CN102579145B - Dental implant and preparation method thereof - Google Patents
Dental implant and preparation method thereof Download PDFInfo
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Abstract
The invention relates to the field of dental implant materials, and discloses a dental implant carrying miRNA (micro-ribonucleic acid) or siRNA (small interfering ribonucleic acid) and a preparation method thereof. The outer surface of the dental implant is a microarc oxidation coating with micro/nano appearance, wherein factors promoting osteogenic differentiation are loaded on the microarc oxidation coating, and are miRNA or siRNA. The preparation method comprises the following steps: processing pure titanium or titanium alloy into an implant matrix; polishing, cleaning and blow-drying; producing the microarc oxidation coating with micro/nano appearance on the outer surface of the implant matrix by adopting a microarc oxidation process, cleaning and blow-drying; mixing liposome and miRNA or siRNA to produce a carrier mixture; immersing the implant matrix into the carrier mixture, freezing with drikold; and loading the carrier mixture to the microarc oxidation coating by adopting a vacuum lyophilisation process, and standing at normal temperature to obtain the dental implant carrying miRNA or siRNA.
Description
Technical field
The present invention relates to dentistry implant material, particularly a kind of dentistry implant that is loaded with miRNA or siRNA and preparation method thereof.
Background technology
Along with planting technology development, dentistry implant has been widely used in clinical defect of dentition and disappearance patient's reparation.But after implantation body implants, synosteosis is of long duration.In general, implantation body implants after 3-6 month just can load reparation, especially some especial patients, as diabetes, osteoporosis patient etc., carry out plantation more difficult, the plantation appearance of case that often leads to the failure reluctantly.This is greatly perplexing patient and odontologist.
Nowadays, in order to improve synosteosis speed, shorten the synosteosis time, existing multiple implant surface pattern occurs and is applied to clinical, mainly to adopt the technology such as differential arc oxidation (MicroarcOxidation, MAO), the spraying of titanium slurry or sandblast acid etching to improve implant surface pattern.Differential arc oxidation is as a kind of implant surface treatment technology of comparative maturity in wide clinical application, and its implant surface has loose structure, can improve synosteosis speed.
In addition, in order to improve synosteosis speed, the existing multiple scientific research report of facilitating bone differentiation factor to be successfully loaded into implant surface, mainly contains bone morphogenetic protein(BMP) (BMP), transforming growth factor-beta (TGF-β), insulin like growth factor (IGF-1) now.Its a kind of loading method is that PDLLA (poly (D, L-lactide), poly-meso lactic acid) is mixed to form to microsphere with TGF-β, IGF-1 albumen, and is wrapped in pure titanium silk surface, after normal temperature drying, and-20 ℃ of preservations in refrigerator.Another kind method, is immersed in implantation body's matrix the mixed solution 48h of biotic factor BMP and calcium phosphate exactly, to be dried after in-80 ℃ of preservations.Microsphere diameter in said method is about 30-100 μ m, covers the original pattern of dentistry implant completely, and need to preserve its activity of guarantee at-20 ℃ or-80 ℃ after having loaded; Above-mentioned steps is loaded down with trivial details in addition, first needs to adopt multi-step to prepare PDLA, PDLLA etc., then could load with after the albumen such as BMP mixes at least 48 hours load times.
Summary of the invention
One object of the present invention is to provide a kind of dentistry implant, and this dentistry implant is the differential arc oxidation coating with micro-nano pattern, is loaded with again and efficiently facilitates bone differentiation factor, can improve more rapidly synosteosis speed, shortens the synosteosis time.
Another object of the present invention is to provide the preparation method of above-mentioned dentistry implant.
In order to achieve the above object, the present invention is achieved by the following technical solutions.
A kind of dentistry implant, is characterized in that, the outer surface of described dentistry implant is the differential arc oxidation coating with micro-nano pattern, and described differential arc oxidation coating is loaded with facilitates bone differentiation factor, described in to facilitate bone differentiation factor be miRNA or siRNA.
The preparation method of above-mentioned dentistry implant, is characterized in that, comprises the following steps:
Step 1, selects pure titanium or titanium alloy to be processed into implantation body's matrix, then by after implantation body's matrix surface polishing, uses successively acetone, dehydrated alcohol and deionized water ultrasonic cleaning, then dries up;
Step 2, adopts micro-arc oxidation process to have the differential arc oxidation coating of micro-nano pattern in implantation body's outer surface of matrix preparation, then uses successively acetone, dehydrated alcohol and deionized water ultrasonic cleaning, then dries up;
Step 3, is mixed into carrier mixture by liposome and miRNA or siRNA;
Step 4, is immersed in implantation body's matrix in carrier mixture, and dry ice is freezing;
Step 5, adopts vacuum freeze-drying technique that carrier mixture is loaded into differential arc oxidation coating, and room temperature is placed, and obtains the dentistry implant that is loaded with miRNA or siRNA.
In technique scheme, preferably, step 1 adopts pure titanium plantation matrix.
Preferably, in the micro-arc oxidation process described in step 2, differential arc oxidation electrolyte solute is prepared by 0.04M sodium β-glycerophosphate and 0.2M calcium acetate, solvent is deionized water, supply frequency 100Hz, and voltage is 300 to 450V, dutycycle 20%, processing time 4-6min.
Preferably, in step 3, in described carrier mixture, miRNA or siRNA concentration are 0.3-0.5 μ M; In step 3, described liposome is one of lipid transfection reagent, comprises commercial Lipofectamine2000, Oligofectamine or TransIT-TKO.
Preferably, the cooling time of dry ice described in step 4 is 3-8min; More preferably, in step 4, dry ice cooling time is 5min.
Preferably, in vacuum freeze-drying technique, in vacuum chamber, force value is equal to or less than 1Pa described in step 5, and temperature is-20 ℃ to-30 ℃, and freeze-drying time is more than or equal to 24h.
Along with the discovery of RNA and the research of function thereof.RNA interferes (RNAi) as a kind of powerful experimental tool, and development rapidly; The RNA that its utilization has a homology acts on the special target gene silence of messenger RNA (mRNA) induced sequence of target gene, blocks rapidly target gene activity, and becomes the effective tool of gene analysis and potential clinical treatment.Wherein, siRNA (small interfering RNA; And microRNA (microRNA siRNA); MiRNA) be two kinds of regulatory factors that sequence-specific posttranscriptional gene is expressed.
With respect to the multiple bone differentiation factor of facilitating in background technology, for example, bone morphogenetic protein(BMP) (BMP), transforming growth factor-beta (TGF-β), insulin-like growth factor-i (IGF-1), RNAi has following characteristic: (1) RNAi has the specificity of height.MiRNA and siRNA can induce mRNA degraded or the translation of sequence homology with it to suppress very specifically, and unaffected with the irrelevant mRNA of sequence.(2) RNAi has high efficiency.Relatively seldom the miRNA of amount and siRNA molecule (quantity is far less than the quantity of endogenous mRNA) just can produce strong RNAi effect, produce the effect of similar gene " rejecting ".(3) there is enlarge-effect in RNAi.A small amount of miRNA and siRNA can make a large amount of target gene silences, even if cell proliferation 50-100 doubly still can keep RNAi effect, this shows that RNAi exists amplification mechanism.(4) RNAi effect extensively also can heredity.MiRNA and siRNA mediate rna i are not only importing position generation depression effect, and can cross over cell boundary and transmit and maintain in the long distance of different iuntercellulars.Research shows, RNAi effect can be propagated in vivo, and can pass to filial generation; And miRNA and siRNA source are easily, and completely external synthetic, immunogenicity is little, can be supplied in a large number clinical.
MiRNA, the siRNA of a lot of short Osteoblast Differentiation, antiinflammatory, insulin secretion accelerating found in research.Research shows, miRNA and siRNA can disturb the expression of posttranscriptional gene specifically, and for example, the miRNA of short Osteoblast Differentiation, antiinflammatory and siRNA can effectively promote synosteosis and improve the plantation success rate of implantation body under inflammatory conditions.
Dentistry implant of the present invention, adopts micro-arc oxidation process to have the differential arc oxidation coating of micro-nano pattern in implantation body's outer surface of matrix preparation, and micro-nano pattern is loose structure, and aperture is micron order; Adopt vacuum freeze-drying technique that carrier mixture is loaded into differential arc oxidation coating, in carrier mixture, the miRNA of liposome or siRNA diameter only have 200nm left and right, can evenly spread to preferably in the hole of differential arc oxidation coating and outside hole, can be because External Force Acting comes off easily, and can not change egregiously the micro-nano pattern of differential arc oxidation coating.But, the multiple bone differentiation factor of facilitating in background technology, its microsphere diameter is 30-100 μ m, has the dentistry implant of differential arc oxidation coating, by original complete covers implants pattern for conventional dentistry implant or outer surface.Therefore, dentistry implant of the present invention has the differential arc oxidation coating of micro-nano pattern, is loaded with again and efficiently facilitates bone differentiation factor, can improve more rapidly synosteosis speed, shortens the synosteosis time, improves plantation success rate.
In addition, the miRNA of liposome is difficult for preserving at normal temperatures, but can stablize and preserve 2 weeks after lyophilizing, and still preserves 70% transfection activity.The vacuum freeze-drying technique adopting in the present invention, the restriction of Bu Shou implantation body shape, can on its complicated irregular surface, realize being uniformly distributed of miRNA or siRNA, and the material that contains moisture is lowered the temperature in advance and is frozen into solid, then under the condition of vacuum, make steam directly from solid, distil out, and in the solid frame of the left ice state freezing of material itself, keep the dry rear constancy of volume of dentistry implant, loose porous.
Accompanying drawing explanation
Below in conjunction with the drawings and specific embodiments, the present invention is described in further detail.
Fig. 1 is the enforcement schematic diagram of micro-arc oxidation process, wherein, and 101, power supply; 102, electrolyte; 103, treat differential arc oxidation sample; 104, stainless steel tank; 105, cathode power supply line; 106, anode power cord; 107, chiller.
Fig. 2 is vacuum freeze-drying machine schematic diagram, wherein, and 201, vacuum pump; 202, vacuometer; 203, treat lyophilizing sample; 204, specimen holder; 205, compressor; 206, vacuum chamber.
Fig. 3 is scanning electron microscope (SEM) the photo figure of dentistry implant differential arc oxidation porous pattern, and wherein (a) be not for carrying the differential arc oxidation porous shape appearance figure of miRNA; (b) for carrying the titanium differential arc oxidation porous shape appearance figure of miRNA.
Fig. 4 is the laser co-focusing figure that dentistry implant differential arc oxidation porous pattern carries miRNA, and wherein redness is the fluorescently-labeled miRNA of cy-3.
Fig. 5 is that dentistry implant differential arc oxidation porous pattern carries the 24h laser co-focusing figure after inoculating cell after miRNA, and green is the dyeing of cell membrane green fluorescence probe (DiO) cell membrane, and redness is the fluorescently-labeled miRNA of cy-3.
The specific embodiment
With reference to Fig. 1, the enforcement schematic diagram of micro-arc oxidation process adopting for the present invention, wherein: the frequency 100Hz of power supply 101, voltage is 400V, dutycycle 20%; The interior splendid attire electrolyte 102 of stainless steel tank 104, treats that differential arc oxidation sample 103, for pure titanium or titanium alloy are processed into implantation body's matrix, inserts in electrolyte 102.Treat that differential arc oxidation sample 103 is electrically connected cathode power supply line 105, stainless steel tank 104 is electrically connected anode power cord 106.The arranged outside of stainless steel tank 104 has chiller 107.
With reference to Fig. 2, for vacuum freeze-drying machine schematic diagram, for realizing the present invention's vacuum freeze-drying technique, wherein in vacuum chamber 206, be provided with specimen holder 204, on specimen holder 204, place and treat lyophilizing sample 203, compressor 205 freezes for vacuum chamber 206, and vacuum pump 201 keeps the vacuum of vacuum chamber 206, and vacuometer 202 detects for vacuum.
The preparation method of the dentistry implant of the miRNA of being loaded with of the present invention or siRNA is described below by embodiment.
Embodiment 1
1) select pure titanium to be processed into dentistry implant matrix, after the polishing of dentistry implant matrix surface, use successively acetone, dehydrated alcohol and deionized water ultrasonic cleaning 30 minutes, dry up stand-by;
2) configuration micro-arc oxidation process electrolyte: deionized water is solvent, and solute is 0.04M sodium β-glycerophosphate and 0.2M calcium acetate;
3) adopt micro-arc oxidation process to there is the differential arc oxidation coating of micro-nano pattern in implantation body's outer surface of matrix preparation: stainless steel tank is negative electrode, dentistry implant matrix is as anode, put into the electrolyte of previous step configuration, concrete technology parameter is: supply frequency 100Hz, voltage is 400V, dutycycle 20%, processing time 5min, reaction temperature is room temperature;
4), after prepared by implantation body's matrix surface titanium dioxide differential arc oxidation coating, clean successively with acetone, dehydrated alcohol and deionized water;
5) configuration carrier mixture: 20nmol miRNA is joined to 100 μ L without RNA enzyme deionized water, form 20 μ M miRNA solution, 15 μ L 20 μ M miRNA got by test tube and 500 μ L serum-free mediums (Opti-MEM) mix; 15 μ L transfection reagents (Oligofectamine) got by test tube and 500 μ L serum-free mediums (Opti-MEM) mix; After the equal room temperature of two pipes is placed 5min, mix, miRNA concentration is 0.3 μ M; Under last room temperature, place again 20min, obtain carrier mixture;
6) the dentistry implant matrix of differential arc oxidation being crossed is immersed in carrier mixture, after the freezing 3min of dry ice; Adopt vacuum freeze-drying technique that carrier mixture is loaded into differential arc oxidation coating, wherein, in vacuum chamber, force value is equal to or less than 1Pa, and temperature is-30 ℃, and freeze-drying time is 24h; Last room temperature is placed, and must be loaded with the dentistry implant of miRNA or siRNA.
Embodiment 2
1) select pure titanium to be processed into dentistry implant matrix, after the polishing of dentistry implant matrix surface, use successively acetone, dehydrated alcohol and deionized water ultrasonic cleaning 30 minutes, dry up stand-by;
2) configuration micro-arc oxidation process electrolyte: deionized water is solvent, and solute is 0.04M sodium β-glycerophosphate and 0.2M calcium acetate;
3) adopt micro-arc oxidation process to there is the differential arc oxidation coating of micro-nano pattern in implantation body's outer surface of matrix preparation: stainless steel tank is negative electrode, dentistry implant matrix is as anode, put into the electrolyte of previous step configuration, concrete technology parameter is: supply frequency 100Hz, voltage is 400V, dutycycle 20%, processing time 4min, reaction temperature is room temperature;
4), after prepared by implantation body's matrix surface titanium dioxide differential arc oxidation coating, clean successively with acetone, dehydrated alcohol and deionized water;
5) configuration carrier mixture: 20nmol miRNA is joined to 100 μ L without RNA enzyme deionized water, form 20 μ M miRNA solution, 20 μ L 20 μ M miRNA got by test tube and 500 μ L serum-free mediums (Opti-MEM) mix; 20 μ L transfection reagents (Lipofectamine2000) got by test tube and 500 μ L serum-free mediums (Opti-MEM) mix; After the equal room temperature of two pipes is placed 5min, mix, miRNA concentration is 0.4 μ M; Under last room temperature, place again 20min, obtain carrier mixture;
6) the dentistry implant matrix of differential arc oxidation being crossed is immersed in carrier mixture, after the freezing 5min of dry ice; Adopt vacuum freeze-drying technique that carrier mixture is loaded into differential arc oxidation coating, wherein, in vacuum chamber, force value is equal to or less than 1Pa, and temperature is-30 ℃, and freeze-drying time is 26h; Last room temperature is placed, and must be loaded with the dentistry implant of miRNA or siRNA.
Embodiment 3
1) select pure titanium to be processed into dentistry implant matrix, after the polishing of dentistry implant matrix surface, use successively acetone, dehydrated alcohol and deionized water ultrasonic cleaning 30 minutes, dry up stand-by;
2) configuration micro-arc oxidation process electrolyte: deionized water is solvent, and solute is 0.04M sodium β-glycerophosphate and 0.2M calcium acetate;
3) adopt micro-arc oxidation process to there is the differential arc oxidation coating of micro-nano pattern in implantation body's outer surface of matrix preparation: stainless steel tank is negative electrode, dentistry implant matrix is as anode, put into the electrolyte of previous step configuration, concrete technology parameter is: supply frequency 100Hz, voltage is 400V, dutycycle 20%, processing time 6min, reaction temperature is room temperature;
4), after prepared by implantation body's matrix surface titanium dioxide differential arc oxidation coating, clean successively with acetone, dehydrated alcohol and deionized water;
5) configuration carrier mixture: 20nmol miRNA is joined to 100 μ L without RNA enzyme deionized water, form 20 μ M miRNA solution, 25 μ L 20 μ M miRNA got by test tube and 500 μ L serum-free mediums (Opti-MEM) mix; 25 μ L transfection reagents (Lipofectamine2000) got by test tube and 500 μ L serum-free mediums (Opti-MEM) mix; After the equal room temperature of two pipes is placed 5min, mix, miRNA concentration is 0.5 μ M; Under last room temperature, place again 20min, obtain carrier mixture;
6) the dentistry implant matrix of differential arc oxidation being crossed is immersed in carrier mixture, after the freezing 5min of dry ice; Adopt vacuum freeze-drying technique that carrier mixture is loaded into differential arc oxidation coating, wherein, in vacuum chamber, force value is equal to or less than 1Pa, and temperature is-30 ℃, and freeze-drying time is 28h; Last room temperature is placed, and must be loaded with the dentistry implant of miRNA or siRNA.
Embodiment 4
1) select titanium alloy to be processed into dentistry implant matrix, after the polishing of dentistry implant matrix surface, use successively acetone, dehydrated alcohol and deionized water ultrasonic cleaning 30 minutes, dry up stand-by;
2) configuration micro-arc oxidation process electrolyte: deionized water is solvent, and solute is 0.04M sodium β-glycerophosphate and 0.2M calcium acetate;
3) adopt micro-arc oxidation process to there is the differential arc oxidation coating of micro-nano pattern in implantation body's outer surface of matrix preparation: stainless steel tank is negative electrode, dentistry implant matrix is as anode, put into the electrolyte of previous step configuration, concrete technology parameter is: supply frequency 100Hz, voltage is 400V, dutycycle 20%, processing time 5min, reaction temperature is room temperature;
4), after prepared by implantation body's matrix surface titanium dioxide differential arc oxidation coating, clean successively with acetone, dehydrated alcohol and deionized water;
5) configuration carrier mixture: 20nmol siRNA is joined to 100 μ L without RNA enzyme deionized water, form 20 μ M siRNA solution, 15 μ L transfection reagents (TransIT-TKO) got by test tube and 500 μ L serum-free mediums (Opti-MEM) mix, room temperature is placed 5min, 2.25 μ L 20 μ M siRNA are added in above-mentioned solution, and siRNA concentration is 0.3 μ M; Under last room temperature, place again 15min, obtain carrier mixture;
6) the dentistry implant matrix of differential arc oxidation being crossed is immersed in carrier mixture, after the freezing 5min of dry ice; Adopt vacuum freeze-drying technique that carrier mixture is loaded into differential arc oxidation coating, wherein, in vacuum chamber, force value is equal to or less than 1Pa, and temperature is-20 ℃, and freeze-drying time is 24h; Last room temperature is placed, and must be loaded with the dentistry implant of miRNA or siRNA.
Embodiment 5
1) select titanium alloy to be processed into dentistry implant matrix, after the polishing of dentistry implant matrix surface, use successively acetone, dehydrated alcohol and deionized water ultrasonic cleaning 30 minutes, dry up stand-by;
2) configuration micro-arc oxidation process electrolyte: deionized water is solvent, and solute is 0.04M sodium β-glycerophosphate and 0.2M calcium acetate;
3) adopt micro-arc oxidation process to there is the differential arc oxidation coating of micro-nano pattern in implantation body's outer surface of matrix preparation: stainless steel tank is negative electrode, dentistry implant matrix is as anode, put into the electrolyte of previous step configuration, concrete technology parameter is: supply frequency 100Hz, voltage is 400V, dutycycle 20%, processing time 4min, reaction temperature is room temperature;
4), after prepared by implantation body's matrix surface titanium dioxide differential arc oxidation coating, clean successively with acetone, dehydrated alcohol and deionized water;
5) configuration carrier mixture: 20nmol siRNA is joined to 100 μ L without RNA enzyme deionized water, form 20 μ M siRNA solution, 20 μ L transfection reagents (TransIT-TKO) got by test tube and 500 μ L serum-free mediums (Opti-MEM) mix, room temperature is placed 5min, 3.00 μ L 20 μ M siRNA are added in above-mentioned solution, and siRNA concentration is 0.4 μ M; Under last room temperature, place again 15min, obtain carrier mixture;
6) the dentistry implant matrix of differential arc oxidation being crossed is immersed in carrier mixture, after the freezing 5min of dry ice; Adopt vacuum freeze-drying technique that carrier mixture is loaded into differential arc oxidation coating, wherein, in vacuum chamber, force value is equal to or less than 1Pa, and temperature is-20 ℃, and freeze-drying time is 26h; Last room temperature is placed, and must be loaded with the dentistry implant of miRNA or siRNA.
Embodiment 6
1) select titanium alloy to be processed into dentistry implant matrix, after the polishing of dentistry implant matrix surface, use successively acetone, dehydrated alcohol and deionized water ultrasonic cleaning 30 minutes, dry up stand-by;
2) configuration micro-arc oxidation process electrolyte: deionized water is solvent, and solute is 0.04M sodium β-glycerophosphate and 0.2M calcium acetate;
3) adopt micro-arc oxidation process to there is the differential arc oxidation coating of micro-nano pattern in implantation body's outer surface of matrix preparation: stainless steel tank is negative electrode, dentistry implant matrix is as anode, put into the electrolyte of previous step configuration, concrete technology parameter is: supply frequency 100Hz, voltage is 400V, dutycycle 20%, processing time 6min, reaction temperature is room temperature;
4), after prepared by implantation body's matrix surface titanium dioxide differential arc oxidation coating, clean successively with acetone, dehydrated alcohol and deionized water;
5) configuration carrier mixture: 20nmol siRNA is joined to 100 μ L without RNA enzyme deionized water, form 20 μ M miRNA solution, 25 μ L transfection reagents (TransIT-TKO) got by test tube and 500 μ L serum-free mediums (Opti-MEM) mix, room temperature is placed 5min, 3.75 μ L 20 μ M siRNA are added in above-mentioned solution, and siRNA concentration is 0.5 μ M; Under last room temperature, place again 15min, obtain carrier mixture;
6) the dentistry implant matrix of differential arc oxidation being crossed is immersed in carrier mixture, after the freezing 5min of dry ice; Adopt vacuum freeze-drying technique that carrier mixture is loaded into differential arc oxidation coating, wherein, in vacuum chamber, force value is equal to or less than 1Pa, and temperature is-20 ℃, and freeze-drying time is 28h; Last room temperature is placed, and must be loaded with the dentistry implant of miRNA or siRNA.
That prepares with embodiment 1 is loaded with miRNA dentistry implant, and in conjunction with relevant drawings, the present invention is further described.With reference to Fig. 3, for surface sweeping Electronic Speculum (SEM) photo of pattern before and after the matrix differential arc oxidization surface loading miRNA of implantation body, comparison diagram, in 3 (a), do not carry the microscopic appearance of the front pattern of miRNA, after the visible lyophilizing through miRNA loads, the porous surface of differential arc oxidation and inside all have one deck liposome particles to cover.Fig. 4 is the laser co-focusing photo that implantation body's matrix differential arc oxidization surface loads miRNA front and back pattern, wherein redness is the fluorescently-labeled miRNA of cy-3, as can be seen from the figure after the lyophilizing of miRNA loads, the implant surface of differential arc oxidation is the deposition of miRNA really.Fig. 5 is that dentistry implant differential arc oxidation porous pattern carries the laser co-focusing figure after inoculating cell 24h after miRNA, green is the dyeing of cell membrane green fluorescence probe (DiO) cell membrane, redness is the fluorescently-labeled miRNA of cy-3, as can be seen from the figure, through lyophilization load after, miRNA still effectively transfection enter into cell interior.
Other embodiment prepare dentistry implant, have similar results.Be after lyophilizing loads, the porous surface of differential arc oxidation and inside all have one deck liposome particles to cover, and truly have the deposition of miRNA or siRNA; After inoculating cell 24h, miRNA or siRNA still effectively transfection enter into cell interior.
Although below by reference to the accompanying drawings embodiment of the present invention are described, the present invention is not limited to above-mentioned specific embodiments and applications field, and above-mentioned specific embodiments is only schematic, guiding, rather than restrictive.Those of ordinary skill in the art, under the enlightenment of this description, in the case of not departing from the scope that the claims in the present invention protect, can also make a variety of forms, and these all belong to the row of the present invention's protection.
Claims (6)
1. a preparation method for dentistry implant, is characterized in that, comprises the following steps:
Step 1, selects pure titanium or titanium alloy to be processed into implantation body's matrix, then by after implantation body's matrix surface polishing, uses successively acetone, dehydrated alcohol and deionized water ultrasonic cleaning, then dries up;
Step 2, adopts micro-arc oxidation process to have the differential arc oxidation coating of micro-nano pattern in implantation body's outer surface of matrix preparation, then uses successively acetone, dehydrated alcohol and deionized water ultrasonic cleaning, then dries up;
Step 3, is mixed into carrier mixture by liposome and miRNA or siRNA;
Step 4, is immersed in implantation body's matrix in carrier mixture, and dry ice is freezing;
Step 5, adopts vacuum freeze-drying technique that carrier mixture is loaded into differential arc oxidation coating, and room temperature is placed, and obtains the dentistry implant that is loaded with miRNA or siRNA.
2. the preparation method of dentistry implant according to claim 1, it is characterized in that, in micro-arc oxidation process described in step 2, differential arc oxidation electrolyte solute is prepared by 0.04M sodium β-glycerophosphate and 0.2M calcium acetate, solvent is deionized water, supply frequency 100Hz, and voltage is 300 to 450V, dutycycle 20%, processing time 4-6min.
3. the preparation method of dentistry implant according to claim 1, is characterized in that, in step 3, in described carrier mixture, miRNA or siRNA concentration are 0.3-0.5 μ M.
4. the preparation method of dentistry implant according to claim 1, is characterized in that, the cooling time of dry ice described in step 4 is 3-8min.
5. the preparation method of dentistry implant according to claim 1, is characterized in that, in step 4, dry ice cooling time is 5min.
6. the preparation method of dentistry implant according to claim 1, is characterized in that, in vacuum freeze-drying technique, in vacuum chamber, force value is equal to or less than 1Pa described in step 5, and temperature is-20 ℃ to-30 ℃, and freeze-drying time is more than or equal to 24h.
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Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1541719A (en) * | 2003-11-07 | 2004-11-03 | 西安交通大学 | Rapid synthesize method for bone phosphorite film |
| CN1658914A (en) * | 2002-04-09 | 2005-08-24 | 艾斯特勒科技公司 | Medical prosthetic devices having improved biocompatibility |
| CN1927410A (en) * | 2006-09-25 | 2007-03-14 | 华南理工大学 | Strontium containing hydroxyapatite biologically active film and preparation method thereof |
| CN101054708A (en) * | 2007-02-10 | 2007-10-17 | 青岛科技大学 | Method of preparing hydroxyl apatite bioceramic film by plasma micro-arc oxidization method |
| CN101292907A (en) * | 2008-06-12 | 2008-10-29 | 马威 | Construction method for dental implant biological activity surface |
| CN101310897A (en) * | 2008-02-20 | 2008-11-26 | 暨南大学 | Titanium material with biological activity and preparation method thereof |
| CN101575726A (en) * | 2008-06-30 | 2009-11-11 | 华南理工大学 | Method for preparing bioactive gradient film of fluor-hydroxyapatite |
| CN101611051A (en) * | 2005-02-16 | 2009-12-23 | 加州大学董事会 | The pharmaceutical composition of treatment or prevention osteopathia |
| CN101914799A (en) * | 2010-07-13 | 2010-12-15 | 中国科学院上海硅酸盐研究所 | Method for preparing calcium fluoride coating on surface of titanium and alloy thereof and application thereof |
-
2012
- 2012-02-23 CN CN201210041330.6A patent/CN102579145B/en active Active
Patent Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1658914A (en) * | 2002-04-09 | 2005-08-24 | 艾斯特勒科技公司 | Medical prosthetic devices having improved biocompatibility |
| CN1541719A (en) * | 2003-11-07 | 2004-11-03 | 西安交通大学 | Rapid synthesize method for bone phosphorite film |
| CN101611051A (en) * | 2005-02-16 | 2009-12-23 | 加州大学董事会 | The pharmaceutical composition of treatment or prevention osteopathia |
| CN1927410A (en) * | 2006-09-25 | 2007-03-14 | 华南理工大学 | Strontium containing hydroxyapatite biologically active film and preparation method thereof |
| CN101054708A (en) * | 2007-02-10 | 2007-10-17 | 青岛科技大学 | Method of preparing hydroxyl apatite bioceramic film by plasma micro-arc oxidization method |
| CN101310897A (en) * | 2008-02-20 | 2008-11-26 | 暨南大学 | Titanium material with biological activity and preparation method thereof |
| CN101292907A (en) * | 2008-06-12 | 2008-10-29 | 马威 | Construction method for dental implant biological activity surface |
| CN101575726A (en) * | 2008-06-30 | 2009-11-11 | 华南理工大学 | Method for preparing bioactive gradient film of fluor-hydroxyapatite |
| CN101914799A (en) * | 2010-07-13 | 2010-12-15 | 中国科学院上海硅酸盐研究所 | Method for preparing calcium fluoride coating on surface of titanium and alloy thereof and application thereof |
Non-Patent Citations (2)
| Title |
|---|
| RNAi技术及其在口腔医学研究中的应用;冯志远,刘磊;《现代生物学进展》;20080228;第8卷(第2期);全文 * |
| 冯志远,刘磊.RNAi技术及其在口腔医学研究中的应用.《现代生物学进展》.2008,第8卷(第2期),全文. |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP4056205A1 (en) * | 2021-03-12 | 2022-09-14 | Vilnius University Hospital Zalgiris Clinic | Micro-rna composition functionalized for dental implants |
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