CN115814168A - Medical titanium alloy device with polydopamine coating as carrier and preparation method thereof - Google Patents
Medical titanium alloy device with polydopamine coating as carrier and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a medical titanium alloy device with a polydopamine coating as a carrier and a preparation method thereof, and relates to the field of medical devices. The method comprises the following steps: s10, preprocessing a titanium alloy device; s20, electroplating a layer of polydopamine on the surface of the titanium alloy device to obtain the polydopamine coated titanium alloy device; and S30, loading vascular endothelial growth factors on the titanium alloy device with the polydopamine coating to obtain the medical titanium alloy device with the polydopamine coating as a carrier. The method overcomes the defects of long time, waste caused by DA self-polymerization in the solution, uneven coating and the like in the method for forming the PDA coating on the surface of the titanium nail by self-polymerization in the alkaline environment. Compared with the traditional preparation method, the preparation method has higher preparation efficiency, can reduce the waste caused by self-polymerization of dopamine in the solution, and has uniform coating distribution and no cracking or peeling phenomenon.
Description
Technical Field
The invention belongs to the field of medical devices, and particularly relates to a medical titanium alloy device taking a polydopamine coating as a carrier and a preparation method thereof.
Background
The titanium alloy device is a medical device commonly used in clinic, for example, titanium nails are commonly used for anastomosis of surgical tissues such as stomach, intestine and lung, but because of the biological inertia of titanium, poor healing of an anastomotic site can be caused under the conditions of bleeding, inflammation and the like of a sutured area, and complications such as anastomotic fistula and the like are easily induced. In the anastomotic healing process, the surface properties of the titanium staples play a critical role. In short, conventional titanium nails provide only physical apposition and are not bioactive, and their efficacy in promoting healing may be compromised in special settings.
In order to make the traditional titanium nail bioactive, the surface of the titanium nail can be loaded with polydopamine coating at present. The Polydopamine (PDA) material formed by polymerization of Dopamine (DA) molecules of bionic mussels has the advantages of good biological/cell compatibility, secondary modification reactivity and the like, and can be loaded on the surface of a titanium nail.
Currently, loading a polydopamine coating on the surface of a titanium nail is mainly based on self-polymerization formation of DA in an alkaline environment. The method can be realized by the following steps: dissolving DA in buffer solution with pH value (pH) of Tris (hydroxymethyl) aminomethane (Tris) of 8.5, soaking titanium nail in the buffer solution, and attaching a layer of PDA coating on the surface of the titanium nail after a period of time. However, the method has the defects that the time for constructing the coating is too long (more than 12 hours), the DA self-polymerization in the solution causes waste, the prepared coating is not uniform, and the polymerization in an alkaline environment is needed.
Disclosure of Invention
Aiming at the defects of the prior art, the invention provides a medical titanium alloy device using a polydopamine coating as a carrier and a preparation method thereof aiming at the defects of long time, waste caused by self-polymerization of DA in a solution, uneven coating and the like in a method for forming the PDA coating on the surface of a titanium nail in an alkaline environment. A preparation method of a medical titanium alloy device with a polydopamine coating as a carrier comprises the following steps:
s10, preprocessing a titanium alloy device;
s20, electroplating a layer of polydopamine on the surface of the titanium alloy device to obtain the polydopamine coated titanium alloy device;
and S30, loading vascular endothelial growth factors on the titanium alloy device with the polydopamine coating to obtain the medical titanium alloy device with the polydopamine coating as a carrier.
Preferably, the titanium alloy device is one of a titanium nail, a titanium clip and a titanium plate.
Preferably, the titanium alloy device is pretreated by adopting an organic solvent through ultrasonic treatment at room temperature for 30min, and after the surface is clean, the titanium alloy device is taken out and dried. The organic solvent is acetone or ethanol.
Preferably, the S20 includes:
taking a medical titanium alloy device as a working electrode;
a saturated calomel electrode is used as a reference electrode;
a platinum sheet is used as a counter electrode;
adding dopamine into the electro-polymerization solution, and electroplating a polydopamine coating on the surface of the medical titanium alloy device by adopting a cyclic voltammetry method.
Preferably, the electropolymerization solution is a TBS solution.
Preferably, the preparation method further comprises:
before dopamine is added into the electropolymerization liquid, high-purity nitrogen is introduced into the electropolymerization liquid, and oxygen dissolved in the electropolymerization liquid is removed.
Preferably, the electropolymerization solution TBS solution contains 25mM Tris,140mM sodium chloride and 3mM potassium chloride, the pH of the TBS solution is 7.4, and the mass-to-volume ratio of the dopamine to the electropolymerization solution TBS solution is 2-15:1.
the specific parameters of electroplating the polydopamine coating on the surface of the medical titanium alloy device by adopting a cyclic voltammetry method are as follows: setting the continuous cyclic voltammetry scanning voltage to-0.8V-0.8V or-0.5V-0.5V or-0.6V-0.6V or-1.0V-1.0V, and performing 50-100 cycles.
Preferably, the S30 includes:
soaking the titanium alloy device with the polydopamine coating in a vascular endothelial growth factor solution;
adding a buffer solution;
soaking at a set temperature for a set time and taking out;
preferably, the concentration of the vascular endothelial growth factor is 0.1-1 mug/mL; the buffer solution is TBS buffer solution, and the flushing solution is phosphate buffer salt solution. The set temperature is 37 ℃, and the set time is 8-12 hours. Too high a temperature can result in inactivation of the loaded functional component, vascular endothelial growth factor, and too low a temperature can result in a decrease in the reaction rate and a decrease in loading.
The medical titanium alloy device which is prepared by any one of the methods and takes the polydopamine coating as a carrier.
The medical titanium alloy device prepared by the method overcomes the defects of long time required by a method for forming a PDA coating on the surface of a titanium nail by self polymerization in an alkaline environment, waste caused by DA self polymerization in a solution, uneven coating and the like. In a three-electrode electrochemical system, a medical titanium alloy device is used as a working electrode, a saturated calomel electrode is used as a reference electrode, a platinum sheet is used as a counter electrode, dopamine is added into an electropolymerization solution, and a polydopamine coating is electroplated on the surface of the medical titanium alloy device by adopting a cyclic voltammetry method. Compared with the traditional preparation method, the preparation method has higher preparation efficiency, can reduce the waste caused by the self-polymerization of the dopamine in the solution, and has uniform distribution of the coating without cracking or peeling.
Compared with the prior art, the method has the following beneficial effects:
1. the poly-dopamine coating is constructed on the surface of the medical titanium alloy device by an electroplating technology, compared with the traditional preparation method, the poly-dopamine coating has higher preparation efficiency, can reduce the waste caused by self-polymerization of dopamine in a solution, and has uniform distribution and no cracking or peeling phenomena.
2. The invention can load the vascular endothelial growth factor on the coating through simple soaking, has the efficacy of promoting angiogenesis, further promotes healing of anastomotic stoma, and reduces the occurrence of complications such as anastomotic fistula and the like.
3. The invention has mild electroplating conditions, is friendly to substrate materials, and has simpler operation and easy process control compared with the traditional preparation method.
Drawings
FIG. 1 is a scanning electron microscope image of a common titanium nail, a titanium nail loaded with a polydopamine coating by a conventional method, and a titanium nail loaded with a polydopamine coating by the method of the present invention, wherein (a) is the common titanium nail; (b) Loading a polydopamine coating titanium nail by adopting a traditional method; (c) The titanium nail loaded with the polydopamine coating by the method is provided.
FIG. 2 is a graph showing the results of ELISA kits for detecting the release of vascular endothelial growth factor from the titanium nails of the present invention (Ti-ePDA-VEGF) and the conventional titanium nails (Ti-PDA-VEGF).
FIG. 3 is a cell compatibility test chart of the titanium nail with the polydopamine coating and the titanium nail loaded with the vascular endothelial growth factor after being loaded with the polydopamine coating, which are prepared by the method 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.
The technical scheme of the invention is to provide a preparation method of a medical titanium alloy device with a polydopamine coating as a carrier, which is characterized in that: in a three-electrode electrochemical system, a medical titanium alloy device is used as a working electrode, a saturated calomel electrode is used as a reference electrode, a platinum sheet is used as a counter electrode, dopamine is added into an electropolymerization solution, and a polydopamine coating is electroplated on the surface of the medical titanium alloy device by adopting a cyclic voltammetry method.
Further, in order to obtain the biological function of promoting healing of the medical titanium alloy device, the method also comprises the step of loading vascular endothelial growth factors: soaking the medical titanium alloy device with the polydopamine coating electroplated on the surface in a vascular endothelial growth factor solution, adding a buffer solution, and soaking at a set temperature for a set time and taking out.
Furthermore, the concentration of the vascular endothelial growth factor in the vascular endothelial growth factor solution is 0.1-1 mu g/ml; the buffer solution is tris (hydroxymethyl) aminomethane buffered saline (TBS), and the washing solution is Phosphate Buffered Saline (PBS).
Further, in order to ensure the activity and certain loading capacity of the vascular endothelial growth factor, the set temperature is 37 ℃, the set time is 8-12 hours, too high temperature can lead to inactivation of the loaded functional component of the vascular endothelial growth factor, and too low temperature can lead to reduction of the reaction rate and reduction of the loading capacity.
Further, before electroplating, the method can also comprise the step of pretreating the medical titanium alloy device: and (3) ultrasonically treating the titanium nail by adopting an organic solvent, taking out and drying after the surface is clean.
Further, the organic solvent is acetone or ethanol.
Further, TBS solution is selected as electropolymerization solution, is a buffer solution which is commonly used at present and simulates the physiological condition of a human body, and is isotonic, nontoxic and biologically friendly.
Further, the TBS solution contains 25mM Tris,140mM sodium chloride and 3mM potassium chloride, and the pH of the TBS solution is 7.4; the content of dopamine in TBS solution is 2-15mg/1ml.
Further, in order to reduce the additional oxidation of dopamine during electroplating, the oxygen dissolved in the electropolymerization liquid needs to be removed before the dopamine is added into the electropolymerization liquid, and the oxygen can be introduced into the electropolymerization liquid in a high-purity nitrogen mode.
Further, the specific parameters of electroplating the polydopamine coating on the surface of the medical titanium alloy device by adopting a cyclic voltammetry method are as follows: setting the scanning voltage of continuous cyclic volt-ampere to-0.8V-0.8V or-0.5V-0.5V or-0.6V-0.6V or-1.0V-1.0V for 50-100 cycles, and the parameter can cover the voltage required by oxidation in the electropolymerization process and avoid the decomposition of electrolyte into hydrogen and oxygen due to overhigh voltage.
Example 1
1) Pretreatment of the titanium nail: ultrasonically treating the titanium nail by using an organic solvent (such as acetone and ethanol) to clean the titanium nail, drying the titanium nail for 30 minutes at 60 ℃ after the titanium nail is cleaned, and taking out the titanium nail;
2) Electroplating a layer of polydopamine on the surface of the titanium nail by using an electrochemical deposition method: standard three-electrode electrochemical polymerization experiments were performed at room temperature using an electrochemical workstation of CHl660 (chenghua, shanghai, china). Saturated Calomel Electrode (SCE) was used as a reference electrode, a platinum sheet was used as a counter electrode, and a titanium nail was used as a working electrode. And constructing a polydopamine coating on the surface of the working electrode by using cyclic voltammetry. In this example, a 20 mM LTBS solution (containing 25mM Tris,140mM sodium chloride and 3mM potassium chloride, pH 7.4) was prepared as an electropolymerization solution. The prepared electropolymerization solution was purged with high-purity nitrogen for 10min to remove oxygen dissolved in the electropolymerization solution. Then, 40-300mg of dopamine is added into 20ml of electropolymerization liquid to carry out cyclic voltammetry electropolymerization scanning, and the continuous cyclic voltammetry scanning voltage is set to be-0.8V-0.8V. After 100 cycles, the working electrode was rinsed with deionized water to obtain a PDA coating, which took about 20 minutes. Of course, in other embodiments, other common electropolymerization liquids may also be used, and the cyclic voltammetry scan voltage may also be set to-0.5V for 50-100 cycles, which may be set according to specific experimental needs.
3) Loading of vascular endothelial growth factor: soaking the titanium nails obtained in the step 2) in a 0.5 mu g/ml vascular endothelial growth factor solution, wherein the solvent is 10mM Tris buffer solution and the pH =7.4. After being soaked for 12h at 37 ℃, the sample is taken out, washed with PBS three times to remove the excessive VEGF adsorbed on the surface, dried and sealed for storage. In other embodiments, the concentration of the VEGF solution may be between 0.1-1 μ g/ml, and the soaking period may be between 8-12 hours.
To verify the properties of the titanium nails prepared in the above examples in terms of coating uniformity, surface morphology, etc., the following property tests were performed on the samples of the examples:
compared with the common titanium nail, the titanium nail prepared in the embodiment obviously has a black polymer coating on the surface, and the poly-dopamine coating can be successfully loaded on the surface of the common titanium nail by using the method.
Fig. 1 shows scanning electron micrographs of a conventional titanium nail, a titanium nail with a polydopamine coating prepared by a conventional method, and a titanium nail with a polydopamine coating prepared in the example. By comparison, it is obvious that compared with the common titanium nail, the titanium nail with the polydopamine coating prepared by the traditional method and the titanium nail with the polydopamine coating prepared in the embodiment have a layer of polymer on the surface, and the polymer on the surface of the titanium nail with the polydopamine coating prepared in the embodiment is more uniform and compact compared with the graph b and the graph c.
The experimental method of the titanium clip or the titanium plate is similar, and the detailed description is omitted here.
Effect example 1
In order to verify the release amount of the vascular endothelial growth factor of the polydopamine-loaded coating and the vascular endothelial growth factor titanium nail prepared in the examples, the following tests were performed:
the experimental scheme is as follows: the coated titanium pins were placed in 12-well plates, covered with 2mL sterile PBS, and incubated at 37 ℃. Samples were taken and the solutions were completely changed after 1 day, 3 days, 5 days, 7 days, 10 days, 12 days, 14 days, respectively. The concentration of VEGF released at each time point was quantified using a Human VEGF ELISA kit according to the manufacturer's instructions.
As shown in fig. 2, the effect of releasing the titanium nails loaded with the polydopamine coating for a long time can be achieved by loading the titanium nails loaded with the polydopamine coating by the conventional method and loading the vascular endothelial growth factor by the electropolymerization method of the invention, and the release amount of the vascular endothelial growth factor loaded by the polydopamine coating by the electropolymerization method is more.
In order to evaluate the biological safety of the prepared coating, a biocompatibility experiment is carried out by co-culturing mouse epithelial fibroblast L929 and the titanium sheet carrying the coating. The experimental procedure is as follows: firstly, preparing a cell suspension, counting cells, inoculating the cells into a 96-well plate, wherein the number of the cells in each well is more than or equal to 5 multiplied by 104; adding 5mm titanium nails respectively loaded with ePDA and ePDA-VEGF coatings into each cell pore plate, and co-culturing for 24h in a 37 ℃ incubator; then, the cell morphology was observed under a microscope, and then, 10ul CCK8 was added thereto for culture for 2 hours, and the cell viability was calculated by measuring the absorbance at a wavelength of 450 nm.
As can be seen from FIG. 3, the titanium nails loaded with the polydopamine coating and loaded with the vascular endothelial growth factor after being loaded with the polydopamine coating by the electropolymerization method have good cell compatibility.
Aiming at the defects that the traditional oxidative polymerization method under alkaline conditions needs too long time (often needs more than 8 hours) for constructing the polydopamine coating, DA self-polymerization in a solution causes waste and the like, the polydopamine coating is constructed on the surface of a medical titanium alloy device through electroplating, the coating construction time is reduced (the construction can be completed in about 20 minutes), and the self-polymerization of DA is overcome. Specifically, in a three-electrode electrochemical system, a medical titanium alloy device is used as a working electrode, a saturated calomel electrode is used as a reference electrode, a platinum sheet is set as a counter electrode, dopamine is added into an electropolymerization solution, and a poly-dopamine coating is electroplated on the surface of the medical titanium alloy device by adopting a cyclic voltammetry method. In addition, the medical titanium alloy device electroplated with the polydopamine coating can be soaked in a vascular endothelial growth factor solution, and vascular endothelial growth factors are loaded on the coating, so that the surface of the medical titanium alloy device has a biological function of promoting healing, healing of anastomotic stoma is promoted, and complications such as anastomotic fistula and the like are reduced.
Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.
Claims (10)
1. A preparation method of a medical titanium alloy device taking a polydopamine coating as a carrier is characterized by comprising the following steps:
s10, preprocessing a titanium alloy device;
s20, electroplating a layer of polydopamine on the surface of the titanium alloy device to obtain the polydopamine coated titanium alloy device;
and S30, loading vascular endothelial growth factors on the titanium alloy device with the polydopamine coating to obtain the medical titanium alloy device with the polydopamine coating as a carrier.
2. The method for preparing a medical titanium alloy device with a polydopamine coating as a carrier in claim 1, wherein the titanium alloy device is one of a titanium nail, a titanium clip and a titanium plate.
3. The method for preparing a medical titanium alloy device with a polydopamine coating as a carrier in claim 1, wherein the pretreatment of the titanium alloy device is ultrasonic treatment with an organic solvent.
4. The method for preparing a medical titanium alloy device with a polydopamine coating as a carrier in claim 1, wherein the step S20 includes:
using a medical titanium alloy device as a working electrode;
a saturated calomel electrode is used as a reference electrode;
a platinum sheet is used as a counter electrode;
adding dopamine into the electro-polymerization solution, and electroplating a polydopamine coating on the surface of the medical titanium alloy device by adopting a cyclic voltammetry method.
5. The method for preparing a medical titanium alloy device with a polydopamine coating as a carrier in claim 4, wherein the electropolymerization solution is tris buffer saline solution.
6. The method of making a medical titanium alloy device with a polydopamine coating as a carrier according to claim 4, wherein the method further comprises:
before dopamine is added into the electropolymerization liquid, high-purity nitrogen is introduced into the electropolymerization liquid, and oxygen dissolved in the electropolymerization liquid is removed.
7. The method for preparing a medical titanium alloy device using polydopamine coating as a carrier in claim 5, wherein the electro-polymerization solution TBS solution contains 25mM tris,140mM NaCl and 3mM KCl, the pH of the TBS solution is 7.4, and the mass-to-volume ratio of dopamine to electro-polymerization solution TBS solution is 2-15:1.
8. the method for preparing a medical titanium alloy device with a polydopamine coating as a carrier in claim 1, wherein the step S30 includes:
soaking the titanium alloy device with the polydopamine coating in a vascular endothelial growth factor solution;
adding a buffer solution;
taking out the soaked materials at the set temperature for the set time.
9. The method for preparing a medical titanium alloy device with a polydopamine coating as a carrier according to claim 8, wherein the concentration of the vascular endothelial growth factor is 0.1-1 μ g/ml; the buffer solution is triethanolamine buffer saline solution, and the flushing solution is phosphate buffer saline solution.
10. Medical titanium alloy devices with a polydopamine coating as carrier prepared by any of the methods of claims 1-9.
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| CN117661063A (en) * | 2024-01-31 | 2024-03-08 | 四川大学 | Lead-free piezoelectric nano coating modified titanium alloy bracket and preparation method thereof |
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