Antibacterial material based on nano-enzyme synergistic sonodynamic curative effect as well as preparation method and application of antibacterial material

Abstract

The invention provides an antibacterial material based on a nano-enzyme synergistic sonodynamic curative effect and a preparation method and application. The antibacterial material comprises BiPt nano-enzyme, the outer side of the BiPt nano-enzyme is coated with BSA, PpIX is connected to the BSA, BPBP nano-particles are formed, the outer side of the BPBP nano-particles is coated with AP hydrogel, and a BPBP (at) AP antibacterial material is obtained. The BPBP (at) AP nano antibacterial material disclosed by the invention is combined with ultrasound, the bacteriostasis rate on helicobacter pylori exceeds 99%, a biological membrane is removed, and the risk caused by antibiotic treatment is avoided.

Description

Technical Field

[0001] The present invention relates to the technical field of antibacterial materials, and in particular to an antibacterial material based on nanoenzyme-enhanced sonodynamic efficacy, a preparation method and an application thereof. Background Art

[0002] Helicobacter pylori (H. pylori) is a Gram-negative, microaerophilic, spiral-shaped bacterium that has been shown to infect more than half of the world's population and is the leading cause of gastric cancer. The efficacy of standard antibiotic-based quadruple therapy is declining due to the development of drug resistance. In addition, the use of high-dose antibiotics may cause serious side effects on commensal bacteria, which are closely related to many other beneficial physiological and metabolic processes. Therefore, strategies that can effectively treat H. pylori infection without harming commensal bacteria are urgently needed.

[0003] Ultrasound-driven antimicrobial sonodynamic therapy (SDT) is a promising treatment for bacterial infections. Compared with light irradiation, SDT has the advantage of excellent tissue penetration of US and shows great feasibility for treating deep infections. Compared with antibiotic treatment, SDT benefits from a completely different bactericidal mechanism and rarely raises concerns about drug resistance. The main reasons that currently affect the therapeutic effect of SDT are traditional sonosensitizers, poor hydrophobicity, low bioavailability, and easy toxic side effects. In addition, bacterial infections are always accompanied by a hypoxic microenvironment, which further inhibits the antibacterial efficacy of oxygen-dependent SDT.

[0004] Chinese patent document CN116350585A discloses an antibacterial material based on polyamino acid-protoporphyrin polymer and liquid perfluorocarbon oil. The light / ultrasound external stimulation time of the antibacterial material is slightly longer, and the bactericidal effect needs to be further improved. Summary of the invention

[0005] The present invention proposes an antibacterial material based on nanozyme-enhanced sonodynamic efficacy, as well as a preparation method and application thereof. The BPBP@AP nano antibacterial material combined with ultrasound has an inhibition rate of more than 99% against Helicobacter pylori, removes biofilms, and avoids the risks brought by antibiotic treatment.

[0006] The technical solution of the present invention is achieved as follows: an antibacterial material based on nanozyme-enhanced sonodynamic efficacy includes a BiPt nanozyme, the outside of the BiPt nanozyme is coated with bovine serum albumin (BSA), and BSA is connected to protoporphyrin (PpIX) to form BiPt@BSA-PpIX (BPBP) nanoparticles, and the outside of the BPBP nanoparticles is coated with ascorbyl palmitate (AP) hydrogel to obtain a BPBP@AP (BiPt@BSA-PpIX@AP) antibacterial material.

[0007] Furthermore, the particle size of BiPt nanozyme is 10-200 nm, and the particle size of BPBP nanoparticles is 100-500 nm.

[0008] The method for preparing an antibacterial material based on nanozyme-enhanced sonodynamic efficacy comprises the following steps:

[0009] (1) Synthesis of BiPt nanozymes;

[0010] (2) BSA was coated on the outside of BiPt nanozyme and connected to PpIX through EDC / NHS biochemical conjugation activation to form BPBP nanoparticles;

[0011] (3) BPBP nanoparticles were encapsulated in AP hydrogel to obtain BPBP@AP antibacterial material.

[0012] Furthermore, in step (2), the specific method for forming BPBP nanoparticles is as follows:

[0013] 1) Disperse 20 mg of BiPt nanozyme in 20 mL of aqueous solution, add BSA, and the mass ratio of BSA to BiPt nanozyme is 1-5:1, then heat to 90°C, keep warm for 30 minutes, cool to room temperature, and stir for 6 hours to obtain solution A;

[0014] 2) Add 20 mg of protoporphyrin to a mixed solvent, then add N-hydroxysuccinimide (NHS) and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDC.HCL), the amount of NHS used is 27 mg, and the molar ratio of NHS to EDC.HCL is 1:1-2:1, then adjust the pH value to 5.5-6.5, stir and react for 2 hours, and obtain solution B, the mixed solvent consists of 2 mL of dimethylformamide (DMF) and 8 mL of PBS buffer solution;

[0015] 3) Solution A and solution B were mixed and stirred at room temperature for 12 h to obtain a BPBP solution.

[0016] Furthermore, in step (4), the specific method for obtaining the BPBP@AP antibacterial material is as follows:

[0017] 250 mg of AP was dissolved in 1 mL of dimethyl sulfoxide (DMSO), and then 4 mL of BPBP solution was added and heated at 40 °C for 5 min to obtain the BPBP@AP antibacterial material.

[0018] Furthermore, in step (1), the specific method for synthesizing BiPt nanozyme is as follows:

[0019] 150 mg of PVP was dissolved in 2 mL of anhydrous ethanol, and then 0.8 mL of deionized water, 0.3-1.2 mL of BiCl3 aqueous solution, 1.6 mL of PtCl4 aqueous solution, and 2 mL of AA (ascorbic acid) aqueous solution were added, and stirred at room temperature until the solution completely turned black, and finally ultrafiltered and washed to obtain BiPt nanozyme;

[0020] The concentration of the BiCl3 aqueous solution is 40 mM, the concentration of the PtCl4 aqueous solution is 15 mM, and the concentration of the AA aqueous solution is 1 M, where M is mol / L.

[0021] Application of antibacterial materials based on nanozyme-enhanced sonodynamic efficacy in the preparation of antibacterial drugs or anti-inflammatory drugs.

[0022] Furthermore, the antibacterial drug is a drug that inhibits Helicobacter pylori, Staphylococcus aureus or Escherichia coli.

[0023] Furthermore, the anti-inflammatory drug is a drug for treating gastritis, periodontitis and osteomyelitis caused by bacterial infection.

[0024] Beneficial effects of the present invention:

[0025] The BPBP@AP antibacterial material of the present invention can exert the enzyme activities of oxidase (POD) and catalase (CAT) in the acidic environment (pH = 4-5) of the bacterial infection site to produce strong and effective bactericidal active oxygen, and can also have a sensitizing effect on sonodynamic therapy, thereby achieving the synergistic killing of bacteria by sonodynamic therapy and nanozymes, overcoming the limitations of the hypoxic environment in the bacterial infection environment, improving the bactericidal effect, and requiring a low concentration for antibacterial treatment. At the same time, it has good biocompatibility and a broad-spectrum bactericidal effect.

[0026] In vitro experiments have shown that BPBP@AP antibacterial material combined with ultrasound can effectively inhibit the growth of -1 Under the same conditions, the inhibition rate of Helicobacter pylori exceeded 99%, and biofilm was removed. Moreover, BPBP@AP nano-antibacterial material showed high efficacy in a mouse model of Helicobacter pylori infection. Unlike triple therapy, BPBP@AP nano-antibacterial material had negligible side effects on normal intestinal microbiota.

[0027] The BPBP@AP antibacterial material of the present invention responds to external ultrasonic stimulation and produces 1 O2 has a killing effect on bacteria, which can cause the disintegration of bacterial biofilm, rupture of cell membrane, protein leakage and ultimate death, avoiding a series of risks brought by antibiotic treatment, and achieving non-invasive treatment of multiple bacterial infection sites (wounds / stomach), realizing a "one-stop" treatment that can promote the improvement of inflammation in the wound site / without affecting the balance of intestinal flora. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0029] Figure 1 Transmission electron microscopy images of antibacterial materials; a is BiPt NPs, b is BiPt@BSA-PpIX NPs; c is the lattice image of BiPt NPs, and d is the optical image of BPBP@AP antibacterial material;

[0030] Figure 2 The enzyme activity performance diagram of the antibacterial material, a is the POD enzyme activity diagram, b is the CAT enzyme activity diagram;

[0031] Figure 3 Ultrasonic generation of ROS for antibacterial materials 1 O2) Performance diagram;

[0032] Figure 4 Figure 2 shows the antibacterial effect of BPBP nanoparticles; a shows the antibacterial effect of BPBP nanoparticles at different concentrations on H.pylori, and b shows the antibacterial effect of BPBP nanoparticles at a concentration of 150 μg mL -1 When, the antibacterial effects of different groups;

[0033] Figure 5 Figure 1 is the antibacterial mechanism diagram of the nanomaterial BPBP; a is the SEM image of H.pylori bacteria after treatment in the same group; b is the removal of H.pylori biofilm after treatment in different groups; c is the protein leakage of H.pylori after treatment in different groups

[0034] Figure 6 is the biosafety diagram of antibacterial materials; a is the cell survival rate bar graph of Ges-1 cells; b is the cell survival rate bar graph of HUVEC cells; c is the cell survival rate bar graph of NIH-3T3 cells

[0035] Figure 7 The figure is a comparison of the effect of the nanomaterial in clearing H. pylori in vivo; a is the statistical analysis of the number of colonies on the gastric tissue homogenate of mice after treatment in different groups; b is the result of mixing the gastric tissue homogenate of mice with the urease indicator after treatment in different groups.

[0036] Figure 8 The antibacterial effect of BPBP nanoparticles at different ultrasound times;

[0037] Fig. 9 TEM image of BiPt nanozyme when the PVP concentration is lower than 2.5 mmol / L. DETAILED DESCRIPTION

[0038] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0039] Embodiment 1:

[0040] The method for preparing an antibacterial material based on nanozyme-enhanced sonodynamic efficacy comprises the following steps:

[0041] (1) Synthesis of BiPt nanozymes

[0042] In a 25mL round-bottom flask, 150mg PVP (Mw = 30000, polyvinyl pyrrolidone) was dissolved in 2mL of anhydrous ethanol to prepare a 2.5mmol / L PVP ethanol solution. Then, 0.8mL of deionized water, 0.6mL of 40mM BiCl3 aqueous solution, 1.6mL of 15mM PtCl4 aqueous solution and 2mL of 1M AA aqueous solution were added to the above solution to form a light yellow solution. The reaction solution was stirred at room temperature for 6 hours until the solution turned completely black. The reaction solution was then concentrated with an ultrafiltration tube with a molecular weight cutoff of 50,000 and washed 3 times. The obtained nanozyme product was stored at 4°C after vacuum drying.

[0043] In this step, if an ethanol solution with a concentration of less than 2.5 mmol / L PVP is used, the BiPt nanozyme obtained by the one-step reduction method has an unstable morphology and a non-uniform particle size. Fig. 9 As shown in the figure, when the concentration of PVP is higher than 2.5mmol / L, the high concentration of PVP inhibits the nucleation of bimetallic and makes it difficult to purify the product by centrifugation. In summary, the concentration of PVP is 2.5mmol / L.

[0044] (2) Synthesis of BiPt@BSA-PpIX (BPBP) nanoparticles

[0045] 20 mL of an aqueous solution containing 20 mg BiPt and 100 mg BSA was heated to 90 °C for 30 min and then kept at room temperature for 6 h to obtain solution A;

[0046] 42.30 mg of EDC.HCL and 27.00 mg of NHS were mixed with 20 mg of PpIX in a mixed solution of 2 mL of DMF and 8 mL of PBS to obtain a reaction mixture, the pH value of the reaction mixture was adjusted to about 5.5, and stirred for 2 h to obtain a solution B;

[0047] Solution A and solution B were mixed and stirred at room temperature for 12 h to obtain a BPBP solution.

[0048] (3) Preparation of BPBP@AP antibacterial materials

[0049] 250 mg AP was dissolved in 1 mL dimethyl sulfoxide (DMSO) until clear; 4 mL of the above-synthesized BPBP solution was slowly added and heated at 40°C for 5 minutes. Rinse with sterile PBS 5 times and store in a sealed container at 4°C.

[0050] Embodiment 2:

[0051] This embodiment is basically the same as Example 1, except that during the synthesis of BiPt nanozyme, the amount of 40mM BiCl3 aqueous solution used is 1.2mL, and the amount of 15mM PtCl4 aqueous solution used is 1.6mL.

[0052] The BiPt nanoparticles prepared in Example 1 were dispersed on a copper mesh and observed using a transmission electron microscope (TEM). The results are as follows: Figure 1 As shown in a, the prepared BiPt nanoparticles have a uniform particle size of about 20 nm. The BPBP nanoparticles are dispersed on a copper mesh and observed using a transmission electron microscope (TEM). Figure 1 As shown in b, BPBP nanoparticles are evenly distributed, regular in shape, and have a particle size of about 200 nm.

[0053] The lattice of BiPt nanoparticles is Figure 1 As shown in (c), the BiPt nanoparticles have good crystallinity, orderly stripe patterns, and lattice spacings of 0.221 and 0.244 nm, corresponding to the planes of Pt and Bi, respectively.

[0054] Figure 1 d is an optical microscopy image of the BPBP@AP material, which shows the lamellar structure in the AP hydrogel, and high-order fiber assembly was observed when BPBP nanoparticles were encapsulated in the hydrophobic core.

[0055] The BiPt nanozyme and BPBP nanoparticles prepared in Example 1 were characterized for the oxidase (POD) enzyme activity and the catalase (CAT) enzyme activity, as follows: The characterization of the POD enzyme activity was performed using the TMB colorimetric method, because the nanozyme with POD enzyme activity can catalyze H2O2 in a physiological environment to generate ·OH, and at the same time, the colorless TMB is converted into blue oxidized TMB (TMBox), which has a maximum characteristic absorbance at 652nm. Figure 2 As shown in a, compared with the control group pure water (ddH2O), TMB in the EP tube of BiPt nanoparticles and BPBP nanoparticles was oxidized to blue, and both showed higher enzyme activity under acidic conditions (pH = 4.0).

[0056] Characterization of CAT enzyme activity The Ru(bpy)3Cl2 oxygen probe was used to evaluate the catalase (CAT) enzyme activity of BPBP nanoparticles. The fluorescent dye Ru(bpy)3Cl2 is an oxygen-sensitive indicator, and the fluorescence intensity decreases with the increase of oxygen content in the system. Figure 2 As shown in b, the luminescence intensity of Ru(bpy)3Cl2 in the control group (pure water) was higher than that in the BiPt and BPBP groups, indicating that a large amount of oxygen was produced in both, and more was produced under acidic conditions (pH = 4.0).

[0057] At a power of 1 W cm -2 , the frequency was 1 MHz, and the duty cycle was 50% (3 seconds for ultrasound and 3 seconds for rest). The antibacterial time of ultrasound was explored, and the concentration of Helicobacter pylori was adjusted to 1×10 6 CFU mL -1 , then with 150 μg mL -1 The mixture was treated with ultrasound for 0, 1, 2, and 3 min, and 10 μL of the mixture was plated for growth. The blood plate was cultured under microaerobic conditions for 2 to 3 days. Figure 8 As shown in the figure, 1 minute cannot completely kill Helicobacter pylori, so the ultrasound time is selected as 2 minutes.

[0058] The sonodynamic properties of the BPBP nanoparticles prepared in Example 1 were characterized as follows: DPBF is a 1 O2-sensitive fluorescent probe, which can be indicated by its absorbance change at 425nm 1 O2 production. A PpIX combined with ultrasound (PpIX+US) group and a BPBP+US group were established for comparison. Figure 3 As shown in Figure 2, the sonodynamic performance of the PpIX+US group was poor, while that of the BPBP+US group was significantly improved. 1 Compared with the neutral environment (pH = 7.4), the production of O2 in the acidic environment (pH = 4.0)1 Higher O2 production.

[0059] In vitro clearance effect of BPBP nanoformulation on H.pylori:

[0060] The concentration of Helicobacter pylori was adjusted to 1×10 6 CFU mL -1 , and then mixed with BPBP nanoparticles of different concentrations in equal volumes. The mixture was ultrasonically treated for 2 min (1 W cm -2 , 1MHz, 50% duty cycle). Finally, 100 μL of the bacterial solution was inoculated onto a Columbia agar plate containing 7% sterile defibrinated sheep blood and counted after three days of incubation. According to the colony plate image and colony counting results, Figure 4 As shown in a, the minimum bactericidal concentration (MBC) of BPBP nanoparticles was determined to be 150 μg mL -1 .

[0061] The concentration of BPBP nanoparticles was 150 μg mL -1 , Helicobacter pylori (1×10 6 CFU mL -1 ) were grouped and treated as follows: control group (bacterial solution plus pH=7 PBS mixed solution), US (ultrasound) group, BPBP group, and BPBP combined with ultrasound group (BPBP+US group). Figure 4 b shows that the BPBP+US group is effective against Helicobacter pylori in vitro, with an inhibition rate of more than 99%, while the use of US or BPBP alone cannot completely inhibit the growth of bacteria, with inhibition rates of approximately 49.77% and 27.14%, respectively. The above experimental results prove that the external stimulation of ultrasound or the enzyme activity of nanomaterials alone is not enough to completely inhibit the growth of Helicobacter pylori, and the BPBP+US group has a better anti-Helicobacter pylori effect, which indirectly confirms the necessity of combining nanomaterials with sonodynamic therapy.

[0062] Antibacterial mechanism of BPBP nanoformulation:

[0063] The effects of BPBP nanoparticles combined with ultrasound on Helicobacter pylori in vitro were investigated using scanning electron microscopy (SEM). Figure 5 The image in a shows that the membrane of Helicobacter pylori in the control group was intact and clear. -1 The membrane of Helicobacter pylori cultured with BPBP nanoparticles began to show wrinkles, and the bacteria in the BPBP+US group were the most severely damaged, with fragmentation, breakage, and blurring. Figure 5 b shows that the protein leakage level of the BPBP+US group was the highest and the bacterial cell membrane was most severely damaged.

[0064] Helicobacter pylori has the ability to form biofilms, which increases the survival rate of bacteria and poses a challenge to antibiotic treatment, so it is crucial to eradicate Helicobacter pylori biofilms. First, the Helicobacter pylori biofilm was cultured statically at 37°C and microaerobic conditions for 4-5 days to successfully culture it, and then treated according to different subgroups. Subsequently, the biofilm was stained with crystal violet and quantified at OD = 590nm. The pictures after staining and the quantitative results of enzyme labeling are shown in Figure 5 As shown in Figure c, the biofilm formed by Helicobacter pylori in the control group was relatively intact and tightly connected. After ultrasound treatment, the biofilm lost its tight junctions and became dispersed, with the membrane content reduced to 70.69±3.59% of the control group. The combined use of ultrasound (US) and BPBP produced even less biofilm, only 29.22±2.48% of the control group. Ultrasound alone destroyed half of the biofilm, which may be attributed to the mechanical dispersion induced by ultrasound. These results illustrate that the antibacterial activity of SDT mediated by BPBP is related to the destruction of biofilm by ROS formation, bacterial cell membrane rupture, and protein leakage.

[0065] Biocompatibility evaluation of BPBP:

[0066] The CCK-8 method was used to evaluate the cytotoxicity of BPBP nanoparticles on mouse gastric epithelial cells (Ges-1), human umbilical vein endothelial cells (HUVEC) and fibroblasts (NIH-3T3). The cells were seeded into 96-well cells (1×10 5 Cells were incubated at 37°C in a 5% CO2 incubator for 24 h and treated with different concentrations of BPBP nanoparticles for 24 and 72 h. After treatment, the cells were washed three times with sterile PBS and fresh culture medium containing CCK8 reagent (100 μL) was added each time. The cells were incubated at 37°C for 4 h and the absorbance at 495 nm was measured for each well. The results are shown in Figure 6 As shown, when the concentration of BPBP nanoparticles reached 200 μg / mL, the survival rate of the three cells remained above 80%, indicating that BPBP nanoparticles have good biocompatibility.

[0067] Evaluation of the efficacy of BPBP@AP antimicrobial preparation in eradicating H.pylori in vivo:

[0068] By plating gastric tissue homogenates of mice treated in different groups and detecting urease indicators, it was found that the antibacterial effect of BPBP@AP+US treatment was similar to that of OAC treatment, and the CFU was reduced by more than 99% compared with that of PBS-treated mice ( Figure 7a). Although the number of bacteria was significantly reduced (>50%), the antibacterial activity of the US alone group was weaker than that of the BPBP@AP+US group and the OAC treatment group, indicating that BPBP@AP has a synergistic antibacterial effect in vivo. In addition, the negative result of the urease test, an important clinical detection indicator of Helicobacter pylori ( Figure 7 b) also showed that Helicobacter pylori was cleared from the stomach of BPBP@AP-treated mice, similar to that of mice treated with OAC group.

[0069] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. Antibacterial material based on nanozyme-enhanced sonodynamic efficacy, characterized by: It includes BiPt nanozyme, the outside of BiPt nanozyme is coated with BSA, BSA is connected to PpIX to form BPBP nanoparticles, and the outside of BPBP nanoparticles is coated with AP hydrogel to obtain BPBP@AP antibacterial material.

2. The method for preparing the antibacterial material based on nanozyme-enhanced sonodynamic efficacy according to claim 1, characterized in that: The following steps are involved: (1) Synthesis of BiPt nanozymes; (2) BSA was coated on the outside of BiPt nanozyme and connected to PpIX through EDC / NHS biochemical conjugation activation to form BPBP nanoparticles; (3) BPBP nanoparticles were encapsulated in AP hydrogel to obtain BPBP@AP antibacterial material.

3. The preparation method according to claim 2, characterized in that: In step (2), the specific method for forming BPBP nanoparticles is as follows: 1) Disperse 20 mg of BiPt nanozyme in 20 mL of aqueous solution, add BSA, and the mass ratio of BSA to BiPt nanozyme is 1-5:1, then heat to 90°C, keep warm for 30 minutes, cool to room temperature, and stir for 6 hours to obtain solution A; 2) Add 20 mg of protoporphyrin to the mixed solvent, then add NHS and EDC.HCL, the amount of NHS is 27 mg, the molar ratio of NHS to EDC.HCL is 1:1-2:1, then adjust the pH value to 5.5-6.5, stir and react for 2 hours to obtain solution B, the mixed solvent consists of 2 mL of dimethylformamide (DMF) and 8 mL of PBS buffer solution; 3) Solution A and solution B were mixed and stirred at room temperature for 12 h to obtain a BPBP solution.

4. The preparation method according to claim 2 or 3, characterized in that: In step (4), the specific method for obtaining the BPBP@AP antibacterial material is as follows: 250 mg of AP was dissolved in 1 mL of DMSO, and then 4 mL of BPBP solution was added and heated at 40 °C for 5 min to obtain the BPBP@AP antibacterial material.

5. The preparation method according to claim 2, characterized in that: In step (1), the specific method for synthesizing BiPt nanozyme is as follows: 150 mg of PVP was dissolved in 2 mL of anhydrous ethanol, and then 0.8 mL of deionized water, 0.3-1.2 mL of BiCl3 aqueous solution, 1.6 mL of PtCl4 aqueous solution and 2 mL of AA aqueous solution were added, and stirred at room temperature until the solution completely turned black, and finally ultrafiltered and washed to obtain BiPt nanozyme; The concentration of the BiCl3 aqueous solution was 40 mM, the concentration of the PtCl4 aqueous solution was 15 mM, and the concentration of the AA aqueous solution was 1 M.

6. Use of the antibacterial material based on nanozyme-enhanced sonodynamic efficacy as claimed in claim 1 in the preparation of antibacterial drugs or anti-inflammatory drugs.

7. The use according to claim 6, characterized in that: The antibacterial drug is a drug that inhibits Helicobacter pylori, Staphylococcus aureus or Escherichia coli.

8. The use according to claim 6, characterized in that: The anti-inflammatory drug is a drug for treating gastritis, periodontitis and osteomyelitis caused by bacterial infection.

Images