Background technology
Canine implant is considered to the first-selection of absence of tooth in the world and repairs means, and it is similar at aspect such as function, form, result of use and dermal tooth, can be referred to as " emulation tooth ".Dental Implant is made up of artificial tooth root and porcelain corona, and pure titanium and its alloy are widely used in the manufacturing of dental implant artificial tooth root.Yet common titanium dioxide layer does not have enough activity and bone directly to form to get in touch, and sometimes is converted into to lack bone and integrate and make long-term graft failure.Implant surfaces pattern and chemical feature modification are proved to be a kind of effective means that bone is integrated that improves, and are a kind of reliable approach of guaranteeing bone contact implantation body stability.
Show that according to the study silver has broad-spectrum antiseptic, germicidal efficiency height, characteristics such as be difficult for developing immunity to drugs, its contact microorganism and with the tropina enzyme-the SH base combines, make the microorganism loss of activity and cause antibacterial death.Silver can combine and form cross-linked with the DNA of pathogenic base in addition, and the hydrogen bond in displacement purine and the pyrimidine between the adjacent nitrogen makes the DNA degeneration and reproducible not, plays and suppresses or the effect of kill bacteria.Carry out the artificial tooth root that surgery is transplanted, all more or less meeting contacts with external environment condition before and after implanting, thereby pollutes.And mix the artificial tooth root material of silver element, and can greatly improve the antibiotic and sterilizing ability of artificial tooth root self, strengthen the safety in its clinical practice.But silver ion easily excretes with the blood of human body circulation, must seek appropriate method and control its slow release.
(bone morphogenetic protein BMP) is the multi-functional somatomedin of a class to bone morphogenetic protein, and induced osteogenesis, the reparation of promotion bone have significant curative effect to union of fracture delay and bone does not connect effectively.BMP degrades rapidly in local organization or with the body fluid diffusion, can not be evenly distributed in the bone defect lastingly, is difficult to give play to ideal bone-forming effect.The effect that the exogenous BMP treatment of simple application bone is damaged is unsatisfactory, thereby must have the suitable carriers material compound with it, plays the effect that makes the BMP slow release.Various carrier systems respectively have pluses and minuses to the slow release of BMP, thereby the more perfect composite slow-released carrier of research and development function, as: inorganic/inorganic compounding, organic/inorganic is compound, organic/organic compound or many materials complex carrier, reaching better slow release even accurate controlled release BMP will become a main direction of research from now on.
The main inanimate matter of human body hard tissue is a calcium phosphorus phase, and the metatitanic acid sodium salt can change hydrated titanium dioxide in body fluid, make the metal surface original position form many Ti-OH groups, thereby induce calcium, phosphorus deposition in the body fluid, form apatite layer (Ceramics International, 1996,79 (24): 1127-1129).Because the crystalline special construction of sodium titanate makes sodium ion that very strong exchange capacity be arranged, can replace with multiple cation, thereby carry out the ion slow release in addition, strengthen the functional of sodium titanate structure sheaf.The special appearance of surface micropore has increased specific surface area simultaneously, makes the sodium titanate structure sheaf be strengthened greatly aspect load capacity.
The common method of the sodium titanate structure of preparation biologically active is a lye hydrothermal synthesis technology.Chinese patent ZL02117866.6 " a kind of process for preparing hydrated titanium acid sodium-salt and serial titanate nanotube " discloses a kind of method for preparing the sodium titanate nanostructured based on lye hydrothermal synthesis technology, promptly in hermetic container, use titanium dioxide powder or metatitanic acid powder body and sodium hydroxide solution to carry out hydro-thermal reaction, finally make dispersive metatitanic acid sodium salt with tubular structure.But this preparation method can only be prepared dispersive titanate structure, does not relate to the preparation of metal base surface.And sodium titanate also seldom is applied to artificial tooth root material technology field as a kind of bio-medical material that has potentiality.Therefore seek suitable technology and have the sodium titanate bioactivity coatings of special construction, and strengthen its functional and load capacity, will more and more be subjected to the attention in artificial tooth root material technology field in the preparation of artificial tooth root metal base surface.
Summary of the invention
Technical problem: the objective of the invention is to propose a kind of antimicrobial artificial tooth root based on nanometer tube array surface and preparation method thereof, construct titania nanotube shape structure by two step anodizings at metal base surface, synthesize through hydro-thermal again and change titania nanotube into sodium titanate nanotubes, in sodium titanate nanotube array layer, mix trace element silver by solution impregnation iron exchange subsequently, pass through the centrifugal load technology at last at nanotube layer internal burden bone morphogenetic protein.Prepare mix silver-colored sodium titanate nanotube array layer and substrate combinating strength is good, preparation technology is simple, and is workable.
Technical scheme: the present invention is realized by the following technical programs:
One deck is arranged through the adulterated sodium titanate nanotube array layer with tubular structure of silver at this artificial tooth root material metal matrix surface, further load has bone morphogenetic protein (BMP) in its layer.
The step of concrete preparation is as follows:
1) at first select for use pure titanium or titanium alloy to process artificial tooth root;
2) artificial tooth root surface preparation: behind the surface grinding, use acetone, deionized water and dehydrated alcohol supersound washing 10-120 minute successively, use HF and HNO subsequently
3And mixed solution (volume ratio is 1: 4: 5) the chemical polishing 20-120 second of water, behind the deionized water cleaning down, dry up stand-by again;
3) two step anodic oxidations prepare titania nanotube: with the artificial tooth root of surface preparation as anode, graphite or platinum are negative electrode, in moisture organic electrolyte solution, carry out the anodic oxidation first time, concrete anodic oxidation parameter is: voltage 20-240V, response time 12-200 hour, the two poles of the earth are apart from 3-10cm, and temperature remains room temperature.After the anodic oxidation for the first time, take out, in percent by volume is in the hydrochloric acid solution of 5-30vol% ultrasonic cleaning 0.5-3 hour, after taking-up dries up, artificial tooth root is carried out the anodic oxidation second time again under aforementioned anodic oxidation first time parameter condition, take out then, with dehydrated alcohol ultrasonic cleaning 10-30 minute, after being to soak 0.5-3 hour in the hydrochloric acid solution of 5-30vol%, percent by volume takes out again, with dehydrated alcohol and deionized water alternately clean 6-10 all over after, at 60-100 ℃ of following drying sample, obtain titanium dioxide nanotube array layer again on the artificial tooth root surface;
4) the synthetic sodium titanate nanotube array layer of hydro-thermal: the artificial tooth root of the surface coverage titanium dioxide nanotube array layer of above-mentioned preparation is carried out hydro-thermal reaction in the teflon-lined autoclave, concrete hydro-thermal reaction parameter is: reaction temperature 90-180 ℃, response time 10-100 hour, concentration of sodium hydroxide solution 5-15molL
-1, the reaction solution volume is the volumetrical 50-80% of inner liner of reaction kettle.After the hydro-thermal reaction, treat natural cooling, take out, centrifugal with deionized water wash, repeatedly for several times, be neutral until the supernatant, taking-up is dry, obtains sodium titanate nanotube array layer on the artificial tooth root surface;
5) artificial tooth root of the surface coverage sodium titanate nanotube array layer of above-mentioned preparation being immersed in concentration is 5 * 10
-2~2 * 10
-1The AgNO of mol/L
3Mix trace element silver by ion exchange in the solution, liquid level of solution should flood artificial tooth root, and the water bath with thermostatic control temperature is 60~100 ℃, soak time 2~12 hours.With washed with de-ionized water several times subsequently, to eliminate remaining soluble ion, sample is drying at room temperature under vacuum, obtains silver-colored adulterated sodium titanate nanotube array layer on the artificial tooth root surface;
6) adopt the centrifugal load technology at the adulterated sodium titanate nanotube array layer internal burden of silver bone morphogenetic protein at last, the artificial tooth root that the surface is had silver-colored adulterated sodium titanate nanotube array layer places the centrifuge container bottom, pour the mixed solution that bone morphogenetic protein and phosphate buffered saline(PBS) are formed again into, liquid level flooded artificial tooth root to half of centrifuge container to 2/3rds height, bone morphogenetic protein concentration is 0.1-50mgL
-1, centrifugal under the room temperature (1500-80000Rpm) soaked 0.5-10 hour, took out, get final product artificial tooth root with bioactivity of the present invention.
It is solvent that moisture organic electrolyte solution of the present invention adopts ethylene glycol, and other effective ingredient is: water 0-10vol% (percent by volume), phosphoric acid 2-10vol% (percent by volume), ammonium fluoride 0.1-0.9wt% (mass percent).
The sodium titanate nanotubes rule oriented of the inventive method preparation is arranged, and sodium titanate nanotubes is producing perpendicular to the matrix direction, and caliber reaches 80-500nm, and pipe range is 1-150 μ m, and specific surface area is bigger.
The artificial tooth root with bioactivity of mixing silver-colored sodium titanate nanotube array layer of the present invention preparation and method has the following advantages and beneficial effect:
1. silver has broad-spectrum antiseptic, germicidal efficiency height, characteristics such as be difficult for developing immunity to drugs.The artificial tooth root material of Ag-carried antibacterial type of the present invention has sterilizing effect and induced osteogenesis effect, can form firm physiology with osseous tissue safely in a short time and combine.
2. compare with common sodium titanate layer, the sodium titanate nanotubes array is porous and aligns, and specific surface area is higher, helps the load of bone morphogenetic protein and to the controlled release of bone morphogenetic protein.
3. the titanium dioxide nanotube array layer of anodic oxidation formation is that matrix is generated in-situ, and is fine with the adhesion of matrix.Bone morphogenetic protein has good induced osteogenesis effect, can form firm physiology with osseous tissue in the short time and combine.Therefore, be expected to increase artificial tooth root stability.
4. anodic oxidation of Cai Yonging and hydro-thermal synthesis process are not subjected to the influence of base shape, can prepare uniform sodium titanate nanotube array layer on complex-shaped matrix.
The specific embodiment
Embodiment 1
(1) at first select for use pure titanium to process artificial tooth root;
(2) artificial tooth root surface preparation: behind the surface grinding, use acetone, deionized water and dehydrated alcohol supersound washing 10-30 minute successively, use HF and HNO subsequently
3And mixed solution (volume ratio is 1: 4: 5) the chemical polishing 20-40 second of water, behind the deionized water cleaning down, dry up stand-by again;
(3) prepare moisture organic electrolyte solution: employing ethylene glycol is solvent, and other effective ingredient is: water 2vol% (percent by volume), phosphatase 24 vol% (percent by volume), ammonium fluoride 0.3wt% (mass percent);
(4) two step anodic oxidations prepare titania nanotube: with the artificial tooth root of surface preparation as anode, graphite or platinized platinum are negative electrode, in moisture organic electrolyte solution, carry out the anodic oxidation first time, concrete anodic oxidation parameter is: voltage 60V, 18 hours response time, the two poles of the earth are apart from 3-10cm, and temperature remains room temperature.After the anodic oxidation for the first time, take out, in percent by volume is in the hydrochloric acid solution of 10vol% ultrasonic cleaning 0.5-3 hour, after taking-up dries up, artificial tooth root is carried out the anodic oxidation second time again under aforementioned anodic oxidation first time parameter condition, take out then, with dehydrated alcohol ultrasonic cleaning 10-30 minute, after being to soak 0.5-3 hour in the hydrochloric acid solution of 10vol%, percent by volume takes out again, with dehydrated alcohol and deionized water alternately clean 6-10 all over after, at 60-100 ℃ of following drying sample, obtain titanium dioxide nanotube array layer again on the artificial tooth root surface;
(5) the synthetic sodium titanate nanotube array layer of hydro-thermal: the artificial tooth root of the surface coverage titanium dioxide nanotube array layer of above-mentioned preparation is carried out hydro-thermal reaction in the teflon-lined autoclave, concrete hydro-thermal reaction parameter is: 110 ℃ of reaction temperatures, 20 hours response time, concentration of sodium hydroxide solution 10molL
-1, the reaction solution volume is the volumetrical 50-80% of inner liner of reaction kettle.After the hydro-thermal reaction, treat natural cooling, take out, centrifugal with deionized water wash, repeatedly for several times, be neutral until the supernatant, taking-up is dry, obtains sodium titanate nanotube array layer on the artificial tooth root surface;
5) artificial tooth root of the surface coverage sodium titanate nanotube array layer of above-mentioned preparation being immersed in concentration is 5 * 10
-2The AgNO of mol/L
3Mix trace element silver by ion exchange in the solution, liquid level of solution should flood artificial tooth root, and the water bath with thermostatic control temperature is 60 ℃, soak time 2~12 hours.With washed with de-ionized water several times subsequently, to eliminate remaining soluble ion, sample is drying at room temperature under vacuum, obtains silver-colored adulterated sodium titanate nanotube array layer on the artificial tooth root surface;
6) adopt the centrifugal load technology at the adulterated sodium titanate nanotube array layer internal burden of silver bone morphogenetic protein at last, the artificial tooth root that the surface is had silver-colored adulterated sodium titanate nanotube array layer places the centrifuge container bottom, pour the mixed solution that the phosphate buffered saline(PBS) of the bone morphogenetic protein of 0.1~50mg/L and 0.1mol/L is formed into, liquid level flooded artificial tooth root half height to centrifuge container, centrifugal speed working time 5~30min with 6000rpm, keep 4~40 ℃ of operating temperatures, take out, get final product the artificial root of the tooth of the present invention.
Embodiment 2
(1) at first select for use titanium to process artificial tooth root;
(2) artificial tooth root surface preparation: behind the surface grinding, use acetone, deionized water and dehydrated alcohol supersound washing 30-60 minute successively, use HF and HNO subsequently
3And mixed solution (volume ratio is 1: 4: 5) the chemical polishing 40-60 second of water, behind the deionized water cleaning down, hair dryer dries up stand-by again;
(3) prepare moisture organic electrolyte solution: employing ethylene glycol is solvent, and other effective ingredient is: water 2vol% (percent by volume), phosphatase 24 vol% (percent by volume), ammonium fluoride 0.5wt% (mass percent);
(4) two step anodic oxidations prepare titania nanotube: with the artificial tooth root of surface preparation as anode, graphite is negative electrode, in moisture organic electrolyte solution, carry out the anodic oxidation first time, concrete anodic oxidation parameter is: voltage 80V, 18 hours response time, the two poles of the earth are apart from 3-10cm, and temperature remains room temperature.After the anodic oxidation for the first time, take out, in percent by volume is in the hydrochloric acid solution of 10vol% ultrasonic cleaning 0.5-3 hour, after taking-up dries up, artificial tooth root is carried out the anodic oxidation second time again under aforementioned anodic oxidation first time parameter condition, take out then, with dehydrated alcohol ultrasonic cleaning 10-30 minute, after being to soak 0.5-3 hour in the hydrochloric acid solution of 10vol%, percent by volume takes out again, with dehydrated alcohol and deionized water alternately clean 6-10 all over after, at 60-100 ℃ of following drying sample, obtain titanium dioxide nanotube array layer again on the artificial tooth root surface;
(5) the synthetic sodium titanate nanotube array layer of hydro-thermal: the artificial tooth root of the surface coverage titanium dioxide nanotube array layer of above-mentioned preparation is carried out hydro-thermal reaction in the teflon-lined autoclave, concrete hydro-thermal reaction parameter is: 130 ℃ of reaction temperatures, 20 hours response time, concentration of sodium hydroxide solution 10molL
-1, the reaction solution volume is the volumetrical 50-80% of inner liner of reaction kettle.After the hydro-thermal reaction, treat natural cooling, take out, centrifugal with deionized water wash, repeatedly for several times, be neutral until the supernatant, taking-up is dry, obtains sodium titanate nanotube array layer on the artificial tooth root surface, and its SEM photo and XRD spectrum are seen Fig. 2 and Fig. 3 respectively;
5) artificial tooth root of the surface coverage sodium titanate nanotube array layer of above-mentioned preparation being immersed in concentration is 2 * 10
-1AgNO
3Mix trace element silver by ion exchange in the solution, liquid level of solution should flood artificial tooth root, and the water bath with thermostatic control temperature is 60 ℃, soak time 2~12 hours.With washed with de-ionized water several times subsequently, to eliminate remaining soluble ion, sample is drying at room temperature under vacuum, obtains silver-colored adulterated sodium titanate nanotube array layer on the artificial tooth root surface;
6) adopt the centrifugal load technology at the adulterated sodium titanate nanotube array layer internal burden of silver bone morphogenetic protein at last, the artificial tooth root that the surface is had silver-colored adulterated sodium titanate nanotube array layer places the centrifuge container bottom, pour the mixed solution that the phosphate buffered saline(PBS) of the bone morphogenetic protein of 0.1~50mg/L and 0.1mol/L is formed into, liquid level flooded 2/3rds the height of artificial tooth root to centrifuge container, centrifugal speed working time 5~30min with 8000rpm, keep 4~40 ℃ of operating temperatures, take out, get final product the artificial root of the tooth of the present invention.
Embodiment 3
(1) at first select for use pure titanium to process artificial tooth root;
(2) artificial tooth root surface preparation: behind the surface grinding, use acetone, deionized water and dehydrated alcohol supersound washing 60-90 minute successively, use HF and HNO subsequently
3And mixed solution (volume ratio is 1: 4: 5) the chemical polishing 60-80 second of water, behind the deionized water cleaning down, hair dryer dries up stand-by again;
(3) prepare moisture organic electrolyte solution: employing ethylene glycol is solvent, and other effective ingredient is: water 4vol% (percent by volume), phosphoric acid 6vol% (percent by volume), ammonium fluoride 0.5wt% (mass percent);
(4) two step anodic oxidations prepare titania nanotube: with the artificial tooth root of surface preparation as anode, graphite or platinized platinum are negative electrode, in moisture organic electrolyte solution, carry out the anodic oxidation first time, concrete anodic oxidation parameter is: voltage 40V, 24 hours response time, the two poles of the earth are apart from 3-10cm, and temperature remains room temperature.After the anodic oxidation for the first time, take out, in percent by volume is in the hydrochloric acid solution of 20vol% ultrasonic cleaning 0.5-3 hour, after taking-up dries up, artificial tooth root is carried out the anodic oxidation second time again under aforementioned anodic oxidation first time parameter condition, take out then, with dehydrated alcohol ultrasonic cleaning 10-30 minute, after being to soak 0.5-3 hour in the hydrochloric acid solution of 20vol%, percent by volume takes out again, with dehydrated alcohol and deionized water alternately clean 6-10 all over after, at 60-100 ℃ of following drying sample, obtain titanium dioxide nanotube array layer again on the artificial tooth root surface;
(5) the synthetic sodium titanate nanotube array layer of hydro-thermal: the artificial tooth root of the surface coverage titanium dioxide nanotube array layer of above-mentioned preparation is carried out hydro-thermal reaction in the teflon-lined autoclave, concrete hydro-thermal reaction parameter is: 150 ℃ of reaction temperatures, 20 hours response time, concentration of sodium hydroxide solution 10molL
-1, the reaction solution volume is the volumetrical 50-80% of inner liner of reaction kettle.After the hydro-thermal reaction, treat natural cooling, take out, centrifugal with deionized water wash, repeatedly for several times, be neutral until the supernatant, taking-up is dry, obtains sodium titanate nanotube array layer on the artificial tooth root surface;
5) artificial tooth root of the surface coverage sodium titanate nanotube array layer of above-mentioned preparation being immersed in concentration is 2 * 10
-1The AgNO of mol/L
3Mix trace element silver by ion exchange in the solution, liquid level of solution should flood artificial tooth root, and the water bath with thermostatic control temperature is 60 ℃, soak time 8 hours.With washed with de-ionized water several times subsequently, to eliminate remaining soluble ion, sample is drying at room temperature under vacuum, obtains silver-colored adulterated sodium titanate nanotube array layer on the artificial tooth root surface;
6) adopt the centrifugal load technology at the adulterated sodium titanate nanotube array layer internal burden of silver bone morphogenetic protein at last, the artificial tooth root that the surface is had silver-colored adulterated sodium titanate nanotube array layer places the centrifuge container bottom, pour the mixed solution that the phosphate buffered saline(PBS) of the bone morphogenetic protein of 0.1~50mg/L and 0.1mol/L is formed into, liquid level flooded half the height of artificial tooth root to centrifuge container, centrifugal speed working time 5~30min with 6000rpm, keep 4~40 ℃ of operating temperatures, take out, get final product the artificial root of the tooth of the present invention.
Embodiment 4
(1) at first select for use pure titanium to process artificial tooth root;
(2) artificial tooth root surface preparation: behind the surface grinding, use acetone, deionized water and dehydrated alcohol supersound washing 90-120 minute successively, use HF and HNO subsequently
3And mixed solution (volume ratio is 1: 4: 5) the chemical polishing 80-120 second of water, behind the deionized water cleaning down, hair dryer dries up stand-by again;
(3) prepare moisture organic electrolyte solution: employing ethylene glycol is solvent, and other effective ingredient is: water 2vol% (percent by volume), phosphoric acid 5vol% (percent by volume), ammonium fluoride 0.5wt% (mass percent);
(4) two step anodic oxidations prepare titania nanotube: with the artificial tooth root of surface preparation as anode, graphite or platinized platinum are negative electrode, in moisture organic electrolyte solution, carry out the anodic oxidation first time, concrete anodic oxidation parameter is: voltage 80V, 12 hours response time, the two poles of the earth are apart from 3-10cm, and temperature remains room temperature.After the anodic oxidation for the first time, take out, in percent by volume is in the hydrochloric acid solution of 20vol% ultrasonic cleaning 0.5-3 hour, after taking-up dries up, artificial tooth root is carried out the anodic oxidation second time again under aforementioned anodic oxidation first time parameter condition, take out then, with dehydrated alcohol ultrasonic cleaning 10-30 minute, after being to soak 0.5-3 hour in the hydrochloric acid solution of 20vol%, percent by volume takes out again, with dehydrated alcohol and deionized water alternately clean 6-10 all over after, at 60-100 ℃ of following drying sample, obtain titanium dioxide nanotube array layer again on the artificial tooth root surface;
(5) the synthetic sodium titanate nanotube array layer of hydro-thermal: the artificial tooth root of the surface coverage titanium dioxide nanotube array layer of above-mentioned preparation is carried out hydro-thermal reaction in the teflon-lined autoclave, concrete hydro-thermal reaction parameter is: 110 ℃ of reaction temperatures, 30 hours response time, concentration of sodium hydroxide solution 10molL
-1, the reaction solution volume is the volumetrical 50-80% of inner liner of reaction kettle.After the hydro-thermal reaction, treat natural cooling, take out, centrifugal with deionized water wash, repeatedly for several times, be neutral until the supernatant, taking-up is dry, obtains sodium titanate nanotube array layer on the artificial tooth root surface;
5) artificial tooth root of the surface coverage sodium titanate nanotube array layer of above-mentioned preparation being immersed in concentration is 5 * 10
-2The AgNO of mol/L
3Mix trace element silver by ion exchange in the solution, liquid level of solution should flood artificial tooth root, and the water bath with thermostatic control temperature is 60 ℃, soak time 2~12 hours.With washed with de-ionized water several times subsequently, to eliminate remaining soluble ion, sample is drying at room temperature under vacuum, obtains silver-colored adulterated sodium titanate nanotube array layer on the artificial tooth root surface;
6) adopt the centrifugal load technology at the adulterated sodium titanate nanotube array layer internal burden of silver bone morphogenetic protein at last, the artificial tooth root that the surface is had silver-colored adulterated sodium titanate nanotube array layer places the centrifuge container bottom, pour the mixed solution that the phosphate buffered saline(PBS) of the bone morphogenetic protein of 0.1~50mg/L and 0.1mol/L is formed into, liquid level flooded 2/3rds the height of artificial tooth root to centrifuge container, centrifugal speed working time 5~30min with 6000rpm, keep 4~40 ℃ of operating temperatures, take out, get final product the artificial root of the tooth of the present invention.