CN115414478A - Preparation method of antibacterial photoresponse composite material - Google Patents
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Abstract
The invention relates to a preparation method of an antibacterial photoresponse composite material, which comprises the steps of adding absolute ethyl alcohol into Tris buffer solution with a certain volume to prepare a solution system, weighing selenium nanoparticles and adding the selenium nanoparticles into the solution system, carrying out ultrasonic treatment to fully disperse the selenium nanoparticles in the solution, adding dopamine hydrochloride after dispersion, slowly stirring at room temperature, centrifuging after reaction is finished to obtain supernatant, dialyzing and purifying the supernatant, and carrying out freeze drying after dialysis to obtain polydopamine-coated selenium nanoparticles; and adjusting the pH value of the polydopamine-coated selenium nanoparticle solution to be acidic, adding an indocyanine green solution, stirring under a dark condition to carry out drug loading, centrifuging after the reaction is finished, discarding the supernatant, and reserving the precipitate for washing to obtain the antibacterial photoresponse composite material. The preparation method of the antibacterial photoresponse composite material is simple and feasible, and has a high photothermal effect, good biocompatibility and an excellent photoresponse sterilization effect.
Description
Technical Field
The invention belongs to the field of antibacterial materials, and relates to a preparation method of an antibacterial photoresponse composite material.
Background
The high morbidity and mortality associated with bacterial infections are of increasing concern to the medical community and the public. The advent of the first class of antibiotics, penicillin, opened the golden age of antibiotics, and various antibiotics were continually investigated to discover that antibiotic therapy is becoming a common method of combating pathogenic infections. However, in decades of years, the use of antibiotics in large quantities accelerates the rise of drug resistance of bacteria, and the evolution and induction behaviors provide the bacteria with the capability of resisting antibiotic drugs, thereby causing great harm to the environment and human beings. Some deadly drug-resistant bacteria, such as drug-resistant mycobacterium tuberculosis and drug-resistant staphylococcus aureus, cause millions of deaths worldwide each year. Therefore, there is an urgent need to develop an effective antibacterial agent as a substitute for antibiotics without causing the development of bacterial resistance.
Researches show that photo-thermal poly-dopamine particles combined with indocyanine green can be used in the antibacterial field, but due to the existence of a polymer shell layer on the outer surface of the material, the photo-thermal conversion efficiency and the bactericidal effect on staphylococcus aureus of the material are not ideal.
Disclosure of Invention
The invention aims to overcome the defects of the prior art and provides a preparation method of an antibacterial photoresponse composite material, and the prepared material has good biocompatibility, photo-thermal conversion efficiency and sterilization effect.
The technical scheme for realizing the purpose of the invention is as follows:
according to the invention, the selenium nano material with various biological effects is introduced into the system, and is in synergistic effect with polydopamine and indocyanine green, so that the photothermal conversion efficiency and the sterilization effect on staphylococcus aureus and escherichia coli are improved.
The selenium nanoparticle is a nano material with various biological effects, and serves as a basic carrier of the composition; the poly-dopamine shell layer can improve the dispersibility and biocompatibility of the nano particles and endow the material with photo-thermal performance; the indocyanine green can improve the photo-thermal effect of the material and enable the material to generate a photodynamic effect. The nano-particle size of the selenium nano-particles and the thickness of the polydopamine shell layer are relatively proper, so that the selenium nano-particles have relatively excellent photo-thermal conversion efficiency; when the material is applied to antibiosis, the material and bacteria are incubated together and then irradiated by near infrared laser, partial PDA shell layer is decomposed, the selenium nano-particles are exposed, and the growth of the bacteria can be inhibited.
A preparation method of an antibacterial photoresponse composite material comprises the following steps:
(1) Adding absolute ethyl alcohol into a certain volume of Tris buffer solution to prepare a solution system, weighing selenium nanoparticles, adding the selenium nanoparticles into the solution system, performing ultrasonic treatment to fully disperse the nanoparticles in the solution, adding dopamine hydrochloride after dispersion, and slowly stirring for 3-5 hours at room temperature. The mass ratio of the poly-dopamine hydrochloride to the selenium nano-particles is 0.5-1.5: 1, centrifuging after the reaction is finished, taking supernatant, dialyzing and purifying the supernatant, and freeze-drying after the dialysis is finished to obtain polydopamine-coated selenium nanoparticles named Se @ PDA;
(2) Adjusting the pH value of the polydopamine-coated selenium nanoparticle solution to be acidic, adding a proper amount of indocyanine green solution, wherein the mass ratio of the polydopamine-coated selenium nanoparticles to the indocyanine green is 1.1-0.8, stirring for 1-3h under a dark condition for carrying out drug loading, centrifuging after the reaction is finished, discarding supernatant, leaving precipitate, washing for three times, and resuspending the solution for later use after the washing is finished.
The concentration of the Tris buffer solution is 5-15mM, and the pH value is 8-9; the centrifugation condition is that the rotating speed is 4000-6000rpm, and the centrifugation time is 10-20min; the freeze drying temperature is-50 to-54 ℃, and the drying time is 1 to 2 days.
The pH value of the polydopamine-coated selenium nanoparticle solution is adjusted to 2-3; the centrifugal washing condition is 8000-15000rpm, and the centrifugal time is 20-30min.
The composition is prepared by firstly feeding dopamine hydrochloride and selenium nanoparticles according to the mass ratio of 1.2-1.5 to coat the polydopamine with the selenium nanoparticles, and then combining the coated nano material and indocyanine green according to the mass ratio of 1.6-0.8 to load the indocyanine green. This ratio is determined by a number of experiments and the combination and ratios provide the photoresponsive composition of the invention with an excellent balance of biosafety and antimicrobial properties.
Further, the preparation method of the selenium nano-particles comprises the following steps:
sodium selenite, glutathione and polyvinylpyrrolidone are mixed according to the mass ratio of 16-18: 1 to 3: 40-60 of the selenium nanoparticles are dissolved in ultrapure water, hydroxylamine solution (the concentration is 50 percent, and the adding amount is 1/30 of the volume of water) is added for uniform mixing, the reaction system is at 70-80 ℃ under the anaerobic condition, the reaction time is 0.5-1.5 hours, after the reaction is finished, the supernatant is obtained by centrifugation, the supernatant is purified by using deionized water for dialysis, and after the purification is finished, the selenium nanoparticles are obtained by freeze drying.
The centrifugation conditions are that the rotating speed is 5000-6000rpm, and the centrifugation time is 10-20min; the freeze drying temperature is-50 to-55 ℃, and the drying time is 1 to 2 days.
The invention has the advantages and beneficial effects that:
the preparation method of the antibacterial photoresponse composite material is simple and feasible, and has high photo-thermal propertyGood effect, good biocompatibility and excellent light response sterilization effect, and 1W/cm is used when the concentration of the composition is 125 mug/mL 2 808nm near-infrared laser irradiation with power density for 10min has good sterilization effect on staphylococcus aureus and escherichia coli.
Drawings
FIG. 1 is a Transmission Electron Microscope (TEM) image of selenium nanoparticles at a reaction temperature of 70 ℃;
FIG. 2 is a Transmission Electron Microscope (TEM) image of selenium nanoparticles at a reaction temperature of 90 ℃;
fig. 3 shows the mass ratio of poly dopamine hydrochloride to selenium nanoparticles is 1.5:1, a Transmission Electron Microscope (TEM) picture of the prepared polydopamine-coated selenium nano-particles;
fig. 4 is a Transmission Electron Microscope (TEM) image of polydopamine coated selenium nanoparticles prepared with a mass ratio of polydopamine hydrochloride to selenium nanoparticles of 0.4;
fig. 5 is an ultraviolet absorption spectrum diagram when the mass ratio of the polydopamine-coated selenium nanoparticle to indocyanine green is 1;
FIG. 6 is a graph of photothermal heating-cooling data of a polydopamine-coated selenium nanomaterial;
FIG. 7 is a graph of cell survival data after co-incubation of a selenium nanomaterial-based photo-responsive composition with L929 and NIH 3T3 cells;
FIG. 8 is a graph of percent hemolysis data for various concentrations of a selenium nanomaterial-based photo-responsive composition after co-incubation with mouse red blood cells;
FIG. 9 is a photograph of a plate with or without 808nm NIR laser irradiation for killing Escherichia coli in Se @ PDA-ICG, se @ PDA, ICG and control group;
FIG. 10 is a photograph of a plate specimen with or without 808nm NIR laser irradiation to kill Staphylococcus aureus in Se @ PDA-ICG, se @ PDA, ICG and control group.
Detailed Description
The present invention will be described in further detail with reference to the following embodiments, which are illustrative only and not limiting, and the scope of the present invention is not limited thereby.
In the following examples and comparative examples, the biocompatibility of the material was evaluated by the cell survival rate and the percentage of hemolysis, the higher the cell survival rate, the lower the percentage of hemolysis, the better the biocompatibility of the material; the antibacterial effect of the material was evaluated by the survival rate of the bacteria, and the lower this value, the better the antibacterial performance of the material.
Example 1
Dissolving 0.173g of sodium selenite, 20mg of glutathione and 0.5g of polyvinylpyrrolidone in 30mL of ultrapure water, adding 1mL of hydroxylamine solution, uniformly mixing for 10 minutes, introducing nitrogen into a reaction system after the system is uniformly mixed, violently stirring for reaction for 1 hour at the temperature of 70 ℃, changing the solution from clear and transparent to orange-red, centrifuging at the rotating speed of 5000rpm after the reaction is finished, taking supernatant, dialyzing and purifying the supernatant by using deionized water, and freeze-drying for 1-2 days after the purification is finished to obtain the selenium nanoparticles. A TEM image of the prepared selenium nanoparticles is shown in fig. 1. As can be seen from the figure, the selenium nanoparticles are in the form of regular spheres, uniformly dispersed, and have an average size of about 90nm.
Adding 1/5 absolute ethyl alcohol into a certain volume of Tris buffer solution (with the concentration of 10mM and the pH value of 8.5) to prepare a solution system, weighing selenium nanoparticles, adding the selenium nanoparticles into the solution system, performing ultrasonic treatment for 1 hour to fully disperse the nanoparticles in the solution, and adding dopamine hydrochloride after the dispersion is finished to ensure that the mass ratio of the poly-dopamine hydrochloride to the selenium nanoparticles is 1.5: the reaction system is slowly stirred at room temperature for 4 hours, and the color of the solution gradually changes to gray brown as the reaction progresses. And after the reaction is finished, 5000rpm is carried out, centrifugation is carried out for 15 minutes, supernatant is left, the supernatant is purified by dialysis, and freeze drying is carried out for 1-2 days after the dialysis is finished, so that the polydopamine-coated selenium nano particle named as Se @ PDA is obtained. A TEM image of the prepared selenium nanoparticles is shown in fig. 3. As can be seen from fig. 3, the outer layer of the selenium nanoparticle is uniformly coated with a poly-dopamine shell layer with a thickness of about 30nm,
adjusting the pH value of a polydopamine-coated selenium nanoparticle solution to 2-3, adding a proper amount of indocyanine green solution, wherein the mass ratio of the polydopamine-coated selenium nanoparticle to the indocyanine green is 1.8, stirring the reaction system for 2 hours under a dark condition for drug loading, centrifuging at 10000rpm for 20 minutes after the reaction is finished, discarding supernatant, leaving precipitate for washing three times, and after washing, re-suspending the solution for later use, wherein the obtained precipitate is an antibacterial photoresponse composite material named as Se @ PDA-ICG.
Ultraviolet absorption spectrum tests were performed on drug-loaded solutions of different mass ratios (mass ratios of polydopamine-coated selenium nanoparticles to indocyanine green of 1. As shown in fig. 5, the drug loading of the material gradually increases with the increase of the mass of indocyanine green, and when the mass ratio of the polydopamine-coated selenium nanoparticle to the indocyanine green is 1.8, the absorption peak at 780nm reaches the maximum, and the drug loading rate at this time is 8.11%.
Application example 1
In order to research the photothermal conversion efficiency of Se @ PDA, a solid sample of Se @ PDA is dissolved in deionized water, the concentration is set to be 200 mug/mL, and the power density of a near-infrared laser with the wavelength of 808nm is set to be 2W/cm 2 . And (3) placing the Se @ PDA solution on a stable platform, enabling the distance between a laser light source and the upper surface of the solution to be 5cm, turning on a laser switch, vertically irradiating the solution for 10min by using near-infrared laser, and observing the temperature rise condition. And after the irradiation is finished, the laser is turned off, and the temperature cooling condition of the liquid sample is observed. And recording temperature change data in the temperature rise-temperature reduction process and calculating the photothermal conversion efficiency of the material. Fig. 6 is a fitting data graph of photo-thermal temperature rise and temperature drop of the polydopamine-coated selenium nanomaterial, and the photo-thermal conversion efficiency of the material can be calculated to be 81.98% according to the data in fig. 6.
Application example 2
The cytotoxicity of the photoresponsive composition was examined using the MTT method. When the cells were cultured to a good condition, the cells were seeded into a 96-well cell culture plate, and the culture was continued until the cells adhered to the wall. Se @ PDA-ICG composition solutions (6.25. Mu.g/mL, 12.5. Mu.g/mL, 50. Mu.g/mL, 100. Mu.g/mL, 200. Mu.g/mL, 300. Mu.g/mL) of different concentrations were prepared as experimental groups, and PBS buffer solution was used as a control group. After adding different sets of solutions to the wells, the well plates were placed in a carbon dioxide incubator for 24h (6 secondary wells per concentration). The well plate was removed and 20. Mu.L of thiazole blue (MTT) solution was added to each well. Incubation was continued for 4 hours, and the liquid was removed from each well using a pipette gun and 150 μ L of dimethyl sulfoxide solution was added. The well plate was shaken for 5 minutes, and the absorbance at 490nm was measured by a microplate reader, and the cell survival rate was calculated from the absorbance value. FIG. 7 is a graph showing the cell survival data of Se @ PDA-ICG after incubation with L929 and NIH 3T3 cells, and it can be seen that the survival rate of both cell lines is maintained above 90% in the concentration range of 6.5-300. Mu.g/mL, indicating that Se @ PDA-ICG has low cytotoxicity.
Application example 3
The in vitro biocompatibility of the co-therapy system was assessed by hemolytic assay. 1mL of mouse blood is taken and placed in a centrifuge for 10 minutes at 8000rpm centrifugal force, and after the centrifugation is finished, supernatant is discarded, and red blood cells are left. The erythrocytes were washed five times with PBS and resuspended in 10mL of PBS solution. Se @ PDA-ICG composition solutions (100. Mu.g/mL, 200. Mu.g/mL, 300. Mu.g/mL, 400. Mu.g/mL, 500. Mu.g/mL, 600. Mu.g/mL) of different concentrations were prepared as experimental groups, PBS was used as a negative control group, and water was used as a positive control group. 200 μ L of the red blood cell suspension was mixed with 800 μ L of each set of solutions, respectively, and incubated at room temperature for 4 hours. After completion of incubation, centrifugation was performed, and the ultraviolet absorbance of the supernatant was measured and the percentage of hemolysis of erythrocytes was calculated. Fig. 8 is a graph of the hemolysis percentage data of the photo-responsive composition based on selenium nanomaterial incubated with mouse red blood cells at various concentrations, and it can be seen from the graph that even if the concentration of the experimental group reaches 600 μ g/mL, the hemolysis percentage of the nanomaterial is only 0.9%, and the amount of hemolysis is negligible, indicating that the material has good biocompatibility.
Application example 4
500 μ L of 10 6 The mixture of the CFU/mL E.coli suspension and 500. Mu.L of the composition sample was incubated for 2 hours in a shaker at a concentration of 125. Mu.g/mL of the photoresponsive composition. After incubation was complete 1W/cm using a 808nm laser (NIR) 2 Is irradiated for 10min. Diluting the treated bacteria liquid of each group by 100 times after irradiation, inoculating 100 mu L of the diluted bacteria liquid on a solid culture medium flat plate, and uniformly smearing the bacteria liquid on the solid culture medium flat plate by using a coating rodThe plate was incubated in a 37 ℃ incubator. Taking out the plate the next day, observing the growth condition of colonies on the plate, counting the colonies, calculating the survival rate of bacteria and evaluating the antibacterial effect of each group of materials. FIG. 9 is a photograph of a plate specimen with or without 808nm NIR laser irradiation to kill Escherichia coli in Se @ PDA-ICG, se @ PDA, ICG and control group. From FIG. 9, it can be seen that the survival rate of the Se @ PDA-ICG + NIR was 85.7% in the case of the laser-irradiated group and 0% in the case of the laser-irradiated group, compared with the number of bacteria in the control group without the material treatment; the survival rate of the Se @ PDA is 94.8 percent of the bacteria in the laser irradiation group and 67.17 percent of the bacteria in the laser irradiation group; the survival rate of the ICG group bacteria is 91.1 percent without the assistance of laser irradiation, and the survival rate of the ICG group bacteria is 3.5 percent with the assistance of laser irradiation.
Application example 5
Then 500. Mu.L of 10 6 The mixture of the CFU/mL Staphylococcus aureus liquid and 500. Mu.L of the composition sample was incubated in a shaker for 2 hours at a concentration of 125. Mu.g/mL of the photoresponsive composition. After incubation was complete 1W/cm using a 808nm laser (NIR) 2 Is irradiated for 10min. After the irradiation is finished, diluting the treated bacterial liquid of each group by 100 times, inoculating 100 mu L of the diluted bacterial liquid on a solid culture medium flat plate, uniformly coating the flat plate by using a coating rod, and placing the flat plate in a constant-temperature incubator at 37 ℃ for incubation. Taking out the plate the next day, observing the growth condition of colonies on the plate, counting the colonies, calculating the survival rate of bacteria and evaluating the antibacterial effect of each group of materials. FIG. 10 is a photograph of a panel showing killing of Staphylococcus aureus in the case of irradiation with or without 808nm near-infrared laser in Se @ PDA-ICG, se @ PDA, ICG and control group. From FIG. 10, it can be seen that the survival rate of the Se @ PDA-ICG + NIR was 47.3% in the case of the laser-irradiated group and 0% in the case of the laser-irradiated group, compared with the number of bacteria in the control group without the material treatment; the survival rate of the Se @ PDA is 96.3 percent in the laser irradiation group and 75.8 percent in the laser irradiation group without assistance; the ICG was not supplemented with laser irradiation group bacterial survival rate of 90.8%, laser irradiation group bacterial survival rate of 14.9%.
Comparative example 1
Dissolving 1mmol of sodium selenite, 20mg of glutathione and 0.5g of polyvinylpyrrolidone in 30mL of ultrapure water, adding 1mL of hydroxylamine solution, uniformly mixing for 10 minutes, introducing nitrogen into a reaction system after the system is uniformly mixed, violently stirring for 1 hour at the temperature of 90 ℃, changing the solution from clear and transparent to orange red, centrifuging at the rotating speed of 5000rpm after the reaction is finished, taking supernatant, dialyzing and purifying the supernatant by using deionized water, and freeze-drying for 1-2 days after the purification is finished to obtain the selenium nanoparticles. A TEM image of the prepared selenium nanoparticles is shown in fig. 2. As can be seen from the figure, selenium nanoparticles having a spherical shape can still be found, but they are not uniform in size and undergo agglomeration phenomenon as compared with the material obtained in example 1.
Comparative example 2
Adding 1/5 absolute ethyl alcohol into a certain volume of Tris buffer solution (the concentration is 10mM, and the pH value is 8.5) to prepare a solution system, weighing selenium nanoparticles, adding the selenium nanoparticles into the solution system, performing ultrasonic treatment for 1 hour to fully disperse the nanoparticles in the solution, and adding dopamine hydrochloride after dispersion is finished to ensure that the mass ratio of the poly-dopamine hydrochloride to the selenium nanoparticles is 0.4: the reaction system was left to stir slowly at room temperature for 4 hours, and the color of the solution gradually changed to gray-brown as the reaction proceeded. And after the reaction is finished, centrifuging for 15 minutes at 5000rpm, taking supernatant, dialyzing and purifying the supernatant, and freeze-drying for 1-2 days after the dialysis is finished to obtain the solid sample coated with the polydopamine. A TEM image of the prepared selenium nanoparticles is shown in fig. 4. As can be seen from the figure, the presence of selenium nanoparticles with spherical shape and poly-dopamine outer layer can still be found, but compared with the material obtained in example 1, poly-dopamine has a large area sheet-like morphology and cannot coat the selenium nanoparticle outer layer well.
Comparative example 3
After the polydopamine nanoparticles and indocyanine green were complexed, PDA-ICG-NPs were encapsulated in stealth polyethylene glycol and pH-sensitive poly (β -aminoester) micelles. The photothermal conversion efficiency of PDA-NPs was determined using a suspension of nanoparticles in PBS (200. Mu.g/mL). The suspension was used at 808nm and 1.3W/cm 2 Near-infrared laser irradiation of power density, recording temperature rise-temperature fall data and calculating photo-thermal conversion efficiency, wherein the photo-thermal conversion efficiency is 33% lower than Se @ PDA.
Comparative example 4
After the polydopamine nanoparticles and indocyanine green were complexed, PDA-ICG-NPs were encapsulated in stealth polyethylene glycol and pH-sensitive poly (β -aminoester) micelles. The cytotoxicity of the material was evaluated using L929 fibroblasts, and when the cells were cultured to a good state, the cells were seeded into a 96-well cell culture plate, and the culture was continued until the cells adhered to the wall. After 12 hours, the growth medium was removed and 100 μ L of the material suspension was added. The suspension was then removed and subsequently washed 3 times in PBS. Add 20. Mu.L of CCK-8 solution and incubate at 37 ℃ for 1.5 hours. The absorbance of the medium and suspension at 450nm was then measured using a microplate reader, and the cell viability was calculated from the absorbance. After the concentration of the material is more than 200 mu g/mL, the survival rate of the L929 cells is about 80 percent and is slightly lower than Se @ PDA-ICG.
Comparative example 5
After the polydopamine nano-particles and indocyanine green are compounded, PDA-ICG-NPs are encapsulated in stealth polyethylene glycol and pH sensitive poly (beta-amino ester) micelles. The in vitro biocompatibility of the synergistic therapeutic system was assessed by hemolysis assay. The mouse red blood cells were harvested by centrifugation, washed 3 times with PBS, and then resuspended in PBS to a volume fraction of 5%. Next, 200. Mu.L of the red blood cell suspension was mixed with 1mL of the material suspension in PBS at various concentrations (up to 0.5 mg/mL) and allowed to stand at room temperature for 3 hours. Water was used as a positive control and PBS without nanoparticles was used as a negative control. Next, the suspension was centrifuged at 2000rpm for 5 minutes, and the absorbance of the supernatant was measured at 540nm using a microplate reader to calculate the percentage of hemolysis. After the concentration of the material is more than 500 mu g/mL, the hemolysis rate is about 1.5 percent and is slightly higher than Se @ PDA-ICG.
Comparative example 6
After the polydopamine nanoparticles and indocyanine green were complexed, PDA-ICG-NPs were encapsulated in stealth polyethylene glycol and pH-sensitive poly (β -aminoester) micelles. To determine the bacterial killing effect of the nanoparticles after micelle encapsulation, a suspension of staphylococcus aureus (3 × 10) was used 8 bacteria/mL) was mixed with 250. Mu.L of the material, incubated at 37 ℃ for 2 hours, and after incubation the mixed suspension was applied with a near infrared laser (808 nm) at 1.3W/cm 2 Is irradiated for 10 minutes. Subsequently, the air conditioner is operated to,the suspension was serially diluted and plated on TSB agar plates. After overnight incubation at 37 ℃, the number of colony forming units was counted and bacterial survival was calculated from the number of colonies. The bacterial survival rate of the material at a concentration of 500. Mu.g/mL at neutral pH is about 40% higher than that of Se @ PDA-ICG.
Comparative example reference is made to the paper "Encapsulation of Photonic Nanoparticles In Steel and pH-reactive Micelles for the administration of Infectious groups Biofilms In Vitro and In Vivo" which has been published. Compared with the comparative example, the Se @ PDA-ICG photoresponse composition has more excellent photo-thermal conversion efficiency, biocompatibility and in-vitro antibacterial performance.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various changes and modifications can be made without departing from the inventive concept, and these changes and modifications are all within the scope of the present invention.
Claims (10)
1. The preparation method of the antibacterial photoresponse composite material is characterized by comprising the following steps of:
(1) Adding absolute ethyl alcohol into a certain volume of Tris buffer solution to prepare a solution system, weighing selenium nanoparticles, adding the selenium nanoparticles into the solution system, performing ultrasonic treatment to fully disperse the selenium nanoparticles in the solution, adding dopamine hydrochloride after dispersion, slowly stirring for 3-5h at room temperature, wherein the mass ratio of the poly-dopamine hydrochloride to the selenium nanoparticles is 0.5-1.5: 1, centrifuging after the reaction is finished, taking supernatant, dialyzing and purifying the supernatant, and freeze-drying after the dialysis is finished to obtain polydopamine-coated selenium nanoparticles;
(2) Adjusting the pH value of the polydopamine-coated selenium nanoparticle solution to be acidic, adding an indocyanine green solution, wherein the mass ratio of the polydopamine-coated selenium nanoparticles to the indocyanine green is 1.1-0.8, stirring for 1-3h under a dark condition for carrying out drug loading, centrifuging after the reaction is finished, discarding supernatant, and reserving precipitate and washing to obtain the antibacterial photoresponse composite material.
2. The preparation method according to claim 1, wherein the mass ratio of the polydopamine hydrochloride to the selenium nanoparticles is 1.2-1.5: 1.
3. the preparation method of claim 2, wherein the mass ratio of the polydopamine-coated selenium nanoparticles to the indocyanine green is 1.
4. The method of claim 1, wherein the selenium nanoparticles are prepared by: sodium selenite, glutathione and polyvinylpyrrolidone are mixed according to the mass ratio of 16-18: 1 to 3: 40-60 of the selenium nanoparticles are dissolved in ultrapure water, hydroxylamine solution is added, the concentration is 50%, the addition amount is 1/30 of the volume of water, the mixture is uniformly mixed, the reaction system is 70-80 ℃ under the anaerobic condition, the reaction time is 0.5-1.5 hours, after the reaction is finished, the supernatant is obtained by centrifugation, the supernatant is purified by using deionized water for dialysis, and after the purification is finished, the selenium nanoparticles are obtained by freeze drying.
5. The method according to claim 1, wherein the Tris buffer solution has a concentration of 5 to 15mM and a pH value of 8 to 9.
6. The method according to claim 1, wherein the freeze-drying temperature is from-50 ℃ to-54 ℃ and the drying time is from 1 to 2 days.
7. The method according to claim 1, wherein the pH adjustment to acidity is performed at a pH of 2 to 3.
8. An antibacterial photoresponsive composite material prepared by the method of any one of claims 1 to 7.
9. Use of the antibacterial photo-responsive composite material of claim 8 as a material for inhibiting escherichia coli or staphylococcus aureus.
10. Use of the antibacterial photoresponse composite material of claim 8 in the preparation of a medicament for the therapeutic inhibition of escherichia coli or staphylococcus aureus.
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