CN113274498A - Photothermal and photodynamic bactericide based on boron alkene and preparation method thereof - Google Patents

Photothermal and photodynamic bactericide based on boron alkene and preparation method thereof Download PDF

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Publication number
CN113274498A
CN113274498A CN202110607219.8A CN202110607219A CN113274498A CN 113274498 A CN113274498 A CN 113274498A CN 202110607219 A CN202110607219 A CN 202110607219A CN 113274498 A CN113274498 A CN 113274498A
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China
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photodynamic
photothermal
bactericide
dimensional
boron alkene
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CN202110607219.8A
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张晗
范涛健
张家宜
杨玲
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Shenzhen University
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Shenzhen University
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    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K41/00Medicinal preparations obtained by treating materials with wave energy or particle radiation ; Therapies using these preparations
    • A61K41/0052Thermotherapy; Hyperthermia; Magnetic induction; Induction heating therapy
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K41/00Medicinal preparations obtained by treating materials with wave energy or particle radiation ; Therapies using these preparations
    • A61K41/0057Photodynamic therapy with a photosensitizer, i.e. agent able to produce reactive oxygen species upon exposure to light or radiation, e.g. UV or visible light; photocleavage of nucleic acids with an agent
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/04Antibacterial agents
    • BPERFORMING OPERATIONS; TRANSPORTING
    • B82NANOTECHNOLOGY
    • B82YSPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
    • B82Y5/00Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery
    • CCHEMISTRY; METALLURGY
    • C01INORGANIC CHEMISTRY
    • C01BNON-METALLIC ELEMENTS; COMPOUNDS THEREOF; METALLOIDS OR COMPOUNDS THEREOF NOT COVERED BY SUBCLASS C01C
    • C01B35/00Boron; Compounds thereof
    • C01B35/02Boron; Borides
    • C01B35/023Boron

Abstract

The invention provides a boroalkene-based photo-thermal and photodynamic bactericide which comprises a two-dimensional boroalkene nanosheet, wherein the thickness of the two-dimensional boroalkene nanosheet is not more than 10 nm. The invention discloses a novel photothermal and photodynamic bactericide based on boron alkene, which is developed by using boron alkene as a photothermal agent and a photosensitizer. The boron alkene can not only efficiently convert light energy into heat energy through photo-thermal action to kill bacteria, but also can be excited to generate singlet oxygen with strong activity, and the singlet oxygen and various adjacent biological macromolecules undergo oxidation reaction to kill the bacteria. The photothermal and photodynamic bactericide based on the boron alkene uses the boron alkene as a photothermal agent, high-efficiency sterilization is realized, the drug resistance defect of the existing therapeutic drugs such as antibiotics can be effectively avoided, the boron alkene is also a degradable photosensitizer, and the degradation process of the boron alkene is harmless to the ecological environment. The invention also provides a preparation method of the photothermal and photodynamic bactericide based on the borane.

Description

Photothermal and photodynamic bactericide based on boron alkene and preparation method thereof
Technical Field
The invention relates to the technical field of antibacterial agents, in particular to a borolene-based photo-thermal and photo-dynamic bactericide, and also relates to a preparation method of the borolene-based photo-thermal and photo-dynamic bactericide.
Background
With the increasing use of antibiotics, bacteria exhibit varying degrees of resistance to some commonly used drugs. For those drugs that are used for longer periods of time and over a wider range of applications, the bacterial resistance tends to be more severe. Bacterial resistance refers to a reduction or ineffectiveness of the therapeutic effect of a drug due to a decrease or even disappearance of the sensitivity of microorganisms to antibacterial or chemotherapeutic drugs when the antibacterial drugs are applied to treat infections.
The two-dimensional material has excellent optical and thermal properties, and can kill bacteria through a physical or chemical way. Among them, photothermal therapy is an effective method for killing bacteria, and photothermal therapy is a treatment method for killing bacteria by converting light energy into heat energy by irradiating a material concentrated on a focal tissue with laser light using a material having a high photothermal conversion efficiency. Because the laser with the wavelength of 808nm has small damage to tissues, and water and protein have weak absorption capacity, so that the laser can penetrate through human tissues deeply, the laser with the wavelength is generally selected as a light source for photothermal therapy. In addition, photodynamic therapy is an effective method of killing bacteria. The conventional organic photosensitizers have poor photostability. Inorganic photosensitizers themselves are difficult to degrade, leading to potential safety issues. The two-dimensional material of the borane has low toxicity and biodegradability, has high extinction coefficient and high active oxygen generation efficiency, and has great application potential in the biomedical field, especially in the aspect of bacterial photodynamic therapy. The treatment method can greatly reduce the generation of bacterial drug resistance.
Disclosure of Invention
In view of the above, the invention provides a borolene-based photo-thermal and photodynamic bactericide and a preparation method of the borolene-based photo-thermal and photodynamic bactericide, and solves the problems of bacterial drug resistance and environmental protection of the existing bactericide by providing a novel bactericide and a preparation method thereof.
In a first aspect, the invention provides a borolene-based photothermal and photodynamic fungicide, which comprises two-dimensional borolene nanosheets, wherein the thickness of the two-dimensional borolene nanosheets is no more than 10 nm.
Preferably, the thickness of the two-dimensional boron alkene nano sheet is 4-6 nm, and the size of the two-dimensional boron alkene nano sheet is 50-300 nm.
Preferably, the concentration of the two-dimensional boroalkene nano-sheet is 25-500 ppm.
Preferably, the boracene-based photothermal and photodynamic bactericide is an aqueous solution containing boracene.
The invention discloses a novel photothermal and photodynamic bactericide based on boron alkene, which is developed by using boron alkene as a photothermal agent. The boron alkene can not only efficiently convert light energy into heat energy to kill bacteria through photothermal action, but also the degradation product boric acid is a widely used antibacterial agent. In addition, the borane has excellent photodynamic effect, can generate a large amount of active free radicals, such as hydroxyl free radicals, under the irradiation of light, and the active free radicals can destroy the structure of proteins and nucleic acids on the surface of bacteria, thereby having the photodynamic sterilization effect. The boron alkene is used as a two-dimensional nano material, and the small size is beneficial to the boron alkene to be rapidly degraded after exerting the photodynamic action and the photothermal action. The ultrathin thickness determines that the boron-alkene nano material has a larger specific surface area, so that the boron-alkene nano material is favorable for the adhesion of the material on the surface of bacteria and is also favorable for the modification and biodegradation of the material. The photothermal and photodynamic bactericide based on the boron alkene uses the boron alkene as a photothermal agent and a photosensitizer, has high-efficiency sterilization, can effectively avoid the drug resistance defect of the existing therapeutic drugs such as antibiotics and the like, has biodegradability, and has no harm to the ecological environment in the degradation process.
In a second aspect, the invention also provides a preparation method of the boroalkene-based photo-thermal and photodynamic bactericide, which comprises the following steps:
providing an isopropanol dispersion liquid of boron powder with the concentration of 1-2 mg/mL, transferring the isopropanol dispersion liquid of the boron powder to a water bath at 10-20 ℃ for ultrasonic treatment for 12-24 h, performing primary centrifugation on the isopropanol dispersion liquid of the boron powder after ultrasonic treatment for 20-30 min, taking a supernatant, performing secondary centrifugation on the supernatant for 5-10 min, collecting precipitates, and drying to obtain the borane, wherein the borane is used for preparing a photothermal and photodynamic bactericide based on the borane;
the rotating speed of the primary centrifugation is 3000-5000 rpm, and the rotating speed of the secondary centrifugation is 10000-18000 rpm.
Preferably, the drying is freeze drying.
The preparation method of the photothermal and photodynamic bactericide based on the boron alkene obtains the boron alkene dispersion liquid containing the two-dimensional boron alkene nanosheets by adopting a one-step water bath ultrasonic method, then obtains the supernatant by a centrifugal method, and further concentrates the supernatant to obtain the two-dimensional boron alkene nanosheets meeting the requirements. And dispersing the prepared two-dimensional boron alkene nanosheet in an aqueous solution or other organic reagents to obtain the corresponding bactericide. The preparation method of the photothermal and photodynamic bactericide based on the boron alkene has the advantages of simple steps, low cost and the like, and can be used for large-scale industrial production.
Advantages of the invention will be set forth in part in the description which follows, and in part will be obvious from the description, or may be learned by practice of embodiments of the invention.
Drawings
In order to more clearly illustrate the contents of the present invention, a detailed description thereof will be given below with reference to the accompanying drawings and specific embodiments.
FIG. 1 is a TEM representation of a borolene nanoplatelet prepared in example 1 of the present invention;
FIG. 2 is an AFM representation of a borolene nanoplatelet prepared in example 1 of the present invention;
FIG. 3 is a graph of the light absorption spectrum of the boracene-based photo-thermal and photodynamic fungicide of the present invention;
FIG. 4 is a graph of photothermal temperature increase for the boronic-ene-based photothermal, photodynamic fungicide of the present invention;
FIG. 5 is a graph of ESR measurements for a boracene-based photo-thermal and photodynamic fungicide of the present invention;
FIG. 6 is a chart showing the results of photothermal antimicrobial tests of the boracene-based photothermal and photodynamic bactericide of the present invention;
FIG. 7 is a graph showing the results of photodynamic antibacterial tests of the boracene-based photothermal and photodynamic bactericide of the present invention;
FIG. 8 is a graph of the degradation color change of the boracene-based photo-thermal and photodynamic fungicide of the present invention;
FIG. 9 is a diagram of the light absorption spectrum during the degradation of the boracene-based photo-thermal and photodynamic bactericide of the present invention.
Detailed Description
While the following is a description of the preferred embodiments of the present invention, it will be understood by those skilled in the art that various changes and modifications may be made without departing from the spirit and scope of the invention.
In a first aspect, the invention provides a borolene-based photothermal and photodynamic fungicide, which comprises two-dimensional borolene nanosheets, wherein the thickness of the two-dimensional borolene nanosheets is no more than 10 nm. More preferably, the thickness of the two-dimensional boron alkene nano-sheet is 1-5 nm, the ratio of the surface area of the body is larger, and the small-size effect and the photothermal conversion efficiency are more obvious.
Preferably, the thickness of the two-dimensional boron alkene nano sheet is 4-6 nm, and the size of the two-dimensional boron alkene nano sheet is 500-300 nm.
Preferably, the concentration of the two-dimensional boroalkene nano-sheet is 25-500 ppm. Specifically, it may be 25ppm, 50ppm, 75ppm, 100ppm, 200ppm, 300ppm, 400ppm or 500 ppm.
Preferably, the boracene-based photothermal and photodynamic bactericide is an aqueous solution containing boracene. In other embodiments, the boracene-based photo-thermal, photodynamic bactericide may also be a boracene-containing ethanol solution, a boracene-containing isopropanol solution, a boracene-containing polyethylene glycol solution, or the like.
In a second aspect, the invention also provides a preparation method of the boroalkene-based photo-thermal and photodynamic bactericide, which comprises the following steps:
providing an isopropanol dispersion liquid of boron powder with the concentration of 1-2 mg/mL, transferring the isopropanol dispersion liquid of the boron powder to a water bath at 10-20 ℃ for ultrasonic treatment for 12-24 h, performing primary centrifugation on the isopropanol dispersion liquid of the boron powder after ultrasonic treatment for 20-30 min, taking a supernatant, performing secondary centrifugation on the supernatant for 5-10 min, collecting precipitates, and drying to obtain the borane, wherein the borane is used for preparing a photothermal and photodynamic bactericide based on the borane;
the rotating speed of the primary centrifugation is 3000-5000 rpm, and the rotating speed of the secondary centrifugation is 10000-18000 rpm. Specifically, the rotation speed of the primary centrifugation may be 3000rpm, 4000rpm or 5000rpm, and the time of the primary centrifugation may be 20min, 25min or 30 min. The rotation speed of the secondary centrifugation can be 10000rpm, 13000rpm, 16000rpm or 18000rpm, and the time of the secondary centrifugation can be 5min, 7min, 8min or 10 min. Specifically, the temperature of the water bath ultrasound may be 10 ℃, 12 ℃, 14 ℃, 15 ℃, 16 ℃, 18 ℃ or 20 ℃. The time of the water bath ultrasound can be 12h, 14h, 16h, 18h, 20h, 22h or 24 h. The power of the water bath ultrasound can be 250W, 280W, 300W, 320W or 350W.
Preferably, the drying is freeze drying. The freeze drying can effectively avoid the degradation and agglomeration of the prepared two-dimensional boron alkene nano-sheets.
The following examples illustrate the preparation of the borane-based photothermal and photodynamic fungicides and the resulting borane-based photothermal and photodynamic fungicides in detail.
Example 1
Providing 500mg of high-purity boron powder and dispersing the high-purity boron powder in isopropanol to obtain isopropanol dispersion liquid of the boron powder with the concentration of 1mg/mL, transferring the isopropanol dispersion liquid of the boron powder to a water bath at 10 ℃ for ultrasonic treatment for 12 hours, and stripping the ultrathin two-dimensional boron alkene nanosheet. Centrifuging the isopropanol dispersion liquid of the boron powder after ultrasonic treatment for 30min at the rotating speed of 3000rpm, taking the supernatant, centrifuging for 8min at the rotating speed of 15000rpm, collecting the precipitate, and freeze-drying to obtain the two-dimensional boron alkene nanosheet. Dispersing the two-dimensional borolene nano-sheets in deionized water to obtain the borolene-based photo-thermal and photo-dynamic bactericide.
Example 2
Providing 500mg of high-purity boron powder and dispersing the high-purity boron powder in isopropanol to obtain isopropanol dispersion liquid of the boron powder with the concentration of 1.5mg/mL, transferring the isopropanol dispersion liquid of the boron powder to a water bath at 12 ℃ for carrying out ultrasonic treatment for 14 hours, and stripping the ultrathin two-dimensional boron alkene nanosheet. And centrifuging the isopropanol dispersion liquid of the boron powder subjected to ultrasonic treatment for 30min at the rotating speed of 4000rpm, taking the supernatant, centrifuging for 10min at the rotating speed of 15000rpm, collecting the precipitate, and freeze-drying to obtain the two-dimensional boron alkene nanosheet. Dispersing the two-dimensional borolene nano-sheets in deionized water to obtain the borolene-based photo-thermal and photo-dynamic bactericide.
Effects of the embodiment
The two-dimensional boron alkene nanosheet prepared in the embodiment 1 is characterized by adopting a TEM, and the result is shown in FIG. 1, the prepared two-dimensional boron alkene nanosheet is flaky, the range of the transverse size of the two-dimensional boron alkene nanosheet is 50-300 nm, and the smaller size is beneficial to rapid degradation of boron alkene after the photo-thermal effect is exerted, so that the embodiment 1 successfully prepares the two-dimensional boron alkene nanosheet. The two-dimensional borolene nano sheet prepared in the embodiment 1 is characterized by adopting AFM, as shown in FIGS. 2a and 2b, the thickness of the prepared two-dimensional borolene nano sheet is 4-6 nm, the transverse dimension is 50-300 nm, and the result is consistent with the TEM characterization result. The ultrathin thickness determines the larger specific surface area of the material, and is beneficial to the adhesion of the material on the surface of bacteria, the modification of the material and the biodegradation.
The two-dimensional borolene nanoplatelets of example 1 were dissolved in deionized water to prepare 25ppm, 50ppm, 75ppm and 100ppm borolene-based photothermal and photodynamic bactericides, and the absorption spectra of the borolene-based photothermal and photodynamic bactericides were measured, and as shown in fig. 3, the four lines at a wavelength of 1000nm correspond to 100ppm, 75ppm, 50ppm and 25ppm in this order from top to bottom. The strong absorption of the photothermal and photodynamic bactericide based on the boron alkene at 808nm ensures the efficient photothermal sterilization capability of the boron alkene; similarly, the strong absorption of the photothermal and photodynamic bactericide based on the boron alkene at 660nm ensures the high-efficiency photodynamic bactericidal capability of the boron alkene.
Photothermal heating test was carried out on the aforesaid 25ppm, 50ppm, 75ppm and 100ppm of the squalene-based photothermal and photodynamic bactericide, the laser wavelength was 808nm, and the laser power was 1W/cm2The irradiation time was 10 min. As shown in FIG. 4, the photo-thermal and photodynamic bactericide based on borolene with each concentration rapidly increases the temperature within 10min of laser irradiation, confirming that the invention is based on boroleneThe photo-thermal and photo-dynamic bactericide has excellent photo-thermal effect and sterilization effect. The boron alkene photosensitive preparation prepared in example 1 is subjected to a photodynamic test, and the test result is shown in fig. 5, and an ESR test shows that boron alkene nanosheets can generate strong and strong hydroxyl radicals under illumination. As a contrast, no signal of hydroxyl radical can be detected under no light condition, and the fact that the photothermal and photodynamic bactericide based on the borane has excellent photodynamic effect is confirmed.
The photothermal antimicrobial test was performed on the aforementioned 25ppm of the boracene-based photothermal, photodynamic bactericide using Staphylococcus aureus and Escherichia coli. The specific operation is as follows: the control group G1 was not treated at all, and the bacteria in the experimental groups G2 and G3 were cultured in 2.5% broth at 37 ℃, the bacteria treated with borane in G2 did not receive laser irradiation, and the bacteria treated with borane in G3 were exposed to 808nm laser irradiation (1W/cm)210 min) and incubated overnight. The bacterial solutions in G1, G2, and G3 were then diluted 1000-fold and 20mL of the dilution was spotted on agar plates. Bacterial density on agar plates was quantified by Image J software. The antibacterial test results are shown in fig. 6, and the antibacterial test results show that the G3 group can effectively inhibit the activity of staphylococcus aureus and escherichia coli by virtue of photothermal and photodynamic bactericide based on borolene and laser irradiation of 808 nm.
The aforementioned 25ppm boracene-based photothermal, photodynamic bactericide was subjected to a photodynamic antibacterial test using staphylococcus aureus and escherichia coli. The specific operation is as follows: the control group G1 was not treated at all, and the bacteria in the experimental groups G2 and G3 were cultured in 2.5% broth at 37 ℃, the bacteria treated with borane in G2 did not receive laser irradiation, and the bacteria treated with borane in G3 were exposed to irradiation with 660nm laser (0.5W/cm)210 min) and incubated overnight. The bacterial solutions in G1, G2, and G3 were then diluted 1000-fold and 20mL of the dilution was spotted on agar plates. Bacterial density on agar plates was quantified by Image J software. The results of the antibacterial tests are shown in fig. 7, and the results of both bacteria indicate that the photodynamic therapy based on the borane can effectively achieve the bactericidal function.
The foregoing 25ppm of the boracene-based photo-thermal, photodynamic fungicide was tested for degradation. The color of the cell containing 25ppm of the borolene-based photothermal, photodynamic bactericide of example 1 was observed every other day, and as can be seen from the change in color and transparency of the cell solution in fig. 8a-8e, the color of the bactericide in the cell gradually decreased and the transparency increased (became clear) by the degradation of the two-dimensional borolene nanoplatelets. As shown in fig. 8 (curves corresponding to the 300nm wavelength are monday, tuesday, wednesday, thursday, and friday in order from top to bottom), it can be found by analyzing the absorbance values of the liquid in the cuvette per week that the longer the number of days, the lower the absorbance value of the liquid in the cuvette, indicating the degradation of the boracene-based photothermal and photodynamic bactericide in the liquid. Further, ICP analysis shows that boric acid exists in the degradation product, and the degradation product of the two-dimensional boroalkene nanosheet is boric acid.
The above-mentioned embodiments only express several embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the present invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present patent shall be subject to the appended claims.

Claims (6)

1. The photothermal and photodynamic bactericide based on the boron alkene is characterized by comprising two-dimensional boron alkene nanosheets, wherein the thickness of the two-dimensional boron alkene nanosheets is not more than 10 nm.
2. The borolene-based photothermal, photodynamic fungicide according to claim 1 wherein the thickness of said two-dimensional borolene nanoplatelets is from 4 to 6nm and the size of said two-dimensional borolene nanoplatelets is from 50 to 300 nm.
3. The borolene-based photothermal, photodynamic fungicide according to claim 1 wherein the concentration of said two-dimensional borolene nanoplatelets is between 25 and 500 ppm.
4. The borane-based photothermal, photodynamic fungicide of claim 1 wherein said borane-based photothermal, photodynamic fungicide is an aqueous solution containing borane.
5. A preparation method of a boroalkene-based photo-thermal and photodynamic bactericide is characterized by comprising the following steps:
providing an isopropanol dispersion liquid of boron powder with the concentration of 1-2 mg/mL, transferring the isopropanol dispersion liquid of the boron powder to a water bath at 10-20 ℃ for ultrasonic treatment for 12-24 h, performing primary centrifugation on the isopropanol dispersion liquid of the boron powder after ultrasonic treatment for 20-30 min, taking a supernatant, performing secondary centrifugation on the supernatant for 5-10 min, collecting precipitates, and drying to obtain the borane, wherein the borane is used for preparing a photothermal and photodynamic bactericide based on the borane;
the rotating speed of the primary centrifugation is 3000-5000 rpm, and the rotating speed of the secondary centrifugation is 10000-18000 rpm.
6. The method of preparing a boracene-based photothermal, photodynamic bactericide as claimed in claim 5, wherein said drying is freeze-drying.
CN202110607219.8A 2021-06-01 2021-06-01 Photothermal and photodynamic bactericide based on boron alkene and preparation method thereof Pending CN113274498A (en)

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