Summary of the invention
The purpose of this invention is to provide nano-particle modified titanio titanate nano wire biologic bracket material of a kind of adsorbed property of porous hydroxyl calcium phosphate and preparation method thereof.
For achieving the above object, the technical solution used in the present invention is: the nano-particle modified titanio titanate nano wire biologic bracket material of this adsorbed property of porous hydroxyl calcium phosphate is to grow in the titanio titanate nano wire that constitutes latticed microcellular structure on the titanium substrate by the titanate nano wire, and the superficial growth of described titanio titanate nano wire has the adsorbed property of porous hydroxyl calcium phosphate nanoparticle coating.
The preparation method of the titanio titanate nano wire biologic bracket material that adsorbed property of porous hydroxyl calcium phosphate of the present invention is nano-particle modified comprises the steps:
(1) with the titanium substrate in ultrasonic cleaning in acetone, ethanol solution successively under the room temperature, the back is transferred to this titanium substrate after clean with distilled water flushing and is contained NaOH and Ba (OH)
2﹒ 8H
2Sealed reaction in the politef reactor of the mixed solution of O makes the titanate nano wire of growing on the titanium substrate obtain titanio titanate nano wire, and this titanio titanate nano wire is latticed microcellular structure; Titanio titanate nano wire is extremely neutral with deionized water rinsing, and at air drying;
(2) under constant current or the constant voltage mode, in three electrode reaction ponds, the titanio titanate nano wire that finally obtains with step (1) is a working electrode, with the pseudo body fluid is that deposit fluid deposits so that the superficial growth adsorbed property of porous hydroxyl calcium phosphate nano-particle of described titanio titanate nano wire obtains the nano-particle modified titanio titanate nano wire biologic bracket material of adsorbed property of porous hydroxyl calcium phosphate.
Further, NaOH of the present invention and Ba (OH)
2﹒ 8H
2The concentration of the mixed solution of O is 0.1~10 mol/L.
Further, the sealed reaction time of step of the present invention (1) is that 1~16 h, reaction temperature are 100~280 ° of C.
Further, in the step of the present invention (2), described pseudo body fluid (SBF) consist of NaCl:8.0g/L, CaCl
2: 0.14 g/L, KCl:0.40 g/L, NaHCO
3: 0.35 g/L, Glucose:1.00 g/L, KH
2PO
4: 0.10 g/L, MgCl
26H
2O:0.10 g/L, Na
2HPO
42H
2O:0.06 g/L, MgSO
47H
2O:0.06 g/L.
Further, in the step of the present invention (2), the pH of described deposit fluid is 8.0~12.6, and depositing temperature is room temperature~90 ° C, deposition current Wei – 50~50 mA, and deposition voltage Wei – 4.0~4.0 V, sedimentation time is 0.5~12 h.
Compared with prior art, the invention has the beneficial effects as follows: the titanio titanate nano wire that the present invention obtained has class bone shape grid microcellular structure, titanio titanate nanowire surface has obtained the modification of adsorbed property of porous hydroxyl calcium phosphate nano-particle, this flourishing microcellular structure in biologic bracket material surface can provide the place for the propagation and the differentiation of cell, the modification of calcium hydroxy phosphate nano-particle helps the propagation and the differentiation of cell more, make that it can be better and the area of new bone combination, excellent biocompatibility is arranged, and adsorbed property of porous hydroxyl calcium phosphate surface and the inner a large amount of pore structures that exist make biologic bracket material of the present invention have the favorable mechanical performance, the loading and the sustained release that help medicine or cell growth factor, can promote further growing into of area of new bone, thereby realize that Drug therapy and bone are transplanted or the double effects of damaged reparation.This material has reached structure and the bionical purpose of functionalization, can be used as the bone transplanting of a new generation or the biologic bracket material of damaged reparation.
Compared with prior art, outstanding advantage of the present invention is: the required equipment investment of preparation method of the present invention is few, raw material is cheap and utilization rate is high, producing cost is low, technology is simple, easy operating, the reaction condition gentleness, be convenient to automatization's mass production and the gained material contamination is few, modern technologies have been reached for requirements of green environmental protection, it is the feasible method of the nanoparticle of a kind of preparation particle diameter that has very much a prospect and controllable shapes, by the nano-particle modified titanio titanate nano wire biologic bracket material of adsorbed property of porous hydroxyl calcium phosphate of the inventive method preparation, be the concentrated reflection of the inventive method advantage.
Description of drawings
Fig. 1 prepares the structure diagram of the device of the nano-particle modified titanio titanate nano wire biologic bracket material of adsorbed property of porous hydroxyl calcium phosphate for electrochemical process among the present invention, wherein, IM6 provides the device of constant current/constant voltage, and WE is a working electrode, RE is a reference electrode, and CE is to the utmost point.
Fig. 2 is scanning electron microscope (SEM) figure of the titanio titanate nano wire with latticed microcellular structure of dissolved salt synthetic method preparation among the present invention.
Fig. 3 is the SEM figure of the nano-particle modified titanio titanate nano wire biological support of the adsorbed property of porous hydroxyl calcium phosphate of electrochemical deposition method preparation among the present invention.
Fig. 4 is X ray energy dispersive (EDX) collection of illustrative plates of the nano-particle modified titanio titanate nano wire biological support of the adsorbed property of porous hydroxyl calcium phosphate of electrochemical deposition method preparation among the present invention.
Fig. 5 is X-ray diffraction (XRD) collection of illustrative plates of the nano-particle modified titanio titanate nano wire biological support of the adsorbed property of porous hydroxyl calcium phosphate of electrochemical deposition method preparation among the present invention.
Fig. 6 for the nano-particle modified titanio titanate nano wire biological support of the adsorbed property of porous hydroxyl calcium phosphate of electrochemical deposition method preparation among the present invention at 450 ° of C(lower curves) and 550 ° of C(upper curves) calcine X-ray diffraction (XRD) collection of illustrative plates behind the 3h.
Fig. 7 is transmission electron microscope (TEM) figure of the nano-particle modified titanio titanate nano wire biological support of the adsorbed property of porous hydroxyl calcium phosphate of electrochemical deposition method preparation among the present invention.
Fig. 8 is high resolution transmission electron microscopy (HRTEM) figure of the nano-particle modified titanio titanate nano wire biological support of the adsorbed property of porous hydroxyl calcium phosphate of electrochemical deposition method preparation among the present invention.
Fig. 9 is Fourier transform infrared spectroscopy-attenuated total reflectance (FTIR-ATR) collection of illustrative plates of the nano-particle modified titanio titanate nano wire biological support of the adsorbed property of porous hydroxyl calcium phosphate of electrochemical deposition method preparation among the present invention.
Figure 10 for the nano-particle modified titanio titanate nano wire biological support of the adsorbed property of porous hydroxyl calcium phosphate of electrochemical deposition method preparation among the present invention at 450 ° of C(lower curves) and 550 ° of C(upper curves) calcine Fourier transform infrared spectroscopy-attenuated total reflectance (FTIR-ATR) collection of illustrative plates behind the 3h.
Figure 11 is scanning electron microscope (SEM) figure of cell culture experiment research for the nano-particle modified titanio titanate nano wire biological support of adsorbed property of porous hydroxyl calcium phosphate of electrochemical deposition method preparation among the present invention.Wherein, A ﹑ B ﹑ C ﹑ D is respectively the MG63 cell and cultivates 1,3,5,7 day SEM figure on this biologic bracket material, and corresponding insertion figure is respectively the growth conditions figure of the cultivation 1,3,5,7 of MG63 cell on the titanate nano wire of micropore and the nano-particle modified mistake of adsorbed property of porous hydroxyl calcium phosphate.
Figure 12 does the shows fluorescent microscopy images of cell culture experiment research for the nano-particle modified titanio titanate nano wire biological support of adsorbed property of porous hydroxyl calcium phosphate of electrochemical deposition method preparation among the present invention.Wherein, A ﹑ B ﹑ C ﹑ D is respectively the MG63 cell and cultivates 1,3,5,7 day MG63 cell growth state figure on this biologic bracket material, and corresponding insertion figure is respectively MG63 1,3,5,7 days nuclear fluorescence microscope pictures of cell culture on this biologic bracket material.
Figure 13 does the rectangular histogram of tetramethyl azo azoles salt (MTT) colorimetric method for determining of cell culture experiment research for the nano-particle modified titanio titanate nano wire biological support of adsorbed property of porous hydroxyl calcium phosphate of electrochemical deposition method preparation among the present invention.
Figure 14 does the rectangular histogram of alkali phosphatase (ALP) determination of activity of cell culture experiment research for the nano-particle modified titanio titanate nano wire biological support of adsorbed property of porous hydroxyl calcium phosphate of electrochemical deposition method preparation among the present invention.
The specific embodiment
Fig. 1 uses electrochemical deposition method to prepare the apparatus structure sketch of the nano-particle modified titanio titanate nano wire biological support of cellular calcium hydroxy phosphate in following examples.As shown in Figure 1, the electrochemical deposition method apparatus structure for preparing the nano-particle modified titanio titanate nano wire of adsorbed property of porous hydroxyl calcium phosphate totally is made up of three parts: computer machine user controls figure software section (for example Time-Controlled DC-Measurements software), core I M6 constant current/constant voltage output, reaction tank part.
The invention will be further described below in conjunction with embodiment.
Embodiment 1: present embodiment carries out according to the following steps:
(1) the solution technique preparation has the titanio titanate nano wire of latticed microcellular structure.Concrete preparation method is: the titanium substrate is in ultrasonic cleaning 20 minutes in acetone and ethanol solution successively under the room temperature, and the back is with distilled water flushing 4 times, this titanium substrate transferred to after air-dry and contains 0.1 mol/L NaOH and Ba (OH)
2﹒ 8H
2Seal in the politef reactor of the mixed solution of O, behind reaction 16 h under 280 ° of C, obtain having the titanio titanate nano wire of latticed microcellular structure, titanio titanate nano wire is extremely neutral with deionized water rinsing, and at air drying.
(2) electrochemical deposition method prepares the nano-particle modified titanio titanate nano wire biological support of adsorbed property of porous hydroxyl calcium phosphate.Concrete preparation process is as follows: in three electrode reaction ponds, adopt IM6 constant current/constant voltage output mode, with the synthetic titanio titanate nano wire with latticed microcellular structure of step (1) solution technique is working electrode, platinum electrode is to the utmost point, saturated calomel electrode is a reference electrode, and pseudo body fluid is as deposit fluid.Pseudo body fluid (SBF) consist of NaCl:8.0g/L, CaCl
2: 0.14 g/L, KCl:0.40 g/L, NaHCO
3: 0.35 g/L, Glucose:1.00 g/L, KH
2PO
4: 0.10 g/L, MgCl
26H
2O:0.10 g/L, Na
2HPO
42H
2O:0.06 g/L, MgSO
47H
2O:0.06 g/L.The temperature of deposit fluid is a room temperature, and the pH of deposit fluid is 8.0, deposition current Wei – 50 mA, and deposition voltage Wei – 4.0 V, sedimentation time is to obtain the nano-particle modified titanio titanate of adsorbed property of porous hydroxyl calcium phosphate nano wire biological support under the 12 h conditions.
Adopt Japanese HITACHI S-4800 scanning electron microscope under 10 kV high pressure, titanio titanate nano wire pattern to be carried out observation analysis.As seen from Figure 2, by the micropore of the titanio titanate nano wire with latticed microcellular structure of solution technique preparation between 5~50 μ m, regular shape and being evenly distributed.
Adopt Japanese HITACHI S-4800 scanning electron microscope under 10 kV high pressure, the pattern of the nano-particle modified titanio titanate nano wire biological support of adsorbed property of porous hydroxyl calcium phosphate to be carried out observation analysis.Fig. 3 is the SEM figure of the nano-particle modified titanio titanate nano wire biological support of adsorbed property of porous hydroxyl calcium phosphate.As seen from Figure 3, the adsorbed property of porous hydroxyl calcium phosphate nanoparticle growth presents the pattern of " pearl necklace " shape on the titanate nano wire, adsorbed property of porous hydroxyl calcium phosphate nano-particle size is evenly distributed between 50~500 nm, and the microcellular structure of regular shape and titanio titanate nano wire is uninfluenced.
Employing is analyzed the element that the nano-particle modified titanio titanate nano wire biological support of adsorbed property of porous hydroxyl calcium phosphate is contained with the Japanese attached X-ray energy spectrum of joining of HITACHI S-4800 scanning electron microscope (EDX).Fig. 4 is the EDX figure of the nano-particle modified titanio titanate nano wire biological support of adsorbed property of porous hydroxyl calcium phosphate.As seen from Figure 4, this biologic bracket material contains sodium, magnesium, calcium, phosphorus, titanium, oxygen element, contains the necessary all elements of calcium hydroxy phosphate and titanate nano wire and forms.
Adopt German Bruker AXS D8 Discover X-ray diffractometer with Cu K alpha ray (wavelength X=0.15405 nm, 0.020 ° of leg speed of scanning/s) at room temperature the nano-particle modified titanio titanate nano wire biological support of adsorbed property of porous hydroxyl calcium phosphate is done X-ray diffraction (XRD) analysis for diffraction light sources.Fig. 5 is the XRD figure of the nano-particle modified titanio titanate nano wire biological support of adsorbed property of porous hydroxyl calcium phosphate.As seen from Figure 5,2 θ have calcium hydroxy phosphate and the corresponding diffraction maximum of titanate nano wire at 10 °~90 ° in the X-ray diffraction spectrogram, wherein, represents the diffraction maximum of titanate nano wire correspondence, ● the diffraction maximum of representation hydroxy calcium phosphate correspondence, but all equal indexs of diffraction maximum of calcium hydroxy phosphate are hexagonal structure, have
P6
3/
mIn the face of claiming structure, the position is all consistent with the result of standard powder diffraction card (JCPDS 72-1243).Fig. 6 is that the nano-particle modified titanio titanate nano wire biological support of adsorbed property of porous hydroxyl calcium phosphate is through 450 ° of C(lower curves) and 550 ° of C(upper curves) X-ray diffraction (XRD) collection of illustrative plates of calcining behind the 3h.As seen from Figure 6, obvious variation does not take place in its XRD figure spectrum, this shows by nano-particle modified its structural behaviour of titanio titanate nano wire biological support of the adsorbed property of porous hydroxyl calcium phosphate of electrochemical process preparation stable, do not need calcining, and the prepared calcium hydroxy phosphate of most existing preparation method all will pass through calcination process basically, compared with prior art this method has been simplified technology to a certain extent, has reduced cost.
Adopt Japanese JEM 2100F transmission electron microscope (TEM) under 200 kV high pressure, the pattern of the nano-particle modified titanio titanate nano wire of its adsorbed property of porous hydroxyl calcium phosphate to be observed.Fig. 7 is the TEM figure of the nano-particle modified titanio titanate nano wire biological support of adsorbed property of porous hydroxyl calcium phosphate.As seen from Figure 7, adsorbed property of porous hydroxyl calcium phosphate nano-particle size is about 100 nm, and presents cellular, and this further provides foundation for the loading and the sustained release of adsorbed property of porous hydroxyl calcium phosphate medicine or cell growth factor.Fig. 8 is the HRTEM figure of the nano-particle modified titanio titanate nano wire biological support of adsorbed property of porous hydroxyl calcium phosphate.What as seen from Figure 8, lattice fringe spacing 0.409 nm of six sides' adsorbed property of porous hydroxyl calcium phosphate nano-particle and 0.345 nm were corresponding respectively is its (200) and (002) crystal face.
Adopt U.S. Nicolet 5700 FTIR spectrogrphs at 4000~400 cm
– 1The scope the inside is made FTIR-ATR to the nano-particle modified titanio titanate nano wire biological support of adsorbed property of porous hydroxyl calcium phosphate and is analyzed.Fig. 9 is the FTIR-ATR figure of the nano-particle modified titanio titanate nano wire biological support of adsorbed property of porous hydroxyl calcium phosphate.As seen from Figure 9,1041 cm
-1The place is PO
4 3-ν
3The vibrations peak, 3698 cm
-1The place is OH
-The absorbing vibration peak of component, 872 cm
-1The place may be CO
3 2-ν
2The vibrations peak, its ν
3The vibrations peak is at 1453 cm
-1And 1417 cm
-1The place, 1540 cm
-1The CO of place
3 2-The disappearance at vibrations peak may show the generation of the HA that the Type B replacement is arranged, i.e. CO
3 2-The PO of substituted hydroxy calcium phosphate the inside
4 3-, this replacement makes this kind biologic bracket material more approach the calcium hydroxy phosphate of organism self, makes this biological support to be accepted by this somatic cell of organism and tissue better.Figure 10 is that the nano-particle modified titanio titanate nano wire biological support of adsorbed property of porous hydroxyl calcium phosphate is through 450 ° of C(lower curves) and 550 ° of C (upper curve) calcining 3h after FTIR-ATR figure.As seen from Figure 10, obvious variation does not take place in its FTIR-ATR collection of illustrative plates, this shows that the nano-particle modified titanio titanate nano wire biological support of cellular calcium hydroxy phosphate by the electrochemical deposition method preparation has good heat stability.
For the biocompatibility of the nano-particle modified titanio titanate nano wire biological support of the adsorbed property of porous hydroxyl calcium phosphate of understanding electrochemical deposition method preparation of the present invention better, further this biological support has also been done cell culture experiments research.Concrete grammar is as follows: with this biologic bracket material, place among the aseptic culture plate, adding volume fraction is that 75% alcoholic solution is disinfected 30 min, PBS buffer (pH 7.4) fully soaks 1 h, the middle liquid that changed every 15 minutes, to guarantee to exchange out all residual ethanol, place irradiation 1.5 h under the uviol lamp then, natural air drying, biologic bracket material after sterilization DMEM culture fluid (20% hyclone, 100 U/mL penicillins and 100 μ g/mL streptomycins) soak 24 h, move into 24 hole aseptic culture plates.Get 500 μ L MG63 cell suspension (1.0 * 10
5Individual/as mL) to be inoculated in the culture plate that contains biological support, at 37 ° of C, saturated humidity, volume fraction 5% CO
2After cultivating 2h in the incubator,,, continue to cultivate 2 h to the cell suspension 500 μ L of opposite side inoculation same concentrations with the upset of rectangle sample.Every then hole adds 1 mL DMEM culture fluid, and culture plate places 37 ° of C, saturated humidity, volume fraction 5% CO
2Cultivate in the incubator, change liquid next day, change culture fluid every other day once later on, cultivated altogether 7 days.
For the growth conditions situation of cell behind the cell culture on the nano-particle modified titanio titanate nano wire biological support of the adsorbed property of porous hydroxyl calcium phosphate of understanding electrochemical deposition method preparation of the present invention, further the cell on this biological support is carried out the scanning electron microscopy study analysis.Concrete grammar is as follows: get the culture plate of cultivating 1 day that contains sample, clean sample 2 times with PBS buffer (pH 7.4), the sucking-off cleanout fluid, with volume fraction is that 10%, 20%, 30%, 50%, 60%, 70%, 80%, 90% the alcoholic solution biological support after to cultured cell carries out the gradient processed, processed is 20 minutes under each volume fraction, uses the dehydrated alcohol processed at last 3~5 times, each 15 minutes, sucking-off ethanol can carry out the scanning of SEM behind the natural air drying.Similarly, the biological support behind 3,5,7 days the cultured cell is also operated according to above-mentioned steps.
For the growth conditions situation of cell behind the cell culture on the nano-particle modified titanio titanate nano wire biological support of the adsorbed property of porous hydroxyl calcium phosphate of understanding electrochemical deposition method preparation of the present invention, further the cell on this biological support is carried out fluorescence microscope and research and analyse.Concrete grammar is as follows: get the culture plate of cultivating 1 day that contains biological support, clean sample 2 times with PBS buffer (pH 7.4), each 5 minutes, back sucking-off cleanout fluid, at room temperature, the glutaraldehyde fixed cell 10 minutes that adds 500 μ l 2.5%, with the PBS washing sample that contains 0.1% Triton X-100 3 times, each 5 minutes, back sucking-off cleaning mixture added 500 μ l Actin-Tracker Green(microfilament green fluorescence probes) in incubator, placed 60 minutes, biological support is washed 3 times with the PBS that contains 0.1% Triton X-100 in the back, each 5 minutes, back sucking-off cleaning mixture added 500 μ l DAPI dyeing liquors, placed 10 minutes under the room temperature, with the PBS washing sample that contains 0.1% Triton X-100 3 times, each 5 minutes, can carry out the shooting of fluorescence microscope after finishing.Similarly, 3,5,7 days sample is also operated according to above-mentioned steps.
In order to understand the cytoactive and the biocompatibility of the nano-particle modified titanio titanate nano wire biological support of adsorbed property of porous hydroxyl calcium phosphate of the present invention better, further this biological support is done tetramethyl azo azoles salt (MTT) colorimetry and alkali phosphatase (ALP) determination of activity analysis.It is as follows that mtt assay is measured process: biological support is respectively after cell inoculation is cultivated 1,3,5,7 day, DMEM cell culture fluid with serum-free washes this biological support, with the not adherent cell of removal, and in the aseptic culture plate that cell/timbering material complex immigration is new.Every hole adds the culture fluid and the 500 μ L MTT stock solutions (5 mg/mL) of 1 mL serum-free, continue to cultivate 4 h in 37 ° of C incubators, in this process, there is the purple material on support, to form, the careful culture fluid of drawing, add 500 μ L DMSO, and constantly vibration, make all dissolvings of purple precipitation, get 200 μ L lysates and change in 24 orifice plates, microplate reader is measured its optical density value at 570 nm places (OD).OD570 when cultivating with pure DMEM culture fluid (being the inanimate object timbering material) is a benchmark, calculates ability of cell proliferation, all makees 3 parallel laboratory test samples at every turn, gets its meansigma methods.
It is as follows that the ALP method is measured process: the biological support of inoculated and cultured after 1,3,5,7 days taken out from culture fluid, with PBS buffer (pH 7.4) flushing 2 times, adding 700 μ L Digestive systems left standstill 5 minutes, blow and beat gently for several times with suction pipe, after add again and contain blood serum medium and stop digestion, fully extrude in the biological support behind the liquid centrifugal collecting cell.With PBS buffer (pH 7.4) rinsing cell, add 500 μ L TritonX-100(2% again) cell pyrolysis liquid, 3 cracking cells of multigelation.After the optical density value (OD) at 520 nm places, calculate the ALP activity with the spectrophotometric determination cell pyrolysis liquid.All make 3 parallel laboratory test samples at every turn, get its meansigma methods.
Wherein, the U/100 mL of ALP(King unit)=(measure pipe OD/ standard pipe OD) * standard pipe contains phenol amount (0.005 mg) * (100 mL/0.05 mL)
Adopt the pattern of the cell on the nano-particle modified titanio titanate nano wire biological support of the adsorbed property of porous hydroxyl calcium phosphate of Japanese HITACHI S-4800 scanning electron microscope after pair cell under the 10 kV high pressure is cultivated to carry out observation analysis.Figure 11 is scanning electron microscope (SEM) figure of cell culture experiment research for the nano-particle modified titanio titanate nano wire biological support of adsorbed property of porous hydroxyl calcium phosphate.A ﹑ B ﹑ C ﹑ D is respectively the SEM figure that the MG63 cell was cultivated 1,3,5,7 day on this timbering material, the growth conditions figure that corresponding insertion figure is the MG63 cell on the nano-particle modified titanate nano wire of the micropore of this biological support and calcium hydroxy phosphate.As seen from Figure 11, what MG63 sprawled on this biologic bracket material after through 7 days cultivation opens very much, and basically that calcium hydroxy phosphate is nano-particle modified titanate nano wire wraps up, and the MG63 cell has good adhesion, multiplication capacity on this timbering material.
Adopt American I X71, the Olympus inverted fluorescence microscope carries out observation analysis to the growth morphology of MG63 cell on this biologic bracket material under blue light (excitation wavelength 420~485 nm) and green glow (excitation wavelength 460~550 nm).Figure 12 is the inverted fluorescence microscope figure of cell culture experiment research for the nano-particle modified titanio titanate nano wire biological support of adsorbed property of porous hydroxyl calcium phosphate.A ﹑ B ﹑ C ﹑ D is respectively the MG63 cell and cultivates 1,3,5,7 day MG63 cell growth state figure on this timbering material, and corresponding insertion figure is the nuclear fluorescence microscope picture of MG63.As seen from Figure 12, MG63 has good propagation and differentiating phenomenon after through 7 days cultivation on this biologic bracket material, and the result is consistent with SEM cell Electronic Speculum picture.
Adopt U.S. Bio-RAD Model680 microplate reader that sample is demarcated, calculate its MTT value and ALP value.The MTT colorimetric method for determining rectangular histogram of Figure 13 cell culture experiments research, Figure 14 is the ALP determination of activity rectangular histogram of cell culture experiments research, wherein, TN represents titanio titanate nano wire, and HA-TN represents the nano-particle modified titanio titanate nano wire biological support of adsorbed property of porous hydroxyl calcium phosphate.By Figure 13 and Figure 14 as seen, compare with simple titanio titanate nano wire, the MG63 cell has good adhesion, multiplication capacity on the nano-particle modified titanio titanate nano wire biological support of adsorbed property of porous hydroxyl calcium phosphate, and can cause early stage bone differentiation.
Embodiment 2: present embodiment carries out according to the following steps:
(1) the titanio titanate nano wire with latticed microcellular structure of solution technique preparation.Concrete preparation method is: the titanium substrate is in ultrasonic cleaning 20 min in acetone and ethanol solution successively under the room temperature, and the back is with distilled water flushing 4 times, this titanium substrate transferred to after air-dry and contains 5.0 mol/L NaOH and Ba (OH)
2﹒ 8H
2Seal in the politef reactor of the mixed solution of O, behind reaction 8 h under 200 ° of C, obtain having the titanio titanate nano wire of latticed microcellular structure, titanio titanate nano wire is extremely neutral with deionized water rinsing, and at air drying.
(2) electrochemical deposition method prepares the nano-particle modified titanio titanate nano wire biological support of adsorbed property of porous hydroxyl calcium phosphate.Preparation process is as follows: in three electrode reaction ponds, adopt IM6 constant current/constant voltage output mode, with the synthetic titanio titanate nano wire with latticed microcellular structure of step (1) solution technique is working electrode, platinum electrode is to the utmost point, saturated calomel electrode is a reference electrode, with pseudo body fluid (SBF) as deposit fluid (composition of pseudo body fluid is identical with embodiment 1), the deposit fluid temperature is 37 ° of C, the pH of deposit fluid is 9.6, deposition current Wei – 10 mA, deposition voltage Wei – 2.0 V, sedimentation time is to obtain the nano-particle modified titanio titanate nano wire biological support of adsorbed property of porous hydroxyl calcium phosphate under the 5 h conditions.
Adopt the scanning electron microscope identical with embodiment 1 to carry out observation analysis to the titanio titanate nano wire with latticed microcellular structure of gained, the result of gained is identical with embodiment 1, its micropore between 5~50 μ m, regular shape and being evenly distributed.
Adopt SEM, XRD, TEM, HRTEM, the FTIR-ATR identical with embodiment 1 to research and analyse to the nano-particle modified titanio titanate nano wire biological support of the adsorbed property of porous hydroxyl calcium phosphate of gained, the result of gained is identical with embodiment 1.
The nano-particle modified titanio titanate nano wire biological support of the adsorbed property of porous hydroxyl calcium phosphate of gained is adopted cell culture experiments research and accordingly SEM, fluorescence microscope and MTT, the ALP experimentation analysis identical with embodiment 1, and the result of gained is identical with embodiment 1.
Embodiment 3: present embodiment carries out according to the following steps:
(1) the titanio titanate nano wire with latticed microcellular structure of solution technique preparation.Concrete preparation method is: the titanium substrate is in ultrasonic cleaning 20 min in acetone and ethanol solution successively under the room temperature, and the back is with distilled water flushing 4 times, this titanium substrate transferred to after air-dry and contains 10 mol/L NaOH and Ba (OH)
2﹒ 8H
2Seal in the politef reactor of the mixed solution of O, behind reaction 1 h under 100 ° of C, obtain having the titanio titanate nano wire of latticed microcellular structure, titanio titanate nano wire is extremely neutral with deionized water rinsing, and at air drying.
(2) electrochemical deposition method prepares the nano-particle modified titanio titanate nano wire biological support of adsorbed property of porous hydroxyl calcium phosphate.Preparation process is as follows: in three electrode reaction ponds, adopt IM6 constant current/constant voltage output mode, with the synthetic titanio titanate nano wire with latticed microcellular structure of step (1) solution technique is working electrode, platinum electrode is to the utmost point, saturated calomel electrode is a reference electrode, with pseudo body fluid (SBF) is deposit fluid (composition of pseudo body fluid is identical with embodiment 1), the deposit fluid temperature is 90 ° of C, the pH of deposit fluid is 12.6, deposition current is 50 mA, deposition voltage is 4.0 V, and sedimentation time is to obtain the nano-particle modified titanio titanate nano wire biological support of adsorbed property of porous hydroxyl calcium phosphate under the 0.5 h condition.
Adopt the scanning electron microscope identical with embodiment 1 to carry out observation analysis to the titanio titanate nano wire with latticed microcellular structure of gained, the result of gained is identical with embodiment 1, its micropore between 5~50 μ m, regular shape and being evenly distributed.
Adopt SEM, XRD, TEM, HRTEM, the FTIR-ATR identical with embodiment 1 to research and analyse to the nano-particle modified titanio titanate nano wire biological support of the adsorbed property of porous hydroxyl calcium phosphate of gained, the result of gained is identical with embodiment 1.
The nano-particle modified titanio titanate nano wire biological support of the adsorbed property of porous hydroxyl calcium phosphate of gained is adopted cell culture experiments research and accordingly SEM, fluorescence microscope and MTT, the ALP experimentation analysis identical with embodiment 1, and the result of gained is identical with embodiment 1.