阅读说明：本技术 一种具有骨诱导和协同抗菌功能的聚吡咯@纳米铜/帕米膦酸二钠复合涂层的制备方法 (Preparation method of polypyrrole @ nano copper/pamidronate disodium composite coating with osteoinduction and synergistic antibacterial functions ) 是由 王英波 武华君 于 2021-05-27 设计创作，主要内容包括：本发明公开了一种具有骨诱导和光热/铜协同抗菌功能的聚吡咯@纳米铜/帕米膦酸二钠复合涂层的制备方法,该方法利用具有螯合能力和药物掺杂作用的聚吡咯对骨诱导剂帕米膦酸二钠和光热协同抗菌剂铜纳米粒子进行双重调控,来制备兼具骨诱导和光热/铜协同抗菌功能的复合涂层,该复合涂层通过脉冲电化学沉积技术能负载到人工骨生物医用材料如钛及其合金基底上,一方面能模拟体内的天然细胞外基质,快速促进成骨和血管生成,另一方面具有快速、持久抗细菌感染的效果,能广泛应用于骨损伤修复和骨缺损治疗领域。该制备方法操作简便,成本低廉,制得的复合涂层作为骨修复材料具有良好的应用前景。(The invention discloses a preparation method of a polypyrrole @ nano copper/disodium pamidronate composite coating with osteoinduction and photothermal/copper synergistic antibacterial functions, the method utilizes polypyrrole with chelation capacity and drug doping effects to carry out double regulation and control on disodium pamidronate serving as an osteoinducer and copper nanoparticles serving as a photothermal synergistic antibacterial agent to prepare the composite coating with osteoinduction and photothermal/copper synergistic antibacterial functions. The preparation method is simple and convenient to operate and low in cost, and the prepared composite coating has a good application prospect as a bone repair material.)

1. A preparation method of a polypyrrole @ nano copper/pamidronate disodium composite coating with osteoinduction and photo-thermal/copper synergistic antibacterial functions is characterized by comprising the following steps:

(1) mechanically polishing the surface of a titanium plate serving as a matrix to remove an oxide layer or an organic layer, putting the titanium plate into an organic solvent for ultrasonic cleaning for not less than 10 min, and then cleaning the titanium plate by using deionized water;

(2) putting the titanium plate obtained in the step (1) into an inorganic acid solution for full acid washing and passivation, and then cleaning the titanium plate by using deionized water;

(3) putting the titanium plate obtained in the step (2) into an inorganic alkali solution for full alkali washing and passivation, then cleaning the titanium plate by using deionized water, and airing for later use;

(4) dissolving pyrrole in deionized water, and mechanically stirring to completely dissolve the pyrrole to obtain a uniform pyrrole solution;

(5) adding the nano-copper precursor into the pyrrole solution obtained in the step (4), and mechanically stirring to completely dissolve the nano-copper precursor to obtain a uniform and stable mixed solution;

(6) adding pamidronate disodium into the mixed solution obtained in the step (5), mechanically stirring to completely dissolve the pamidronate disodium, and adjusting the pH value of the solution to be between 5.0 and 5.1 by using hydrochloric acid;

(7) and (4) forming a three-electrode system by taking the titanium plate obtained in the step (3) as a working electrode, placing the three-electrode system in the mixed solution obtained in the step (6) for pulse electrochemical deposition to obtain the polypyrrole @ nano copper/pamidronate disodium composite coating, and naturally drying a coating sample.

2. The preparation method of the polypyrrole @ nanocopper/disodium pamidronate composite coating of claim 1, wherein the preparation method comprises the following steps: the organic solvent in the step (1) is at least one of acetone, acetonitrile, ethyl acetate, N-dimethylformamide or trichloromethane.

3. The preparation method of the polypyrrole @ nanocopper/disodium pamidronate composite coating of claim 1, wherein the preparation method comprises the following steps: the inorganic acid solution in the step (2) is sulfuric acid, hydrochloric acid or a water solution mixed by the sulfuric acid and the hydrochloric acid in any proportion, the pickling temperature is 50-80 ℃, and the pickling time is 3-5 hours.

4. The preparation method of the polypyrrole @ nanocopper/disodium pamidronate composite coating of claim 1, wherein the preparation method comprises the following steps: the inorganic alkali solution in the step (3) is a saturated aqueous solution of at least one of sodium hydroxide, potassium hydroxide and lithium hydroxide, the alkali washing temperature is 50-80 ℃, and the alkali washing time is 1-2 hours.

5. The preparation method of the polypyrrole @ nanocopper/disodium pamidronate composite coating of claim 1, wherein the preparation method comprises the following steps: the adding concentration of the pyrrole in the step (4) is 2.0-2.2 mL/L, the adding concentration of the nano-copper precursor in the step (5) is 0.2-0.24g/L, and the adding concentration of the pamidronate disodium in the step (6) is 0.8-1.2 mg/L.

6. The preparation method of the polypyrrole @ nanocopper/disodium pamidronate composite coating of claim 1, wherein the preparation method comprises the following steps: the nano-copper precursor in the step (5) is at least one of water-soluble inorganic copper salts.

7. The preparation method of the polypyrrole @ nanocopper/disodium pamidronate composite coating of claim 1, wherein the preparation method comprises the following steps: and (3) the auxiliary electrode in the three-electrode system in the step (7) is a titanium, platinum or carbon electrode, and the reference electrode is a saturated calomel electrode or a silver | silver chloride electrode.

8. The preparation method of the polypyrrole @ nanocopper/disodium pamidronate composite coating of claim 1, wherein the preparation method comprises the following steps: the pulse electrochemical deposition parameters in the step (7) comprise an oxidation potential of 0.8V, an oxidation time of 80-120 s, a reduction potential of-1.5V, a reduction time of 400-600 s, cycle times of 5-8 times and a total time of 0.5-1.5 h.

9. The method for preparing a polypyrrole @ nanocopper/disodium pamidronate composite coating according to any one of claims 1 to 8, wherein the method comprises the following steps: the nano copper is a copper simple substance nano particle.

Technical Field

The invention relates to a preparation method of a medical polymer composite material, in particular to a preparation method of a polypyrrole @ nano copper/pamidronate disodium composite coating with osteoinduction and photo-thermal/copper synergistic antibacterial functions.

Background

The development of bone repair materials is a difficult problem to solve, and the ideal artificial bone synthetic biomaterial should have good biocompatibility, bioactivity, osteoinductivity and antibacterial property.

Polypyrrole (PPy) is a conductive polymer which can be prepared by an electrochemical method and has good biocompatibility, can be used as a stabilizer of metal ions, and the positive charge carried by oxidation can generate electrostatic interaction with charged ions in an electrolyte to enhance the stability of the Polypyrrole (PPy), so that the Polypyrrole (PPy) is widely applied to drug delivery. By utilizing the oxidation-reduction state transformation of polypyrrole in an electrochemical state, in the polymerization process of a pyrrole monomer, an anionic drug is incorporated into a polymer to neutralize positive charges caused by oxidation, and the purpose that the drug is doped into a polypyrrole main chain as a dopant can be achieved.

Pamidronate disodium (Pamidronate disodium) is a bisphosphonate, which acts as a potent inhibitor of osteoclasts, regulating bone metabolism, and is widely used in the treatment of a variety of diseases of excessive bone resorption. Because pamidronate disodium has a high affinity for calcium ions, new bone can be formed on the surface of the bone tissue adsorbing the bisphosphonate, thereby actually accelerating the regeneration of bone.

Bone infection is a difficult problem to overcome in clinical bone repair process, and since bone infection caused by bacteria damages healing and repairing capability of bone tissue, resulting in implant failure, research and development of implant materials with antibacterial property become one of current research hotspots. Bone infection is mainly caused by bacterial infection, and the key point of inhibiting infection and promoting bone tissue healing is to kill bacteria on the tissue surface in a timely and effective manner and inhibit the formation of bacterial biofilms. The current common methods of treating bone infections are debridement and systemic antibiotic therapy, but chronic or excessive use of antibiotics increases surgical risk and creates resistance. Therefore, the improvement of the bacterial drug resistance and the bone infection resistance of the biological material becomes a difficult problem to be solved urgently. Copper is a trace element necessary for a human body, Copper nanoparticles (Cu-NPs) have wide antibacterial property and antibacterial resistance, and have responsiveness to near infrared light, and are clinically applied to killing bacteria on biological materials. It is noteworthy that copper can be liberated from dead bacteria and the above cycle repeated, enabling a lasting antibacterial action; in addition, the copper nanoparticles can stimulate the formation of bones and promote angiogenesis, thereby further promoting the repair of bone tissues. Therefore, the copper nano-particles are introduced into the bone repair composite material to endow the bone repair composite material with quick and durable antibacterial performance and osteogenic performance.

The Pulse Electrochemical Deposition (PED) technology can construct a uniform composite coating on a complex substrate surface by switching between oxidation-reduction potentials, and can precisely control the coating morphology by adjusting electrochemical parameters. The ideal biomimetic composite coating is designed for cell adhesion and proliferation, promotes new tissue growth, and prevents the occurrence of early bacterial infections. Has important application value in bone tissue engineering.

Based on the characteristics of the polypyrrole on metal ions, the chelating capacity and the drug doping effect of the polypyrrole, the broad-spectrum antibacterial property, the antibacterial drug resistance and the photothermal effect of copper nanoparticles, the osteoinduction of pamidronate disodium and the like, and the unique advantages of a pulse electrochemical deposition technology in constructing a composite coating structure with a controllable appearance, the invention provides the preparation method of the polypyrrole @ nano copper/pamidronate disodium composite coating with the osteoinduction and photothermal/copper synergistic antibacterial functions.

Disclosure of Invention

The invention aims to provide a preparation method of a polypyrrole @ nano copper/pamidronate disodium composite coating with osteoinduction and photothermal/copper synergistic antibacterial functions, wherein polypyrrole with chelation capacity and drug doping effects is used for carrying out double regulation and control on an osteoinducer pamidronate disodium and photothermal synergistic antibacterial agent copper nanoparticles to prepare the composite coating with osteoinduction and photothermal/copper synergistic antibacterial functions.

The invention is realized by the following technical scheme:

a preparation method of a polypyrrole @ nano copper/pamidronate disodium composite coating with osteoinduction and photo-thermal/copper synergistic antibacterial functions comprises the following steps:

(1) mechanically polishing the surface of a titanium plate serving as a matrix to remove an oxide layer or an organic layer, putting the titanium plate into an organic solvent for ultrasonic cleaning for not less than 10 min, and then cleaning the titanium plate by using deionized water;

(2) putting the titanium plate obtained in the step (1) into an inorganic acid solution for full acid washing and passivation, and then cleaning the titanium plate by using deionized water;

(3) putting the titanium plate obtained in the step (2) into an inorganic alkali solution for full alkali washing and passivation, then cleaning the titanium plate by using deionized water, and airing for later use;

(4) dissolving pyrrole in deionized water, and mechanically stirring to completely dissolve the pyrrole to obtain a uniform pyrrole solution;

(5) adding the nano-copper precursor into the pyrrole solution obtained in the step (4), and mechanically stirring to completely dissolve the nano-copper precursor to obtain a uniform and stable mixed solution;

(6) adding pamidronate disodium into the mixed solution obtained in the step (5), mechanically stirring to completely dissolve the pamidronate disodium, and adjusting the pH value of the solution to be between 5.0 and 5.1 by using hydrochloric acid;

(7) and (4) forming a three-electrode system by taking the titanium plate obtained in the step (3) as a working electrode, placing the three-electrode system in the mixed solution obtained in the step (6) for pulse electrochemical deposition to obtain the polypyrrole @ nano copper/pamidronate disodium composite coating, and naturally drying a coating sample.

As a further optimization of the technical scheme of the invention, the organic solvent in the step (1) is at least one of acetone, acetonitrile, ethyl acetate, N-dimethylformamide or trichloromethane.

As a further optimization of the technical scheme of the invention, the inorganic acid solution in the step (2) is sulfuric acid, hydrochloric acid or a water solution mixed by the sulfuric acid and the hydrochloric acid in any proportion, the acid washing temperature is 50-80 ℃, and the acid washing time is 3-5 h.

As a further optimization of the technical scheme of the invention, the inorganic alkali solution in the step (3) is a saturated aqueous solution of at least one of sodium hydroxide, potassium hydroxide and lithium hydroxide, the alkali washing temperature is 50-80 ℃, and the alkali washing time is 1-2 h.

As further optimization of the technical scheme of the invention, the adding concentration of the pyrrole in the step (4) is 2.0-2.2 mL/L, the adding concentration of the nano-copper precursor in the step (5) is 0.2-0.24g/L, and the adding concentration of the pamidronate disodium in the step (6) is 0.8-1.2 mg/L.

As a further optimization of the technical scheme of the invention, the nano-copper precursor in the step (5) is at least one of water-soluble inorganic copper salts.

As a further optimization of the technical scheme of the invention, the auxiliary electrode in the three-electrode system in the step (7) is a titanium, platinum or carbon electrode, and the reference electrode is a saturated calomel electrode or a silver | silver chloride electrode.

As a further optimization of the technical scheme of the invention, the pulse electrochemical deposition parameters in the step (7) comprise an oxidation potential of 0.8V, an oxidation time of 80-120 s, a reduction potential of-1.5V, a reduction time of 400-600 s, a cycle time of 5-8 times and a total time of 0.5-1.5 h.

As further optimization of the technical scheme of the invention, the nano copper is a copper simple substance nano particle.

In conclusion, the preparation method of the polypyrrole @ nano copper/pamidronate disodium composite coating with the osteoinduction and photothermal/copper synergistic antibacterial functions has the advantages that: the polypyrrole with chelating ability and drug doping function is used for carrying out dual regulation and control on the bone inducer pamidronate disodium and the photo-thermal synergistic antibacterial agent copper nanoparticles to prepare the composite coating with bone induction and photo-thermal/copper synergistic antibacterial functions. The preparation method is simple and convenient to operate and low in cost, and the prepared composite coating has a good application prospect as a bone repair material.

Drawings

FIG. 1 is a Scanning Electron Microscope (SEM) image of a composite coating prepared according to a first embodiment of the invention;

FIG. 2 is a qualitative graph of Escherichia coli and Staphylococcus aureus of a composite coating prepared according to a first embodiment of the present invention;

FIG. 3 is a graph of the quantification of Escherichia coli and Staphylococcus aureus in a composite coating prepared according to a first embodiment of the present invention;

FIG. 4 is a graph of the UV absorption spectrum of the composite coating and the titanium substrate according to the first embodiment of the present invention.

Detailed Description

The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

Example one

The invention provides a preparation method of a polypyrrole @ nano copper/pamidronate disodium composite coating with osteoinduction and photo-thermal/copper synergistic antibacterial functions, which comprises the following steps:

(1) mechanically polishing the surface of a titanium plate serving as a matrix to remove an oxide layer or an organic layer, putting the titanium plate into acetone for ultrasonic cleaning for 10 min, and cleaning with deionized water;

(2) putting the titanium plate obtained in the step (1) into a mixed acid solution (V)_H2SO4：V_HCl：V_H2O= 1: 1: 1) pickling and passivating for 5 hours at the constant temperature of 60 ℃, and then cleaning by using deionized water;

(3) putting the titanium plate obtained in the step (2) into a saturated sodium hydroxide solution, carrying out alkali washing and passivation treatment for 1 h at a constant temperature of 60 ℃, then cleaning the titanium plate by using deionized water, and airing for later use;

(4) dissolving 2.0 mL of pyrrole in 1000 mL of deionized water, and mechanically stirring to completely dissolve the pyrrole to obtain a uniform pyrrole solution;

(5) adding 0.2 g of copper nitrate into the pyrrole solution obtained in the step (4), and mechanically stirring to completely dissolve the copper nitrate to obtain a uniform and stable mixed solution of pyrrole and copper nitrate;

(6) adding 1.0mg of pamidronate disodium into the mixed solution obtained in the step (5), mechanically stirring to completely dissolve the pamidronate disodium, and adjusting the pH value of the solution to be between 5.0 and 5.1 by using hydrochloric acid;

(7) and (3) forming a three-electrode system by using the titanium plate obtained in the step (3) as a working electrode, the titanium foil as an auxiliary electrode and the saturated calomel electrode as a reference electrode, and placing the three-electrode system in the mixed solution obtained in the step (6) for pulse electrochemical deposition, wherein the relevant parameters comprise: and the oxidation potential is 0.8V, the oxidation time is 90 s, the reduction potential is-1.5V, the reduction time is 450 s, the cycle times are 5 times, and the total time is 0.75 h, so that the polypyrrole @ nano copper/pamidronate disodium composite coating is obtained, and the coating sample is naturally dried.

As shown in fig. 1-4, fig. 1 is an SEM image of polypyrrole @ nano copper/disodium pamidronate composite coating, and it can be seen that the copper nanoparticles on the composite coating are distributed more uniformly; fig. 2 is a qualitative diagram of escherichia coli and staphylococcus aureus of the polypyrrole @ nano copper/pamidronate disodium composite coating, and it can be seen that the escherichia coli and staphylococcus aureus grow more on the surface of the substrate titanium and form colonies, because the substrate titanium does not have antibacterial property, and no obvious escherichia coli and aurora staphylococcus colonies are observed on the surface of the polypyrrole @ nano copper/pamidronate disodium composite coating, because the copper nanoparticles in the composite coating have significant antibacterial property and antibacterial resistance; FIG. 3 is a bacterial quantitative diagram of the polypyrrole @ nano copper/pamidronate disodium composite coating, and it can be seen from the diagram that the antibacterial rates of the substrate titanium to Escherichia coli and Staphylococcus aureus are both 0%, and the antibacterial rates of the polypyrrole @ nano copper/pamidronate disodium composite coating to the two bacteria are both 100%, which indicates that the composite coating has excellent antibacterial ability; fig. 4 is an ultraviolet absorption spectrum diagram of the polypyrrole @ nano copper/disodium pamidronate composite coating and the substrate titanium, and it can be known from the diagram that the absorbance of the substrate titanium coating at 808nm is 1.141, and the absorbance of the polypyrrole @ nano copper/disodium pamidronate composite coating at 808nm is 1.45, and compared with the two, the absorbance of the polypyrrole @ nano copper/disodium pamidronate composite coating at 808nm is improved, because the copper nanoparticles have a surface plasma resonance effect, and the light absorption performance of the composite coating is enhanced.

Example two

The invention provides a preparation method of a polypyrrole @ nano copper/pamidronate disodium composite coating with osteoinduction and photo-thermal/copper synergistic antibacterial functions, which comprises the following steps:

(1) mechanically polishing the surface of a titanium plate serving as a matrix to remove an oxide layer or an organic layer, putting the titanium plate into ethyl acetate for ultrasonic cleaning for 15 min, and cleaning with deionized water;

(2) placing the titanium plate obtained in the step (1)Adding hydrochloric acid solution (V)_HCl：V_H2O= 3: 1) pickling and passivating for 4 hours at a constant temperature of 70 ℃, and then cleaning with deionized water;

(3) putting the titanium plate obtained in the step (2) into a saturated potassium hydroxide solution, carrying out alkali washing and passivation treatment for 1.5 h at a constant temperature of 70 ℃, then cleaning the titanium plate by using deionized water, and airing the titanium plate for later use;

(4) dissolving 2.1 mL of pyrrole in 1000 mL of deionized water, and mechanically stirring to completely dissolve the pyrrole to obtain a uniform pyrrole solution;

(5) adding 0.22 g of copper sulfate into the pyrrole solution obtained in the step (4), and mechanically stirring to completely dissolve the copper sulfate and the pyrrole to obtain a uniform and stable mixed solution of pyrrole and copper sulfate;

(6) adding 0.8mg of pamidronate disodium into the mixed solution obtained in the step (5), mechanically stirring to completely dissolve the pamidronate disodium, and adjusting the pH value of the solution to be between 5.0 and 5.1 by using hydrochloric acid;

(7) and (3) forming a three-electrode system by using the titanium plate obtained in the step (3) as a working electrode, a platinum sheet as an auxiliary electrode and a silver/silver chloride electrode as a reference electrode, and placing the three-electrode system in the mixed solution obtained in the step (6) for pulse electrochemical deposition, wherein the related parameters comprise: the polypyrrole @ nano copper/pamidronate disodium composite coating is obtained by oxidizing at 0.8V for 100s, reducing at-1.5V for 550 s, circulating for 7 times and totally 1.25h, and the coating sample is naturally dried.

EXAMPLE III

The invention provides a preparation method of a polypyrrole @ nano copper/pamidronate disodium composite coating with osteoinduction and photo-thermal/copper synergistic antibacterial functions, which comprises the following steps:

(1) mechanically polishing the surface of a titanium plate serving as a matrix to remove an oxide layer or an organic layer, putting the titanium plate into a mixed solution of acetonitrile and trichloromethane in equal proportion, performing ultrasonic cleaning for 20 min, and cleaning with deionized water;

(2) putting the titanium plate obtained in the step (1) into a sulfuric acid solution (V)_H2SO4：V_H2O= 2: 1) pickling and passivating at constant temperature of 80 deg.C for 3 hr, thenThen cleaning with deionized water;

(3) putting the titanium plate obtained in the step (2) into a saturated lithium hydroxide solution, carrying out alkali washing and passivation treatment for 2 hours at a constant temperature of 80 ℃, then cleaning the titanium plate by using deionized water, and airing the titanium plate for later use;

(4) dissolving 2.2 mL of pyrrole in 1000 mL of deionized water, and mechanically stirring to completely dissolve the pyrrole to obtain a uniform pyrrole solution;

(5) adding 0.24g of copper chloride into the pyrrole solution obtained in the step (4), and mechanically stirring to completely dissolve the copper chloride to obtain a uniform and stable mixed solution of pyrrole and copper chloride;

(6) adding 1.2mg of pamidronate disodium into the mixed solution obtained in the step (5), mechanically stirring to completely dissolve the pamidronate disodium, and adjusting the pH value of the solution to be between 5.0 and 5.1 by using hydrochloric acid;

(7) and (3) forming a three-electrode system by using the titanium plate obtained in the step (3) as a working electrode, the carbon electrode as an auxiliary electrode and the saturated calomel electrode as a reference electrode, and placing the three-electrode system in the mixed solution obtained in the step (6) for pulse electrochemical deposition, wherein the relevant parameters comprise: and the oxidation potential is 0.8V, the oxidation time is 110 s, the reduction potential is-1.5V, the reduction time is 600 s, the cycle times are 8 times, and the total time is 1.5 h, so that the polypyrrole @ nano copper/pamidronate disodium composite coating is obtained, and the coating sample is naturally dried.

The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the scope of the present invention, and any modifications, equivalents, improvements and the like made by those skilled in the art without departing from the spirit and principle of the present invention should be included in the protection scope of the present invention.