pure titanium differential arc oxidation coating and titanium sheet.Wherein, A is scanning electron microscope diagram spectrum; B is X-ray energy spectrum.
Fig. 3 is the statistics of pure titanium differential arc oxidation coating and titanium plate surface cell adhesion rate.
Fig. 4 be pure titanium differential arc oxidation coating and titanium plate surface MC3T3-E1Subclone14 cell attachment 4h time scanning electron microscope diagram spectrum.Wherein, A is titanium sheet; B is pure titanium differential arc oxidation coating.
Fig. 5 is the statistics of pure titanium differential arc oxidation coating and titanium plate surface cell proliferation rate.
Embodiment
The experimental technique used in following embodiment if no special instructions, is ordinary method.
Material used in following embodiment, reagent etc., if no special instructions, all can obtain from commercial channels.
The preparation of embodiment 1, pure titanium differential arc oxidation coating and qualification
One, the preparation of pure titanium differential arc oxidation coating
Electrolytic solution: 14.2g water glass, 20g sodium hydroxide, 8.8g lime acetate, 15g disodium ethylene diamine tetraacetate, be dissolved in the deionized water of 9L, pH7.2.
Adopt business medical pure titanium, dimensions is the titanium sheet of 10 × 10 × 1mm.Specimen surface adopts 300# respectively successively, the sand papering (often organize when making coating and all utilize this three kinds of different sand paperings) of 800# and 1500#, then uses ultrasonic (ultrasound parameter power 100 hertz) cleaning, the 50 DEG C of oven dry successively of acetone, dehydrated alcohol, deionized water.Using the titanium sheet after cleaning as anode, insert in electrolytic solution (formula as above), stainless steel is negative electrode.Oxidation voltage is 200V, frequency 600Hz, dutycycle 8%, oxidization time 5min.Obtain pure titanium differential arc oxidation coating.
Two, the qualification of pure titanium differential arc oxidation coating
With the identical titanium sheet of dimensions for contrast, X-ray diffractometer (XRD, D/max-γ B, Japan) is utilized to carry out material phase analysis to pure titanium differential arc oxidation coating prepare by step one, use CuK
αx-ray source, tube voltage 40KV, tube current is 40mA, continuous sweep pattern, sweep velocity 4 °/min, and diffraction angle 2 θ is 10 ~ 90 °.Pure titanium differential arc oxidation coating surface topography prepared by step one is observed and is utilized scanning electronic microscope (SEM, Quanta200, FEICo., American), surface-element content detection adopts the X-ray energy spectrometer (EDS, EDAX, American) of scanning electron microscope system disposition.
As shown in Figure 1, as can be seen from the figure, after 200V differential arc oxidation process in step one, there is anatase octahedrite diffraction peak in pure titanium to X ray diffracting spectrum, in figure, the diffraction peak of titanium mainly comes from Titanium base.In addition, between 2 θ=25 ~ 35 °, there is wider back end peak, illustrate in coating have non-crystalline state thing phase.And XRD figure spectrum does not find the new thing phase of containing calcium silicon element, thus in coating, containing calcium silicon product is non-crystalline state.
Scanning electron microscope diagram spectrum and X-ray energy spectrum thereof are as shown in Figure 2, as can be seen from scanning electron microscope diagram spectrum (in Fig. 2 A), pure titanium is after 200V differential arc oxidation process in step one, and Titanium base surface forms typical differential arc oxidation vesicular structure, and the size of porous diameter is about 2 μm.As can be seen from X-ray energy spectrum (in Fig. 2 B), pure titanium differential arc oxidation coating prepared by step one is also containing Si, Ca element except Ti, O element, and the mass percentage that wherein mass percentage of Si is about 2.2%, Ca is about 1.5%.This illustrates in the pure titanium differential arc oxidation coating prepared in step one and successfully introduces Si, Ca element.
Embodiment 2, pure titanium differential arc oxidation coating cytoactive are tested
Experimental cell: MC3T3-E1Subclone14(mouse calvarium prebone cell subclone 14), Shanghai Inst. of Life Science, CAS.
Experiment reagent: modified form α-MEMHyclone substratum (Sai Mo flies generation that biological chemistry goods Beijing company limited); FBSGibco foetal calf serum; CouCountingkit-8(CCK8, green skies biotechnology research institute); CO222 Pen .-Strep solution (green skies biotechnology research institute); PBS damping fluid (pH value is 7.2-7.6, Wuhan Boster Biological Technology Co., Ltd.); Glutaraldehyde (Chemical Reagent Co., Ltd., Sinopharm Group); Paraformaldehyde (Tianjin Ke Miou chemical reagent company limited); The trimethyl carbinol; Osmic acid; Dehydrated alcohol (volume fraction is 50%, 70%, 90%, 95% and 100%).
Experiment grouping: the pure titanium differential arc oxidation coating (experimental group) that in embodiment 1, step one prepares; The titanium sheet (control group) that dimensions is identical with pure titanium differential arc oxidation coating.
One, cell adhesion test
1, cell cultures
When MC3T3-E1Subclone14 cell converges completely, outwell old nutrient solution, after PBS buffer solution, add Digestive system (final concentration is the trypsinase+final concentration of 0.25g/100mL is the EDTA of 0.03g/100mL) digestion, basis of microscopic observation is shown in that intercellular substance increases, retraction becomes circle, when a little cell has taken off wall, (FBS of volume content l0% in modified form α-MEMHyclone substratum, is added with complete culture solution, and final concentration is 20%(20g/100ml) CO222 Pen .-Strep) in and trypsinase stop its digestion, single cell suspension is made in piping and druming, wash centrifugal 5min(1000 and turn/min), again hanged with complete culture solution and made single cell suspension.Be placed in 37 DEG C, 5%CO
2and cultivate in carbon dioxide cell incubator under saturated humidity condition.Change liquid after 24h, within later every 3 days, change liquid once, when cell to cover with at the bottom of culture dish about 70%, go down to posterity (ratio of going down to posterity is for 1:2) with Digestive system.Inverted microscope observation of cell growing state, period is noted having pollution-free.
2, cell adhesion test
The pure titanium differential arc oxidation coating (experimental group) that step one in embodiment 1 is prepared, and the dimensions titanium sheet (control group) identical with pure titanium differential arc oxidation coating is placed in 24 well culture plates respectively, cell with after Digestive system digestion by 5 × 10
5the density in individual/hole is inoculated in culture plate, every hole adds complete culture solution 1.0ml, insert in incubator and cultivate, stop cultivating at 0.5h, 1h, 2h, 4h respectively after cell inoculation, test specimen (coating or titanium sheet) is rinsed gently 3 times with PBS damping fluid, after washing away the MC3T3-E1Subclone14 cell failing to attach, test specimen is proceeded in another 24 well culture plate, every hole adds 0.3ml Digestive system and digests 3 minutes, add complete culture solution and stop digestion, blow and beat test specimen gently, make single cell suspension, meter cell count, calculates cell adhesion rate Ln by formula (1).In experiment, for each of point, two groups all arrange 3 repetitions detection time, and result gets 3 mean values repeated, and adopts SAS6.0 software package to carry out one-way analysis of variance to data.
Ln=(Tn/Jn)×(Sp/Ss)×100%(1)
In formula, Tn is the cell count sticked on test specimen; Jn is inoculating cell number; Sp is culture hole floorage; Ss is that surface of test piece is amassed.
In addition, when cell cultures 4h, scanning electronic microscope is adopted to carry out morphologic observation to the MC3T3-E1Subclone14 cell on test specimen (coating or titanium sheet) surface.Specific as follows: cell adopt successively the PBS damping fluid of pH7.2 configure volume fraction be 2.5% glutaraldehyde, volume fraction be that 1% osmic acid is fixed, the set time is 30 minutes.Then dewater successively with the ethanol that volume fraction is respectively 50%, 70%, 90%, 100%, with pure trimethyl carbinol displacement ethanol, then sample is put into the refrigerator freezing 30min of-20 DEG C, with ES-2030(HITACHI) type freeze drier carries out critical drying to sample.Scanning electronic microscope (SEM, Quanta200, FEICo., American) observation of cell form is utilized after sample metal spraying after process.
The statistics of cell adhesion rate as shown in table 1 and Fig. 3, cell attachment 0.5,1, P<0.05 between two groups of data of 2h, 4h, there is statistical significance.Along with the increase cell pure titanium differential arc oxidation coating adhesion rate that is surperficial and titanium plate surface that step one prepares in embodiment 1 of time all improves, after inoculation 4h, titanium plate surface cell adhesion rate is about 38.5%, and the pure titanium differential arc oxidation coating superficial cell adherence rate that in embodiment 1, step one prepares is about 29.6%, gap obviously diminishes between the two.In addition, during cell attachment 4h, scanning electronic microscope is adopted to carry out the result of morphologic observation as shown in Figure 4 to the MC3T3-E1Subclone14 cell on test specimen (coating or titanium sheet) surface, as can be seen from the figure, after adherent 4h, the scleroblast form of titanium plate surface is plentiful, occurred filopodia and lamellipodia, cell is attached on the surface of titanium completely, and the area of stretching, extension is large; And the pure titanium differential arc oxidation coating surface scleroblast globulate that in embodiment 1, step one prepares, not yet full extension.
The statistics (unit: %) of cell adhesion rate in table 1 different time points two groups
Two, cell proliferation test
1, cell cultures
With in step one described in 1.
2, cell proliferation test
The pure titanium differential arc oxidation coating (experimental group) that step one in embodiment 1 is prepared, and the dimensions titanium sheet (control group) identical with pure titanium differential arc oxidation coating is placed in 24 well culture plates respectively, arrange the hole not adding test specimen (coating or titanium sheet) to contrast as without test specimen, cell is to press 5 × 10 after Digestive system digestion simultaneously
4the density in individual/hole is inoculated in culture plate, every hole adds complete culture solution 1.0mL, inserts in incubator and cultivates, after cultivating 1,4,7,10 day respectively, after PBS wash buffer 3 times, every hole adds 0.3mL Digestive system and digests 3 minutes, adds complete culture solution and stops digestion, blow and beat into single cell suspension gently, wash centrifugal 5min(1000 and turn/min), again hanged with 200 μ L complete culture solutions and made single cell suspension, and added CCK-8 liquid 20 μ L, at 5%CO
2, saturated humidity, continue under 37 DEG C of environment to cultivate 4h, after 4h, in each hole, sucking-off 100 μ L is placed in 96 orifice plates, absorbancy OD value is measured at 450nm wavelength with microplate reader, arrange simultaneously and only add 200 μ L complete culture solutions and 20 μ LCCK-8 liquid, but do not add the blank well of cell, carry out the detection of OD450, and then calculate cell proliferation rate ODa according to formula (2).In experiment, for each of point, two groups all arrange 3 repetitions detection time, and result gets 3 mean values repeated, and adopts SAS6.0 software package to carry out one-way analysis of variance to data.
ODa=(ODs-ODb)/(ODc-ODb)×100%(2)
In formula, ODs is the OD450 value of experimental group or control group (containing test specimen, cell, complete culture solution and CCK-8 liquid); ODc is the OD450 value without test specimen control group (containing cell, complete culture solution and CCK-8 liquid); ODb is the OD450 value of blank well (containing complete culture solution and CCK-8 liquid).
The statistics of cell proliferation rate is as shown in table 2 and Fig. 5, as can be seen from the table, in identical incubation time, in embodiment 1, the multiplication capacity of the pure titanium differential arc oxidation coating superficial cell that step one prepares contrasts significantly better than titanium sheet, inoculating cell after 1 day in embodiment 1 cell proliferation rate on the pure titanium differential arc oxidation coating surface that step one prepares reach more than 0.593, along with the growth of cell cultures time, in titanium sheet and embodiment 1, the proliferation rate of the pure titanium differential arc oxidation coating superficial cell that step one prepares all improves, when cell cultures 7 days, titanium plate surface proliferation rate was about 0.713, the pure titanium differential arc oxidation coating surface growth rate that in embodiment 1, step one prepares reaches about 1.172.Cell cultures 10 days, cell proliferation rate substantially no longer increases, and osteoclast number declines to some extent, and when showing cell cultures 10 days, vigor declines.Experimental result shows that the pure titanium differential arc oxidation coating that in embodiment 1, step one prepares can promote that scleroblast is at its surface growth.Due in the cell proliferation stage, only have when cell reaches certain quantity, three-dimensional extra-cellular matrix structure is formed in guarantee subsequent process, and then promote that scleroblast breaks up and functionating further in implant surface, so contrast relative to titanium sheet, the pure titanium differential arc oxidation coating that in embodiment 1, step one prepares can accelerate cell proliferation rate, advantageously in forming three-dimensional cell epimatrix structure fast.
The statistics of cell proliferation rate in table 2 different time points two groups
The experimental result of cumulated volume embodiment, the cell adhesion ability on pure titanium differential arc oxidation coating surface provided by the present invention is a little less than pure titanium plate surface, but ability of cell proliferation is better than pure titanium plate surface significantly.Experimental result shows that pure titanium differential arc oxidation coating provided by the present invention has good cell compatibility.
Comparative example 1, utilize different electrolytes prepare pure titanium differential arc oxidation coating and cytoactive experiment
One, the preparation of pure titanium differential arc oxidation coating
Its preparation method is compared with embodiment 1, and difference is only that the formula of electrolytic solution is different, and in this comparative example, electrolyte prescription is as follows:
Electrolytic solution: 11.2g water glass, 15g sodium hydroxide, 8.0g lime acetate, 10g disodium ethylene diamine tetraacetate, be dissolved in the deionized water of 9L, pH7.2.
Two, pure titanium differential arc oxidation coating cytoactive experiment
In this comparative example, the pure titanium differential arc oxidation coating of gained is prepared as a control group to utilize electrolyte prescription in embodiment 1, the pure titanium differential arc oxidation coating that in this comparative example prepared by step one is as experimental group, adopt method described in embodiment 2 step 2, cell proliferation test is carried out to above two groups of pure titanium differential arc oxidation coatings.
Result is as shown in table 3, as can be seen from the table, in identical incubation time, the multiplication capacity of control group (the pure titanium differential arc oxidation coating that in embodiment 1, step one prepares) superficial cell is significantly better than experimental group (the pure titanium differential arc oxidation coating that in this comparative example, step one prepares).Visible, the multiplication capacity of the pure peptide differential arc oxidation coating superficial cell utilizing the electrolytic solution of different ingredients to prepare is differentiated.
The statistics of cell proliferation rate in table 3 different time points two groups
Claims (6)
1. pure titanium differential arc oxidation coating, by matrix, and the oxide film of the porous surface be combined with described matrix forms;
Described matrix is titanium; Described oxide film is the anatase titanium dioxide containing element silicon and calcium constituent;
The mass percentage of described element silicon in described oxide film is 2.2%; The mass percentage of described calcium constituent in described oxide film is 1.5%.
2. pure titanium differential arc oxidation coating according to claim 1, is characterized in that: the diameter in described hole is 2 μm.
3. prepare the method for pure titanium differential arc oxidation coating described in claim 1 or 2, titanium is comprised the steps: to be placed in electrolytic solution, with described titanium for anode, take stainless steel as negative electrode, voltage be 200V, under frequency is 600Hz, dutycycle is the condition of 8%, oxidation 5min, obtains described pure titanium differential arc oxidation coating;
The solute of described electrolytic solution is water glass, sodium hydroxide, lime acetate, disodium ethylene diamine tetraacetate, and solvent is water; The proportioning of described water glass, described sodium hydroxide, described lime acetate, described disodium ethylene diamine tetraacetate and described water is 14.2g:20g:8.8g:15g:9L; The pH of described electrolytic solution is 7.2.
4. pure titanium differential arc oxidation coating described in claim 1 or 2 is as the application in animal body hard tissue alternate material or implant.
5. pure titanium differential arc oxidation coating described in claim 1 or 2 is as the application in cell cultures base material.
6. pure titanium differential arc oxidation coating described in claim 1 or 2 promotes the application in the product of cell proliferation in preparation.
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| CN104846414B (en) * | 2015-04-17 | 2017-07-18 | 江苏大学 | A kind of TiO2The differential arc oxidation preparation method of semiconductor optical anode |
| CN106176249B (en) * | 2016-07-25 | 2018-11-02 | 中国人民解放军第四军医大学 | A kind of plantation base station and preparation method thereof with antimicrobial effect |
| CN107829123B (en) * | 2017-10-09 | 2020-05-12 | 深圳市中科摩方科技有限公司 | Aluminum alloy with double-layer coating on surface and preparation method and application thereof |
| CN110180020B (en) * | 2019-05-29 | 2021-01-12 | 中国科学院上海硅酸盐研究所 | Nitrogen-doped titanium oxide coating and preparation method and application thereof |
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Inventor after: Zou Zhiqun Inventor after: Zhao Xiulan Inventor after: Hu Zhongbo Inventor after: Liu Yun Inventor before: Zou Zhiqun Inventor before: Hu Zhongbo Inventor before: Wang Meng Inventor before: Jiang Quanchun Inventor before: Liu Nannan Inventor before: Wu Xiaonan Inventor before: Qu Pingchuan Inventor before: Liu Xiaojing Inventor before: Wang Chunna Inventor before: Liu Miao Inventor before: Chen Yizi Inventor before: Li Yuanzhong Inventor before: Zhou Hongzhi Inventor before: Zhang Rui Inventor before: Zhao Xiulan Inventor before: Li Dechao Inventor before: Liu Yun Inventor before: Zhu Jianhua Inventor before: Wang Hongyan Inventor before: Pan Pan Inventor before: Hei Xiaoxia |
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| COR | Change of bibliographic data | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant |