CN103203039A - Preparation method of coating with bone induction and antibiosis functions on surface of medical metal - Google Patents
Preparation method of coating with bone induction and antibiosis functions on surface of medical metal Download PDFInfo
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Abstract
The invention discloses a method for preparing a nanoparticle coating with the bone induction and antibiosis functions on the surface of a medical metal. According to the method, nanoparticles with different electric properties as used as a carrier, the nanoparticles are formed by mutual crosslinking of high-molecular polymers, and growth factors and antibiotics are respectively loaded in the process of preparing the nanoparticles with dissimilar electric properties, wherein factors and factor-friendly polyanions are sufficiently mixed to react first and are then immobilized into the nanoparticles through ionic crosslinking; and amino-containing antibiotics are grafted to a polyaldehyde anionic polymer through chemical crosslinking first and are then loaded into the nanoparticles through ionic crosslinking. The two types of nanoparticles are alternately assembled on the surface of the medical metal through electrostatic adsorption, covalent crosslinking is also formed among the particles, and the stability of the coating is enhanced. The coating prepared by using the method has the function of controlling adsorption and release of the growth factors and the antibiotics under the normal physiological environment, the activity of the growth factors are keep well and a strong bone induction function is achieved; and in addition, the antibiotics can be slowly released for a long time, and a good antibiosis effect is achieved.
Description
Technical field
The present invention relates to a kind of medical metal surface has bone and induces preparation method with antibiotic property effect coating.
Background technology
Medical metal material becomes important osseous tissue alternate material because its excellent biological compatibility and mechanical property successfully have been applied to the bone defect repair.Medical metal surface is because its biologically inert makes new bone slow in its superficial growth, and the mechanically link on the new bone of growing into and surface a little less than.Somatomedin (as BMPs, VEGF, FGF etc.) plays important regulation in the bone repair process.Can it improve biological activity at medical metal area load BMP-2. yet somatomedin medicine high price is expensive, and the half-life is short, and degeneration easily is easy to dissipation in tissue behind the topical, and excessive input may have potential toxicity and carcinogenecity.Realize that the immobilized and slow release of somatomedin on the medical metal surface is a difficult problem.Existing method mainly is the direct Covalent Immobilization factor in the metal surface, but factor active may be damaged because of covalent reaction.
On the other hand, the bio-medical material surface causes that easily microorganism such as antibacterial adheres at it, behind the implant into body, can cause infecting and take place, and must not no longer carry out second operation, takes out the implant that has infected, makes patient's sup sorrow, seriously threat to life often.Can immobilized antibiotic be reduce infecting again at material surface, but existing solid support method is the physical absorption method, and antibiotic is discharging easily in a short time, and the time of its antibacterial action is short, and antibacterial effect is poor.
Summary of the invention
The purpose of this invention is to provide a kind of medical metal surface has bone and induces preparation method with antibiotic property effect coating, the coating of this method preparation can be regulated and control somatomedin and antibiotic absorption and release under bad border of normal physiological, the activity of somatomedin keeps, and bone inductive effect is strong; And antibiotic can be realized long-term slowly release, good anti-bacterial effect.
The present invention realizes its goal of the invention, and the technical scheme that adopts is, a kind of medical metal surface has bone and induces preparation method with antibiotic property effect coating, and its concrete steps are as follows:
The oxidation of A, polysaccharide polymer
To show electronegative poly-polysaccharide and be mixed with the solution that concentration is 20mg/ml in solution, and add sodium metaperiodate, lucifuge again and stir 6h in this solution, the mol ratio of the sodium metaperiodate of adding and poly-monomers and polysaccharide unit is 1: 1-2.5; Add subsequently and the equimolar glycol reaction 2h of sodium metaperiodate, after the reaction solution dialysed, gets the liquid freezing drying in the bag filter, obtain many aldehyde radicals polysaccharide polymer;
The preparation of B, load antibiotic nano-particle:
B1, configuration polycation concentration are the solution of 0.5-2.5mg/ml;
B2, will contain amino antibiotic be dissolved in the oxidation of polysaccharides polymer solution that concentration is 0.5-2.5mg/ml mixed solution, antibiotic concentration is 0.1-1mg/ml in the mixed solution;
B3, mixed solution that said polycation solution and the B2 in B1 step gone on foot are by 2-5: 1 volume ratio is mixed, and stirring, centrifugal, lyophilization, must be loaded with antibiotic electropositive nano-particle; Obtain the electropositive nanoparticles solution that concentration is 0.5-5mg/ml with the electropositive nano-particle is soluble in water subsequently.
The preparation of C, growth factor-loaded nano-particle:
C1, configuration concentration are the said polycation solution of 0.5-1mg/ml;
C2, will to osteogenesis have the somatomedin of facilitation be dissolved in the polyanion solution that concentration is 2.5-10mg/ml mixed solution, the concentration of somatomedin is 0.5-10 μ g/ml in the mixed solution;
C3, the mixed solution in C2 step is mixed by 1: 1 volume ratio with the said polycation solution that C1 goes on foot, and stirring, centrifugal, lyophilization, prepare the elecrtonegativity nano-particle that is loaded with somatomedin; Obtain the elecrtonegativity nanoparticles solution that concentration is 0.5-5mg/ml with the elecrtonegativity nano-particle is soluble in water subsequently.
D, in medical metal surface grafting dopamine coating, it is immersed in the glutaraldehyde solution of volume fraction 0.5%-3% again, make its surface grafting aldehyde radical;
What E, the medical metal after will handling were immersed in the B step was loaded with in the antibiotic electropositive nanoparticles solution reaction 5-30 minute, took out, flushing, namely at the positively charged nano-particle of medical metal surface-assembled last layer;
F, the medical metal after E step handled were immersed in the C elecrtonegativity nanoparticles solution that is loaded with somatomedin in step reaction 5-30 minute, took out, flushing, namely at the electronegative nano-particle of medical metal surface-assembled last layer;
G, repeat operation 0-100 time in above E-F step.
Compared with prior art, the invention has the beneficial effects as follows:
1. somatomedin is wrapped in respectively in the different nano-particle with antibiotic, nano-particle can reach the purpose of site-specific delivery of drugs, realizes topical, improves bioavailability in somatomedin and the antibiotic body, reduce poisonous side effect of medicine, reach the effect that improves therapeutic effect.
2. somatomedin fully mixes with the polyanion of the close factor earlier, and immobilized in nano-particle by mode rather than the covalently bound mode of ionomer again, growth factor activity keeps, bone inductive effect is strong.
3. the amino antibiotic of band is grafted to earlier on the poly-polysaccharide polymer chain of aldehyde radicalization, and is immobilized in nano-particle with polycation generation ionomer again, slowly discharges antibiotic long action time, good anti-bacterial effect with the slow degraded of nano-particle.
4. the nano-particle layer alternate group of carrying somatomedin and two kinds of medicines of antibiotic is contained in the medical metal surface, by selecting different somatomedin and antibiotic for use, can obtain different somatomedin/antibiotic assembly, avoid producing drug resistance, realize that effectively the bone of implantation material surface is induced and antibacterial action.
The above-mentioned polysaccharide polymer of A in the step is sodium alginate, chondroitin sulfate or hyaluronic acid.
These several polysaccharide polymers can be made aldehyde radical on its molecular structure band by sodium periodate oxidation, and by the western not alkali reaction antibiotic generation covalence graft amino with band, by ionomer antibiotic are wrapped in the nano-particle.
The antibiotic that contain amino of above-mentioned B in the step is vancomycin, gentamycin, streptomycin, kanamycin, cefradine, tobramycin, amikacin, neomycin, ribostamycin, micronomicin or A Si mycin.
All have different number amino on these several antibiotic molecule chains, polysaccharide polymer easy and oxidation reacts, thereby antibiotic is wrapped in the nano-particle.
The polyanion of above-mentioned B in the step is heparin, sodium alginate, chondroitin sulfate or hyaluronic acid.
These several materials all are natural polysaccharide polymers, show elecrtonegativity in aqueous solution, can form nano-particle under temperate condition by ionomer mode and polycation.
The somatomedin of above-mentioned C in the step is bone morphogenesis protein-2 (BMP-2), bone morphogenesis protein-7 (BMP-7), VEGF (VEGF) or fibroblast growth factor (FGF).
These several somatomedin all have good bone inducing function, show electropositive under neutrallty condition, after fully mixing with polyanion solution, can effectively be fixed on the polysaccharide polyanion.
Above-mentioned B step and the polycation of C in the step are polylysine or chitosan.
This is two kinds of electropositive polysaccharide polymers, and good biocompatibility easily and under the anion stirring forms nano-particle.
The medical metal of above-mentioned D in the step is titanium, titanium alloy or rustless steel.
These several medical metal alloys are because excellent biological compatibility and mechanical property are widely used in the osseous tissue alternate material, and the surface easily is activated, and can promote the bone quickly-healing.
The invention will be further described below in conjunction with the specific embodiment.
The specific embodiment
Embodiment 1
A kind of medical metal surface has bone and induces preparation method with antibiotic property effect coating, and its concrete steps are as follows:
The oxidation of A, sodium alginate:
Sodium alginate is mixed with the solution that concentration is 20mg/ml, in this solution, add sodium metaperiodate, lucifuge again and stir 6h, the sodium metaperiodate that adds and the mol ratio of sodium alginate monomeric unit are 1: 1, add and the equimolar glycol reaction 2h of sodium metaperiodate subsequently; After the reaction solution dialysed, gets the liquid freezing drying in the bag filter, obtain many aldehyde radicals sodium alginate.
The preparation of B, load vancomycin nano-particle:
B1, configuration polylysine concentration are the solution of 2.5mg/ml;
B2, vancomycin is dissolved in the oxidized sodium alginate solution that concentration is 0.5mg/ml, obtains mixed solution, vancomycin concentration is 0.5mg/ml in the mixed solution;
B3, the polylysine solution in B1 step is mixed by 3: 1 volume ratio with the mixed solution that B2 goes on foot, and stirring, centrifugal, lyophilization, prepare the electropositive nano-particle that is loaded with vancomycin; Obtain the electropositive nanoparticles solution that concentration is 2.5mg/ml with the electropositive nano-particle is soluble in water subsequently;
The preparation of C, load BMP-2 nano-particle:
C1, configuration concentration are the polylysine solution of 0.5mg/ml;
C2, will have the bone morphogenesis protein-2 (BMP-2) of facilitation to be dissolved in the chondroitin sulfate cellulose solution that concentration is 5mg/ml to osteogenesis, obtain mixed solution, in the mixed solution in the concentration of somatomedin be 5 μ g/ml;
C3, the polylysine solution in C1 step is mixed by 1: 1 volume ratio with the mixed solution that C2 goes on foot, and stirring, centrifugal, lyophilization, prepare the elecrtonegativity nano-particle that is loaded with somatomedin; Obtain the elecrtonegativity nanoparticles solution that concentration is 0.5mg/ml with the elecrtonegativity nano-particle is soluble in water subsequently;
D, in pure titanium surface grafting dopamine coating, again it is immersed in the glutaraldehyde solution of volume fraction 2% and soaks 24h, make its surface grafting aldehyde radical;
E, the pure titanium after will handling were immersed in the B electropositive nanoparticles solution that is loaded with vancomycin in step reaction 15 minutes, took out, flushing, namely at the positively charged nano-particle of pure titanium surface-assembled last layer;
F, the pure titanium after E step handled were immersed in the C elecrtonegativity nanoparticles solution that is loaded with bone morphogenesis protein-2 (BMP-2) in step reaction 30 minutes, took out, flushing, namely at the electronegative nano-particle of pure titanium surface-assembled last layer;
G, repeat the operation 10 times in above E-F step.
Embodiment 2
The operation of this example is substantially the same manner as Example 1, just the antibiotic that will use among the embodiment 1 by vancomycin change cefradine into, medical metal changes titanium alloy into by titanium.
Embodiment 3
The operation of this example is substantially the same manner as Example 1, just changes the antibiotic that uses among the embodiment 1 into kanamycin by vancomycin.
Embodiment 4
A kind of medical metal surface has bone and induces preparation method with antibiotic property effect coating, and its concrete steps are as follows:
A, hyaluronic oxidation
Hyaluronic acid is mixed with the solution that concentration is 20mg/ml, in this solution, add sodium metaperiodate, lucifuge again and stir 6h, the sodium metaperiodate that adds and the mol ratio of hyaluronic acid monomeric unit are 1: 2, add and the equimolar glycol reaction 2h of sodium metaperiodate subsequently; After the reaction solution dialysed, gets the liquid freezing drying in the bag filter, obtain many aldehyde radicals hyaluronic acid.
The preparation of B, load bearing chain mycin nano-particle:
B1, configuration chitosan concentration are the solution of 0.5mg/ml;
B2, streptomycin is dissolved in the oxidation chondroitin sulfate solution that concentration is 1.0mg/ml, obtains mixed solution, streptomycin concentration is 0.1mg/ml in the mixed solution;
B3, the chitosan solution in B1 step is mixed by 2: 1 volume ratio with the mixed solution that B2 goes on foot, and stirring, centrifugal, lyophilization, prepare the electropositive nano-particle that is loaded with streptomycin; Obtain the electropositive nanoparticles solution that concentration is 0.5mg/ml with the electropositive nano-particle is soluble in water subsequently;
The preparation of C, load BMP-7 nano-particle:
C1, configuration concentration are the chitosan solution of 0.7mg/ml;
C2, will have the bone morphogenesis protein-7 (BMP-7) of facilitation to be dissolved in the broad liquid of hyaluronic acid that concentration is 10mg/ml to osteogenesis, obtain mixed solution, in the mixed solution in the concentration of somatomedin be 10 μ g/ml;
C3, the chitosan solution in C1 step is mixed by 1: 1 volume ratio with the mixed solution that C2 goes on foot, and stirring, centrifugal, lyophilization, prepare the elecrtonegativity nano-particle that is loaded with somatomedin; Obtain the elecrtonegativity nanoparticles solution that concentration is 5mg/ml with the elecrtonegativity nano-particle is soluble in water subsequently;
D, in pure titanium surface grafting dopamine coating, again it is immersed in the glutaraldehyde solution of volume fraction 0.5% and soaks 24h, make its surface grafting aldehyde radical;
E, the pure titanium after will handling were immersed in the B electropositive nanoparticles solution that is loaded with streptomycin in step reaction 30 minutes, took out, flushing, namely at the positively charged nano-particle of pure titanium surface-assembled last layer;
F, the pure titanium after E step handled were immersed in the C elecrtonegativity nanoparticles solution that is loaded with bone morphogenesis protein-7 (BMP-7) in step reaction 5 minutes, took out, flushing, namely at the electronegative nano-particle of pure titanium surface-assembled last layer;
Embodiment 5
The operation of this example is substantially the same manner as Example 4, just changes the antibiotic that uses among the embodiment 4 into tobramycin by streptomycin.
Embodiment 6
The operation of this example is substantially the same manner as Example 4, just changes the antibiotic that uses among the embodiment 4 into amikacin by streptomycin.
Embodiment 7
A kind of medical metal surface has bone and induces preparation method with antibiotic property effect coating, and its concrete steps are as follows:
The oxidation of A, chondroitin sulfate
Chondroitin sulfate is mixed with the solution that concentration is 20mg/ml, in this solution, add sodium metaperiodate, lucifuge again and stir 6h, the sodium metaperiodate that adds and the mol ratio of chondroitin sulfate monomeric unit are 1: 2.5, add and the equimolar glycol reaction 2h of sodium metaperiodate subsequently; After the reaction solution dialysed, gets the liquid freezing drying in the bag filter, obtain many aldehyde radicals chondroitin sulfate.
The preparation of B, load gentamycin nano-particle:
B1, configuration polylysine concentration are the solution of 2mg/ml;
B2, gentamycin is dissolved in the oxidation chondroitin sulfate solution that concentration is 2.5mg/ml, obtains mixed solution, gentamicin concentration is 1mg/ml in the mixed solution;
B3, the polylysine solution in B1 step is mixed by 5: 1 volume ratio with the mixed solution that B2 goes on foot, and stirring, centrifugal, lyophilization, prepare the electropositive nano-particle that is loaded with gentamycin; Obtain the electropositive nanoparticles solution that concentration is 5.0mg/ml with the electropositive nano-particle is soluble in water subsequently;
The preparation of C, supported V EGF nano-particle:
C1, configuration concentration are the chitosan solution of 1.0mg/ml;
C2, will have the VEGF of facilitation to be dissolved in the sodium alginate soln that concentration is 2.5mg/ml to osteogenesis, obtain mixed solution, in the mixed solution in the concentration of somatomedin be 0.5 μ g/ml;
C3, the chitosan solution in C1 step is mixed by 1: 1 volume ratio with the mixed solution that C2 goes on foot, and stirring, centrifugal, lyophilization, prepare the elecrtonegativity nano-particle that is loaded with somatomedin; Obtain the elecrtonegativity nanoparticles solution that concentration is 2.0mg/ml with the elecrtonegativity nano-particle is soluble in water subsequently;
D, in stainless steel surfaces grafting dopamine coating, again it is immersed in the glutaraldehyde solution of volume fraction 3% and soaks 24h, make its surface grafting aldehyde radical;
E, the rustless steel after will handling were immersed in the B electropositive nanoparticles solution that is loaded with gentamycin in step reaction 5 minutes, took out, flushing, namely at the positively charged nano-particle of stainless steel surfaces assembling last layer;
F, the rustless steel after E step handled were immersed in the C elecrtonegativity nanoparticles solution that is loaded with VEGF in step reaction 20 minutes, took out, flushing, namely at the electronegative nano-particle of stainless steel surfaces assembling last layer;
G, repeat the operation 100 times in above E-F step.
Embodiment 8
This example is substantially the same manner as Example 7, different is with the growth among the embodiment 7 because of long by VEGF (VEGF) change into fibroblast growth factor (FGF), antibiotic by gentamycin change the A Si mycin into, polyanion changes heparin into by sodium alginate.
Embodiment 9
This example is substantially the same manner as Example 7, and different is to change the antibiotic among the embodiment 7 into neomycin by gentamycin.
Embodiment 10
This example is substantially the same manner as Example 7, and different is to change the antibiotic among the embodiment 7 into ribostamycin by gentamycin.
Embodiment 11
This example is substantially the same manner as Example 7, and different is to change the antibiotic among the embodiment 7 into micronomicin by gentamycin.
Claims (7)
1. a medical metal surface has bone and induces preparation method with antibiotic property effect coating, and its concrete steps are as follows:
The oxidation of A, polysaccharide polymer:
To show electronegative poly-polysaccharide and be mixed with the solution that concentration is 20mg/ml in solution, and add sodium metaperiodate, lucifuge again and stir 6h in this solution, the mol ratio of the sodium metaperiodate of adding and poly-monomers and polysaccharide unit is 1: 1-2.5; Add subsequently and the equimolar glycol reaction 2h of sodium metaperiodate, after the reaction solution dialysed, gets the liquid freezing drying in the bag filter, obtain many aldehyde radicals polysaccharide polymer;
The preparation of B, load antibiotic nano-particle:
B1, configuration polycation concentration are the solution of 0.5-2.5mg/ml;
B2, will contain amino antibiotic be dissolved in the oxidation of polysaccharides polymer solution that concentration is 0.5-2.5mg/ml mixed solution, antibiotic concentration is 0.1-1mg/ml in the mixed solution;
B3, mixed solution that said polycation solution and the B2 in B1 step gone on foot are by 2-5: 1 volume ratio is mixed, and stirring, centrifugal, lyophilization, must be loaded with antibiotic electropositive nano-particle; Obtain the electropositive nanoparticles solution that concentration is 0.5-5mg/ml with the electropositive nano-particle is soluble in water subsequently;
The preparation of C, growth factor-loaded nano-particle:
C1, configuration concentration are the said polycation solution of 0.5-1mg/ml;
C2, will to osteogenesis have the somatomedin of facilitation be dissolved in the polyanion solution that concentration is 2.5-10mg/ml mixed solution, the concentration of somatomedin is 0.5-10 μ g/ml in the mixed solution;
C3, the mixed solution in C2 step is mixed by 1: 1 volume ratio with the said polycation solution that C1 goes on foot, and stirring, centrifugal, lyophilization, prepare the elecrtonegativity nano-particle that is loaded with somatomedin; Obtain the elecrtonegativity nanoparticles solution that concentration is 0.5-5mg/ml with the elecrtonegativity nano-particle is soluble in water subsequently;
D, in medical metal surface grafting dopamine coating, it is immersed in the glutaraldehyde solution of volume fraction 0.5%-3% again, make its surface grafting aldehyde radical;
What E, the medical metal after will handling were immersed in the B step was loaded with in the antibiotic electropositive nanoparticles solution reaction 5-30 minute, took out, flushing, namely at the positively charged nano-particle of medical metal surface-assembled last layer;
F, the medical metal after E step handled were immersed in the C elecrtonegativity nanoparticles solution that is loaded with somatomedin in step reaction 5-30 minute, took out, flushing, namely at the electronegative nano-particle of medical metal surface-assembled last layer;
G, repeat operation 0-100 time in above E-F step.
2. a kind of medical metal according to claim 1 surface has bone and induces preparation method with antibiotic property effect coating, and it is characterized in that: the polysaccharide polymer of described A in the step is sodium alginate, chondroitin sulfate or hyaluronic acid.
3. a kind of medical metal according to claim 1 surface has bone and induces preparation method with antibiotic property effect coating, it is characterized in that: described B in the step to contain amino antibiotic be vancomycin, gentamycin, streptomycin, kanamycin, cefradine, tobramycin, amikacin, neomycin, ribostamycin, micronomicin or A Si mycin.
4. a kind of medical metal according to claim 1 surface has bone and induces preparation method with antibiotic property effect coating, and it is characterized in that: the polyanion of described B in the step is heparin, sodium alginate, chondroitin sulfate or hyaluronic acid.
5. a kind of medical metal according to claim 1 surface has bone and induces preparation method with antibiotic property effect coating, and it is characterized in that: the somatomedin of described C in the step is bone morphogenesis protein-2 (BMP-2), bone morphogenesis protein-7 (BMP-7), VEGF (VEGF) or fibroblast growth factor (FGF).
6. a kind of medical metal according to claim 1 surface has bone and induces preparation method with antibiotic property effect coating, and it is characterized in that: the polycation during described B step and C go on foot is polylysine or chitosan.
7. a kind of medical metal according to claim 1 surface has bone and induces preparation method with antibiotic property effect coating, and it is characterized in that: the medical metal of described D in the step is titanium, titanium alloy or rustless steel.
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| CN107661544A (en) * | 2017-09-29 | 2018-02-06 | 北京大学第三医院 | Antibacterial facilitates porous orthopaedics implant of bone complex function and preparation method thereof |
| CN108355169A (en) * | 2018-03-01 | 2018-08-03 | 吉林大学 | A kind of dopamine-heparin-hyaluronic acid coatings material and preparation method thereof carrying and be sustained growth factor |
| CN111138940A (en) * | 2019-12-23 | 2020-05-12 | 普施耐（苏州）工业技术有限公司 | High-adhesion and high-toughness protective agent and preparation method thereof |
| CN111138940B (en) * | 2019-12-23 | 2021-08-17 | 普施耐（苏州）工业技术有限公司 | High-adhesion and high-toughness protective agent and preparation method thereof |
| CN111939317A (en) * | 2020-07-14 | 2020-11-17 | 温州医科大学附属第二医院、温州医科大学附属育英儿童医院 | Method for constructing bone morphogenetic protein sustained-release system |
| CN111939317B (en) * | 2020-07-14 | 2021-12-17 | 温州医科大学附属第二医院、温州医科大学附属育英儿童医院 | Method for constructing bone morphogenetic protein sustained-release system |
| CN115785718A (en) * | 2022-11-25 | 2023-03-14 | 四川大学 | Metal coordination antibacterial coating with multiple substrate surfaces and preparation method thereof |
| CN115785718B (en) * | 2022-11-25 | 2023-08-22 | 四川大学 | Metal coordination antibacterial coating on multi-substrate surface and preparation method thereof |
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