An antibacterial biological coating and its preparation method and application

Abstract

The present invention belongs to the field of antimicrobial medical devices, and specifically relates to an antimicrobial biocoating, its preparation method, and its application. The present invention provides an antimicrobial biocoating comprising a porous photocrosslinked hydrogel and Saccharomyces boulardii, wherein the Saccharomyces boulardii aggregates and grows in the pores of the hydrogel. The antimicrobial biocoating provided by the present invention significantly inhibits the adhesion of Candida albicans to the surface of medical devices; under adverse conditions, the survival rate of Saccharomyces boulardii is higher; it reduces the risk of fungal infection caused by endotracheal intubation intervention, and also has an inhibitory effect on Escherichia coli and Staphylococcus aureus, and has a wide range of applications.

Description

Technical Field

[0001] The present invention belongs to the field of antibacterial medical devices, and specifically relates to an antibacterial biological coating and a preparation method and application thereof. Background Art

[0002] Tracheal intubation improves ventilation and increases ventilation, making it a common supportive treatment for critically ill patients. However, once placed in the patient's airway, the tube can easily damage the airway mucosa through compression, friction, and irritation, disrupting the airway barrier and internal environment, and promoting the growth and reproduction of pathogens (Chen X et al. Antimicrobial coating: tracheal tube application. International Journal of Nanomedicine, 2022, 17:1483). In addition, endotracheal intubation provides conditions for the adhesion of pathogens and the formation of biofilms, which can easily cause intubation complications such as pathogen infection, seriously endangering the patient's life safety (Wang Y et al. A novel antibacterial and antifouling nanocomposite coated endotracheal tube to prevent ventilator-associated pneumonia. Journal of nanobiotechnology, 2022, 20 (1): 1-19; Thorarinsdottir HR et al. Biofilm formation on three different endotracheal tubes: a prospective clinical trial. Critical Care, 2020, 24 (1): 1-12). Pathogenic infections caused by endotracheal intubation include bacterial and fungal infections. Bacterial infections can be treated with drugs, but pathogenic fungi belong to eukaryotic organisms. The use of antifungal drugs can easily damage host eukaryotic cells and produce a series of toxic side effects. Therefore, preventing pathogenic fungal infections caused by endotracheal intubation has become an urgent problem to be solved in clinical treatment.

[0003] Among pathogenic fungal infections, the mortality rate of candidemia caused by Candida albicans is as high as 40% (Poulain D. et al. Candida albicans, plasticity and pathogenesis. Critical reviews in microbiology, 2015, 41(2): 208-217). This is mainly due to: (I) endotracheal intubation disrupts the balance of airway flora, leading to the massive growth and reproduction of Candida albicans normally present on the mucosa (Li H et al. Interactions between Candida albicans and the resident microbiota. Frontiers in Microbiology, 2022, 13: 930495); (II) Candida albicans has strong adhesion properties and easily colonizes on the surface of medical devices to form dense biofilms, which are highly tolerant to external environmental pressures (Wall G et al. Candida albicans biofilm growth and dispersal: contributions to pathogenesis. Current Opinion in Microbiology, 2019, 52: 1-6); (III) Candida albicans has a strong yeast-hyphae morphogenesis ability, which can change its surface antigen characteristics and evade the attack of the host immune system, resulting in an enhanced ability to infect the host (Xie J et al. White-opaque switching in natural MTL a / α isolates of Candida albicans: evolutionary implications for roles in host adaptation, pathogenesis, and sex. PLOS Biology, 2013, 11(3):e1001525).

[0004] With the widespread use of antifungal drugs, Candida albicans has gradually developed strong drug resistance. Chinese patent application 201911166967.6 discloses a pharmaceutical composition for treating invasive infections caused by Candida albicans resistant to echinocandin drugs and its use. The active ingredients of the pharmaceutical composition are composed of caspofungin and compound L-269289. In in vitro inhibition of echinocandin-resistant strains of Candida albicans, the mass ratio of caspofungin to compound L-269289 is 1:3-1:36. In the treatment of invasive infections of Candida albicans resistant to echinocandin drugs in mice, the mass ratio of caspofungin to compound L-269289 is 1:10. A Chinese patent application discloses an antifungal drug composition, which is composed of Xinbai Biyuan mixture and antifungal drugs in a mass ratio of 2077.5:1, wherein the antifungal drugs include but are not limited to: fluconazole, voriconazole, itraconazole, etc.; the antifungal drug composition can be used to prepare a dosage form for treating Candida albicans infection.

[0005] All of the above drugs are anti-Candida albicans compounds and are prone to drug resistance. Therefore, preventing C. albicans from adhering and multiplying on the surface of endotracheal tubes can effectively prevent fungal infections caused by C. albicans and reduce the risk of death in critically ill patients, while preventing drug resistance, and has become a scientific problem that urgently needs to be solved in clinical treatment. Summary of the Invention

[0006] In order to overcome the defects of the prior art, the present invention proposes an antibacterial biological coating.

[0007] In their research, the inventors discovered that Saccharomyces boulardii (S. boulardii) can effectively inhibit the adhesion of Candida albicans, but its practical application presents numerous difficulties. Free Saccharomyces boulardii cannot colonize on the surface of medical devices, preventing its beneficial function against pathogens and making it difficult to recover. Encapsulating microorganisms in biomaterials does not completely colonize them. Microorganisms have strong reproductive capacity and are extremely prone to escaping from biomaterials, potentially disrupting the surrounding microbial population.

[0008] The inventors surprisingly discovered that confining the growth of Saccharomyces boulardii in a biological coating can solve the problem of its difficulty in aggregating and growing on the surface of medical devices, prevent its escape, and effectively reduce Candida albicans infections caused by medical devices.

[0009] In order to achieve the above technical objectives, the technical solutions proposed by the present invention are as follows:

[0010] In one aspect, the present invention provides an antimicrobial bio-coating comprising a porous photo-crosslinked hydrogel and Saccharomyces boulardii, wherein the Saccharomyces boulardii aggregates and grows in the pores of the hydrogel (microorganisms settle together to form a tightly connected whole).

[0011] The antibacterial biological coating provided by the present invention realizes the confined growth of Saccharomyces boulardii and does not overflow from the hydrogel during the growth process, thereby enabling it to better exert its antibacterial function.

[0012] In a preferred embodiment, the hydrogel is GelMA or a derivative thereof.

[0013] In another aspect, the present invention provides a method for preparing the antibacterial biological coating, the method comprising the following steps:

[0014] (1) dissolving an initiator in a solvent and heating until dissolved to obtain an initiator solution;

[0015] (2) dissolving the reaction monomer in the initiator solution and heating until dissolved to obtain a premixed solution;

[0016] (3) uniformly mixing the premixed solution with a water-soluble porogen and a Saccharomyces boulardii solution to obtain a mixed solution, and cooling the solution until a gel-like mixture is formed;

[0017] (4) The gel-like mixture is photocrosslinked to obtain an antibacterial biocoating.

[0018] In a preferred embodiment, the solvent may be sterile water.

[0019] In one embodiment, the initiator is lithium phenyl (2,4,6-trimethylbenzoyl) phosphate or 2-hydroxy-2-methyl-1-phenyl-1-propanone.

[0020] In one embodiment, the reactive monomer is gelatin methacryloyl (GelMA) or a derivative thereof.

[0021] In one embodiment, the final concentration of the reactive monomer in the mixed solution may be about 5% by weight to volume, where the weight is measured in g and the volume is measured in mL.

[0022] In one embodiment, the water-soluble porogen is polyethylene oxide (PEO).

[0023] In one embodiment, the final concentration of the water-soluble porogen in the mixed solution is 0.8% by mass to volume, with the mass being measured in g and the volume being measured in mL.

[0024] In one embodiment, the photocrosslinking is ultraviolet light or blue light crosslinking.

[0025] In a preferred embodiment, the ultraviolet light is 365 nm ultraviolet light.

[0026] In one embodiment, the photocrosslinking time is 15-30 s.

[0027] In one embodiment, the concentration of the Saccharomyces boulardii solution is 0.1×10 6 -0.1×10 8 cells / mL.

[0028] In one embodiment, the heating conditions in steps (1) and (2) are 50-60°C.

[0029] In one embodiment, the Saccharomyces boulardii bacterial solution is obtained by culturing Saccharomyces boulardii in a yeast extract peptone dextrose agar medium.

[0030] In a preferred embodiment, the culture conditions are:

[0031] 1) Temperature is 27-40°C;

[0032] 2) The rotation speed is 160-220rpm.

[0033] In yet another aspect, the antibacterial biological coating or the antibacterial biological coating prepared by the method is used in the antibacterial treatment of medical devices.

[0034] In one embodiment, the fungi include fungi and bacteria.

[0035] In a preferred embodiment, the fungus comprises Candida albicans;

[0036] And / or, the bacteria include Escherichia coli and Staphylococcus aureus.

[0037] In one embodiment, the medical device can be a basic surgical knife, such as a lancet, a scalpel; a disposable sterile medical device, such as a disposable sterile endotracheal tube; an orthopedic implant medical device, such as a surgical implant joint prosthesis, etc.

[0038] In a preferred embodiment, the medical device may be a disposable sterile endotracheal tube.

[0039] The antibacterial biological coating provided by the present invention has at least the following beneficial effects:

[0040] (1) The microorganisms are aggregated and grown in the hydrogel, preventing them from escaping from the hydrogel;

[0041] (2) The antibacterial bio-coating significantly inhibited the adhesion of Candida albicans to the surface of medical devices, and compared with the control group, the adhesion of Candida albicans was reduced by 60%;

[0042] (3) In 2 mg / mL copper ion solution and 5 mmol / mL H2O2 solution, the survival rate of Saccharomyces boulardii increased by 20%, significantly improving its ability to resist stress;

[0043] (4) The risk of fungal infection caused by endotracheal intubation was reduced. The number of CFU colonies in mice was reduced by 70% compared with the control group, and no obvious damage was found in the kidney tissue of the mice.

[0044] (5) The antibacterial coating also has an inhibitory effect on Escherichia coli and Staphylococcus aureus, significantly inhibiting the activity of Escherichia coli and Staphylococcus aureus. Compared with the control group, the survival rate of Escherichia coli after 4 hours of co-culture was significantly reduced by 30%, and the survival rate of Staphylococcus aureus was significantly reduced by 20%. BRIEF DESCRIPTION OF THE DRAWINGS

[0045] Figure 1 Schematic diagram of Saccharomyces boulardii growth within hydrogel cavities for 6 and 36 hours. The GelMA+Saccharomyces boulardii group represents the direct mixing of GelMA and Saccharomyces boulardii followed by solidification and subsequent incubation. The GelMA+PEO+Saccharomyces boulardii group represents the direct mixing of GelMA, PEO, and Saccharomyces boulardii followed by solidification and subsequent incubation.

[0046] Figure 2 is the OD of Saccharomyces boulardii grown in the hydrogel pores at 0 h, 18 h, and 36 h 600 Statistical chart.

[0047] Figure 3 Schematic diagram of the distribution of Saccharomyces boulardii in the hydrogel. BF represents bright field, CFW (Calcofluor white) represents fluorescence imaging of Saccharomyces boulardii labeled with CFW fluorescent dye, and the Before group represents cells not cultured in YPD medium; the After group represents cells cultured in YPD medium for 36 hours.

[0048] Figure 4 The top and side views of the 3D reconstructed Saccharomyces boulardii distribution within the hydrogel. Fluorescence microscopy was used to observe the distribution of Saccharomyces boulardii within the biocoating. Saccharomyces boulardii was stained with CFW and observed using an all-in-one fluorescence microscope. (A) The top view and B the side view of the 3D reconstructed Saccharomyces boulardii.

[0049] Figure 5The following is a scanning electron micrograph of the antimicrobial biocoating. The aggregation of Saccharomyces boulardii within the biocoating was observed using a scanning electron microscope. A is a scanning electron micrograph of Saccharomyces boulardii within the biocoating, B is a magnified view of a section of A, C is a magnified view of a section of A, and D is a magnified view of a section of C.

[0050] Figure 6 Schematic diagram of PI staining of free Saccharomyces boulardii (free) and antibacterial bio-coating (encapsulation) after culture in copper ion solution.

[0051] Figure 7 Schematic diagram of PI staining of free Saccharomyces boulardii (free) and antibacterial bio-coating (encapsulation) after culture in H2O2 solution.

[0052] Figure 8 Fluorescence was used to observe the survival of Candida albicans after treatment with Candida albicans alone, Candida albicans + S. boulardii, Candida albicans + GelMA hydrogel (the hydrogel does not contain S. boulardii), and Candida albicans + S. boulardii bioactive material. The Candida albicans used was a GFP-labeled modified strain. BF stands for bright field, and the fluorescent marker is Csp37-GFP.

[0053] Figure 9 This is the survival status of Escherichia coli and Staphylococcus aureus after the antibacterial bio-coating was co-incubated with Escherichia coli and Staphylococcus aureus respectively. The cell survival was determined by Calcein-AM / PI live cell / dead cell double staining kit.

[0054] Figure 10 The CFU colony counts of Candida albicans in mouse kidneys, where a is a tracheal cannula; b is a tracheal cannula + GelMA hydrogel coating; c is a tracheal cannula + Saccharomyces boulardii antibacterial biological coating.

[0055] Figure 11 HE staining of mouse kidneys. (a) Intubation group; (b) Intubation + GelMA hydrogel coating group; (c) Intubation + Saccharomyces boulardii antimicrobial biocoating group. Scale bars: 100 μm. DETAILED DESCRIPTION

[0056] Unless otherwise defined, all technical and scientific terms used in the present invention have the same meaning as commonly used in the field to which the present invention belongs. For the purpose of interpreting this specification, the following definitions will apply, and where appropriate, terms used in the singular will also include the plural form, and vice versa.

[0057] Unless the context clearly dictates otherwise, as used herein, the expressions "a" and "an" include plural references. For example, reference to "a cell" includes a plurality of such cells and equivalents thereof known to those skilled in the art, and so forth.

[0058] As used herein, the term "about" refers to a range of ±20% of the value that follows. In some embodiments, the term "about" refers to a range of ±10% of the value that follows. In some embodiments, the term "about" refers to a range of ±5% of the value that follows.

[0059] The term "medical device" as used herein refers to instruments, equipment, appliances, in vitro diagnostic reagents and calibrators, materials and other similar or related items that are used directly or indirectly on the human body.

[0060] The term "aggregate growth" used herein refers to the initial growth of microorganisms within a specific area. As the culture time increases, the microorganisms aggregate in the specific area (microorganisms grow in clusters or piles).

[0061] To make the objectives, technical solutions, and advantages of the present invention more clearly understood, the present invention will be further described in detail below with reference to the following examples. The specific embodiments described herein are intended only to illustrate the present invention and are not intended to limit the present invention in any way. In addition, in the following description, descriptions of known structures and technologies are omitted to avoid unnecessary confusion about the concepts of the present disclosure. Such structures and technologies are also described in many publications.

[0062] Example 1 Preparation method of antibacterial biological coating

[0063] The present invention constructs an antibacterial biological coating for realizing the aggregated growth of Saccharomyces boulardii through a one-step photocrosslinking method.

[0064] (1) Photoinitiator activation: 10 mL of sterile water was mixed with 25 mg of lithium phenyl (2, 4, 6-trimethylbenzoyl) phosphinate (LAP), and the mixture was heated in a 50°C water bath for 10 minutes to dissolve the mixture. During the heating, the mixture was shaken several times to ensure that the solution was fully mixed to obtain an initiator solution. The mixture was cooled to room temperature and stored at 4°C in the dark.

[0065] (2) Saccharomyces boulardii culture: Saccharomyces boulardii (ura3 / ura3) was cultured overnight in Yeast Peptone Dextrose Agar (YPD) medium (1% yeast extract, 2% peptone, and 2% glucose) at 30°C and 160 rpm. Before use, the Saccharomyces boulardii was centrifuged and collected in PBS buffer to obtain a Saccharomyces boulardii culture solution.

[0066] (3) Photocrosslinking: 10% GelMA, 0.25% LAP (W / V) and 1.6% PEO (W / V) were mixed with 10 μL of Saccharomyces boulardii culture (culture concentration was 0.1×10 6 cells / mL) were mixed to obtain a mixed solution, which was then frozen at 4°C for 30 minutes to convert from a sol-like state to a gel-like state, thereby obtaining a gel-like mixture. The mixture was then irradiated under a 365 nm UV lamp for 30 seconds to achieve polymerization of the GelMA monomer. The final concentration of GelMA in the mixed solution was 5%, and the final concentration of PEO was 0.8%.

[0067] The mixing step (3) includes: dissolving 100 mg of GelMA solid in 1 mL of 0.5% (W / V) LAP solution, and shaking to fully soak the GelMA; placing it in a 60°C water bath in the dark and heating to dissolve for 30 minutes, shaking it several times during the process to ensure that the GelMA is completely dissolved (the GelMA solution is sterilized by filtering it through a 0.22 μm filter membrane to prevent GelMA from gelling at low temperatures), and obtaining a 10% (W / V) GelMA solution (premixed solution); dissolving 16 mg of PEO powder in 1 mL of 0.5% (W / V) LAP solution to obtain 1.6% PEO (W / V); mixing the obtained 10% GelMA solution (premixed solution) with a 1.6% PEO solution in a volume ratio of 1:1 to obtain a prepolymer solution; and finally mixing the prepolymer solution with 10 μL of Saccharomyces boulardii liquid to obtain a mixed solution.

[0068] Example 2 Characterization of the survival of yeast in the antibacterial biological coating

[0069] In order to clarify the survival of Saccharomyces boulardii in the hydrogel, the inventors first used a microscope to observe the growth of the encapsulated hydrogel and found that as time went on, single Saccharomyces boulardii in the pores gradually aggregated and formed aggregates ( Figure 1 The cells were further lysed using GelMA lysis buffer at 37°C for 1 hour, and the Saccharomyces boulardii content of the bio-coatings at different incubation times was determined. It was found that the number of Saccharomyces boulardii in the bio-coatings increased by 5-6 times after 18 hours of incubation, and by 12-14 times after 36 hours of incubation ( Figure 2In the experimental group without porogen, almost all the Saccharomyces boulardii bacteria overflowed, and no aggregation growth was observed ( Figure 1 The above results indicate that Saccharomyces boulardii can reproduce and grow normally in the antibacterial bio-coating, but cannot reproduce and grow normally in the non-porous hydrogel.

[0070] Example 3 Morphological Characterization of Antibacterial Biocoating

[0071] To further clarify the distribution and population status of Saccharomyces boulardii within the biomaterial, the inventors used CFW (purchased from Shanghai Yuanye Biotechnology Co., Ltd., CAS No. 4404-43-7) staining to fluorescently label Saccharomyces boulardii and observed the smears before and after the antibacterial biocoating was placed in YPD medium using an inverted fluorescence microscope. The results showed that after culturing in YPD medium, Saccharomyces boulardii in the hydrogel appeared as blue aggregates ( Figure 3 ), and the formed colony balls are evenly distributed inside the biomaterial ( Figure 4 In addition, the inventors used a scanning electron microscope to observe the state of the Saccharomyces cerevisiae in the antibacterial bio-coating. The results showed that the Saccharomyces cerevisiae in the pores of the antibacterial bio-coating were aggregated ( Figure 5 In summary, this embodiment shows that the inventors have successfully constructed a confined Saccharomyces boulardii bio-coating and achieved the aggregated growth of Saccharomyces boulardii in the hydrogel.

[0072] Example 4 Characterization of the Anti-Adversity Ability of Antibacterial Bio-Coating

[0073] The free Saccharomyces boulardii and the antibacterial coating were placed in a 2mg / mL copper ion solution and a 5mmol / mL H2O2 solution, respectively. The survival of the free Saccharomyces boulardii and the Saccharomyces boulardii in the antibacterial coating was determined by PI staining (purchased from Beijing Solebau Technology Co., Ltd., catalog number IP5030). The results showed that under the adverse conditions of oxidative stress and heavy metal ions, the number of cell death in the free Saccharomyces boulardii increased by 20%, while the survival rate of the Saccharomyces boulardii cells in the antibacterial coating increased by 20% ( Figure 6 and Figure 7 ).

[0074] Example 5 Antagonistic Effect of Antimicrobial Biocoating on Bacteria / Fungi

[0075] 5.1 Antagonistic effect of antibacterial bio-coating on Candida albicans

[0076] This example measures the adhesion of Candida albicans to the surface of the tracheal tube coated with the antibacterial biological coating and the control group. First, the inventors used the pUC18-GFP-URA3 plasmid (wherein the pUC18-GFP-URA3 plasmid is a modified plasmid, which can be obtained through the research group of Wei Yunyun of Shandong First Medical University) as a template to amplify the Csp37-GFP gene marker box with primers (the primers used are consistent with the primers in Function of Atg11in non-selective autophagy and selective autophagy of Candidaalbicans. Biochemical and Biophysical Research Communications 516 (4), 1152–1158), and transferred the Candida albicans (ura3 / ura3) strain through PCR-mediated homologous recombination technology to obtain GFP-labeled Candida albicans. Based on this, the inventors tested the adhesion of the biomaterial to Candida albicans and found that compared with the pure GelMA coating group, the number of green fluorescent-labeled Candida albicans on the surface of the antibacterial biological coating was reduced by 60% ( Figure 8 ). The results showed that the antibacterial bio-coating could significantly inhibit the adhesion of Candida albicans.

[0077] 5.2 Antagonistic effects of antimicrobial organisms on Escherichia coli and Staphylococcus aureus

[0078] Escherichia coli and Staphylococcus aureus were cultured in LB medium, and then Escherichia coli and Staphylococcus aureus were collected and transferred to YPD medium. The bacterial count was adjusted to 1×10 7 cells / mL, then add 1 mL 3 After solidification, the Saccharomyces boulardii bio-coating was co-cultured at 30°C and 160 rpm for 4 hours. The survival of Escherichia coli and Staphylococcus aureus was then observed using the Calcein-AM / PI live / dead cell double staining kit (Beijing Solebeau Technology Co., Ltd., Cat. No. CA1630). The results showed that the antibacterial coating significantly inhibited the activity of Escherichia coli and Staphylococcus aureus. Compared with the control group (Control), the survival rate of Escherichia coli in the co-culture group (Co-culture) after 4 hours was significantly reduced by 30%, and the survival rate of Staphylococcus aureus was significantly reduced by 20% ( Figure 9 ).

[0079] Example 6 Verification of the antibacterial effect of antibacterial biological coating in a mouse model

[0080] ICR female mice (purchased from Jinan Pengyue Experimental Animal Breeding Co., Ltd.) were randomly divided into three groups (10 mice in each group) and then fed for 7 consecutive days. The tracheal cannula, the tracheal cannula covered with hydrogel and Saccharomyces boulardii active material, was placed in a Candida albicans solution (1×10 7 cells / mL) for 24 h; the treated endotracheal tubes were cut into 1 cm; the endotracheal tubes were transplanted into the abdominal cavity of mice in a sterile room; the kidneys of each group of mice were collected after 7 days; tissue sections were made and HE staining was performed; and the CFU load of Candida albicans in the mouse kidneys was counted.

[0081] First, the inventors constructed an antibacterial biological coating coated on the tracheal cannula based on the method of Example 1. The results showed that the antibacterial biological coating was evenly coated on the surface of the tracheal cannula. In addition, the inventors co-incubated the tracheal cannula, the hydrogel-coated tracheal cannula, and the tracheal cannula coated with the antibacterial biological coating with Candida albicans for 24 hours, and then transferred them into the abdominal cavity of mice using abdominal transplantation surgery. After 7 consecutive days of feeding, the inventors counted the CFU of Candida albicans colonies in the kidneys of mice in different treatment groups (a: tracheal cannula group; b: hydrogel-coated tracheal cannula group; c: antibacterial biological coating-coated tracheal cannula group) and found that the number of CFU colonies of Candida albicans in the kidneys of mice in the tracheal cannula group coated with the antibacterial biological coating was reduced by 70% ( Figure 10 In addition, HE staining (Hematoxylin-eosin staining kit purchased from Beyotime, item number C0105S) revealed that significant damage occurred in the kidneys of mice in the tracheal intubation group without antibacterial biological coating and the tracheal intubation group coated with hydrogel ( Figure 11 ). The above results show that the endotracheal tube coated with antibacterial biological coating can reduce the chance of Candida albicans infection in mice by 70%.

[0082] Finally, it should be noted that the above content is only used to illustrate the technical solution of the present invention, rather than to limit the scope of protection of the present invention. Simple modifications or equivalent substitutions of the technical solution of the present invention by ordinary technicians in this field do not deviate from the essence and scope of the technical solution of the present invention.

Claims

1. An antibacterial biological coating, characterized in that: The invention is composed of a porous photo-crosslinked hydrogel and Saccharomyces boulardii, wherein the Saccharomyces boulardii aggregates and grows in the pores of the hydrogel. The preparation method of the antibacterial biological coating comprises the following steps: (1) Take 10 mL of sterile water and mix it with 50 mg of lithium phenyl (2,4,6-trimethylbenzoyl) phosphate, heat it in a 50 ° C water bath, dissolve it for 10 minutes, shake it several times during the period to ensure that the solution is fully mixed, and obtain a LAP solution with a concentration of 0.5%. Cool it to room temperature and store it at 4 ° C in the dark. (2) Saccharomyces boulardii was cultured in a yeast extract peptone glucose agar medium overnight at 30°C and 160 rpm; before use, the Saccharomyces boulardii was centrifuged and collected in PBS buffer to obtain a Saccharomyces boulardii liquid; (3) Dissolve 100 mg of GelMA solid in 1 mL of 0.5% LAP solution and shake to fully soak the GelMA; place in a 60°C water bath in the dark and heat to dissolve for 30 min, shaking several times during the process to ensure that the GelMA is completely dissolved. Filter the GelMA solution through a 0.22 μm filter membrane to sterilize it to prevent low-temperature gelation of the GelMA, and obtain a 10% GelMA solution; dissolve 16 mg of PEO powder in 1 mL of 0.5% LAP solution to obtain a 1.6% PEO solution; mix the obtained 10% GelMA solution with the 1.6% PEO solution in a volume ratio of 1:1 to obtain a prepolymer solution; Finally, the prepolymer solution was mixed with 10 μL of bacterial solution with a concentration of 0.1×10 6 cells / mL of Saccharomyces boulardii liquid was mixed to obtain a mixed solution, which was frozen at 4°C for 30 minutes to transform from a sol state to a gel state to obtain a gel mixture; the mixture was irradiated under a 365nm ultraviolet lamp for 30s to achieve a polymerization reaction of the GelMA monomer.

2. Use of the antibacterial biological coating according to claim 1 in the preparation of antibacterial medical device products.

3. The use according to claim 2, characterized in that The fungi include fungi and bacteria.

4. The use according to claim 3, characterized in that The fungus is Candida albicans, and the bacteria include Staphylococcus aureus and Escherichia coli.

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