WO2021167219A1 - Antibacterial titanium implant and manufacturing method therefor - Google Patents

Abstract

The present invention relates to an antibacterial titanium implant using chitosan. Accordingly, the titanium implant can minimize or prevent inflammation caused by metal ions that are eluted from the implant after being implanted in bone tissue or by bacteria that invade and inhabit an interface between the implant and the bone. In addition, the antibacterial titanium implant can be widely used in clinical applications such as dentistry and orthopedic surgery, since the antibacterial titanium implant can continuously maintain anti-inflammatory performance such as antibacterial property, which has been retained until the continuation of treatment or correction post implantation, thereby improving the treatment success rate of the implant.

Description

Titanium implant having antibacterial properties and manufacturing method thereof

The present invention relates to a titanium implant, and more particularly to a titanium implant having antibacterial properties.

In general, bioimplants used for medical purposes are used to permanently implant spinal fixation prostheses, interspecies correction prostheses, artificial joints, and the like, and require a biocompatible material that is very stable for human tissue. In addition, there should be no side effects or other chemical and biochemical reactivity, and the mechanical strength should be very high so as not to be deformed and destroyed even when repeated loads and instantaneous pressures are applied, and for medical use, the bonding strength with biological tissues, especially bone tissues, is very high. it is an instrument

These implants are removed from the bone tissue after being placed in the bone tissue to function permanently or only for a period of time performing the intended function. It is common to perform processing. Examples of such processing include roughening processing to increase a specific surface area with bone tissue, or coating a component similar to bone tissue, for example, a component such as hydroxyapatite on the implant surface.

However, even when the surface of the implant is processed so as to have a high bonding strength with the bone tissue, the solution to side effects such as inflammation occurring in the surrounding tissue after the implant is placed is insufficient until now. In particular, the act of generating orthodontic force, such as pulling an orthodontic wire as a support for orthodontic implants after being placed like orthodontic micro-implants, forms a gap between the implant and the alveolar bone or surrounding tissues. The causative agent penetrates and causes inflammation, which can lead to the failure of the implant placement procedure.

In addition, implant materials are generally implemented with biocompatible materials such as titanium to minimize in vivo side effects. It contains other elements, and these other elements sometimes cause side effects such as inflammation. Specifically, an orthodontic micro-implant as an example of an implant ^{contains a trace amount of vanadium (V ++} ), and after implantation, vanadium ions are released from the implant to cause tissue inflammation, and this tissue inflammation is caused by the implant and the implanted bone and/or By creating a gap between the gums and making the gap more spaced apart, it can cause inflammation due to additional bacterial penetration.

The present invention has been devised in view of the above points, and it is possible to minimize the inflammatory reaction caused by metal ions or bacteria eluted from the implant after implantation in an in vivo tissue, for example, pubic tissue by imparting antibacterial activity to the titanium implant. An object of the present invention is to provide a titanium implant having antibacterial properties and a method for manufacturing the same.

In addition, another object is to provide a titanium implant having antibacterial properties that can improve the treatment success rate by maintaining the antibacterial power imparted to the titanium implant for a treatment or correction period, and a method for manufacturing the same.

The present invention has been studied with the support of the following national R&D project, and detailed information of the national R&D project is as follows.

[Project unique number] 20001590

[Name of Ministry] All ministries (Ministry of Industry, Ministry of Science and Technology, Ministry of Welfare)

[Research Management Specialized Institution] Korea Institute of Industrial Technology Evaluation and Planning

[Research Project Name] Artificial Intelligence Bio-Robot Medical Convergence Project

[Research project name] Development of an artificial intelligence-based osseolithic arm capable of holding more than 4 types of hand movements and precision objects through the development of patient-customized osseointegration implants

[Contribution rate] 1/1

[Organizer] Jeil Medical Corporation

[Research Period] 2018.05.01 ~ 2020.12.31

In order to solve the above problems, the present invention provides a titanium implant having antibacterial properties including chitosan fixed through a chemical bond on the surface of the titanium implant.

According to an embodiment of the present invention, a spacer for forming a covalent bond with each of the titanium implant and the chitosan may be further provided between the titanium implant and the chitosan.

In addition, the surface of the titanium implant is modified to include an amine group, and the spacer may be a compound having at least two carboxyl groups to form an amide bond with each of the amine group of the titanium implant and the amine group of chitosan.

In addition, the spacer may include at least one compound selected from the group consisting of polyacrylic acid, sulcinic acid, glutamic acid, and asphaltic acid. In this case, more preferably, the spacer may be sulcinic acid.

In addition, the chitosan may be 75-85% deacetylated, and the viscosity may be 20-800 cP. In addition, the chitosan may be dissolved in a 1% aqueous solution of acetic acid or an aqueous solution of pH 6.5 or less.

In addition, the chitosan may be provided to cover 30% or more of the surface area of the titanium implant.

In addition, the titanium implant surface may be modified through a 3-aminopropyltriethoxysilane (APTES) compound.

In addition, the present invention relates to the steps of (1) preparing a titanium implant having an amine group on the surface, (2) treating the surface of the micro-implant with a first solution having a spacer having at least two carboxyl groups on the surface of the carboxyl group and the implant. Inducing a chemical bond between the amine groups, and (3) treating a second solution containing chitosan to induce a chemical bond between the amine group of chitosan and the unreacted carboxyl group of the spacer. A method for manufacturing a titanium implant is provided.

The titanium implant according to the present invention can minimize or prevent inflammation caused by metal ions eluted from the implant after being placed in the bone tissue, or bacteria that infiltrate and inhabit the interface between the implant and bone. In addition, since it is possible to continuously maintain antibacterial and anti-inflammatory properties such as antibacterial until treatment or correction is continued after implantation, the treatment success rate of the implant can be improved, so that it can be widely used in clinics such as dentistry and orthopedics.

1 is a schematic diagram of a process for manufacturing a micro-implant according to an embodiment of the present invention.

2 is an untreated titanium implant (Ti), an APTES-treated titanium implant (Ti-A), a titanium implant in which chitosan is introduced by acrylic acid (Ti-AA-Ch), and succinic acid according to an embodiment of the present invention. This is the XPS analysis spectrum performed on the titanium implant (Ti-SA-Ch) bound to chitosan.

3 is a photograph showing the change of the water contact angle according to the surface modification process of the titanium implant.

4 and 5 are chitosan-coupled titanium implants (FIG. 4, Ti-SA-Ch) and chitosan-coupled titanium implants via polyacrylic acid (FIG. 5, Ti) according to different embodiments of the present invention. -AA-Ch) is an image obtained by binding FITC fluorescent material.

6 is a FE-SEM photographed after evaluating the adhesion of osteoblasts performed with respect to the titanium implant (TI-SA-Ch) and untreated titanium implant (TI) combined with chitosan according to an embodiment of the present invention; It's a photo.

7 is a photograph of the result of performing Live/dead staining staining evaluation on osteoblasts cultured for 1 day and 3 days for the titanium implant coupled with chitosan and the untreated titanium implant according to an embodiment of the present invention; am.

8 and 9 are photographs of the results of culturing E. coli for each titanium implant and untreated titanium implants combined with chitosan according to an embodiment of the present invention.

10 and 11 are photographs of the results of crystal violet staining after inoculation and culture of S. mutans for each titanium implant and untreated titanium implants combined with chitosan according to an embodiment of the present invention.

12 and 13 are photographs of the results of crystal violet staining after inoculation and culture of S. sobrinus for each of the titanium implant bound with chitosan and the untreated titanium implant according to an embodiment of the present invention.

14 is a chitosan-coupled titanium implant according to an embodiment of the present invention, and for each of the untreated titanium implants, S. mutans and S. sobrinus are inoculated and cultured, respectively, a graph for absorbance evaluation results for the formed biofilm. am.

Hereinafter, embodiments of the present invention will be described in detail so that those of ordinary skill in the art to which the present invention pertains can easily implement them. The present invention may be embodied in many different forms and is not limited to the embodiments described herein.

The titanium implant according to the present invention includes chitosan immobilized on the implant surface through chemical bonds.

The titanium implant can be used without limitation in the case of implants placed in vivo, such as for fixing fractures, artificial spine, artificial prostheses, orthodontic treatment, orthodontics. In addition, the implant may be a material of a conventional implant used in a living body based on titanium, and for example, may be an alloy containing trace amounts of metals such as vanadium, aluminum, nickel, iron, and chromium in addition to titanium as a main component. The size of the titanium implant may be, for example, about 1 to 2 mm in diameter and about 6 to 10 mm in length.

In addition, the chitosan is a material that provides antibacterial activity against bacteria that can penetrate the bone tissue in which the titanium implant is placed. In addition, by forming a physical barrier on the surface of the titanium implant, it is possible to prevent or delay the elution of metal ions from the implant, thereby minimizing the inflammatory reaction caused by a trace metal element such as vanadium eluted. The chitosan is 75 to 85% deacetylated, and may have a viscosity of 20-800 cP, and may be used that dissolves in 1% aqueous acetic acid solution or an aqueous solution of pH 6.5 or less, through which it is introduced into the titanium implant surface. It can be more advantageous, and the desired effect can be remarkably expressed through chitosan, and desorption or decomposition after chitosan is introduced into the titanium implant is minimized.

The above-described chitosan is fixed to the titanium implant surface through a chemical bond, and preferably, the chemical bond may be a covalent bond, through which it is possible to minimize the desorption of chitosan from the titanium implant surface.

According to an embodiment of the present invention, a spacer that forms a covalent bond with the chitosan and the titanium implant surface may be further provided. The spacer is a medium for mediating chemical bonding, and materials capable of forming covalent bonds with each of chitosan and titanium implants may be used without limitation. In this case, preferably, the spacer may have a carboxyl group capable of forming a covalent bond with an amine group provided in chitosan.

On the other hand, a hydroxyl group can be easily provided on the titanium surface, and the hydroxyl group has weak reactivity with a carboxyl group or other functional groups, so that even when a covalent bond is formed between chitosan and the spacer, it may be difficult to form a covalent bond between the spacer and the titanium implant surface. Accordingly, according to an embodiment of the present invention, the titanium implant may have a surface modified to include a carboxyl group or an amine group, and the spacer may have an amine group or a carboxyl group to form a covalent bond with a carboxyl group or an amine group provided on the titanium implant surface. can be provided Eventually, the spacer may have at least one carboxyl group capable of forming an amide bond with the amine group of chitosan and at least one carboxyl group or an amine group capable of forming an amide bond with an amine group or carboxyl group provided on the surface of the titanium implant. have.

In this case, a carboxyl group or an amine group provided on the surface of the titanium implant may be introduced into the titanium implant through a known method and a known material. However, in consideration of the use of the implant, a biocompatible material that does not cause cytotoxicity and is harmless to the human body is preferable.

For example, if the functional group provided on the surface of the titanium implant is an amine group, the amine group may be introduced through surface modification of the implant. The amine group may be introduced using a method known in the art to introduce the amine group to the surface of the titanium implant, so the present invention is not particularly limited thereto. However, since the hydroxyl group present on the typical titanium implant surface has very low reactivity, an amine group can be introduced using a silane compound having a high hydroxyl group and reactive group. The silane compound may be an aminosilane-based compound having an amino group, for example, a 3-aminopropyltriethoxysilane (APTES) compound. The titanium implant surface modification through 3-aminopropyltriethoxysilane (APTES) compound is more advantageous to introduce an amine group into the titanium implant surface in an increased content, and the reaction with the carboxyl group provided in the spacer is easy. In addition, it may be more advantageous to increase the content of chitosan to be fixed through this.

In addition, when the functional group provided on the surface of the titanium implant is an amine group, the spacer may be a compound having at least two carboxyl groups, preferably at least one selected from the group consisting of polyacrylic acid, sulcinic acid, glutamic acid, and asphaltic acid. compounds may be included. More preferably, the spacer may be sulcinic acid. Compared to other materials, it may be more advantageous in terms of increasing the content of chitosan introduced into the titanium implant and maintaining the fixing force after the chitosan is introduced.

On the other hand, chitosan can be introduced to the implant surface so as to cover more than 30% of the titanium implant surface, through which it is possible to improve the antibacterial activity against bacteria, there is an advantage that can minimize the elution of metal ions from the titanium implant.

The implant according to an embodiment of the present invention described above may be implemented through a manufacturing method described later, but is not limited thereto.

Specifically, the titanium implant comprises the steps of (1) preparing a titanium implant having an amine group on the surface, (2) treating the micro-implant surface with a first solution having a spacer having at least two carboxyl groups to form the carboxyl group and the implant. Inducing a chemical bond between the amine groups, and (3) treating a second solution containing chitosan to induce a chemical bond between the amine group of chitosan and the unreacted carboxyl group of the spacer.

First, as step (1) of the present invention, a step of manufacturing a titanium implant having an amine group on the surface is performed. Methods known in the art for introducing an amine group to the surface of the titanium implant may be used without limitation. For example, an amine group may be introduced by treating the titanium implant surface with a solution containing an aminosilane-based compound, more specifically, 3-aminopropyltriethoxysilane (APTES). In addition, the aminosilane-based compound may be included in the solution in an amount of 0.01 to 10% by weight, more preferably 1 to 5% by weight. If it is included in the solution in an amount of less than 0.01% by weight, the amount of amine groups bonded to the surface of the micro-implant is small, so the content of chitosan that can be fixed may be significantly reduced, and if it exceeds 5% by weight, aminosilane added An increase in the amount of the introduced amine group relative to the amount of the based compound may be insignificant.

As a solvent included in the solution including the aminosilane-based compound, any conventional solvent capable of dissolving the aminosilane-based compound may be used without limitation, and may be, for example, water or ethanol, but is not limited thereto.

In addition, the method of treating the surface of the titanium implant with the solution containing the aminosilane-based compound may be, for example, an immersion method, but is not limited thereto.

In addition, after the solution containing the aminosilane-based compound is treated on the titanium implant, it is reacted at 60 to 90° C. for 1 to 3 hours, and then washed to remove the physically bound aminosilane-based compound.

Next, as step (2) according to the present invention, a first solution having a spacer having at least two carboxyl groups is treated on the surface of the titanium implant to induce a chemical bond between the carboxyl group of the spacer and the amine group of the titanium implant. carry out

The first solution contains a spacer having at least two carboxyl groups, for example, succinic acid. The first solution further includes a solvent capable of dissolving the spacer, and the solvent may be, for example, water, but is not limited thereto, and may be appropriately changed according to the specific type of the spacer. In this case, the concentration of the spacer in the first solution may be 0.1 to 5% by weight, but is not limited thereto.

In addition, chemical bonding between the carboxyl group of the spacer and the amine group of the micro-implant can be performed through a known coupling reaction, specifically, a couple used in an activation method using water-soluble carbodiimide (WSC) known in the art. The reaction may be through a ring agent, and for example, NHS (Nhydroxysuccinimide) and EDC (1-Ethyl-3-(3-dimethylaminopropyl)-carbodiimide) may be used. Coupling reaction through EDC / NHS is 0.1 to 0.5 parts by weight of EDC (N-ethyl-N'-(3-dimethylaminopropyl) carbodiimide), 0.1 to 0.5 parts by weight of NHS (N-hydroxysuccinimide) and spacer with respect to 100 parts by weight of water. After immersing the titanium implant in an aqueous solution containing After the reaction is completed, it may be washed one or more times using deionized water or the like, and through this, a titanium implant to which a spacer containing a non-reacted carboxyl group is bonded can be manufactured.

On the other hand, after fixing the spacer to the chitosan of step (3) to be described later, it may be fixed on the titanium surface, but it is very advantageous in terms of reactivity to perform step (3) after performing step (2) first. There is an advantage in that chitosan can be introduced to the titanium surface more easily and with high efficiency.

Next, as step (3) according to the present invention, a step of inducing a chemical bond between the amine group of chitosan and the unreacted carboxyl group of the spacer is performed by treating the second solution containing chitosan.

The second solution includes chitosan, and may further include a solvent. The solvent may be used without limitation in the case of a known biocompatible solvent capable of dissolving chitosan, for example, may be an acidic aqueous solution. The chitosan may be included in a concentration of 0.1 to 5% by weight, preferably 0.5 to 2% by weight in the second solution. On the other hand, the second solution may have a pH of 5 to 6, which may be advantageous for dissolution of chitosan.

The spacer-coupled micro-implant may be treated with the second solution for 6 to 10 hours, and may be impregnated, for example. Thereafter, it may be treated with an acidic solution of 0.1 to 2 N concentration for 1 to 20 minutes, and the acidic solution may be, for example, hydrochloric acid. After the acid solution treatment, the titanium implant may be subjected to a washing process one or several times more, and then it may be implemented as a titanium implant in which chitosan is immobilized on the surface through a further drying process.

The present invention will be described in more detail through the following examples, but the following examples are not intended to limit the scope of the present invention, which should be construed to aid understanding of the present invention.

<Example 1>

Among titanium implants, micro-implants for orthodontics were prepared. The micro implant is made of Ti6Al4V alloy material (Al 5.930wt%, Be 0.001wt%, Cd 0.001wt%, Co 0.001wt%, Cr 0.033wt%, Cu 0.001wt%, Fe 0.016wt%, Hf 0.001wt%, Mn 0.004 wt%, Mo 0.002wt%, Ni 0.031wt%, Pd 0.001wt%, V 3.880wt%, balance Ti) was prepared. Thereafter, the micro-implant was impregnated in a solution mixed with 3-aminopropyltriethoxysilane (APTES) to 2% by weight in 95% ethyl alcohol, and then left at 70° C. for 4 hours. Thereafter, the micro-implant was washed with ethanol once and then washed twice more with deionized water to remove APTES physically adsorbed on the surface, thereby preparing a micro-implant in which an amine group was introduced to the surface through APTES.

After preparing a first solution, which is an aqueous solution having a pH of 5.6 containing 0.25% by weight of succinic acid, EDC and NHS were further added and stirred, and then a micro-implant in which an amine group was introduced into the surface was added and stirred again for 6 hours. After that, after washing with deionized water three times and drying the micro-implants to which succinic acid is bound, were prepared.

Then, the second solution prepared by adding 0.5% by weight of chitosan to an aqueous solution having a pH of 5.6 containing 2% by weight of acetic acid and stirring for 4 hours was impregnated with succinic acid-bound micro-implants and reacted for 8 hours. . After treatment with 1N hydrochloric acid for 10 minutes, washed with deionized water and dried to prepare a micro-implant (Ti-SA-Ch) to which the final chitosan was fixed.

<Example 2>

It was prepared in the same manner as in Example 1, but polyacrylic acid was added to the first solution instead of succinic acid to prepare a micro-implant (Ti-AA-Ch) in which chitosan was fixed.

<Experimental Example 1>

XPS analysis was performed on untreated titanium implants (Ti), APTES-treated micro-implants (Ti-A), and micro-implants according to Examples 1 (Ti-SA-Ch) and Example 2 (Ti-AA-Ch). Thus, the results are shown in FIG. 2, and the composition of the elements for which the area ratio of these peaks is calculated is shown in Table 1 below.

	Untreated micro-implant atom (wt%)	APTES-treated micro-implant atom (wt%)	Example 1 (Ti-SA-Ch) Atom (wt%)	Example 2 (Ti-AA-Ch) Atom (wt%)
Ti	15.8	4.9	4.1	4.0
Al	1.8	< 0.1	< 0.1	0.8
V	0.4	< 0.1	< 0.1	< 0.1
C	26.9	59.4	60.8	62.5
O	53.7	32.1	30.7	30.7
N	1.4	3.6	5.0	2.0
Si	-	11.4	5.5	7.9

As can be seen from Table 1, it can be seen that the APTES-treated micro-implant having an amine group introduced onto the surface had a Si content of 11.4 wt%, confirming that the silane coupling agent containing the amine group was well introduced into the micro-implant. In addition, it can be confirmed that chitosan was introduced into the micro-implant through the decrease in the Si content in Examples 1 and 2. On the other hand, in Example 1 and Example 2, Example 1 (5.0%) of nitrogen was much higher than Example 2 (2.0%), so it can be seen that a large amount of chitosan was efficiently introduced when succinic acid was used as a medium.

<Experimental Example 2>

In order to investigate the change in wettability according to the surface modification of the titanium micro-implant, the water contact angle was measured and shown in FIG. 3 . The contact angle of the surface of the untreated titanium implant (Neat Ti) was 71 ^o , and when the aminosilane compound was introduced, the hydrophobicity increased to ^{82 o.} When succinic acid reacted ^{, the hydrophilicity was greatly reduced to 65 o} , and when chitosan was combined, it became ^{more hydrophilic (57 o} ) due to the -OH group present in the sugar chain. As described above, it can be seen that the modification reaction proceeded well from the change in the wettability of the surface according to the surface modification process.

<Experimental Example 3>

In order to confirm the chitosan introduced to the surface of the micro-implant, FITC fluorescent material was bound to the amine group of chitosan through a conventional method, followed by fluorescence microscopy, and the results are shown in FIGS. 4 (Example 1) and 5 (Example 1), respectively. Example 2).

As can be seen in FIGS. 4 and 5 , it can be confirmed that the chitosan is fixed to the surface of the titanium implant from the observation of the green image. On the other hand, it can be seen that Example 1 (FIG. 4) using succinic acid as a spacer shows a darker green color compared to Example 2 (FIG. 5) using polyacrylic acid, which is carried out at the content of chitosan introduced into the micro-implant As in Example 1, it can be seen that the content of chitosan significantly increased when succinic acid was used as a spacer.

<Experimental Example 4>

The biocompatibility of the chitosan-coupled titanium implant and untreated implant according to Example 1 was performed by evaluating the cell adhesion of osteoblasts in vitro.

Specifically, after placing each of these in a 4-well dish, a culture medium (DMEM) having a concentration of osteoblast cells (MC3T3-E1) of 3 x 10 ³ cell/ml was prepared and seeded by 1 ml each. After incubation for 1 day and 3 days in an incubator at 37 ° C., 5% CO ₂ , the culture medium was removed, washed carefully with PBS, fixed with 2.5% (w/v) glutaldehyde, and FE- The adhesion behavior was observed through the SEM image, and the results taken after incubation for 1 day and 3 days are shown in FIG. 6 .

As can be seen from Figure 6, compared to the untreated titanium implant (Ti), in the titanium implant in which chitosan according to Example 1 (Ti-SA-Ch) was introduced, cells were well spread and adhered for 1 day and 3 days in culture. It can be confirmed, and it can be confirmed that the titanium implant (TI-SA-Ch) of Example 1 has a good effect on the initial adhesion of osteoblasts and cell spreading.

<Experimental Example 5>

Live staining staining evaluation was performed on cells cultured for 1 day and 3 days in Experimental Example 4, and after staining using the Live/dead staining staining method, confocal images were taken, and the results are shown in FIG. 7 .

Calcein-AM, a reagent used for staining, is a non-fluorescent material that, when it enters the cell, reacts with an intracellular esterase to convert it to a fluorescent material, giving it green fluorescence, and propidium iodide (PI) is a living cell membrane Since it cannot pass through the cell membrane, it enters only dead cells with damaged cell membranes and ionic bonds with the cell nucleus, which is an anion, resulting in red fluorescence.

7, the chitosan according to Example 1 was combined with the result that green fluorescence was significantly higher in the micro implant (TI-SA-Ch) to which the chitosan according to Example 1 was fixed compared to the untreated micro implant (TI) through FIG. 7 . It can be confirmed that the proliferation of osteoblasts is greater in the titanium implant, and through this, a more improved bond with the tooth/gum is expressed, preventing bacterial penetration through the interface between the micro-implant and the tooth/gum, and thus the occurrence of inflammation is further increased. expected to decrease.

In addition, in the titanium implant (TI-SA-Ch) combined with chitosan according to Example 1 through FIG. 7, no dead cells appearing as a red image were observed, so the titanium implant combined with chitosan used in this study was It can be seen that the biocompatibility is very high.

<Experimental Example 6>

To evaluate the antibacterial properties of titanium implants using E. coli, the experiment was performed as follows. A small amount of E. coli was added with a platinum tip to a liquid medium prepared by putting a nutrient medium in distilled water and cultured for 24 hours. After that, the strain solution was diluted to 1/200,000, put into a vial containing untreated titanium implant (Ti) and chitosan-coupled titanium implant (Ti-SA-Ch), and cultured for 24 hours. After taking a small amount of the solution from the vial and smearing it on the solid medium in a culture dish, the image of E. coli grown by standing still for 24 hours was taken and shown in FIGS. 8 (Example 1) and 9 (untreated titanium implant).

As can be seen through FIGS. 8 and 9 , it can be confirmed that the titanium implant combined with chitosan according to Example 1 has an effect on inhibiting the proliferation of E. coli compared to the untreated implant.

Two strains of S. mutans and S. sobrinus, which are representative oral bacteria that frequently appear in corroded teeth, were inoculated with chitosan-conjugated titanium implants and untreated titanium implants according to Example 1 at 2 x 10 ⁵ cells, respectively. After incubation at 37° C. for 8 hours, it was stained with crystal violet to evaluate the presence and extent of biofilm formed due to the strain, and photographs were taken and shown in FIGS. 10 to 13 . In addition, after removing the biofilm for quantitative evaluation of the degree of biofilm formation, absorbance was measured at 595 nm using a microplate reader, and the results are shown in FIG. 13 .

10 and 11 are antimicrobial evaluation results for S. mutans, when the case of FIG. 10 (Example 1, Ti-SA-Ch) is compared to FIG. 11 (untreated micro-implant, Ti), the biofilm is almost You can check that it didn't happen.

On the other hand, FIGS. 12 and 13 are results of evaluation of antibacterial properties against S. sobrinus, and the biofilm was significantly improved in the case of FIG. 12 (Example 1, Ti-SA-Ch) compared to FIG. 13 (untreated titanium implant, Ti). It can be seen that a small amount occurred.

As can be seen from FIGS. 10 to 13 in the antibacterial experiment using S. mutans and S. sobrinus as described above, the titanium implant (Ti-SA-Ch) to which chitosan according to Example 1 is coupled is S. mutans and S. sobrinus. It can be seen that the antibacterial effect on the strain is significantly superior to that of the untreated titanium implant (Ti).

Although one embodiment of the present invention has been described above, the spirit of the present invention is not limited to the embodiments presented herein, and those skilled in the art who understand the spirit of the present invention can add components within the scope of the same spirit. , changes, deletions, additions, etc. will be able to easily suggest other embodiments, but this will also fall within the scope of the present invention.

Claims (9)

1. A titanium implant having antibacterial properties comprising chitosan fixed through a chemical bond on a titanium (Ti) implant surface.
2. According to claim 1,

   A titanium implant having antibacterial properties, characterized in that it further comprises a spacer for forming a covalent bond with each of the titanium implant and the chitosan between the titanium implant and the chitosan.
3. 3. The method of claim 2,

   The titanium implant has a modified surface to have an amine group,

   Titanium implant having antibacterial properties, characterized in that the spacer is a compound having at least two carboxyl groups to form an amide bond with each of the amine group of the titanium implant and the amine group of chitosan.
4. 4. The method of claim 3,

   The spacer is a titanium implant having antibacterial properties comprising at least one compound selected from the group consisting of polyacrylic acid, succinic acid, glutamic acid and aspartic acid.
5. According to claim 1,

   The chitosan is 75-85% deacetylated, and the titanium implant having antibacterial properties, characterized in that the viscosity is 20-800cP.
6. According to claim 1,

   The chitosan is a titanium implant having antibacterial properties, characterized in that it is provided to cover at least 30% of the surface area of the titanium implant.
7. 5. The method of claim 4,

   The spacer is a titanium implant having antibacterial properties, characterized in that sulcinic acid.
8. According to claim 1,

   The titanium implant has antibacterial properties, characterized in that the titanium implant surface is modified with 3-aminopropyltriethoxysilane (APTES).
9. (1) manufacturing a titanium implant having an amine group on the surface;

   (2) inducing a chemical bond between the carboxyl group and the amine group of the implant by treating the surface of the titanium implant with a first solution having a spacer having at least two carboxyl groups; and

   (3) inducing a chemical bond between the amine group of chitosan and the unreacted carboxyl group of the spacer by treating the second solution containing chitosan;

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