CN106729989A - A kind of tooth implant preparation method of the bioactivity surface containing antibacterial peptide - Google Patents
A kind of tooth implant preparation method of the bioactivity surface containing antibacterial peptide Download PDFInfo
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- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/20—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices containing or releasing organic materials
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- A61L2300/40—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
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- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/60—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a special physical form
- A61L2300/606—Coatings
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- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2400/00—Materials characterised by their function or physical properties
- A61L2400/18—Modification of implant surfaces in order to improve biocompatibility, cell growth, fixation of biomolecules, e.g. plasma treatment
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- A61L2420/02—Methods for coating medical devices
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- A61L2430/00—Materials or treatment for tissue regeneration
- A61L2430/12—Materials or treatment for tissue regeneration for dental implants or prostheses
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Abstract
The present invention discloses a kind of tooth implant preparation method of the bioactivity surface containing antibacterial peptide, specifically with the tooth implant of different number of plies coatings.Its coating relies primarily on cation and anion electrostatic attraction each other, the molecular coatings of ordered arrangement is formed in the planting body surface of solids, because shitosan and AMP composite layers are the biomaterials with good biodegradability properties and biocompatibility.Due to the presence of top layer chondroitin sulfate layer antibacterial peptide layer lamination layer structure, with lasting, efficient sterilized, anti-infection ability, its antibacterial, anti-apoptotic activities, adhesion, survival, growth and the differentiation of various kinds of cell (such as Gegenbaur's cell, cartilage cell) can be supported, is had broad application prospects.
Description
Technical field
The invention belongs to tooth implant technical field, and in particular to a kind of Dental implantion of the bioactivity surface containing antibacterial peptide
Preparation.
Background technology
With continuing to develop for tooth implant technology, and widely approved by doctor patient, be also to send out in dentistry in recent years
A most fast specialty of exhibition, constantly makes a breakthrough, for more tooth disease patients provide specialty help.However, not
It is that existing intervention device is easy to infect after implanting to obtain another present situation not recognized.Main infection mechanism is
Material surface is sticked to first into internal bacterium, and due to nutrient environment abundant in vivo, bacterium breeds, and forms one
The biomembrane of layer protectiveness.The presence of biomembrane causes that the infection that medical apparatus trigger is particularly refractory.
Roos.Jansaker etc. carries out follow-up for many years to plantation postoperative patient, and bleeding is examined while not occurring there is spy
Deossification carries out clinical case statistics for diagnostic criteria, as a result shows, the incidence of disease of peri-implant mucositis is about 79%；Very
To there are some researches show the planting body for about having more than 90% spy occurred and examines bleeding.And peri-implantitiss can cause peri-implant
The defect of bone tissue, forfeiture, such as treatment can cause the loose or dislocation of planting body not in time.
Biologically active peptide be in protein 20 natural amino acids with different compositions and arrangement mode constituted from dipeptides to
The general name of complicated linear, loop configuration different peptides, is derived from the multi-functional compounds of protein.Active peptide has various
Body metabolism and physiological regulation function, absorption easy to digest have immune promotion, hormone control, antibacterial, antiviral, hypotensive, drop blood
Fat etc. is acted on.
Because the main component of dental implant surface is generally processed as negatively charged state.And will be poly- containing electropositive groups
Compound such as shitosan has preferable adhesion with dental implant surface.In addition containing with hydroxyl hydrogen-bond donor/acceptor each other
The polymer of group, such as sodium alginate, hydroxypropyl cellulose are likely to have certain adhesion with dental implant surface.
So, surface nature requirement of the tooth implant to material is very high.Therefore, new dental implant surface is explored to change
Property method has been increasingly becoming the direction of scientific rersearch of field of planting.
The content of the invention
For the technical scheme that solution above-mentioned technical problem is used is：A kind of bioactivity surface containing antibacterial peptide is disclosed
Tooth implant preparation method, its described antibacterial peptide layer is the Dental implantion body surface of titanium dioxide nanotube array layer to be covered in surface
Face, and alternately assembled by positive charge chitosan layer and negative electrical charge AMP compounds, outer layer load chondroitin sulfate layer-antibacterial peptide
Layer.
For above-mentioned technical proposal, it is preferred in the case of, the tooth implant of the described bioactivity surface containing antibacterial peptide
Preparation method, wherein, the preparation method of the antibacterial peptide layer includes following operating procedure：
(1) by surface for the tooth implant cleaning of titanium dioxide nanotube array layer is standby；
(2) chitosan solution of 0.5~5mg/ml is prepared：Concentration is 1mg/mlAMP complex solutions；
(3) by the tooth implant described in step (1) be immersed in step (2) gained chitosan solution in 10~20min so that
Dental implant surface becomes positively charged lotus；Fully cleaned with deionized water again；
(4) it is 5~10min in 1mg/mlAMP complex solutions the tooth implant described in step (3) to be immersed in into concentration,
So that negative electrical charge on dental implant surface band；Fully cleaned with deionized water again；
The number of plies for assembling as needed, repeat step (3), (4) and (3) is that can obtain the tooth kind with different number of plies coatings
Implant；
(5) tooth implant described in step (4) is soaked in concentration is for the chondroitin sulfate sodium sulfate salt solution of 2mg/mL
5~15min of bubble, solution system control ph is 6, then is spontaneously dried after fully being cleaned with deionized water；Sulfuric acid is coated with again
It is 7 that the tooth implant of chondroitin is soaked in pH value, and concentration is 5~15min in the antibacterial peptide solution of 0.5mg/mL, then uses deionization
Water is spontaneously dried after fully cleaning, and acquisition is coated with the material of chondroitin sulfate layer-antibacterial peptide layer composite bed.
In above-mentioned technical proposal, the AMP is cationic antimicrobial small peptide, English name:cationic
antimicrobial peptides。
For above-mentioned technical proposal, it is preferred in the case of, the tooth implant of the described bioactivity surface containing antibacterial peptide
Preparation method, wherein, described cleaning refers to deionized water, and ethanol and acetone are cleaned by ultrasonic, and after being dried up with nitrogen, are put in vacuum
In drying box, dried 24 hours under conditions of 60 DEG C.
For above-mentioned technical proposal, it is preferred in the case of, the tooth implant of the described bioactivity surface containing antibacterial peptide
Preparation method, wherein, the deacetylation of described shitosan is that 50~98%, Mw is 100~1500kDa.
For above-mentioned technical proposal, it is preferred in the case of, the tooth implant of the described bioactivity surface containing antibacterial peptide
Preparation method, wherein, during described chitosan solution is the acetic acid solution for dissolve chitosan in 0.3~3%, it is made into 0.5~
The chitosan solution of 5mg/ml, resulting solution is mixed with atoleine, and volume ratio is 0.1~0.4, and adds the volumetric concentration to be
0.5~1% sorbester p17, while adding dispersant magnesium stearate, sorbester p17 is 2~4: 1 with the mass ratio of magnesium stearate, stirring
Gained water-in-oil emulsion, is added thereto to the crosslinking agent vanillic aldehyde 1ml of 5~10mg/ml, is reacted under 40 DEG C~60 DEG C stirrings
After 4~6 hours；Standing takes supernatant；The chitosan particle of gained is added into a small amount of glacial acetic acid solution, magnetic with aqueous suspension again
Under power agitator stirring condition, NaCl is added, its final concentration is reached 0.15M；The solution is positively charged.
For above-mentioned technical proposal, it is preferred in the case of, the tooth implant of the described bioactivity surface containing antibacterial peptide
Preparation method, wherein, the standing takes supernatant and refers to, stands more than 12 hours oil-water separation phases, takes lower aqueous solution, adjusts
PH value is 8~9, is centrifuged off sediment, takes supernatant and obtains chitosan particle suspension.
For above-mentioned technical proposal, it is preferred in the case of, the tooth implant of the described bioactivity surface containing antibacterial peptide
Preparation method, wherein, the concentration is for the collocation method of the AMP complex solutions of lmg/ml：Weigh 50mg AMP compounds
Powder is added to after the sterilizing of 50ml in deionized water, and fully to being completely dissolved, it is 5.0, this solution band that acetic acid on the rocks adjusts pH for stirring
There is positive charge.
Character of innovation of the invention is：
The tooth implant with different number of plies coatings prepared by the present invention.Its coating relies primarily on cation and anion
Electrostatic attraction each other, the molecular coatings of ordered arrangement are formed in the planting body surface of solids, because shitosan and AMP are compound
Nitride layer is the biomaterial with good biodegradability properties and biocompatibility.Due to top layer chondroitin sulfate layer-antibacterial peptide layer
The presence of lamination layer structure, with lasting, efficient sterilized, anti-infection ability, its antibacterial, anti-apoptotic activities, it would be preferable to support many
Adhesion, survival, growth and the differentiation of cell (such as Gegenbaur's cell, cartilage cell) are planted, is had broad application prospects.
Specific embodiment
Following non-limiting examples can make one of ordinary skill in the art be more fully understood the present invention, but not with
Any mode limits the present invention.Any one skilled in the art in the technical scope of present disclosure, according to
Technical scheme and its inventive concept carry out equivalent or change belongs to protection category of the present invention.
Configuration concentration is the AMP complex solutions of lmg/ml；
50mg AMP composite powders are weighed to add to after the sterilizing of 50ml in deionized water, stirring fully to being completely dissolved,
It is 5.0 that acetic acid on the rocks adjusts pH, and this solution carries positive charge.AMP compounds configuration concentration is 0.5mg/mL antibacterial peptide solutions
5mg antibacterial peptides are weighed, is dissolved in 10mL deionized waters, it is 7 to be prepared into pH value, and concentration is anti-for 0.5mg/mL
Bacterium peptide solution.
Embodiment 1
(1) it is the tooth implant of titanium dioxide nanotube array layer by surface, successively with deionized water, ethanol and acetone are super
Sound is cleaned, and after being dried up with nitrogen, is put in vacuum drying chamber, is dried 24 hours under conditions of 60 DEG C, is placed standby；
(2) dissolve chitosan in 0.3% acetic acid solution, be made into the chitosan solution of 0.5mg/ml, described shell
The deacetylation of glycan is 1000kDa for 50%, Mw.
(3) step (2) resulting solution is mixed with atoleine, volume ratio is 0.1, and it is 0.5% to add volumetric concentration
Sorbester p17, while adding dispersant magnesium stearate, the mass ratio of sorbester p17 and magnesium stearate is 2: 1, is stirred 1 hour at room temperature
More than, form uniform water-in-oil emulsion；
(4) the crosslinking agent vanillic aldehyde 1ml of 5mg/ml, 40 DEG C of stirring shapes will be being added in step (3) gained water-in-oil emulsion
Reacted 4 hours under state, obtain chitosan particle；
(5) step (4) products therefrom is stood into more than 12 hours oil-water separation phases, takes lower aqueous solution, regulation pH value is
8, sediment is centrifuged off, take supernatant and obtain chitosan particle suspension
(6) to the chitosan particle suspension added in deionized water obtained by step (5), the glacial acetic acid solution of lml is added,
Under the conditions of magnetic stirrer, NaCl is added, its final concentration is reached 0.15M；The solution is positively charged；
Embodiment 2
(1) it is the tooth implant of titanium dioxide nanotube array layer by surface, successively with deionized water, ethanol and acetone are super
Sound is cleaned, and after being dried up with nitrogen, is put in vacuum drying chamber, is dried 24 hours under conditions of 60 DEG C, is placed standby；
(2) dissolve chitosan in 3% acetic acid solution, be made into the chitosan solution of 5mg/ml, described shitosan
Deacetylation for 98%, Mw be 1500kDa.
(3) step (2) resulting solution is mixed with atoleine, volume ratio is 0.4, and it is 1% to add volumetric concentration
Sorbester p17, while adding dispersant magnesium stearate, the mass ratio of sorbester p17 and magnesium stearate is 4: 1, stir at room temperature 1 hour with
On, form uniform water-in-oil emulsion；
(4) the crosslinking agent vanillic aldehyde 1ml of 10mg/ml, 60 DEG C of stirring shapes will be being added in step (3) gained water-in-oil emulsion
Reacted 6 hours under state, obtain chitosan particle；
(5) step (4) products therefrom is stood into more than 12 hours oil-water separation phases, takes lower aqueous solution, regulation pH value is
9, sediment is centrifuged off, take supernatant and obtain chitosan particle suspension
(6) to the chitosan particle suspension added in deionized water obtained by step (5), the glacial acetic acid solution of lml is added,
Under the conditions of magnetic stirrer, NaCl is added, its final concentration is reached 0.15M；The solution is positively charged；
Embodiment 3
The tooth implant that step (1) treatment in embodiment 1 is obtained is immersed in the shitosan described in example 1 above and 2
10~20min in solution, the lotus so that dental implant surface becomes positively charged；Fully cleaned with deionized water again；
By the tooth implant after above-mentioned cleaning be immersed in concentration mentioned above for 10 in 1mg/mlAMP complex solutions~
20min, so that negative electrical charge on dental implant surface band；Fully cleaned with deionized water again；
The tooth implant that treatment is obtained is immersed in 10~20min in the chitosan solution described in example 1 above and 2, with
Dental implant surface is set to become positively charged lotus；Fully cleaned with deionized water again；
The step of number of plies for assembling as needed, repetition embodiment 3 more than 2 times, you can obtain with different number of plies coatings
Tooth implant.
Embodiment 4
The tooth implant of different number of plies coatings will be as needed assembled in embodiment 3, in concentration for the sulfuric acid of 2mg/mL is soft
10~20min of immersion in ossein sodium sulfate salt solution (polysaccharide solution with negative electrical charge), solution system control ph is 6, then
Spontaneously dried after fully being cleaned with deionized water；
The material for having chondroitin sulfate will be coated and be soaked in pH value is 7, and concentration is in the antibacterial peptide solution of 0.5mg/mL
15min, then spontaneously dry after fully being cleaned with deionized water, acquisition is coated with chondroitin sulfate layer-antibacterial peptide layer composite bed
Material.
For the tooth implant that the above method is prepared, surface contact angle analysis is carried out, its result is from hydrophily and profit
The presence of coating is also demonstrate that on moist.Understood according to analysis, the Dental implantion with different number of plies coatings prepared by the present invention
Body.Its coating relies primarily on cation and anion electrostatic attraction each other, and row in order is formed in the planting body surface of solids
The molecular coatings of row, because shitosan and AMP compounds are the biomaterials with good biodegradability properties and biocompatibility.
Due to the presence of top layer chondroitin sulfate layer-antibacterial peptide layer lamination layer structure, with lasting, efficient sterilized, anti-infection ability,
Its antibacterial, anti-apoptotic activities, it would be preferable to support the adhesion of various kinds of cell (such as Gegenbaur's cell, cartilage cell), survival, growth with point
Change, have broad application prospects.
Claims (6)
1. a kind of tooth implant preparation method of the bioactivity surface containing antibacterial peptide, it is characterised in that：Antibacterial peptide layer is
The dental implant surface that surface is titanium dioxide nanotube array layer is covered in, and by positive charge chitosan layer and negative electrical charge AMP
Composite layer is alternately assembled, outermost layer load chondroitin sulfate layer-antibacterial peptide layer；
The preparation method of the antibacterial peptide layer includes following operating procedure：
(1) by surface for the tooth implant cleaning of titanium dioxide nanotube array layer is standby；
(2) chitosan solution of 0.5~5mg/ml is prepared：Concentration is 1mg/mlAMP complex solutions；
(3) by the tooth implant described in step (1) be immersed in step (2) gained chitosan solution in 10~20min so that tooth kind
Implant surface becomes positively charged lotus；Fully cleaned with deionized water again；
(4) it is 5~10min in 1mg/mlAMP complex solutions the tooth implant described in step (3) to be immersed in into concentration, so that
Negative electrical charge on dental implant surface band；Fully cleaned with deionized water again；
The number of plies for assembling as needed, repeat step (3), (4) and (3) is that can obtain the Dental implantion with different number of plies coatings
Body；
(5) by the tooth implant described in step (4) concentration for 2mg/mL chondroitin sulfate sodium sulfate salt solution in immersion 5~
15min, solution system control ph is 6, then is spontaneously dried after fully being cleaned with deionized water；Chondroitin sulfate is coated with again
Tooth implant be soaked in pH value for 7, concentration is 5~15min in the antibacterial peptide solution of 0.5mg/mL, then abundant with deionized water
Spontaneously dried after cleaning, acquisition is coated with the material of chondroitin sulfate layer-antibacterial peptide layer composite bed.
2. the tooth implant preparation method of the bioactivity surface containing antibacterial peptide according to claim 1, it is characterised in that：
Described cleaning refers to deionized water, and ethanol and acetone are cleaned by ultrasonic, after being dried up with nitrogen, are put in vacuum drying chamber, 60
Dried 24 hours under conditions of DEG C.
3. the tooth implant preparation method of the bioactivity surface containing antibacterial peptide according to claim 1, it is characterised in that：
The deacetylation of described shitosan is that 50~98%, Mw is 100~1500kDa.
4. the tooth implant preparation method of the bioactivity surface containing antibacterial peptide according to claim 1, it is characterised in that：
During described chitosan solution is the acetic acid solution for dissolve chitosan in 0.3~3%, the shitosan of 0.5~5mg/ml is made into
Solution, resulting solution is mixed with atoleine, and volume ratio is 0.1~0.4, and adds the sapn that volumetric concentration is 0.5~1%
80, while adding dispersant magnesium stearate, sorbester p17 is 2~4: 1 with the mass ratio of magnesium stearate, stirring gained water in oil emulsion
Liquid, is added thereto to the crosslinking agent vanillic aldehyde 1ml of 5~10mg/ml, after being reacted 4~6 hours under 40 DEG C~60 DEG C stirrings；
Standing takes supernatant；The chitosan particle of gained is added into a small amount of glacial acetic acid solution, magnetic stirrer with aqueous suspension again
Under the conditions of, NaCl is added, its final concentration is reached 0.15M；The solution is positively charged.
5. the tooth implant preparation method of the bioactivity surface containing antibacterial peptide according to claim 4, it is characterised in that：
The standing takes supernatant and refers to, stands more than 12 hours oil-water separation phases, takes lower aqueous solution, and regulation pH value is 8~9, from
The heart removes sediment, takes supernatant and obtains chitosan particle suspension.
6. the tooth implant preparation method of the bioactivity surface containing antibacterial peptide according to claim 1, it is characterised in that：
The concentration is that the collocation method of the AMP complex solutions of lmg/ml is：Weigh 50mg AMP composite powders and add to 50ml's
After sterilizing in deionized water, fully to being completely dissolved, it is 5.0 that acetic acid on the rocks adjusts pH, and this solution carries positive charge for stirring.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN111484123A (en) * | 2020-05-06 | 2020-08-04 | 北京工业大学 | Efficient nitrifying embedded bioactive filler and preparation method thereof |
Citations (8)
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