阅读说明：本技术 钛基表面兼具抗氧化及自生氧功能的纳米棒阵列构形化涂层及其制备方法和应用 (Nanorod array-configured coating with anti-oxidation and self-oxygen generation functions on titanium-based surface and preparation method and application thereof ) 是由 李博 刘福立 憨勇 于 2021-02-05 设计创作，主要内容包括：本发明公开了一种钛基表面兼具抗氧化及自生氧功能的纳米棒阵列构形化涂层及其制备方法和应用。首先采用微弧氧化在钛或其合金表层制备含磷和钙的多孔二氧化钛涂层,然后用水热处理法原位生长出羟基磷灰石纳米棒构形化涂层,再用氧化自聚合法制备聚多巴胺包覆的羟基磷灰石纳米棒阵列,最后通过聚多巴胺吸附金属离子、水热法最终得到兼具抗氧化及自生氧涂层的生物医用材料。本发明工艺稳定、可控性强；制得的涂层表层同时具有抗氧化和自生氧的双重功效；作为植入体能够调控RA病况下不良的骨微环境、促进骨组织愈合及再生能力,避免由于不良的骨微环境所导致的植入体失效和膜/基结合力差诱发的涂层剥落及由其诱发的骨溶解或过度炎性等副反应的发生。(The invention discloses a nanorod array formation coating with antioxidant and self-oxygen generation functions on a titanium-based surface, and a preparation method and application thereof. Firstly, preparing a porous titanium dioxide coating containing phosphorus and calcium on the surface layer of titanium or an alloy thereof by adopting micro-arc oxidation, then growing a hydroxyapatite nanorod-structured coating in situ by using a hydrothermal treatment method, preparing a poly-dopamine-coated hydroxyapatite nanorod array by using an oxidation auto-polymerization method, and finally obtaining the biomedical material with the oxidation resistance and the self-oxygen generation coating by using a poly-dopamine adsorption metal ion and hydrothermal method. The invention has stable process and strong controllability; the surface layer of the prepared coating has double effects of oxidation resistance and self-generated oxygen; the implant can regulate and control the poor bone microenvironment in the RA condition, promote the healing and regeneration capacity of bone tissues, and avoid the occurrence of side reactions such as implant failure caused by the poor bone microenvironment, coating spalling induced by poor membrane/base binding force, osteolysis or excessive inflammation induced by the coating spalling, and the like.)

1. A preparation method of a nanorod array-structured coating with a titanium-based surface having anti-oxidation and self-oxygen generation functions is characterized by comprising the following steps:

step 1: micro-arc oxidation is carried out on a pure titanium or titanium alloy matrix in electrolyte containing phosphorus ions and calcium ions, so that a microporous titanium dioxide coating is formed on the surface of the matrix;

step 2: preparing a hydroxyapatite nanorod-structured coating on the microporous titanium dioxide coating obtained in the step 1 by adopting a hydrothermal treatment method;

and step 3: preparing a poly-dopamine-coated hydroxyapatite nanorod-structured coating on the hydroxyapatite nanorod-structured coating obtained in the step 2 by adopting an oxidative self-polymerization method;

and 4, step 4: and (3) placing the hydroxyapatite nanorod structured coating coated with the polydopamine prepared in the step (3) into a mixed salt solution of iron acetate and manganese acetate at normal temperature, stirring and heating to 60 ℃, then placing the product into a sodium hydroxide solution, and performing hydrothermal treatment for 1-2 hours at 90-100 ℃ to obtain the nanorod array structured coating with the titanium-based surface having the functions of oxidation resistance and self-generating oxygen.

2. The method for preparing the nanorod array-shaped coating with the titanium-based surface having the functions of resisting oxidation and generating oxygen automatically as claimed in claim 1, wherein the step 1 specifically comprises:

taking a pure titanium or titanium alloy matrix as an anode and a stainless steel electrolytic tank as a cathode, performing micro-arc oxidation at the temperature of 300-310K, and cleaning and drying by using alcohol and deionized water to form a microporous titanium dioxide coating on the surface of the titanium matrix; the parameters of the micro-arc oxidation are as follows: the arc frequency is 90-110 Hz, the positive pressure is 300-400V, and the duty ratio is 0-10%; the electrolyte includes: 0.005-0.01 mol/L of sodium hydroxide, 0.2-0.3 mol/L of calcium acetate and 0.02-0.03 mol/L of beta-phosphoglyceride disodium salt pentahydrate.

3. The method for preparing the nanorod array-shaped coating with the titanium-based surface having the functions of resisting oxidation and generating oxygen automatically as claimed in claim 1, wherein the step 2 specifically comprises:

step 2.1: putting the microporous titanium dioxide coating obtained in the step 1 into 12-15 mL of a 0.01-0.03 mol/L sodium hydroxide aqueous solution, and sealing for carrying out primary hydrothermal treatment for 1-2 h;

step 2.2: adding calcium sodium ethylene diamine tetraacetate, a beta-phosphoglyceride disodium salt pentahydrate and sodium hydroxide into water, uniformly stirring, and sealing the product obtained in the step 2.1 for secondary hydrothermal treatment, wherein the concentration of the calcium sodium ethylene diamine tetraacetate is 0.05-0.1 mol/L, the concentration of the beta-phosphoglyceride disodium salt pentahydrate is 0.01-0.03 mol/L, and the concentration of the sodium hydroxide is 0.1-0.2 mol/L; and cleaning and drying the product for later use.

4. The method for preparing the nanorod array-configured coating with the titanium-based surface having the functions of resisting oxidation and generating oxygen automatically as claimed in claim 3, wherein the temperature of the primary hydrothermal treatment is 360-380K, and the time is 1-2 h; the temperature of the secondary hydrothermal treatment is 380-400K, and the time is 18-22 h.

5. The method for preparing the nanorod array-shaped coating with the titanium-based surface having the functions of resisting oxidation and generating oxygen automatically as claimed in claim 1, wherein the step 3 specifically comprises: and (3) immersing the product obtained in the step (2) in a 1-3 mg/mL dopamine trihydroxymethylaminomethane solution, wherein the concentration of trihydroxymethylaminomethane in the dopamine trihydroxymethylaminomethane solution is 10mmol/L, stirring at normal temperature in a dark place at the rotating speed of 400-800 r/min for 10-15 h, and cleaning and drying after stirring.

6. The method for preparing the nanorod array-configured coating with the titanium-based surface having the functions of resisting oxidation and generating oxygen automatically as claimed in claim 1, wherein in the step 4, the concentration of iron acetate in the mixed salt solution is 0.5-1.5 mmol/L, and the concentration of manganese acetate is 0.25-0.75 mmol/L; the concentration of the sodium hydroxide solution is 0.01-0.03 mmol/L.

7. The method for preparing the nanorod array-configured coating with the titanium-based surface having both the oxidation resistance and the self-oxygen generation functions of claim 6, wherein the concentration of iron acetate in the mixed salt solution is 1.0mmol/L, and the concentration of manganese acetate in the mixed salt solution is 0.5 mmol/L.

8. The method for preparing the nanorod array-configured coating with the titanium-based surface having both the oxidation resistance and the self-oxygen generation function according to claim 1, wherein in the step 4, after stirring and heating to 60 ℃, the product is placed in a sodium hydroxide solution within 0-45 min.

9. The nanorod array-shaped coating with the oxidation resistance and the self-oxygen generation functions on the titanium-based surface, prepared by the preparation method of any one of claims 1 to 8.

10. The use of the nanorod array-configured coating of claim 9, having both oxidation-resistant and self-generating oxygen functionality, as an implant coating material.

Technical Field

The invention belongs to the technical field of biomedical materials, and particularly relates to a nanorod array-shaped coating with an antioxidant and self-generating oxygen function on a titanium-based surface, and a preparation method and application thereof.

Background

Rheumatoid Arthritis (RA) is a chronic systemic immune disease, in the diseased joint region, macrophages in synovial fluid mostly present M1 proinflammatory phenotype, so that the expression of Reactive Oxygen Species (ROS) and inflammatory factors in the region is remarkably increased and transported to the whole body along with blood circulation; the high expression of ROS and inflammatory factors in serum can induce the anoxic state in bones and promote the maturation and differentiation of osteoclast cells by activating NF-kappa b passage, thereby causing the systemic bone loss of RA patients and increasing the fracture risk. Even more feared, once a RA patient is fractured, the high ROS content in the serum and the hypoxic microenvironment in the bone will cause the macrophages at the fracture site to be over-polarized towards the M1 phenotype, inducing a strong inflammatory response, even though the artificial material implant with high osteointegration in normal bone cannot achieve osteointegration at the fracture site.

Accordingly, there is a need for the targeted design and construction of an endosteal implant to alleviate the high ROS and hypoxic microenvironment in the bone in the RA condition, and in turn to regulate the polarization of macrophages towards the M2 phenotype, i.e. to confer the implant with an immunomodulatory capacity in the RA condition to exhibit high osteointegration in this condition.

Disclosure of Invention

Aiming at the fracture under the RA condition, the invention aims to solve the problem of surface modification of the titanium-based implant, and provides a nanorod array formation coating with the functions of oxidation resistance and self-generated oxygen on the titanium-based surface, and a preparation method and application thereof, wherein the nanorod array formation coating is stable in process and strong in controllability; the surface layer of the prepared biomedical material has double effects of oxidation resistance and self-generated oxygen; and can be used as an implant to promote the healing and regeneration capacity of bone tissues in time after regulating ROS and an anoxic bone microenvironment under an RA condition, and avoid the occurrence of side reactions such as coating spalling induced by poor membrane/base binding force and osteolysis or excessive inflammation induced by the coating spalling.

The invention is realized by the following technical scheme:

the invention discloses a preparation method of a nanorod array-structured coating with both antioxidant and self-generating oxygen functions on a titanium-based surface, which comprises the following steps:

step 1: carrying out micro-arc oxidation on a pure titanium or titanium alloy matrix in an electrolyte containing phosphorus ions and calcium ions to form a microporous titanium dioxide coating on the surface of the titanium-based matrix;

step 2: preparing a hydroxyapatite nanorod-structured coating on the microporous titanium dioxide coating obtained in the step 1 by adopting a hydrothermal treatment method;

and step 3: preparing a poly-dopamine-coated hydroxyapatite nanorod-structured coating on the hydroxyapatite nanorod-structured coating obtained in the step 2 by adopting an oxidative self-polymerization method;

and 4, step 4: and (3) placing the hydroxyapatite nanorod structured coating coated with the polydopamine prepared in the step (3) into a mixed salt solution of iron acetate and manganese acetate at normal temperature, stirring and heating to 60 ℃, then placing the product into a sodium hydroxide solution, and performing hydrothermal treatment for 1-2 hours at 90-100 ℃ to obtain the nanorod array structured coating with the titanium-based surface having the functions of oxidation resistance and self-generating oxygen.

Preferably, step 1 is specifically:

taking a pure titanium or titanium alloy matrix as an anode and a stainless steel electrolytic tank as a cathode, performing micro-arc oxidation at the temperature of 300-310K, and cleaning and drying by using alcohol and deionized water to form a microporous titanium dioxide coating on the surface of the titanium matrix; the parameters of the micro-arc oxidation are as follows: the arc frequency is 90-110 Hz, the positive pressure is 300-400V, and the duty ratio is 0-10%; the electrolyte includes: 0.005-0.01 mol/L of sodium hydroxide, 0.2-0.3 mol/L of calcium acetate and 0.02-0.03 mol/L of beta-phosphoglyceride disodium salt pentahydrate.

Preferably, step 2 specifically comprises:

step 2.1: putting the microporous titanium dioxide coating obtained in the step 1 into 12-15 mL of a 0.01-0.03 mol/L sodium hydroxide aqueous solution, and sealing for carrying out primary hydrothermal treatment for 1-2 h;

step 2.2: adding calcium sodium ethylene diamine tetraacetate, a beta-phosphoglyceride disodium salt pentahydrate and sodium hydroxide into water, uniformly stirring, and sealing the product obtained in the step 2.1 for secondary hydrothermal treatment, wherein the concentration of the calcium sodium ethylene diamine tetraacetate is 0.05-0.1 mol/L, the concentration of the beta-phosphoglyceride disodium salt pentahydrate is 0.01-0.03 mol/L, and the concentration of the sodium hydroxide is 0.1-0.2 mol/L; and cleaning and drying the product for later use.

Further preferably, the temperature of the primary hydrothermal treatment is 360-380K, and the time is 1-2 h; the temperature of the secondary hydrothermal treatment is 380-400K, and the time is 18-22 h.

Preferably, step 3 is specifically: and (3) immersing the product obtained in the step (2) in a 1-3 mg/mL dopamine trihydroxymethylaminomethane solution, wherein the concentration of trihydroxymethylaminomethane in the dopamine trihydroxymethylaminomethane solution is 10mmol/L, stirring at normal temperature in a dark place at the rotating speed of 400-800 r/min for 10-15 h, and cleaning and drying after stirring.

Preferably, in the step 4, the concentration of the iron acetate in the mixed salt solution is 0.5-1.5 mmol/L, and the concentration of the manganese acetate is 0.25-0.75 mmol/L; the concentration of the sodium hydroxide solution is 0.01-0.03 mmol/L.

Further preferably, the concentration of iron acetate in the mixed salt solution is 1.0mmol/L and the concentration of manganese acetate is 0.5 mmol/L.

Preferably, in the step 4, after stirring and heating to 60 ℃, the product is placed into a sodium hydroxide solution within 0-45 min.

The invention also discloses a nanorod array formation coating with the titanium base surface having the functions of oxidation resistance and self-generated oxygen, which is prepared by the preparation method.

The invention also discloses application of the nanorod array formation coating with the titanium-based surface having the functions of oxidation resistance and self-generated oxygen as an implant coating material.

Compared with the prior art, the invention has the following beneficial technical effects:

the invention relates to a preparation method of a nanorod array-structured coating with a titanium-based surface having anti-oxidation and self-oxygen generation functions, which comprises the steps of firstly preparing a phosphorus and calcium-containing porous titanium dioxide coating on a titanium or titanium alloy surface by a micro-arc oxidation process, then growing a Hydroxyapatite (HA) nanorod-structured coating on the phosphorus and calcium-containing porous titanium dioxide coating in situ by a hydrothermal treatment method, finally preparing a Polydopamine (PDA) -coated hydroxyapatite nanorod array (PDA @ HA) by an oxidative self-polymerization method, and finally obtaining a biomedical material with the nanorod-arrayed anti-oxidation and self-oxygen generation coating by a polydopamine adsorption metal ion hydrothermal method. In-situ preparation of microporous TiO by micro-arc oxidation₂A layer providing a strong interfacial bond between the coating and the substrate; and the micro-arc oxidation electrolyte and the hydrothermal solution have simple components, do not contain easily decomposed components, and have stable and controllable process and strong repeatability. Dopamine with reducing catechol groups is selected as an antioxidant and a photosensitizer, and a PDA film layer is coated on the surface of the HA nanorod by utilizing the oxidative autopolymerization behavior of the dopamine in a weak alkaline environment. Finally, the manganese ferrite and polydopamine co-modified hydroxyapatite nanorod coating is obtained by utilizing the capability of a catechol group in polydopamine for adsorbing metal ions and further carrying out hydrothermal crystallization treatment in an alkaline solution. Wherein the manganese ferrite is distributed on the surface of the nano-rod in the form of nano-particles, and can be used for treating hypoxia and ROS (H) rich₂O₂) Oxygen is generated by fenton reaction in the case of (1). Constructing a PDA @ HA nanorod array surface layer with double effects of oxidation resistance and self-generated oxygen.

In the preparation method, the micro-arc oxidation electrolyte and the hydrothermal solution have simple components, do not contain easily decomposed components, and have stable and controllable process and strong repeatability; the polydopamine has strong oxidative self-polymerization capability, can effectively adsorb metal ions by utilizing self functional groups, and has extremely simple and convenient dipping and coating mode and stable process, so that the process can be used for large-scale production and preparation. The raw materials used for preparing the coating, such as NaOH, beta-GP, EDTA-Ca, dopamine hydrochloride and the like, are nontoxic and safe, have no side effect on human bodies, can be directly purchased in the market and are easily obtained, so that the popularization and the application of the technology are ensured.

Further, a 1-3 mg/mL dopamine trihydroxymethyl aminomethane solution is adopted, and the HA nanorod array coating is completely covered by polydopamine due to excessive concentration; too low a concentration may result in failure to form polydopamine on the HA nanorods, which is strongly and uniformly bonded. The stirring speed is 400-800 r/min, and the excessive high speed can cause incomplete oxidation autopolymerization reaction of dopamine on the HA nanorods and insufficient firm combination of polydopamine formed on the HA nanorods; too slow a rate may result in uneven polymeric coating of dopamine on the HA nanorods.

Further, the concentration of iron acetate in the mixed salt solution is 0.5-1.5 mmol/L, the concentration of manganese acetate is 0.25-0.75 mmol/L, the concentration of sodium hydroxide solution is 0.01-0.03 mmol/L, and the uniformly coated manganese ferrite and polydopamine co-modified hydroxyapatite nano-rod can be formed under the concentration. If the concentration is too low, hydrothermal crystallization is insufficient, and iron and manganese cannot be sufficiently converted into manganese ferrite; if the concentration is too high, a large amount of manganese ferrite products are generated to completely cover the surface of the nanorod, and the effects of promoting bone formation and resisting oxidation cannot be achieved.

Furthermore, when the concentration of iron acetate in the mixed salt solution is 1.0mmol/L and the concentration of manganese acetate is 0.5mmol/L, the coating retains HA, PDA and MnFe₂O₄On the basis of the phase, the nano-rod structure is changed into the nano-tube structure, so that the drug loading can be further carried out under the action of realizing the double-layer functions of oxidation resistance and self-generation of oxygen under the condition, and the treatment of RA pathology is facilitated. The concentration is too low, the contents of iron and manganese are low, and the nano rod is not corroded sufficiently in the hydrothermal crystallization reaction process; if the concentration is too high, excessive manganese ferrite is generated and is further filled into the tube after being hydrothermally crystallized into the tube.

Further, in the step 4, after stirring and heating to 60 ℃, the product is placed into a sodium hydroxide solution within 0-45 min, and the time is too long, so that excessive iron and manganese ions can be adsorbed on the surface of the polydopamine, and a large amount of manganese ferrite nanoparticles are generated after further hydrothermal crystallization treatment, so that the polydopamine-coated hydroxyapatite nanorods can be completely wrapped.

The nanorod array-shaped coating with the titanium-based surface having the functions of oxidation resistance and self-generation of oxygen, which is prepared by the preparation method, has a double-layer structure, the surface layer is a nanorod array shaped like a bone matrix, and the inner layer (adjacent to the matrix) is microporous TiO containing phosphorus and calcium₂And (4) coating. The HA nanorod array surface layer coated with the PDA HAs osteogenesis promoting and antioxidant effects. On the basis of the functions of effectively removing free radicals and promoting osteogenesis, the hydroxyapatite nanorod subjected to co-modification by the manganese ferrite and the polydopamine can generate oxygen through Fenton reaction, so that the microenvironment of hypoxia at the fracture part under RA pathology is relieved. And the coating is tightly combined with the titanium-based matrix. The adopted raw materials are nontoxic and safe, have no side effect on human bodies, are easy to obtain and are easy to popularize and apply.

When the nanorod array formation coating with the titanium-based surface having the functions of oxidation resistance and self-generation of oxygen is used as the implant coating material, the inner layer (adjacent to the substrate) is microporous TiO containing phosphorus and calcium₂A coating, which is capable of significantly promoting osteogenesis-related cellular functions, and, in addition, is MnFe-coated₂O₄The surface layer of the PDA co-modified HA nanorod array HAs double effects of oxidation resistance and self-generated oxygen, and can effectively remove excessive ROS in a bone microenvironment under an RA condition and utilize hydrogen peroxide in the ROS to catalyze and generate oxygen to relieve the bone microenvironment with hypoxia. Thereby greatly improving the capability of the titanium-based implant to promote the healing and regeneration of bone tissues. In addition, the coating can be tightly combined with the titanium substrate, and the occurrence of side reactions such as coating stripping induced by poor membrane/base binding force and osteolysis or excessive inflammation induced by the coating stripping is avoided.

Drawings

FIG. 1 is an SEM photograph of the surface morphology of the HA nanorod structured coating prepared by the micro-arc oxidation-hydrothermal treatment in example 1;

FIG. 2 shows MnFe prepared in example 1₂O₄-SEM photograph of surface topography of PDA @ HA coated titanium implants;

FIG. 3 shows MnFe prepared in example 2₂O₄SEM photograph of surface topography of titanium implant coated with PDA @ HASlicing;

FIG. 4 shows MnFe prepared in example 2₂O₄-HRTEM photographs of PDA @ HA coated titanium implants;

FIG. 5 shows MnFe-containing samples prepared in examples 1 and 2₂O₄-fourier ir spectroscopy profile of a titanium implant coated with PDA @ HA;

FIG. 6 shows a titanium implant with HA coating and PDA @ HA coating and examples 2, 4 with MnFe₂O₄-XRD characterization pattern of titanium implants coated with PDA @ HA.

Detailed Description

The present invention will now be described in further detail with reference to the following figures and specific examples, which are intended to be illustrative, but not limiting, of the invention.

Example 1

The preparation method of the biomedical material with the nanorod-arrayed thermal control immunity and oxidation-resistant coating on the titanium-based surface comprises the following steps:

step 1, micro-arc oxidation of titanium and titanium alloy:

the micro-arc oxidation parameters are set as follows: the frequency of the micro-arc oxidation arc is 110Hz, the power supply is positive voltage of 400V, and the duty ratio is 7.5 percent. In the micro-arc oxidation process, pure titanium or titanium alloy is taken as an anode, a stainless steel electrolytic tank is taken as a cathode, and the components and the concentration of the electrolyte are as follows: 0.005mol/L of sodium hydroxide (NaOH), calcium acetate (Ca (CH)₃COO)₂)0.2mol/L, 0.02mol/L of beta-phosphoglyceride disodium salt pentahydrate (beta-GP). In the preparation process, a cooling system is adopted to control the temperature of the micro-arc oxidation electrolyte at 300K. And (3) obtaining a sample coated with the microporous titanium dioxide coating after micro-arc oxidation, and putting the prepared sample into a drying oven for later use after the sample is cleaned by alcohol and deionized water.

Step 2, adding porous TiO containing phosphorus and calcium₂Carrying out hydrothermal treatment on the coating to obtain the HA nanorod structured coating:

step 2.1, primary hydrothermal treatment

Firstly, preparing a solution required by primary hydrothermal, wherein the solution is a sodium hydroxide aqueous solution with the concentration of 0.01mol/L, putting a titanium sheet coated with a titanium dioxide coating after micro-arc oxidation into reaction kettles, then adding 15mL of the sodium hydroxide aqueous solution into each reaction kettle, screwing down the reaction kettles, putting the reaction kettles into a drying oven, and finally setting temperature and time parameters. The temperature was adjusted to 360K and the time was set to 2 h.

Step 2.2, Secondary hydrothermal treatment

Adding calcium sodium ethylene diamine tetraacetate, a beta-phosphoglyceride disodium salt pentahydrate and sodium hydroxide into water, uniformly stirring, and sealing the product obtained in the step 2.1 for secondary hydrothermal treatment, wherein the concentration of the calcium sodium ethylene diamine tetraacetate is 0.02mol/L, the concentration of the beta-phosphoglyceride disodium salt pentahydrate is 0.02mol/L, and the concentration of the sodium hydroxide is 0.125 mol/L; the secondary water heating temperature is set to 385K, and the time is set to 20 h. And (3) obtaining a titanium sheet coated with the HA nanorod-structured coating after two times of hydrothermal treatment, taking the sample out of the reaction kettle, washing the sample with deionized water, and putting the sample into a drying oven for later use.

Step 3, preparing a poly-dopamine (PDA) -coated Hydroxyapatite (HA) nanorod array (PDA @ HA)

Immersing the product obtained in the step 2 in a trihydroxymethylaminomethane solution of dopamine with the concentration of 2mg/mL, wherein the trihydroxymethylaminomethane concentration in the trihydroxymethylaminomethane solution of dopamine is 10mmol/L, and stirring for 12 hours at normal temperature in a dark place at the rotating speed of 600 r/min. And finally, taking out the sample, washing with deionized water, and drying in an oven for later use.

Step 4, preparing manganese ferrite (MnFe)₂O₄) -Polydopamine (PDA) co-modified Hydroxyapatite (HA) nanorod array (MnFe)₂O₄[email protected])

Placing the product obtained in the step (3) in a mixed salt solution of iron acetate and manganese acetate at normal temperature, gently stirring and gradually heating to 60 ℃, wherein the concentration of the iron acetate is 0.5mmol/L and the concentration of the manganese acetate is 0.25 mmol/L; then taking out a sample to perform hydrothermal treatment, wherein the hydrothermal solution is a sodium hydroxide aqueous solution with the concentration of 0.01mol/L, the hydrothermal temperature is set to 98 ℃, and the time is 2 hours. And after the hydrothermal process is finished, taking out the sample, cleaning and drying to obtain the nanorod array formation coating with the titanium-based surface having the functions of oxidation resistance and self-generated oxygen.

Example 2

The preparation method of the biomedical material with the nanorod-arrayed thermal control immunity and oxidation-resistant coating on the titanium-based surface comprises the following steps:

step 1, micro-arc oxidation of titanium and titanium alloy:

the micro-arc oxidation parameters are set as follows: the frequency of the micro-arc oxidation arc is 110Hz, the power supply is positive voltage of 400V, and the duty ratio is 7.5 percent. In the micro-arc oxidation process, pure titanium or titanium alloy is taken as an anode, a stainless steel electrolytic tank is taken as a cathode, and the components and the concentration of the electrolyte are as follows: 0.005mol/L of sodium hydroxide (NaOH), calcium acetate (Ca (CH)₃COO)₂)0.2mol/L, 0.02mol/L of beta-phosphoglyceride disodium salt pentahydrate (beta-GP). In the preparation process, a cooling system is adopted to control the temperature of the micro-arc oxidation electrolyte at 300K. And (3) obtaining a sample coated with the microporous titanium dioxide coating after micro-arc oxidation, and putting the prepared sample into a drying oven for later use after the sample is cleaned by alcohol and deionized water.

Step 2, adding porous TiO containing phosphorus and calcium₂Carrying out hydrothermal treatment on the coating to obtain the HA nanorod structured coating:

step 2.1, primary hydrothermal treatment

Firstly, preparing a solution required by primary hydrothermal, wherein the solution is a sodium hydroxide aqueous solution with the concentration of 0.01mol/L, putting a titanium sheet coated with a titanium dioxide coating after micro-arc oxidation into reaction kettles, then adding 15mL of the sodium hydroxide aqueous solution into each reaction kettle, screwing down the reaction kettles, putting the reaction kettles into a drying oven, and finally setting temperature and time parameters. The temperature was adjusted to 360K and the time was set to 2 h.

Step 2.2, Secondary hydrothermal treatment

Adding calcium sodium ethylene diamine tetraacetate, a beta-phosphoglyceride disodium salt pentahydrate and sodium hydroxide into water, uniformly stirring, and sealing the product obtained in the step 2.1 for secondary hydrothermal treatment, wherein the concentration of the calcium sodium ethylene diamine tetraacetate is 0.02mol/L, the concentration of the beta-phosphoglyceride disodium salt pentahydrate is 0.02mol/L, and the concentration of the sodium hydroxide is 0.125 mol/L; the secondary water heating temperature is set to 385K, and the time is set to 20 h. And (3) obtaining a titanium sheet coated with the HA nanorod-structured coating after two times of hydrothermal treatment, taking the sample out of the reaction kettle, washing the sample with deionized water, and putting the sample into a drying oven for later use.

Step 3, preparing a poly-dopamine (PDA) -coated Hydroxyapatite (HA) nanorod array (PDA @ HA)

Immersing the product obtained in the step 2 in a trihydroxymethylaminomethane solution of dopamine with the concentration of 2mg/mL, wherein the trihydroxymethylaminomethane concentration in the trihydroxymethylaminomethane solution of dopamine is 10mmol/L, and stirring for 12 hours at normal temperature in a dark place at the rotating speed of 600 r/min. And finally, taking out the sample, washing with deionized water, and drying in an oven for later use.

Step 4, preparing manganese ferrite (MnFe)₂O₄) -Polydopamine (PDA) co-modified Hydroxyapatite (HA) nanorod array (MnFe)₂O₄[email protected])

Placing the product obtained in the step (3) in a mixed salt solution of iron acetate and manganese acetate at normal temperature, gently stirring and gradually heating to 60 ℃, wherein the concentration of the iron acetate is 1mmol/L and the concentration of the manganese acetate is 0.5 mmol/L; then taking out a sample to perform hydrothermal treatment, wherein the hydrothermal solution is a sodium hydroxide aqueous solution with the concentration of 0.01mol/L, the hydrothermal temperature is set to 98 ℃, and the time is 2 hours. And after the hydrothermal process is finished, taking out the sample, cleaning and drying to obtain the nanorod array formation coating with the titanium-based surface having the functions of oxidation resistance and self-generated oxygen.

Example 3

The preparation method of the biomedical material with the nanorod-arrayed thermal control immunity and oxidation-resistant coating on the titanium-based surface comprises the following steps:

step 1, micro-arc oxidation of titanium and titanium alloy:

the micro-arc oxidation parameters are set as follows: the frequency of the micro-arc oxidation arc is 110Hz, the power supply is positive voltage of 400V, and the duty ratio is 7.5 percent. In the micro-arc oxidation process, pure titanium or titanium alloy is taken as an anode, a stainless steel electrolytic tank is taken as a cathode, and the components and the concentration of the electrolyte are as follows: 0.005mol/L of sodium hydroxide (NaOH), calcium acetate (Ca (CH)₃COO)₂)0.2mol/L, 0.02mol/L of beta-phosphoglyceride disodium salt pentahydrate (beta-GP). In the preparation process, a cooling system is adopted to control the temperature of the micro-arc oxidation electrolyte at 300K. Coating obtained after micro-arc oxidationAnd cleaning the sample coated with the microporous titanium dioxide coating by using alcohol and deionized water, and putting the sample into a drying oven for later use.

Step 2, adding porous TiO containing phosphorus and calcium₂Carrying out hydrothermal treatment on the coating to obtain the HA nanorod structured coating:

step 2.1, primary hydrothermal treatment

Firstly, preparing a solution required by primary hydrothermal, wherein the solution is a sodium hydroxide aqueous solution with the concentration of 0.01mol/L, putting a titanium sheet coated with a titanium dioxide coating after micro-arc oxidation into reaction kettles, then adding 15mL of the sodium hydroxide aqueous solution into each reaction kettle, screwing down the reaction kettles, putting the reaction kettles into a drying oven, and finally setting temperature and time parameters. The temperature was adjusted to 360K and the time was set to 2 h.

Step 2.2, Secondary hydrothermal treatment

Adding calcium sodium ethylene diamine tetraacetate, a beta-phosphoglyceride disodium salt pentahydrate and sodium hydroxide into water, uniformly stirring, and sealing the product obtained in the step 2.1 for secondary hydrothermal treatment, wherein the concentration of the calcium sodium ethylene diamine tetraacetate is 0.02mol/L, the concentration of the beta-phosphoglyceride disodium salt pentahydrate is 0.02mol/L, and the concentration of the sodium hydroxide is 0.125 mol/L; the secondary water heating temperature is set to 385K, and the time is set to 20 h. And (3) obtaining a titanium sheet coated with the HA nanorod-structured coating after two times of hydrothermal treatment, taking the sample out of the reaction kettle, washing the sample with deionized water, and putting the sample into a drying oven for later use.

Step 3, preparing a poly-dopamine (PDA) -coated Hydroxyapatite (HA) nanorod array (PDA @ HA)

Immersing the product obtained in the step 2 in a trihydroxymethylaminomethane solution of dopamine with the concentration of 2mg/mL, wherein the trihydroxymethylaminomethane concentration in the trihydroxymethylaminomethane solution of dopamine is 10mmol/L, and stirring for 12 hours at normal temperature in a dark place at the rotating speed of 600 r/min. And finally, taking out the sample, washing with deionized water, and drying in an oven for later use.

Step 4, preparing manganese ferrite (MnFe)₂O₄) -Polydopamine (PDA) co-modified Hydroxyapatite (HA) nanorod array (MnFe)₂O₄[email protected])

Placing the product obtained in the step (3) in a mixed salt solution of iron acetate and manganese acetate at normal temperature, gently stirring and gradually heating to 60 ℃, wherein the concentration of the iron acetate is 1.5mmol/L and the concentration of the manganese acetate is 0.75 mmol/L; then taking out a sample to perform hydrothermal treatment, wherein the hydrothermal solution is a sodium hydroxide aqueous solution with the concentration of 0.01mol/L, the hydrothermal temperature is set to 98 ℃, and the time is 2 hours. And after the hydrothermal process is finished, taking out the sample, cleaning and drying to obtain the nanorod array formation coating with the titanium-based surface having the functions of oxidation resistance and self-generated oxygen.

Example 4

The preparation method of the biomedical material with the nanorod-arrayed thermal control immunity and oxidation-resistant coating on the titanium-based surface comprises the following steps:

step 1, micro-arc oxidation of titanium and titanium alloy:

the micro-arc oxidation parameters are set as follows: the frequency of the micro-arc oxidation arc is 90Hz, the power supply is positive pressure 300V, and the duty ratio is 0%. In the micro-arc oxidation process, pure titanium or titanium alloy is taken as an anode, a stainless steel electrolytic tank is taken as a cathode, and the components and the concentration of the electrolyte are as follows: 0.01mol/L of sodium hydroxide (NaOH), calcium acetate (Ca (CH)₃COO)₂)0.3mol/L, 0.03mol/L of beta-phosphoglyceride disodium salt pentahydrate (beta-GP). In the preparation process, a cooling system is adopted to control the temperature of the micro-arc oxidation electrolyte at 310K. And (3) obtaining a sample coated with the microporous titanium dioxide coating after micro-arc oxidation, and putting the prepared sample into a drying oven for later use after the sample is cleaned by alcohol and deionized water.

Step 2, adding porous TiO containing phosphorus and calcium₂Carrying out hydrothermal treatment on the coating to obtain the HA nanorod structured coating:

step 2.1, primary hydrothermal treatment

Firstly, preparing a solution required by primary hydrothermal, wherein the solution is a sodium hydroxide aqueous solution with the concentration of 0.03mol/L, putting a titanium sheet coated with a titanium dioxide coating after micro-arc oxidation into reaction kettles, then adding 12mL of the sodium hydroxide aqueous solution into each reaction kettle, screwing down the reaction kettles, putting the reaction kettles into a drying oven, and finally setting temperature and time parameters. The temperature was adjusted to 380K and the time was set to 1 h.

Step 2.2, Secondary hydrothermal treatment

Adding calcium sodium ethylene diamine tetraacetate, a beta-phosphoglyceride disodium salt pentahydrate and sodium hydroxide into water, uniformly stirring, and sealing the product obtained in the step 2.1 for secondary hydrothermal treatment, wherein the concentration of the calcium sodium ethylene diamine tetraacetate is 0.05mol/L, the concentration of the beta-phosphoglyceride disodium salt pentahydrate is 0.03mol/L, and the concentration of the sodium hydroxide is 0.2 mol/L; the secondary hydrothermal temperature was set to 380K and the time was set to 22 h. And (3) obtaining a titanium sheet coated with the HA nanorod-structured coating after two times of hydrothermal treatment, taking the sample out of the reaction kettle, washing the sample with deionized water, and putting the sample into a drying oven for later use.

Step 3, preparing a poly-dopamine (PDA) -coated Hydroxyapatite (HA) nanorod array (PDA @ HA)

Immersing the product obtained in the step 2 in a trihydroxymethylaminomethane solution of dopamine with the concentration of 3mg/mL, wherein the trihydroxymethylaminomethane concentration in the trihydroxymethylaminomethane solution of dopamine is 10mmol/L, and stirring for 15 hours at normal temperature in a dark place, wherein the stirring speed is 800 r/min. And finally, taking out the sample, washing with deionized water, and drying in an oven for later use.

Step 4, preparing manganese ferrite (MnFe)₂O₄) -Polydopamine (PDA) co-modified Hydroxyapatite (HA) nanorod array (MnFe)₂O₄[email protected])

Placing the product obtained in the step (3) in a mixed salt solution of iron acetate and manganese acetate at normal temperature, gently stirring and gradually heating to 60 ℃, wherein the concentration of the iron acetate is 1mmol/L and the concentration of the manganese acetate is 0.5 mmol/L; then taking out a sample to perform hydrothermal treatment, wherein the hydrothermal solution is a sodium hydroxide aqueous solution with the concentration of 0.02mol/L, the hydrothermal temperature is set to be 90 ℃, and the hydrothermal time is 2 hours. And after the hydrothermal process is finished, taking out the sample, cleaning and drying to obtain the nanorod array formation coating with the titanium-based surface having the functions of oxidation resistance and self-generated oxygen.

Example 5

The preparation method of the biomedical material with the nanorod-arrayed thermal control immunity and oxidation-resistant coating on the titanium-based surface comprises the following steps:

step 1, micro-arc oxidation of titanium and titanium alloy:

the micro-arc oxidation parameters are set as follows: the frequency of the micro-arc oxidation arc is 100Hz, the power supply is positive pressure 350V, and the duty ratio is 10%. In the micro-arc oxidation process, pure titanium or titanium alloy is taken as an anode, a stainless steel electrolytic tank is taken as a cathode, and the components and the concentration of the electrolyte are as follows: 0.008mol/L of sodium hydroxide (NaOH), calcium acetate (Ca (CH)₃COO)₂)0.25mol/L, 0.025mol/L of beta-phosphoglyceride disodium salt pentahydrate (beta-GP). In the preparation process, a cooling system is adopted to control the temperature of the micro-arc oxidation electrolyte at 300K. And (3) obtaining a sample coated with the microporous titanium dioxide coating after micro-arc oxidation, and putting the prepared sample into a drying oven for later use after the sample is cleaned by alcohol and deionized water.

Step 2, adding porous TiO containing phosphorus and calcium₂Carrying out hydrothermal treatment on the coating to obtain the HA nanorod structured coating:

step 2.1, primary hydrothermal treatment

Firstly, preparing a solution required by primary hydrothermal, wherein the solution is a sodium hydroxide aqueous solution with the concentration of 0.02mol/L, putting a titanium sheet coated with a titanium dioxide coating after micro-arc oxidation into reaction kettles, then adding 14mL of the sodium hydroxide aqueous solution into each reaction kettle, screwing down the reaction kettles, putting the reaction kettles into a drying oven, and finally setting temperature and time parameters. The temperature was adjusted to 370K and the time was set to 1.5 h.

Step 2.2, Secondary hydrothermal treatment

Adding calcium sodium ethylene diamine tetraacetate, a beta-phosphoglyceride disodium salt pentahydrate and sodium hydroxide into water, uniformly stirring, and sealing the product obtained in the step 2.1 for secondary hydrothermal treatment, wherein the concentration of the calcium sodium ethylene diamine tetraacetate is 0.01mol/L, the concentration of the beta-phosphoglyceride disodium salt pentahydrate is 0.02mol/L, and the concentration of the sodium hydroxide is 0.1 mol/L; the secondary hydrothermal temperature was set to 400K and the time was set to 18 h. And (3) obtaining a titanium sheet coated with the HA nanorod-structured coating after two times of hydrothermal treatment, taking the sample out of the reaction kettle, washing the sample with deionized water, and putting the sample into a drying oven for later use.

Step 3, preparing a poly-dopamine (PDA) -coated Hydroxyapatite (HA) nanorod array (PDA @ HA)

Immersing the product obtained in the step 2 in a 1mg/mL dopamine trihydroxymethylaminomethane solution, wherein the trihydroxymethylaminomethane concentration in the dopamine trihydroxymethylaminomethane solution is 10mmol/L, and stirring for 10 hours at normal temperature in a dark place at the rotating speed of 400 r/min. And finally, taking out the sample, washing with deionized water, and drying in an oven for later use.

Step 4, preparing manganese ferrite (MnFe)₂O₄) -Polydopamine (PDA) co-modified Hydroxyapatite (HA) nanorod array (MnFe)₂O₄[email protected])

Placing the product obtained in the step (3) in a mixed salt solution of iron acetate and manganese acetate at normal temperature, gently stirring and gradually heating to 60 ℃, wherein the concentration of the iron acetate is 1.5mmol/L and the concentration of the manganese acetate is 0.75 mmol/L; then taking out a sample to perform hydrothermal treatment, wherein the hydrothermal solution is a sodium hydroxide aqueous solution with the concentration of 0.03mol/L, the hydrothermal temperature is set to be 100 ℃, and the time is 1 h. And after the hydrothermal process is finished, taking out the sample, cleaning and drying to obtain the nanorod array formation coating with the titanium-based surface having the functions of oxidation resistance and self-generated oxygen.

The obtained product has MnFe₂O₄-PDA @ HA coated titanium alloy samples comprising TiO located in sequence from the substrate surface outwards₂Coating, MnFe₂O₄-PDA @ HA coating. Fig. 1 is a scanning picture of the hydroxyapatite nanorod coating obtained in examples 1 and 2 after micro-arc oxidation and hydrothermal treatment. As shown in FIG. 2, the MnFe obtained in example 1 is₂O₄Scanning picture of PDA @ HA coating, we can see that the configuration of the nanorods is transformed from regular hexagonal prisms to circles. Referring to FIG. 3, under the preparation conditions of example 2, the coating retains HA, PDA and MnFe₂O₄On the basis of the phase, the nano-rod structure is changed into the nano-tube structure, so that the drug loading can be further carried out under the action of realizing the double-layer functions of oxidation resistance and self-generation of oxygen under the condition, and the treatment of RA pathology is facilitated. FIG. 4 shows MnFe obtained in example 2₂O₄HRTEM picture of the PDA @ HA coating, we can see the crystal plane (210) representing HA and the crystal plane (222) representing manganese ferrite. As shown in FIG. 5, the hydroxyapatite can be visually seen in the related Fourier Infrared characterization chartThe related characteristic peaks comprise phosphate radicals, carbonate radicals and the like, the characteristic peaks of carbon-carbon double bonds and nitrogen-hydrogen bonds of polydopamine, the iron-oxygen bonds and manganese-oxygen bonds of manganese ferrite nanoparticles and the mark MnFe₂O₄Characteristic peaks of spinel structure. As shown in fig. 6, it can be seen from the XRD characterization chart that the titanium alloy with the HA coating is obtained after micro-arc oxidation and hydrothermal treatment, and as dopamine is an amorphous substance, it can be seen that the intensity of the HA-related characteristic peak is lower and lower with the coating of poly-dopamine, which indicates that PDA is successfully coated on the surface of the HA nanorod to a certain extent. In addition, MnFe is clearly seen after hydrothermal crystallization₂O₄Characteristic peak of (2), also to some extent, indicates MnFe₂O₄Successful coating. The surface-related SEM photograph shows that MnFe is contained₂O₄The titanium alloy implant with the PDA @ HA coating bioactive coating is successfully prepared.

Examples are given by way of illustration and not by way of limitation, and in summary, within the scope of the invention, a bilayer structure coating can be obtained on the surface of titanium and its alloys by micro-arc oxidation, hydrothermal treatment and oxidative autopolymeric deposition and further hydrothermal crystallization treatment after adsorption of ferromanganese ions: the inner layer is microporous TiO containing phosphorus and calcium₂The layer is an oxide film containing calcium and phosphorus elements growing on the titanium surface in situ, and can improve the biocompatibility of the titanium and the titanium alloy implant to a certain extent; the surface layer is a nano rod-shaped hydroxyapatite configuration coating which is modified by the ferrite and the polydopamine together, the components of the configuration coating are closer to the components of human bones, and the specific surface area in contact with cells can be improved due to the unique hexagonal prism-shaped nanorod configuration, so that the biocompatibility is further improved, and the bone conduction and the cell adhesion can be promoted to a certain extent; furthermore, via MnFe₂O₄The surface layer of the PDA modified HA nanorod array HAs double effects of oxidation resistance and self-generated oxygen, and can effectively regulate and control inflammatory and anoxic microenvironments of bones in a fracture area under RA pathology, so that the capability of the titanium-based implant for promoting bone tissue healing and regeneration is greatly improved. After the double coating is coated on the surfaces of titanium and titanium alloy, the titanium and titanium alloy has high bonding strength and good biological activity,and because the titanium and titanium alloy implant has good oxidation resistance and oxygen production capacity in the analogous body fluid, the capability of promoting the healing and regeneration of bone tissues of the titanium and titanium alloy implant under the specific pathological condition of RA can be obviously improved.

The above-mentioned contents are only for illustrating the technical idea of the present invention, and the protection scope of the present invention is not limited thereby, and any modification made on the basis of the technical idea of the present invention falls within the protection scope of the claims of the present invention.