CN115414478B - Preparation method of antibacterial light response composite material - Google Patents
Preparation method of antibacterial light response composite material Download PDFInfo
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- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/04—Antibacterial agents
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- B82Y40/00—Manufacture or treatment of nanostructures
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
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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 a Tris buffer solution with a certain volume to prepare a solution system, weighing selenium nano particles, adding the selenium nano particles into the solution system, fully dispersing the selenium nano particles in the solution by ultrasonic, adding dopamine hydrochloride after the dispersing is finished, slowly stirring the mixture at room temperature, centrifuging the mixture after the reaction is finished to obtain supernatant, dialyzing and purifying the supernatant, and freeze-drying the supernatant after the dialyzing is finished to obtain polydopamine coated selenium nano particles; and (3) regulating the pH value of the polydopamine coated selenium nanoparticle solution to be acidic, adding the indocyanine green solution, stirring under dark condition for drug loading, centrifuging after the reaction is finished, discarding the supernatant, and collecting and washing the precipitate to obtain the antibacterial light response composite material. The preparation method of the antibacterial light response composite material is simple and easy to implement, and has higher photo-thermal effect, good biocompatibility and excellent light response sterilization effect.
Description
Technical Field
The invention belongs to the field of antibacterial materials, and relates to a preparation method of an antibacterial light response composite material.
Background
The high morbidity and mortality caused by bacterial infections is of increasing interest to the medical community and the public. The advent of the first class of antibiotics, penicillin, has opened the golden age of antibiotics, and various antibiotics have been continuously investigated to find antibiotic treatment as a common method for combating pathogenic bacterial infections. However, the use of antibiotics in large numbers for decades has led to an accelerated rise in bacterial resistance, and this evolution and induction behavior provides bacteria with the ability to fight antibiotic drugs, causing a great environmental and human hazard. 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 bacterial resistance.
The research has found that the photo-thermal poly-dopamine particles combined with indocyanine green can be used in the field of antibiosis, but the photo-thermal conversion efficiency and the sterilization effect on staphylococcus aureus of the material are not ideal due to the existence of a polymer shell layer on the outer surface of the material.
Disclosure of Invention
The invention aims to overcome the defects of the prior art and provide a preparation method of an antibacterial light response 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 comprises the following steps:
according to the invention, selenium nano materials with various biological effects are introduced into the system, and the selenium nano materials are synergistic with polydopamine and indocyanine green, so that the photo-thermal conversion efficiency and the sterilization effect on staphylococcus aureus and escherichia coli are improved.
Selenium nanoparticles are a nanomaterial with multiple biological effects, which serve as the basic carrier of the composition; the polydopamine shell layer can improve the dispersibility and biocompatibility of the nano particles and endow the material with photo-thermal properties; indocyanine green can improve the photo-thermal effect of the material and can 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 proper, so that the selenium nano particles have excellent photo-thermal conversion efficiency; when the material is applied in antibiosis, after the material is incubated with bacteria, the material is irradiated by near infrared laser, partial PDA shell layer is decomposed, and selenium nano particles are exposed, so that the growth of the bacteria can be inhibited.
The preparation method of the antibacterial light response 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 nano particles, adding the selenium nano particles into the solution system, fully dispersing the nano particles in the solution by ultrasonic, adding dopamine hydrochloride after dispersing, and slowly stirring for 3-5h at room temperature. The mass ratio of the polydopamine hydrochloride to the selenium nano-particles is 0.5-1.5: 1, centrifuging after the reaction is finished, collecting supernatant, dialyzing and purifying the supernatant, and freeze-drying after the dialysis is finished to obtain polydopamine coated selenium nano particles which are named as 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, stirring the polydopamine coated selenium nanoparticle and indocyanine green for 1-3 hours under a dark condition for carrying out drug loading, centrifuging after the reaction is finished, discarding supernatant, taking out precipitate, washing for three times, and re-suspending 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 centrifugal condition is that the rotating speed is 4000-6000rpm, and the centrifugal time is 10-20min; the freeze drying temperature is-50 ℃ to-54 ℃ and the drying time is 1-2 days.
The pH value of the polydopamine coated selenium nanoparticle solution is adjusted to 2-3; the centrifugal washing condition is that the rotating speed is 8000-15000rpm, and the centrifugal time is 20-30min.
The composition is prepared by firstly feeding dopamine hydrochloride and selenium nano particles in a mass ratio of 1.2-1.5:1, so that the poly-dopamine coats the selenium nano particles, and then combining the coated nano material with indocyanine green in a mass ratio of 1:0.6-0.8 to load indocyanine green. This ratio is determined by multiple experiments, and the combination and ratio described above provides the photoresponsive composition of the invention with an excellent balance of biosafety and antimicrobial properties.
Further, the preparation method of the selenium nanoparticle comprises the following steps:
sodium selenite, glutathione and polyvinylpyrrolidone are mixed according to the mass ratio of 16-18: 1 to 3: dissolving 40-60 parts in ultrapure water, adding hydroxylamine solution (the concentration is 50%, the adding amount is 1/30 of the volume of water), uniformly mixing, reacting for 0.5-1.5 hours at 70-80 ℃ under the anaerobic condition of a reaction system, centrifuging after the reaction is finished, collecting supernatant, dialyzing and purifying the supernatant by using deionized water, and freeze-drying after the purification is finished to obtain selenium nano particles.
The centrifugal condition is that the rotating speed is 5000-6000rpm, and the centrifugal time is 10-20min; the freeze drying temperature is-50 ℃ to-55 ℃ and the drying time is 1-2 days.
The invention has the advantages and beneficial effects that:
the preparation method of the antibacterial light response composite material is simple and easy to implement, and has higher photo-thermal effect, good biocompatibility and excellent performanceIs used at a composition concentration of 125. Mu.g/mL at 1W/cm 2 The near infrared laser with the power density of 808nm irradiates for 10min, and has good sterilization effect on staphylococcus aureus and escherichia coli sterilization.
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 is a mass ratio of polydopamine hydrochloride to selenium nanoparticle of 1.5:1, a Transmission Electron Microscope (TEM) image of the prepared polydopamine coated selenium nanoparticle;
FIG. 4 is a Transmission Electron Microscope (TEM) image of the prepared polydopamine coated selenium nanoparticle with a mass ratio of polydopamine hydrochloride to selenium nanoparticle of 0.4:1;
FIG. 5 is an ultraviolet absorption spectrum of polydopamine coated selenium nanoparticle to indocyanine green with mass ratio of 1:0.1, 1:0.2, 1:0.4, 1:0.8, respectively;
FIG. 6 is a photo-thermal warming-cooling data graph of polydopamine coated selenium nanomaterial;
FIG. 7 is a graph of cell survival data after incubation of selenium nanomaterial-based photoresponsive compositions with L929 and NIH 3T3 cells;
FIG. 8 is a graph of percent hemolysis data after incubation of various concentrations of selenium nanomaterial-based photoresponsive composition with mouse red blood cells;
FIG. 9 is a photograph of a plate object of Se@PDA-ICG, se@PDA, ICG and control group killed E.coli after irradiation with or without 808nm near infrared laser;
FIG. 10 is a photograph of a plate object of Se@PDA-ICG, se@PDA, ICG and control groups killing Staphylococcus aureus after irradiation with or without 808nm near infrared laser.
Detailed Description
The invention will now be described in further detail by way of specific examples, which are given by way of illustration only and not by way of limitation, with reference to the accompanying drawings.
In the following examples and comparative examples, the biocompatibility of the material was evaluated by using the cell viability and the percent hemolysis, the higher the cell viability, the lower the percent hemolysis, the better the biocompatibility of the material; the antibacterial effect of the material was evaluated by bacterial viability, and the lower this value, the better the antibacterial performance of the material was demonstrated.
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 uniform mixing, vigorously stirring and reacting for 1 hour at the temperature of 70 ℃, changing the solution from clear and transparent to orange red, centrifuging at the rotation speed of 5000rpm after the reaction is finished, collecting supernatant, dialyzing and purifying the supernatant by using deionized water, and freeze-drying for 1-2 days after the purification is finished to obtain selenium nano particles. A TEM image of the selenium nanoparticle prepared is shown in fig. 1. As can be seen from the figure, the selenium nanoparticles are in the form of regular spheres, dispersed uniformly, and have an average size of about 90nm.
Adding 1/5 absolute ethyl alcohol into a certain volume of Tris buffer solution (the concentration is 10mM, the pH value is 8.5) to prepare a solution system, weighing selenium nano-particles, adding the selenium nano-particles into the solution system, carrying out ultrasonic treatment for 1 hour to fully disperse the nano-particles in the solution, and adding dopamine hydrochloride after the dispersing is finished to ensure that the mass ratio of polydopamine hydrochloride to the selenium nano-particles is 1.5:1, the reaction system was slowly stirred at room temperature for 4 hours, and the color of the solution gradually became grey brown as the reaction proceeded. After the reaction is finished, the mixture is centrifuged for 15 minutes at 5000rpm, supernatant is obtained, the supernatant is dialyzed and purified, and after the dialysis is finished, the polydopamine coated selenium nanoparticle is obtained after freeze drying for 1-2 days, and the obtained product is named as Se@PDA. A TEM image of the selenium nanoparticle prepared is shown in fig. 3. As can be seen from fig. 3, the selenium nanoparticle is uniformly coated with a polydopamine shell layer with a thickness of about 30nm,
the pH value of the polydopamine coated selenium nanoparticle solution is adjusted to be 2-3, a proper amount of indocyanine green solution is added, the mass ratio of the polydopamine coated selenium nanoparticle to indocyanine green is 1:0.8, the reaction system is stirred for 2 hours under dark condition to carry out drug loading, centrifugation is carried out at 10000rpm for 20 minutes after the reaction is finished, supernatant fluid is discarded, precipitate is left for three times of washing, the solution is resuspended for standby after the washing is finished, and the obtained precipitate is the antibacterial photoresponse composite material named Se@PDA-ICG.
And carrying out ultraviolet absorption spectrum tests on drug-carrying solutions with different mass ratios (the mass ratio of the polydopamine coated selenium nano particles to the indocyanine green is 1:0.1, 1:0.2, 1:0.4 and 1:0.8 respectively), wherein the higher the absorption peak value of the absorption spectrum at 780nm is, the more indocyanine green is loaded. As shown in the test result in FIG. 5, with the increase of indocyanine green mass, the drug loading of the material is gradually increased, and when the mass ratio of polydopamine coated selenium nanoparticle to indocyanine green is 1:0.8, the absorption peak value of 780nm is the maximum, and the drug loading rate is 8.11%.
Application example 1
To study the photothermal conversion efficiency of Se@PDA, a solid sample of Se@PDA was dissolved in deionized water at a concentration of 200. Mu.g/mL and a power density of 808nm near infrared laser was set at 2W/cm 2 . Placing 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, using near infrared laser to vertically irradiate the solution for 10min, and observing the temperature rising condition. After the irradiation is completed, the laser is turned off, and the temperature cooling condition of the liquid sample is observed. And recording temperature change data in the heating-cooling process and calculating the photo-thermal conversion efficiency of the material. Fig. 6 is a graph of photo-thermal heating-cooling fitting data 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 detected using the MTT method. When the cells were cultured to a good state, the cells were inoculated into 96-well cell culture plates, and the culture was continued until the cells were adhered to the wall. Different concentrations of 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) were prepared as experimental groups, and PBS buffer solution was used as control group. After adding different sets of solutions to the wells, the well plates were placed in a carbon dioxide incubator for 24h (6 sub-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, liquid was removed from each well using a pipette and 150 μl of dimethyl sulfoxide solution was added. The well plate was shaken for 5 minutes, and absorbance at 490nm was measured by a microplate reader, and cell viability was calculated from the absorbance values. FIG. 7 is a graph of cell survival data after incubation of Se@PDA-ICG with L929 and NIH 3T3 cells, showing that the survival rate of both cell lines remained above 90% at a concentration ranging from 6.5 μg/mL to 300 μg/mL, indicating that Se@PDA-ICG has low cytotoxicity.
Application example 3
The in vitro biocompatibility of the co-therapeutic system was assessed by hemolysis assays. Taking 1mL of mouse blood, placing the mouse blood into a centrifugal machine, centrifuging for 10 minutes at 8000rpm, and discarding the supernatant after centrifugation, so as to leave red blood cells. Erythrocytes were resuspended in 10mL of PBS solution after five washes with PBS. Different concentrations of 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) were prepared as experimental groups, PBS as negative control group, and water as positive control group. 200. Mu.L of the red blood cell suspension was mixed with 800. Mu.L of each set of solutions, respectively, and incubated at room temperature for 4 hours. After incubation is completed, centrifugation, the supernatant ultraviolet absorbance is measured and the percent hemolysis of the erythrocytes is calculated. FIG. 8 is a graph of percent hemolysis data after incubation of selenium nanomaterial-based light-responsive compositions at various concentrations with mouse red blood cells, and it can be seen from the graph that even if the concentration of the experimental group reaches 600 μg/mL, the percent hemolysis 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 mu L of the mixture was concentrated to 10 6 The CFU/mL E.coli bacterial solution and 500. Mu.L of the composition sample were mixed and placed in a shaker for incubation for 2 hours, wherein the incubation concentration of the light-responsive composition was 125. Mu.g/mL. After incubation was completed, a 808nm laser (NIR) was used at 1W/cm 2 Is irradiated for 10min. Diluting the treated bacterial solutions of each group 100 times after irradiation, taking 100 mu L, inoculating on a solid culture medium plate, uniformly coating with a coating rod, and placing the plate in a 37 ℃ constant temperature incubatorAnd (5) internal incubation. The next day the plates were removed, colony growth on the plates was observed, colony counts were performed, and bacterial viability was calculated to evaluate the antibacterial effect of each group of materials. FIG. 9 is a photograph of a plate object of Se@PDA-ICG, se@PDA, ICG and control group killed E.coli after irradiation with or without 808nm near infrared laser. From fig. 9 it can be seen that se@pda-icg+nir is not supplemented with 85.7% of the laser irradiated group bacteria viability, 0% of the laser irradiated group bacteria viability, compared to the number of bacteria of the control group without material treatment; se@PDA is not supplemented with laser irradiation group bacterial survival rate 94.8%, and the laser irradiation group bacterial survival rate 67.17%; ICG was not supplemented with 91.1% bacterial viability in the laser irradiated group, which was 3.5%.
Application example 5
Then 500. Mu.L of the mixture was concentrated to 10 6 The CFU/mL staphylococcus aureus bacterial liquid and 500 mu L of the composition sample are mixed and placed on a shaking table for incubation for 2 hours, wherein the incubation concentration of the light response composition is 125 mu g/mL. After incubation was completed, a 808nm laser (NIR) was used at 1W/cm 2 Is irradiated for 10min. After the irradiation is completed, diluting the bacterial liquid after each group of treatment by 100 times, taking 100 mu L of bacterial liquid to inoculate on a solid culture medium flat plate, uniformly smearing the bacterial liquid by using a coating rod, and placing the flat plate in a constant temperature incubator at 37 ℃ for incubation. The next day the plates were removed, colony growth on the plates was observed, colony counts were performed, and bacterial viability was calculated to evaluate the antibacterial effect of each group of materials. FIG. 10 is a photograph of a plate of Se@PDA-ICG, se@PDA, ICG and control groups with or without 808nm near infrared laser irradiation followed by killing of Staphylococcus aureus. From fig. 10 it can be seen that se@pda-icg+nir is not supplemented with 47.3% of the laser irradiated group bacteria viability, 0% of the laser irradiated group bacteria viability, compared to the number of bacteria of the control group without material treatment; se@PDA is not supplemented with laser irradiation group bacterial survival rate of 96.3%, and the laser irradiation group bacterial survival rate is 75.8%; ICG was not supplemented with 90.8% bacterial viability in the laser irradiated group, 14.9% bacterial viability in the laser irradiated group.
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 uniform mixing, vigorously stirring for 1 hour at the temperature of 90 ℃, changing the solution from clear and transparent to orange red, centrifuging at the rotation speed of 5000rpm after the reaction is finished, collecting supernatant, dialyzing and purifying the supernatant by using deionized water, and freeze-drying for 1-2 days after the purification is finished to obtain selenium nano-particles. A TEM image of the selenium nanoparticle prepared is shown in fig. 2. As can be seen from the figure, the selenium nanoparticle having a spherical shape can be found, but it is not uniform in size and aggregation phenomenon occurs compared to 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, the pH value is 8.5) to prepare a solution system, weighing selenium nano-particles, adding the selenium nano-particles into the solution system, carrying out ultrasonic treatment for 1 hour to fully disperse the nano-particles in the solution, and adding dopamine hydrochloride after the dispersing is finished to ensure that the mass ratio of polydopamine hydrochloride to the selenium nano-particles is 0.4:1, the reaction system was slowly stirred at room temperature for 4 hours, and the color of the solution gradually became grey brown as the reaction proceeded. And (3) centrifuging at 5000rpm after the reaction is finished for 15 minutes, collecting supernatant, dialyzing and purifying the supernatant, and freeze-drying for 1-2 days after the dialyzing is finished to obtain the solid sample coated with polydopamine. A TEM image of the selenium nanoparticle prepared is shown in fig. 4. As can be seen from the figure, the presence of spherical selenium nanoparticles and an outer layer of polydopamine could still be found, but compared with the material obtained in example 1, polydopamine had a large-area sheet-like morphology and could not be well coated on the selenium nanoparticle outer layer.
Comparative example 3
After the polydopamine nanoparticle and indocyanine green are compounded, PDA-ICG-NPs are encapsulated in invisible polyethylene glycol and pH sensitive poly (beta-amino ester) micelle. The photothermal conversion efficiency of PDA-NPs was determined using nanoparticle suspensions (200. Mu.g/mL) in PBS. The suspension was used at 808nm and at 1.3W/cm 2 Near infrared laser irradiation with power density records heating-cooling data to calculate the photo-thermal conversion efficiency, wherein the photo-thermal conversion efficiency is 33% lower than Se@PDA.
Comparative example 4
After the polydopamine nanoparticle and indocyanine green are compounded, PDA-ICG-NPs are encapsulated in invisible polyethylene glycol and pH sensitive poly (beta-amino ester) micelle. The cytotoxicity of the material was assessed using L929 fibroblasts, and when the cells were cultured to good condition, the cells were inoculated into 96-well cell culture plates and continued to culture until the cells adhered to the wall. After 12 hours, the growth medium was removed and 100. Mu.L of the material suspension was added. The suspension was then removed and then washed 3 times in PBS. mu.L of CCK-8 solution was added and incubated for 1.5 hours at 37 ℃. The absorbance of the medium and suspension at 450nm was then measured using a microplate reader, and cell viability was calculated from the absorbance. The viability of L929 cells was about 80% after the material had been concentrated at a concentration of greater than 200. Mu.g/mL, slightly below Se@PDA-ICG.
Comparative example 5
After the polydopamine nanoparticle and indocyanine green are compounded, PDA-ICG-NPs are encapsulated in invisible polyethylene glycol and pH sensitive poly (beta-amino ester) micelle. The in vitro biocompatibility of the co-therapeutic system was assessed by hemolysis assays. Mouse red blood cells were harvested by centrifugation, washed 3 times with PBS, and then red blood cells were 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 at various concentrations (up to 0.5 mg/mL) in PBS and allowed to stand at room temperature for 3 hours. Water was used as positive control and PBS without nanoparticles was used as 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, and the percent hemolysis was calculated. After the concentration of the material is more than 500 mug/mL, the hemolysis rate is about 1.5 percent and is slightly higher than that of Se@PDA-ICG.
Comparative example 6
After the polydopamine nanoparticle and indocyanine green are compounded, PDA-ICG-NPs are encapsulated in invisible polyethylene glycol and pH sensitive poly (beta-amino ester) micelle. To determine the bacterial killing effect of the micelle-encapsulated nanoparticles, a staphylococcus aureus suspension (3×10 8 bacteria/mL) was mixed with 250 μl of material, incubated at 37deg.C for 2 hours, and after incubation was completed the mixed suspension was irradiated with near infrared laser (808 nm) at 1.3W/cm 2 Is irradiated for 10 minutes. Subsequently, the suspension was serially diluted and spread on TSBOn agar plates. After overnight incubation at 37 ℃, the number of colony forming units was counted and bacterial viability was calculated from the number of colonies. The bacterial viability of the material at a concentration of 500. Mu.g/mL at neutral pH was about 40% higher than that of Se@PDA-ICG.
Reference is made to the comparative examples from the published research paper Encapsulation of Photothermal Nanoparticles in Stealth and pH-Responsive Micelles for Eradication of Infectious Biofilms In Vitro and In Vivo. Compared with the comparative example, the Se@PDA-ICG photoresponsive composition has more excellent photothermal conversion efficiency, biocompatibility and in-vitro antibacterial property.
The foregoing is merely a preferred embodiment of the present invention, and it should be noted that it will be apparent to those skilled in the art that variations and modifications can be made without departing from the scope of the invention.
Claims (4)
1. The preparation method of the antibacterial light response composite material for preparing the medicine for inhibiting escherichia coli or staphylococcus aureus is characterized by comprising 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, fully dispersing the selenium nanoparticles in the solution by using ultrasound, adding dopamine hydrochloride after dispersing, and slowly stirring for 3-5 hours under the condition of room temperature, wherein the mass ratio of the polydopamine hydrochloride to the selenium nanoparticles is 1.2-1.5: centrifuging after the reaction is finished to obtain a supernatant, dialyzing and purifying the supernatant, and freeze-drying after the dialysis is finished to obtain the polydopamine coated selenium nanoparticle;
(2) Adjusting the pH value of the polydopamine coated selenium nanoparticle solution to be acidic, adding indocyanine green solution, stirring the polydopamine coated selenium nanoparticle and indocyanine green for 1-3 hours under a dark condition for carrying out drug loading, centrifuging after the reaction is finished, discarding the supernatant, and collecting and washing the precipitate to obtain the antibacterial light response composite material; after the antibacterial light response composite material is incubated with bacteria, partial polydopamine shell is decomposed by near infrared laser irradiation, and selenium nano particles are exposed to inhibit the growth of the bacteria;
the preparation method of the selenium nanoparticle comprises the following steps: sodium selenite, glutathione and polyvinylpyrrolidone are mixed according to the mass ratio of 16-18: 1 to 3: dissolving 40-60% of the selenium-enriched nano-particles in ultrapure water, adding hydroxylamine solution with the concentration of 50%, adding 1/30 of the volume of water, uniformly mixing, reacting for 0.5-1.5 hours at the temperature of 70-80 ℃ under the anaerobic condition of a reaction system, centrifuging after the reaction is finished, collecting supernatant, dialyzing the supernatant by using deionized water, purifying, and freeze-drying after the purification is finished to obtain the selenium-enriched nano-particles.
2. The method of claim 1, wherein the Tris buffer has a concentration of 5 to 15mM and a pH of 8 to 9.
3. The method according to claim 1, wherein the freeze-drying temperature is-50 ℃ to-54 ℃ and the drying time is 1-2 days.
4. The method according to claim 1, wherein the pH is adjusted to be acidic and the pH is adjusted to be 2 to 3.
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