CN111840635A

CN111840635A - Preparation method of polyphenol nano apatite coating on surface of titanium-based nanotube

Info

Publication number: CN111840635A
Application number: CN202010706395.2A
Authority: CN
Inventors: 黄棣; 林巧霞; 魏延; 胡银春; 赵丽琴; 连小洁; 陈维毅
Original assignee: Taiyuan University of Technology
Current assignee: Taiyuan University of Technology
Priority date: 2020-07-21
Filing date: 2020-07-21
Publication date: 2020-10-30

Abstract

The preparation method of the polyphenol nano apatite coating on the surface of the titanium-based nano tube comprises the following steps: (1) preparing a pure titanium-based nanotube: preparing a layer of titanium oxide nanotube on the surface of a pure titanium material, and carrying out high-temperature annealing treatment; (2) preparation of polyphenol-calcium complex coating: soaking a sample in a mixed solution of polyphenol and calcium chloride to perform a crosslinking reaction; (3) and (3) carrying out phosphate hydrothermal treatment: the sample is vertically soaked in phosphate solution, and the reaction temperature and the reaction time are controlled, so that the polyphenol nano hydroxyapatite crystal coating is prepared on the surface of the pure titanium-based nanotube. The invention has low cost and simple preparation process, and can realize safe, high-efficiency and large-scale production. The invention can eliminate active oxygen in implantation environment and promote osseointegration of implantation interface, and has wide application value in the aspect of surface modification of bone implantation material and dental implant.

Description

Preparation method of polyphenol nano apatite coating on surface of titanium-based nanotube

Technical Field

The invention belongs to the surface modification technology of bone implants and dental implants, relates to a preparation method of a polyphenol nano apatite crystal coating on the surface of a titanium-based nanotube, belongs to the field of biomedical engineering, and can be used for surface modification of biological materials. Can endow the implant with oxidation resistance and high biological activity and promote the osseointegration of the implant interface.

Background

Titanium metal is often used as a bone implant material under a load condition due to its excellent corrosion resistance and mechanical properties, but because it is biologically inert, it is slowly integrated with surrounding bone tissues, so that the implantation process is easily accompanied by a series of surgical complications such as infection, inflammation, etc. It has been found that in patients with poor physical condition (e.g., diabetes, osteoporosis, etc.), a significant amount of reactive oxygen species is present at the implant implantation interface during implantation. The generation and accumulation of reactive oxygen species can severely impair the biological function of osteoblasts, thus hindering new bone formation, resulting in poor bone healing and implant failure. In order to solve the problems of the damaged osteogenesis capacity and the insufficient early bone healing caused by active oxygen at an implanted interface, the surface modification of titanium metal is important for endowing the titanium metal with certain oxidation resistance so as to eliminate the active oxygen at the interface.

At present, a plurality of methods for endowing a titanium implant with antioxidant capacity are available, wherein an antioxidant coating such as a cerium oxide coating and the like is mainly prepared on the surface of the implant, or substances with antioxidant capacity, including N-acetylcysteine and chitosan, are loaded on the surface of the implant, the antioxidant capacity of the substances can improve adverse implantation microenvironment caused by active oxygen accumulation, and meanwhile, some antioxidant substances have the functions of resisting bacteria, diminishing inflammation and the like, and are beneficial to improving osseointegration at an interface. In order to better ensure the success rate of implantation under the condition that active oxygen exists at the host implantation interface, hydroxyapatite with very good biological activity and an antioxidant substance are compounded and coated on the surface of titanium, which is more favorable for improving the osseointegration efficiency of the coating. Polyphenol is one of typical antioxidant substances, has a polyphenol structure which enables the polyphenol to have strong antioxidant capacity, is easy to complex with metal ions, forms stable bonding effect with other substances, and is often used as a coating material of teeth. The complex effect of phenolic hydroxyl of polyphenol and calcium ions is utilized to induce and form the polyphenol nano apatite composite coating, so that the titanium implant can be endowed with oxidation resistance and biological activity, and the osseointegration performance of the implant is greatly improved.

Disclosure of Invention

Under mild conditions, the polyphenol nano apatite coating with oxidation resistance and biological activity is prepared on the surface of the pure titanium-based nanotube by a safe and simple method.

The invention relates to a preparation method of a polyphenol nano apatite coating on the surface of a titanium-based nanotube, which comprises the following steps:

(1) preparing a titanium-based nanotube: and ultrasonically cleaning the pure titanium material with a smooth surface in acetone, ethanol and deionized water for 10 min in sequence, and naturally airing for later use. Taking a pure titanium material as an anode and a platinum electrode as a cathode, anodizing for 1-3 h in fluorine-containing electrolyte under the condition of stable voltage of 20-40V, taking out a sample, washing with deionized water, and naturally airing. Placing the mixture in a muffle furnace for high-temperature annealing treatment, raising the temperature from room temperature to 450 ℃ at the heating rate of 0.5-10 ℃/min, preserving the temperature for 2h, naturally cooling, and taking out for later use.

(2) Preparation of polyphenol-calcium complex coating: soaking the sample in a mixed solution of polyphenol and calcium chloride, placing the mixture in an oven at the temperature of 40-80 ℃ for 1-5 h, and then crosslinking the mixture at the temperature of 10-25 ℃ for 30 min to 1 h. The sample was removed from the solution, rinsed with deionized water to remove the loosely adherent floats, and dried in an oven at 80-200 ℃.

(3) And (3) carrying out phosphate hydrothermal treatment: and vertically soaking the titanium-based nanotube material with the polyphenol-calcium complex coating in a phosphate solution, carrying out water bath at 37-90 ℃ for 2-5 h, taking out a sample, washing with deionized water, and drying in an electrothermal blowing drying oven at 37 ℃ to obtain the polyphenol nano apatite crystal coating.

The pure titanium material in the step (1) is commercial pure titanium, the fluorine-containing electrolyte is a glycerol/deionized water/ammonium fluoride system, the mass ratio of glycerol to deionized water is 6:5, and the content of ammonium fluoride is 0.9 wt%.

The polyphenol in the step (2) is at least one of tannic acid, dopamine and catechol, preferably tannic acid, the concentration is 10-50 g/L, and the concentration of calcium chloride is 0.1-1M.

The concentration of the phosphate solution in the step (3) is 0.01-0.1M, and the phosphate is at least one of sodium phosphate, sodium hydrogen phosphate, ammonium hydrogen phosphate, potassium phosphate and potassium hydrogen phosphate.

Through the process steps of the preparation method, the polyphenol nano apatite formed on the surface of the pure titanium-based nano tube is in a rod-shaped form, the shape is regular, and the diameter is 10-100 nm.

The invention firstly prepares a layer of titanium oxide nanotube buffer layer on the surface of a pure titanium material by an anodic oxidation method, and the formed tubular structure and the increased specific surface area are favorable for promoting the formation of the polyphenol nano apatite coating and the stable combination with the substrate. The mixed solution of polyphenol and calcium chloride is soaked, and calcium ions are fixed on the surface and inside of the nanotube by utilizing the adhesion effect of polyphenol on the surface of titanium oxide and the complexing effect of polyphenol and calcium ions at low temperature. The scanning electron microscope shows that a layer of membranous substance is formed on the surface of the nanotube. After being immersed in phosphate solution, hydroxyapatite is continuously nucleated and grows at the nucleation site of the interface, and finally a uniform polyphenol nano apatite coating is formed on the surface of the titanium nano tube. The total oxidation resistance detection test shows that the sample has good oxidation resistance.

The foregoing and other objects, features, and advantages of the invention will be apparent from the following more particular description of the embodiments of the invention, as illustrated in the accompanying drawings. This should not be understood as limiting the scope of the above-described subject matter of the present invention to the following examples. Various substitutions and alterations according to the general knowledge and conventional practice in the art are intended to be included within the scope of the present invention without departing from the technical spirit of the present invention as described above.

Drawings

FIG. 1 is an SEM image of titanium oxide nanotubes on a titanium-based surface;

FIG. 2 is an SEM image of the surface of a sample treated by the tannin calcium chloride mixed solution;

FIG. 3 is an SEM image of a tannin nano-apatite coating formed on the surface of a titanium nanotube;

FIG. 4 is a graph showing the results of the total antioxidant capacity test.

Detailed Description

Example 1

Taking 1cm multiplied by 1cm pure titanium foil, sequentially and respectively ultrasonically cleaning the pure titanium foil for 10min by absolute ethyl alcohol and deionized water in an ultrasonic cleaning machine, taking out the pure titanium foil, and putting the pure titanium foil in a blast drying oven for drying for later use. 30 g of glycerol and 25 mL of deionized water are uniformly mixed, 0.5 g of ammonium fluoride is added, and the mixture is magnetically stirred until the mixture is uniform and clear to prepare the electrolyte. At room temperature, titanium foil is used as anode, platinum electrode

And (3) anodizing for 1 h at constant voltage of 30V for the cathode, taking out a sample, washing with deionized water, and naturally airing. And (3) calcining the dried sample at high temperature in a muffle furnace, setting the heating rate to be 5 ℃/min, heating the sample to 450 ℃ from room temperature, preserving the temperature for 2 h, and naturally cooling the sample to room temperature along with the furnace. The surface structure was observed by SEM to show that a layer of tubular structure perpendicular to the substrate surface was formed on the smooth titanium foil surface (FIG. 1). And (3) placing the treated sample in a mixed solution with the concentration of tannic acid of 30 g/L and the concentration of calcium chloride of 0.5M, placing the sample in an oven at 40 ℃ for soaking for 3 h, taking out the sample, and crosslinking for 1 h at room temperature. The sample was removed from the solution, washed free of surface-bound, and dried in a forced air oven at 80 ℃. And observing the change of the surface topography after the treatment by using an SEM (appended figure 2), and thus, film-shaped substances are formed on the surface of the sample. The treated sample was vertically immersed in a 0.1M sodium phosphate solution, water-washed at 90 ℃ for 3 hours, then the sample was taken out, washed with deionized water, and dried at room temperature. When the surface morphology was observed by SEM, it was found that rod-like nano apatite was formed on the surface (FIG. 3). The total antioxidant capacity of the sample was measured using FRAP method (bph biotechnology limited) (fig. 4), using deionized water as negative control and Trolox as positive control.

Example 2

And (3) anodizing for 1 h at constant voltage of 30V for the cathode, taking out a sample, washing with deionized water, and naturally airing. And (3) calcining the dried sample at high temperature in a muffle furnace, setting the heating rate to be 5 ℃/min, heating the sample to 450 ℃ from room temperature, preserving the temperature for 2 h, and naturally cooling the sample to room temperature along with the furnace. And (3) placing the treated sample in a mixed solution with the tannin concentration of 10 g/L and the calcium chloride concentration of 0.5M, placing the mixed solution in an oven at 40 ℃ for soaking for 3 h, taking out and crosslinking at room temperature for 1 h. The sample was removed from the solution, washed free of surface-bound, and dried in a forced air oven at 100 ℃. And observing the change of the surface appearance after treatment by using the SEM, and thus, forming a film-shaped substance on the surface of the sample. The treated sample was vertically immersed in a 0.1M sodium phosphate solution, and after 2 hours of water bath at 90 ℃, the sample was taken out, washed with deionized water, and dried at room temperature. The surface morphology was observed by SEM to show that rod-like nano apatite was formed on the surface. The total antioxidant capacity of the sample is detected by using an antioxidant capacity detection kit (FRAP method) of Biyuntian biotechnology limited company, deionized water is used as a negative control, and Trolox is used as a positive control.

Example 3

And (3) anodizing for 1 h at constant voltage of 30V for the cathode, taking out a sample, washing with deionized water, and naturally airing. And (3) calcining the dried sample at high temperature in a muffle furnace, setting the heating rate to be 5 ℃/min, heating the sample to 450 ℃ from room temperature, preserving the temperature for 2 h, and naturally cooling the sample to room temperature along with the furnace. The surface structure of the titanium foil is observed by SEM, and a layer of tubular structure vertical to the substrate surface is formed on the smooth surface of the titanium foil. And (3) placing the treated sample in a mixed solution with the tannin concentration of 50 g/L and the calcium chloride concentration of 0.5M, placing the mixed solution in a 50 ℃ oven for soaking for 4 h, taking out and crosslinking for 1 h at room temperature. The sample was removed from the solution, washed free of surface-bound, and dried in a forced air oven at 80 ℃. When observed by SEM, the film-like substance was formed on the surface of the sample. The treated sample was vertically immersed in a 0.1M sodium phosphate solution, water-washed at 90 ℃ for 3 hours, then the sample was taken out, washed with deionized water, and dried at room temperature. As a result of SEM observation, it was found that rod-like nano apatite was formed on the surface. The total antioxidant capacity of the sample is detected by using an antioxidant capacity detection kit (FRAP method) of Biyuntian biotechnology limited company, deionized water is used as a negative control, and Trolox is used as a positive control.

Example 4

And (3) anodizing for 1 h at constant voltage of 30V for the cathode, taking out a sample, washing with deionized water, and naturally airing. And (3) calcining the dried sample at high temperature in a muffle furnace, setting the heating rate to be 5 ℃/min, heating the sample to 450 ℃ from room temperature, preserving the temperature for 2 h, and naturally cooling the sample to room temperature along with the furnace. When the surface of the titanium foil is observed by SEM, a layer of tubular structure vertical to the substrate surface is formed on the smooth surface of the titanium foil. And (3) placing the treated sample in a mixed solution with the concentration of tannic acid of 30 g/L and the concentration of calcium chloride of 1M, placing the mixed solution in a drying oven at 40 ℃ for soaking for 2 hours, taking out the sample, and then crosslinking for 1 hour at room temperature. The sample was removed from the solution, washed free of surface-bound, and dried in a forced air oven at 80 ℃. And observing the change of the surface appearance after treatment by using the SEM, and thus, forming a film-shaped substance on the surface of the sample. The treated sample was vertically immersed in a 0.1M sodium phosphate solution, water-washed at 90 ℃ for 5 hours, then the sample was taken out, washed with deionized water, and dried at room temperature. The surface morphology was observed by SEM to show that rod-like nano apatite was formed on the surface. The total antioxidant capacity of the sample is detected by using an antioxidant capacity detection kit (FRAP method) of Biyuntian biotechnology limited company, deionized water is used as a negative control, and Trolox is used as a positive control.

Example 5

And (3) anodizing for 1 h at constant voltage of 30V for the cathode, taking out a sample, washing with deionized water, and naturally airing. And (3) calcining the dried sample at high temperature in a muffle furnace, setting the heating rate to be 5 ℃/min, heating the sample to 450 ℃ from room temperature, preserving the temperature for 2 h, and naturally cooling the sample to room temperature along with the furnace. By SEM observation, a layer of tubular structure perpendicular to the base surface is formed on the smooth surface of the titanium foil. And (3) placing the treated sample in a mixed solution with the tannin concentration of 10 g/L and the calcium chloride concentration of 1M, placing the mixed solution in a drying oven at 40 ℃ for soaking for 3 h, taking out and crosslinking at room temperature for 1 h. The sample was removed from the solution, washed free of surface-bound, and dried in a forced air oven at 80 ℃. And observing the change of the surface appearance after treatment by using the SEM, and thus, forming a film-shaped substance on the surface of the sample. The treated sample was vertically immersed in a 0.05M sodium phosphate solution, water-washed at 90 ℃ for 4 hours, then the sample was taken out, washed with deionized water, and dried at room temperature. As a result of SEM observation, it was found that rod-like nano apatite was formed on the surface. The total antioxidant capacity of the sample is detected by using an antioxidant capacity detection kit (FRAP method) of Biyuntian biotechnology limited company, deionized water is used as a negative control, and Trolox is used as a positive control.

Claims

1. The preparation method of the polyphenol nano apatite coating on the surface of the titanium-based nanotube is characterized by comprising the following steps:

(1) preparing a pure titanium-based nanotube: ultrasonically cleaning a pure titanium material with a smooth surface in acetone, ethanol and deionized water for 10 min in sequence, and naturally airing for later use; taking pure titanium as an anode and a platinum electrode as a cathode, anodizing in a fluorine-containing electrolyte for 1-3 h under the condition of stable voltage of 20-40V, taking out a sample, washing with deionized water, and naturally airing; placing the sample in a muffle furnace for high-temperature annealing treatment, heating the sample from room temperature to 450 ℃ at the heating rate of 0.5-10 ℃/min, preserving the temperature for 2 h, naturally cooling, and taking out the sample for later use; the fluorine-containing electrolyte is a glycerol/deionized water/ammonium fluoride system, wherein the mass ratio of glycerol to deionized water is 6:5, and the content of ammonium fluoride is 0.9 wt%;

(2) preparation of polyphenol-calcium complex coating: soaking a sample in a mixed solution of polyphenol and calcium chloride, placing the sample in an oven at the temperature of 40-80 ℃ for 1-5 h, and crosslinking the sample at the temperature of 10-25 ℃ for 30 min to 1 h; taking out the sample from the solution, washing away the floating objects which are not firmly bonded by deionized water, and drying in an oven at 80-200 ℃; the polyphenol is at least one of tannic acid, dopamine and catechol, and the concentration of calcium chloride is 0.1-1M;

(3) And (3) carrying out phosphate hydrothermal treatment: preparing a phosphate solution according to the concentration of 0.01-0.1M, vertically soaking the titanium-based nanotube material with the polyphenol-calcium complex coating in the phosphate solution, carrying out water bath at 37-90 ℃ for 2-5 h, taking out a sample, washing with deionized water, and drying in an electrothermal blowing drying oven at 37 ℃ to obtain a polyphenol nano apatite crystal coating; the phosphate is at least one of sodium phosphate, sodium hydrogen phosphate, ammonium hydrogen phosphate, potassium phosphate and potassium hydrogen phosphate.

2. The method for preparing polyphenol nano apatite coating on the surface of titanium-based nanotube as claimed in claim 1, wherein the polyphenol nano apatite formed on the surface of pure titanium-based nanotube has rod shape, regular appearance and diameter of 10-100 nm.

3. A preparation method of a polyphenol nano apatite coating on the surface of a titanium-based nanotube is characterized in that 1cm multiplied by 1cm pure titanium foil is taken to be sequentially subjected to ultrasonic cleaning for 10 min by absolute ethyl alcohol and deionized water respectively, and then the pure titanium foil is fished out and placed in a blast drying oven to be dried for later use; uniformly mixing 30 g of glycerol and 25 mL of deionized water, adding 0.5 g of ammonium fluoride, stirring by magnetic force until the mixture is uniform and clear, preparing an electrolyte, carrying out anodic oxidation for 1 h at constant voltage of 30V by using a pure titanium foil as an anode and a platinum electrode as a cathode under the condition of room temperature, taking out a sample, washing by using the deionized water, and naturally drying; calcining the dried sample at high temperature in a muffle furnace, setting the heating rate to be 5 ℃/min, heating the sample from room temperature to 450 ℃, preserving the temperature for 2 h, and naturally cooling the sample to room temperature along with the furnace; observing the surface structure by using an SEM (scanning electron microscope), and forming a layer of tubular structure vertical to the substrate surface on the smooth surface of the titanium foil; placing the treated sample in a mixed solution with tannic acid concentration of 30 g/L and calcium chloride concentration of 0.5M, soaking in an oven at 40 ℃ for 3 h, taking out, and crosslinking at room temperature for 1 h; taking out the sample from the solution, washing away floating objects with weak surface bonding, and drying in a forced air drying oven at 80 ℃; observing the change of the surface appearance after treatment by using an SEM (scanning Electron microscope), and thus forming a film-shaped substance on the surface of the sample; vertically immersing the treated sample in 0.1M sodium phosphate solution, carrying out water bath at 90 ℃ for 3 h, taking out the sample, washing with deionized water, and drying at room temperature; observing the surface appearance by using SEM, and thus forming rod-shaped nanometer apatite on the surface; the total antioxidant capacity of the sample is detected by using an antioxidant capacity detection kit of Biyuntian biotechnology limited, deionized water is used as a negative control, and Trolox is used as a positive control.