KR20110052355A - Method of inactivating a biofilm using magnetic particles - Google Patents

Abstract

PURPOSE: A method of inactivating a biofilm is provided to inactivate the biofilm through improved sterilization power for a short time. CONSTITUTION: A method of inactivating a biofilm comprises: a step of preparing an object having a biofilm(5) on the surface thereof; a step of adding magnetic particles(4) to the biofilm; a step of sanctioning AC magnetic field from the outside of the object; and a step of inactivating the biofilm by generating heat from magnetic particles through the AC magnetic field applied from the outside.

Description

Method of inactivation of biofilm using magnetic particles {METHOD OF INACTIVATING A BIOFILM USING MAGNETIC PARTICLES}

The present invention relates to a method of inactivating a biofilm using a property that magnetic particles (M.P.) generate heat when an alternating magnetic field is applied. More specifically, the present invention relates to a method of inactivating a biofilm, which can effectively inactivate a biofilm in a short time by using the property of generating an alternating current by applying an alternating magnetic field to magnetic particles (M.P.).

The biofilm is a smooth mucous layer formed by combining microorganisms on the surface and extracellular polymeric substances (EPS) secreted by the microorganisms. In other words, the biofilm is a microbial colony attached to the surface, and each microbial community is surrounded by mucous membranes composed of substances occurring in the metabolic process of the microorganisms or the surrounding environment, and is in contact with water regardless of the surface properties or the surrounding environment. If it exists anywhere. Biofilms are very familiar to our daily lives and live in close contact with them. Biofilms can exist on any surface with sufficient moisture and nutrients. Biofilms are slippery membranes on riverside stones, in flower jars with flowers, and on washbasins and toilet floors. Biofilms are also commonly observed on the surface of living organisms, which actively secrete extracellular macromolecules from the plaque of the teeth and the surface of intestinal tissue cells, forming microbial communities.

The binding material of the biofilm is made of organic polymer material produced and discharged by the microorganisms that make up the biofilm and is called extracellular polymer sustances (EPS). The chemical structure of EPS depends on the type of microorganism and the environmental conditions. The biofilm is composed mostly of water (80 ~ 95%) and the rest is composed of EPS (about 85 ~ 98% of organic matter) and organic matter, microorganisms, and humic substances adsorbed on EPS. In addition, mud, inorganic salts, and corrosion products derived from the outside are often included in the biofilm. As a result, microbial cells account for only 1 to 2% of the total biofilm.

Research into the biofilm structure has been ongoing since the early 1980s, when biofilm research began. The overall structure of the biofilm is that the microbial community in the form of mushrooms surrounded by EPS is separated into water channels, and there is an empty space such as a passage inside the biofilm. Biofilms are sometimes uniformly layered, but in most cases they have a non-uniform distribution of zirconiums. The biofilms found in nature vary in thickness from 50 to 150 μm on average, to several centimeters of water formed in sewage treatment plants. The interior of the biofilm is often divided into layers of various properties to form different environments. For example, biofilms with both aerobic and anaerobic layers are often observed.

Biofilms are like two sides of a coin, so depending on the environment they are created, they can be harmful to humans and can be useful. If biofilms are formed on unwanted surfaces, industrial facilities can cause corrosion, biofouling, and water contamination, resulting in economic losses, as well as secondary sources of tap water and a hotbed of pathogenic microorganisms. In addition, when formed in medical equipment, such as artificial organs that are implanted in the body can have a fatal effect on the human body. In this respect, control of biofilm formation has been an important concern in many fields, and research on it has been actively conducted in the United States and Europe since the 80s. Conversely, studies on the use of biofilms to promote biofilm formation have been ongoing. For example, bioremediation of biofilms to remove contaminants from industrial wastewater or groundwater has been under way for a long time. Biofilms are also used to obtain biochemicals used in food additives and chemical additives.

On the other hand, most bacteria are attached to any surface and reaches a certain concentration to grow in the form of a biofilm through quorum sensing. The biofilm thus formed serves as a habitat and a barrier for the bacteria, thereby increasing tens to thousands of times more resistance to general disinfectants such as chlorine, hydrogen peroxide or ozone than when present as suspended bacteria. The microbial community in the form of the biofilm threatens the safety of medical tools used in the medical industry recently, and a safe disinfection method capable of completely disinfecting and sterilizing the biofilm is required.

Conventionally, as a standard disinfection method for disinfecting biofilms, autoclaving corresponding to the wet method and disinfection using ethylene oxide, gamma irradiation, and hydrogen peroxide plasma corresponding to the dry method are used. Has been used. However, the disinfection methods have a relatively good disinfection and sterilization power, but also has limitations in using in various industrial fields including medical field. High pressure steam sterilization is unsuitable for disinfecting materials susceptible to high temperature and high pressure, and may form an oxide film on the metal surface by water vapor, thereby degrading the performance of the material. In addition, disinfection with ethylene oxide not only destroys specific polymers, but also has strong toxicity and requires secondary purification after disinfection. Gamma rays also weaken the properties of medical polymers, and hydrogen peroxide plasma can produce excess free radicals to corrode alloy metal surfaces.

In order to overcome the problems of conventional disinfection methods that can remove the conventional biofilms are required to develop an alternative disinfection method to compensate for this, in particular, the need for disinfection methods that have excellent cell permeability, no toxicity and no persistence is urgently required. have.

Recently, a lot of researches using magnetic materials for bio-related research materials and biomedical applications have been attempted. In particular, there are many researches on the use of biomaterial separation materials and medical materials, such as separation and purification of DNA and RNA, separation and purification of proteins and amino acids, biosensors, drug delivery systems, and magnetic resonance imaging (MRI). Functional silane-coated magnetic particles have been studied to coat the magnetic particles with organic functional compounds so that they can be used for contrast agents and local thermotherapy.

Magnetic particles have a feature that can be manipulated using an external magnetic field and can be usefully used in various fields. Among magnetic particles, paramagnetic or superparamagnetic particles respond to an external magnetic field. In particular, superparamagnetic particles are applied to various biomedical fields such as gene transfer, magnetic resonance imaging (MRI), tissue repair, and detoxification.

Magnetic particles are characterized by generating self-heating by loss of magnetization in alternating magnetic fields. As another example of application of the magnetic particles to the biological treatment using the properties of such magnetic particles, high temperature treatment using magnetic spin energy is available (US 6,530,944 B2, US 5,411,730). Magnetic particles emit heat through a spin flipping process when an AC current of a radio frequency flows from the outside. At this time, when the temperature around the particle is more than 40 ℃ cells are killed by high heat can selectively kill the diseased cells.

Therefore, as a method for developing a disinfection method that is harmless to a human body or the environment and has no residual property while exhibiting a faster disinfection effect than conventional disinfection methods, heat is generated by magnetic loss caused by an external magnetic field to which magnetic particles are applied. To inactivate the biofilm. In addition, the microorganisms that form the biofilms are relatively easy to inactivate the cell membranes of the microorganisms compared to the floating microorganisms, and show only low inactivation effects. In the present invention, biofilms as well as floating microorganisms are formed using the exothermic properties of the magnetic particles. One attempt was to inactivate one microorganism and solve the above problems.

It is an object of the present invention to provide a method of inactivating a biofilm with improved disinfection power in a short time using magnetic particles.

The present invention comprises the steps of preparing an object in which a biofilm (biofilm) is formed on the surface to achieve the above object; Adding magnetic particles to the biofilm; Applying an alternating magnetic field outside the object on which the biofilm to which the magnetic particles are added is formed; And inactivating the biofilm by exothermic magnetic particles by the alternating magnetic field applied from the outside.

The magnetic particles according to the present invention are Fe ₃ O _{4 ,} Fe ₂ O ₃ , aluminum (Al), cobalt (Co), nickel (Ni), manganese (Mn), tin, zinc, cadmium, magnesium, copper, barium, It may be selected from the group consisting of oxides of lithium and yttrium, preferably Fe ₃ O ₄ Or Fe ₂ O _{3 ,} more preferably Fe ₃ O ₄ can be selected.

In the present invention, the diameter of the magnetic particles is characterized in that 5 nm ~ 1 ㎛.

The biofilm to which the magnetic particles are added according to the present invention is characterized by applying an alternating magnetic field to the magnetic particles from the outside for a time within 10 minutes.

The biofilm according to the present invention is Pseudomonas aeruginosa, Vibrio harveyi, Agrobacterium tumefaciens, Escherichia coli or Candida albicans by Candida albicans. It is characterized by being formed.

In order to achieve the above another object, the present invention coats the magnetic particles on the antibody or the enzyme in an antigen-antibody or enzyme-substrate and transfers them to the biofilm in the body, and applies magnetic alternating current by applying an alternating magnetic field. It is characterized in that the biofilm is inactivated by using the property of generating particles.

Hereinafter, the present invention will be described in more detail.

The present invention relates to a method of inactivating a biofilm using a property in which magnetic particles generate heat by themselves when an alternating magnetic field is applied from the outside. More specifically, preparing a subject having a biofilm (surface) is formed on the surface; Adding magnetic particles to the biofilm; Applying an alternating magnetic field outside the object on which the biofilm to which the magnetic particles are added is formed; And inactivating the biofilm by generating the magnetic particles by the alternating magnetic field applied from the outside.

Here, inactivation of a biofilm means inhibiting proliferation of microorganisms constituting the biofilm or killing or removing microorganisms to eliminate infectivity. According to the method of inactivating the biofilm using the magnetic particles of the present invention described above, the biofilm that is difficult to inactivate by conventional chlorine disinfection or ozone disinfection can be effectively disinfected and sterilized in a short time to inactivate the biofilm.

In addition, by simply adding magnetic particles to the biofilm without undergoing a complicated process, and by applying an alternating magnetic field from the outside, the magnetic particles generate heat due to magnetic loss, and the heat generated thereby inactivates the biofilm with excellent disinfection power. can do. The present invention also has the advantage that it is possible to locally inactivate only the portion where the biofilm is formed because only the portion where the magnetic particles are present is exothermic.

In addition, the present invention is non-toxic because there is no chemical that may be harmful to the human body or the environment, there is an advantage that can change the degree of heat generation by adjusting the concentration and amount of magnetic particles, the application time of the alternating magnetic field.

The magnetic particles refer to a material which does not have a magnetic field but forms a magnetic dipole when exposed to the magnetic field. The physicochemical properties of the magnetic particles are characterized by their size, magnetization degree, surface shape, and the like. The magnetic particles used in the biofilm deactivation method using the magnetic particles of the present invention are iron oxide (Fe ₃ O ₄ Or Fe ₂ O ₃ ) and aluminum (Al), cobalt (Co), nickel (Ni), manganese (Mn), tin (Sn), zinc (Zn), cadmium (Cd), magnesium (Mg), copper (Cu ), Barium (Ba), lithium (Li) and yttrium may be selected from the group consisting of, preferably Fe ₃ O ₄ Or Fe ₂ O ₃ Can be selected. More preferably Fe ₃ O ₄ may be selected. Iron oxide particles such as magnetite (Fe ₂ O ₃ ) or magnetite (Fe ₂ O ₃ ) are ferromagnetic when micrometer-sized, but nanometer-sized particles are superparamagnetic. These particles are easy to synthesize and can be easily adjusted in size, but preferably the particles have an average particle diameter of 100 nm or less, preferably 50 nm or less, particularly preferably 30 nm or less. Preferably the average particle diameter is in the range from 1 to 40 nm, more preferably from 3 to 30 nm, particularly preferably from 5 to 25 nm.

According to an embodiment of the present invention, after the magnetic particles are added to the biofilm, an external alternating magnetic field may be applied, and the biofilm may be inactivated by using the property of generating heat by an external alternating magnetic field to which the magnetic particles are applied. In addition, the biofilm can inactivate the biofilm simply by applying the alternating magnetic field for a time within about 10 minutes, so that the biofilm can be sterilized by a simple method within a short time compared to the conventional disinfection method.

The biofilm according to the present invention is Pseudomonas aeruginosa, Vibrio harveyi, Agrobacterium tumefaciens, Escherichia coli or Candida albicans by Candida albicans. It is characterized by being formed.

In order to achieve the above another object, the present invention coats the magnetic particles on the antibody or the enzyme in an antigen-antibody or enzyme-substrate, transfers them to a biofilm in the body, and applies an alternating magnetic field, thereby applying the applied alternating magnetic field. It is characterized in that the biofilm is inactivated by using the property that the magnetic particles generate heat.

In general, magnetic particles need surface modification to stabilize particles and improve biocompatibility. There is a method of modifying a magnetic particle with specific binding groups that biomolecules can bind to. For example, a carboxyl or amino group capable of covalently binding to biomolecules, and strapavidin / avidine capable of specifically binding to a molecule modified with biotin may be modified in a magnetic particle. These methods are usually used to immobilize specific receptors for the desired particles.

In order to coat the magnetic particles on the antibody or enzyme, the magnetic particles are activated by treating them with 5 to 20% glutaraldehyde. After reacting by rotating at room temperature for 3 hours, glutaraldehyde which did not react with magnetic particles is removed. The antibody or enzyme is dissolved in a buffer for binding reaction to make an antibody or enzyme solution. The antibody or enzyme solution is added to magnetic particles previously activated with glutaraldehyde, and the reaction is performed at room temperature for 16-24 hours while shaking well. After the reaction is completed, the Quenching solution is added to terminate the reaction between the antibody or enzyme and the magnetic particles.

As described above, the antibody or enzyme coated with the magnetic particles is transferred to the organ or tissue in which the biofilm is formed in the body, and an alternating magnetic field is applied to generate the biofilm formed in the body by using the characteristic that the magnetic particles generate heat by the applied alternating magnetic field. By activating it can suppress the infection of bacteria in tissues or human body and induce the death of microorganisms.

According to the deactivation method of the biofilm using the above-described characteristics that the magnetic particles of the present invention generate heat by an alternating magnetic field applied from the outside, the biofilm which is difficult to deactivate by conventional chlorine disinfection or ozone disinfection is very effectively disinfected and Sterilization can inactivate the biofilm. In addition, it is possible to generate heat only in the region where the magnetic particles are present, so that only the portion where the biofilm is formed can be locally inactivated.

In addition, the present invention is non-toxic because there is no chemical that may be harmful to the human body or the environment, there is an advantage that can change the degree of heat generation by adjusting the concentration and amount of magnetic particles, the application time of the alternating magnetic field.

In addition, the present invention may be applied to inactivate only the biofilm-forming part by moving the antibody and the enzyme coated with magnetic particles to the biofilm-forming part in the body based on antigen-antibody specificity and enzyme-substrate specificity. will be.

Hereinafter, a method of inactivating a biofilm using magnetic particles according to exemplary embodiments of the present invention will be described in detail with reference to the accompanying drawings.

1 is used to perform a method of inactivating a biofilm by adding Fe ₃ O ₄ particles as magnetic particles to a polycarbonate coupon as an object on which a biofilm is formed, and then applying an alternating magnetic field outside the polycarbonate coupon. It is a schematic diagram of the whole system to explain the disinfection device.

Referring to FIG. 1, the AC magnetic field induction apparatus 1 is connected to the coil 2 and generates an magnetic field by controlling an output control terminal 1a located in the AC magnetic field induction apparatus 1. A magnetic field is formed around the coil 2 which is connected to the AC magnetic field induction device 1 through (1).

The biofilm specimen in this embodiment has a structure in which a biofilm 5 is formed on a polycarbonate coupon 6 and magnetic particles 4 are added to the formed biofilm 5. The biofilm specimen is placed in the magnetic field region formed around the coil 6. In this case, the biofilm specimen placed in the region in which the magnetic field is formed may locally inactivate only a portion where the magnetic particles are generated by the alternating magnetic field applied from the outside.

In the embodiments of the present invention, the biofilm formed by using the polycarbonate coupon 6 as an object on which the biofilm is formed is inactivated. However, the biofilm that is inactivated according to the present invention may include all kinds of biofilms. In addition, the biofilm may include a medical device or various life tools. When bacteria adhere to a certain surface and reach a certain concentration, they grow in the form of a biofilm through quorum sensing. This kind of biofilm can also be inactivated according to the present invention. ,

The biomembrane acts as a habitat and a protective film of bacteria, and exhibits properties that are not easily killed due to an increase in resistance to a general disinfectant, for example, chlorine, hydrogen peroxide or ozone, by tens to thousands of times. Therefore, the present invention can effectively disinfect the biofilm specimen in a short time by the magnetic particles are generated by the alternating magnetic field applied from the outside of the biofilm specimen. The biofilm deactivation method using the characteristic that the magnetic particles generate heat by the alternating magnetic field applied from the outside of the present invention is very effective to disinfect and sterilize a biofilm which is difficult to inactivate by a conventional chlorine disinfection or ozone disinfection in a short time. Can be inactivated. In addition, by heating only the site where the magnetic particles are present, only the part where the biofilm is formed can be inactivated locally, and there is no toxicity because there is no chemical which may be harmful to human body or the environment.

Hereinafter, the present invention will be described in more detail with reference to the following examples.

Example 1 Exothermic Change with Time and Concentration of Magnetic Particles

Pseudomonas aeruginosa (PA01) on the polycarbonate coupons with the biofilm formed by adding 0.15 ml of magnetic particles Fe ₃ O ₄ by concentration and applying a range of alternating magnetic fields from the outside to change the temperature of the magnetic particles with time The measurement is shown in FIG. 2.

As shown in FIG. 2, an alternating magnetic field is applied to the biofilm to which the magnetic particles are added, and heat is generated due to magnetic loss of the magnetic particles, thereby showing the highest temperature within 8 minutes, and as the concentration of the magnetic particles increases, It was confirmed that the temperature rise was also large.

In FIGS. 3A, 3B, and 3C, 0.15 ml of distilled water, 0.15 ml of 40 mg / ml magnetic particles, and 0.15 ml of 60 mg / ml magnetic particles were respectively added to Pseudomonas aeruginosa to clarify the temperature difference depending on the presence or absence of magnetic particles. , PA01) was added to the biofilm using the strain, and the alternating temperature was applied to the biofilm from the outside using an infrared thermography camera. The applied alternating magnetic field was constant regardless of the magnetic particle concentration and applied to all samples for 8 minutes.

When only distilled water was added as shown in FIG. 3A, even though an external alternating magnetic field was applied to the biofilm, there was no obvious temperature rise or temperature change. On the other hand, in the case of adding 0.15 ml of 40 mg / ml magnetic particles Fe ₃ O ₄ to the biofilm in FIG. 3b, it can be seen that the temperature at which the biofilm exists is light green, and only the temperature at which the biofilm exists is increased. . Compared to FIG. 3B, 60 mg / ml of magnetic particles Fe ₃ O ₄ having 1.5 times increased in concentration of the magnetic particles added to the biofilm is added to FIG. 3C to show a deep light green color in the region where the biofilm is present. It can be seen that the temperature of the magnetic particles has increased.

Accordingly, based on the results of FIGS. 2 and 3, when the magnetic particles were added to the biofilm, the magnetic particles generated heat by magnetic disappearance due to the alternating magnetic field applied thereto. The extent of the rise was also greater.

Example 2 Evaluation of Inactivation of Biofilm Using Magnetic Particles

Example 2-1 Change of Population in Biofilm

In order to confirm the inactivation of the Pseudomonas aeruginosa biofilm according to the concentration of the magnetic particles, 0.15 ml of distilled water, 50 mg / ml, and 60 mg / ml, respectively, were added to the biofilm. Then, the inactivation evaluation of the Pseudomonas aeruginosa biofilm according to the concentration of the magnetic particles by applying an alternating magnetic field outside the biofilm was confirmed by a spread plate method and is shown in FIG. 4.

Figure 4 is a graph showing the change in the number of individuals in the biofilm for the biofilm treated with only distilled water and the addition of magnetic particles. Referring to FIG. 4, the biofilm added with only distilled water did not show any difference from the initial population and did not kill microorganisms even when an alternating magnetic field was applied from the outside. On the contrary, when an alternating magnetic field was applied for 8 minutes from the outside after adding 50 mg / ml of magnetic particles, the population decreased by about 1/10 ² to 1/10 ^2.5 compared to the initial population. At this time, the colony forming unit capacity was almost 99%. That is, even when the external magnetic field is applied without adding the magnetic particles, the killing rate of the microorganisms was significantly increased as compared with the biofilm containing only the distilled water without heat generation.

In addition, when the concentration of the magnetic particles added to the biofilm is 60 mg / ml, when the external magnetic field is applied for 8 minutes, the population was reduced by about 1/10 ^4.5 compared to the initial population. At this time, the decrease in colony forming unit capacity was about 99.99% or more. Therefore, when the concentration of the added magnetic particles is 60 mg / ml, the concentration of the magnetic particles was found to be more than about 100 times when the alternating magnetic field is applied externally for 8 minutes compared to 50 mg / ml.

Therefore, it can be seen that the disinfecting power of the biofilm increases much as the concentration of the magnetic particles added to the biofilm increases through this example.

<Example 2-2> Cell membrane damage effect of the biofilm

To determine whether the Pseudomonas aeruginosa biofilm is effectively inactivated to the inside of the biofilm by an applied external alternating magnetic field, 0.15 ml of distilled water and 0.15 ml of 60 mg / ml magnetic particles Fe ₃ O ₄ are added and an external alternating magnetic field is applied. The biofilms were then stained and observed using Confocal Laser Scanning Microscopy (CLSM). Biofilm staining was performed using a LIVE / DEAD BacLight bacterial viability kit (Molecular Probes). In confocal microscopy, green means live cells and red means dead cells.

In the present embodiment, after adding distilled water and magnetic particles to the biofilm formed on the polycarbonate coupon, an alternating magnetic field was applied for 8 minutes from the outside, and the results of observing the biofilm are shown in FIGS. 5A and 5B. 5A is a confocal micrograph showing a biofilm to which only distilled water is added, and FIG. 5B is a confocal micrograph showing a biofilm to which magnetic particles are added.

Referring to FIGS. 5A and 5B, when the alternating magnetic field was added with only distilled water, as shown in FIG. 5A, the biofilm cell membrane was observed to be green due to no damage. However, when the alternating magnetic field was added after the magnetic particles were added, FIG. 5B. As shown in, most of the microorganisms forming the biofilm were killed and observed in red color. Therefore, it can be seen that in the biofilm to which the magnetic particles are added, the magnetic particles generate heat by applying an external alternating magnetic field, and the cell membrane of the microorganisms is damaged by the generated heat, and most of the microorganisms forming the biofilm are killed within 8 minutes.

As described above, when the biofilm is inactivated using the magnetic particles, it was confirmed that the bacteria inside the biofilm can be effectively inactivated with respect to the biofilm in which the bacteria are thickly formed in a very short time. This could disinfect the biofilm by disinfecting and sterilizing the biofilm which is difficult to inactivate by conventional chlorine disinfection or ozone disinfection very effectively in a short time. In addition, magnetic particles are non-toxic and non-permanent, so that biofilms on medical instruments and biomaterials can be sterilized in an environmentally friendly manner. Furthermore, the biofilm could be inactivated by a simple method such as applying an alternating magnetic field to the biofilm containing the magnetic particles.

1 is an overall system schematic diagram showing that a biofilm can be inactivated by applying an alternating magnetic field after adding magnetic particles to a polycarbonate coupon having a biofilm formed thereon.

2 is a graph showing the temperature change of the magnetic particles according to the concentration of the magnetic particles and the application time of the alternating magnetic field.

Figures 3a, 3b and 3c is a thermal infrared camera photograph showing the temperature change after applying distilled water, 40 mg / ml magnetic particles and 0.15 ml of 60 mg / ml magnetic particles to the polycarbonate coupon formed biofilm and applying an alternating magnetic field This is the result.

4 is a graph showing the change in the number of individuals in the biofilm with time after adding distilled water, 50 mg / ml of magnetic particles and 60 mg / ml of magnetic particles.

5A and 5B are confocal micrographs showing biofilms by adding 0.15 ml of distilled water and 60 mg / ml of magnetic particles and applying an alternating magnetic field.

<Description of Symbols for Main Parts of Drawings>

1: AC magnetic field induction device

1a: Output control terminal

1b: monitor panel

2: chiller

3: coil

4: magnetic particles

5: biofilm

6: polycarbonate coupon

Claims (7)

Preparing a subject having a biofilm formed on a surface thereof; Adding magnetic particles to the biofilm; Applying an alternating magnetic field outside the object on which the biofilm to which the magnetic particles are added is formed; And And deactivating the biofilm by heating the magnetic particles by an alternating magnetic field applied from the outside. The method of claim 1, The magnetic particles include Fe ₃ O ₄ , Fe ₂ O ₃ , aluminum (Al), cobalt (Co), nickel (Ni), manganese (Mn), tin, zinc, cadmium, magnesium, copper, barium, lithium, and yttrium. Inactivation method of a biofilm using magnetic particles, characterized in that selected from the group consisting of oxides. The method of claim 1, The magnetic particles are Fe ₃ O ₄ Or Fe ₂ O ₃ Inactivation method of a biofilm using magnetic particles. The method of claim 1, The diameter of the magnetic particles is 5 nm ~ 1 ㎛ characterized in that the biofilm inactivation method using magnetic particles. The method of claim 1, The biofilm to which the magnetic particles are added inactivates a biofilm using magnetic particles, wherein an alternating magnetic field is externally applied to the magnetic particles for a time within 10 minutes. The method of claim 1, The biofilm is formed by Pseudomonas aeruginosa, Vibrio harveyi, Agrobacterium tumefaciens, Escherichia coli or Candida albicans. Inactivation method of a biofilm using magnetic particles. In the antigen-antibody or enzyme-substrate, magnetic particles are coated on the antibody or the enzyme and transferred to the biofilm in the body, and an alternating magnetic field is applied to inactivate the biofilm by using the characteristic that the magnetic particles generate heat by the applied alternating magnetic field. Inactivation method of a biofilm using magnetic particles, characterized in that.

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