CN114681481A - Chiral two-dimensional sulfur nanosheet for selectively resisting gram-positive bacteria and preparation method and application thereof - Google Patents
Chiral two-dimensional sulfur nanosheet for selectively resisting gram-positive bacteria and preparation method and application thereof Download PDFInfo
- Publication number
- CN114681481A CN114681481A CN202111168521.4A CN202111168521A CN114681481A CN 114681481 A CN114681481 A CN 114681481A CN 202111168521 A CN202111168521 A CN 202111168521A CN 114681481 A CN114681481 A CN 114681481A
- Authority
- CN
- China
- Prior art keywords
- chiral
- sulfur
- nanosheet
- dimensional
- dimensional sulfur
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K33/00—Medicinal preparations containing inorganic active ingredients
- A61K33/04—Sulfur, selenium or tellurium; Compounds thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
- A61K47/54—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic compound
- A61K47/542—Carboxylic acids, e.g. a fatty acid or an amino acid
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
- A61K47/54—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic compound
- A61K47/545—Heterocyclic compounds
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/04—Antibacterial agents
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y40/00—Manufacture or treatment of nanostructures
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y5/00—Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Life Sciences & Earth Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Veterinary Medicine (AREA)
- Nanotechnology (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Epidemiology (AREA)
- Crystallography & Structural Chemistry (AREA)
- Organic Chemistry (AREA)
- Oncology (AREA)
- General Chemical & Material Sciences (AREA)
- Biophysics (AREA)
- Biotechnology (AREA)
- General Engineering & Computer Science (AREA)
- Medical Informatics (AREA)
- Molecular Biology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Inorganic Chemistry (AREA)
- Communicable Diseases (AREA)
- Physics & Mathematics (AREA)
- Condensed Matter Physics & Semiconductors (AREA)
- General Physics & Mathematics (AREA)
- Manufacturing & Machinery (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
The invention discloses a chiral two-dimensional sulfur nanosheet for selectively resisting gram-positive bacteria, and a preparation method and application thereof. The chiral two-dimensional sulfur nanosheet has good stability, biocompatibility and biodegradability, and can be used for selectively resisting gram-positive bacteria infection.
Description
Technical Field
The invention belongs to the technical field of nano materials, and particularly relates to a chiral two-dimensional sulfur nanosheet for selectively resisting gram-positive bacterial infection, and a preparation method and application thereof.
Background
The long-term and excessive use of antibiotics has led to stronger resistance of bacteria, multi-drug resistant bacterial infections have become one of the most urgent public health threats in the world, and the development of novel antibacterial agents is urgently needed to solve the problem. Nanomaterials have unique chemical and physical properties, such as extremely high specific surface area and abundant surface active sites, and have been extensively explored for combating multi-drug resistant bacteria. To date, a variety of metal nanoparticles (e.g., gold, silver, and copper), have been successfully synthesized for use as strong antimicrobial agents. However, cost effectiveness and unknown biological safety limit further clinical applications. The noble metal has degradation difficulty due to the biological inertia of the noble metal, and certain biological safety risk exists after long-term retention. In addition, the antibacterial activity of such noble metal nanoparticles is mostly broad-spectrum, and can kill almost all bacteria including beneficial bacteria. However, some nonpathogenic bacteria present in the body play an important role in the health of the body, and the inaccuracy of broad-spectrum therapy may cause death of beneficial bacteria, thereby affecting physiological functions. Therefore, designing a nano antibacterial agent capable of selectively killing specific bacteria has important clinical value.
Sulfur (S) is a non-metallic chalcogen, atomic number 16. Elemental sulfur is one of the most widely used elements in the biomedical field. Sulfur is important for human body because it is a constituent of various amino acids such as methionine, cysteine, cystine, homocysteine, homocystine, taurine, etc., and is an absolute dietary element. Has strong medicinal background historically, is widely used for treating various skin disease infections, rashes, sterilization, agricultural fungicide and other aspects in the biological medicine industry, and is also an antidote which is acutely exposed to radioactive substances. When sulfur is absorbed by the skin, it is metabolized to inorganic sulfides or some organic sulfides, which gives it the possibility of biodegradation. So far, although a series of sulfur nanoparticles are synthesized and applied to the field of biological medicines by carrying out various material designs based on sulfur element, the nanoparticles still lack effective targeting property and cannot realize accurate selective sterilization. Meanwhile, the antibacterial application of the two-dimensional sulfur nanosheet is not researched. Compared with the conventional nanometer material, the two-dimensional sulfur nanometer sheet has higher specific surface area and more active sites. Therefore, the antibacterial application of the two-dimensional sulfur nanosheet is explored, the specific bacteria targeted killing characteristic is endowed through chiral modification, and the medical problem of selectively killing the drug-resistant bacteria at present is hopefully overcome.
Disclosure of Invention
In order to solve the problems of bacterial drug resistance caused by antibiotic abuse, high cost, difficult metabolism, difficult guarantee of biological safety and difficult specific killing of bacteria in the traditional noble metal nanoparticles, the invention constructs the chiral two-dimensional sulfur nanosheet for selectively killing gram-positive bacteria and the preparation method thereof, and applies the chiral two-dimensional sulfur nanosheet to the targeted inhibition of gram-positive bacterial infection.
In order to solve the technical problem, the invention adopts the following technical scheme:
the invention firstly discloses a chiral two-dimensional sulfur nanosheet for selectively resisting gram-positive bacteria, which is characterized in that: the chiral two-dimensional sulfur nanosheet takes a two-dimensional sulfur nanosheet as a core, and chiral amino acid molecules are modified on the surface of the two-dimensional sulfur nanosheet. The thickness of the chiral two-dimensional sulfur nanosheet is 3-6 nm, and the diameter of the chiral two-dimensional sulfur nanosheet is 200-300 nm.
Furthermore, the chiral amino acid molecule is dextrorotatory amino acid, such as D-histidine, D-glutamic acid and the like.
The preparation method of the chiral two-dimensional sulfur nanosheet comprises the following steps: adding chiral amino acid into the aqueous solution of the pure two-dimensional sulfur nanosheets, ultrasonically dispersing the chiral amino acid uniformly, and then stirring the mixture at room temperature for 12-24 h: and carrying out ultrafiltration centrifugal separation on the obtained product to obtain the chiral two-dimensional sulfur nanosheet.
Further, the mass ratio of the chiral amino acid to the pure two-dimensional sulfur nanosheets is 10-100: 1.
furthermore, the power of the ultrasonic wave is 300W, and the time is 0.5-1 h.
Further, the rotating speed of the centrifugation is 2500rpm, and the centrifugation time is 15-30 min.
The chiral two-dimensional sulfur nanosheet has the function of killing gram-positive bacteria in a targeted manner, has no obvious toxicity to mammalian cells, has good biological safety, can be used for preparing a selective antibacterial agent for resisting the gram-positive bacteria,
the mechanism for realizing the chiral two-dimensional sulfur nanosheet used for targeted recognition of bacteria and selective killing of gram-positive bacteria is as follows: there are significant differences in the envelope of gram-positive bacteria, which have specialized lipopolysaccharides on the outer membrane surface, and gram-negative bacteria, which have a surface layer of LPS that contributes to the strict permeability characteristics of the outer membrane and is resistant to the penetration of many compounds, including sulfur. In addition, bacteria take up D-amino acids by transpeptidase during growth and reproduction, and participate in peptidoglycan synthesis, while mammalian cells generally take up L-amino acids. Thus, D-amino acid modified two-dimensional sulfur nanoplates can be selectively enriched on the surface of gram-positive bacteria. When the sulfur nanosheets are attached to the surface of a bacterial cell, on one hand, pits are formed by strong interaction between the sulfur nanosheets (with negative charges) and biological target molecules such as enzymes and proteins existing on the surface of the cell, so that the integrity of a membrane, the membrane potential and depolarization are changed, and the content leaks, so that the bacterial membrane is dissolved or the bacteria die; on the other hand, due to the ultrathin characteristic of the two-dimensional sulfur nanosheet, irreversible mechanical damage can be caused to microbial cells, so that bacteria are killed.
The invention has the beneficial effects that:
1. the chiral two-dimensional sulfur nanosheet has good stability, biocompatibility and biodegradability. The sulfur nanomaterials undergo partial reduction at their surface and partial deprotonation to form sulfide species, such as inorganic sulfides (S)2-) And polysulfides (S)X 2-) When the sulfur nanosheets contact the skin, the anions can be metabolized into inorganic sulfides or organic sulfides, and D-type amino acid molecules on the surface are not taken up by cells, so that the safety of clinical application is improved; meanwhile, the specific uptake of the D-type amino acid by the bacteria realizes the specific target enrichment of the material to the bacteria.
2. The chiral two-dimensional sulfur nanosheet is used for selectively resisting gram-positive bacteria, so that the antibacterial performance of the traditional sulfur powder is obviously improved, and the material consumption is obviously reduced.
3. The preparation process of the chiral two-dimensional sulfur nanosheet is simple, the conditions are mild, the possibility of large-scale production is realized, and the potential of industrial and practical application is realized.
4. The material used in the invention has good biocompatibility, no direct or indirect toxic action on human body and no potential toxicity.
5. The two-dimensional nanosheet disclosed by the invention has good dispersibility and stability, and is beneficial to clinical use.
Drawings
FIG. 1 is a schematic synthesis of the present invention.
Fig. 2 is a transmission electron micrograph of the chiral two-dimensional sulfur nanosheets prepared in example 1.
Fig. 3 is an atomic force microscope image of chiral two-dimensional sulfur nanoplates prepared in example 1.
Fig. 4 is a Zeta potential diagram of chiral two-dimensional sulfur nanosheets prepared in example 1.
FIG. 5 is a Raman spectrum of the chiral two-dimensional sulfur nanosheet, the pure two-dimensional sulfur nanosheet, and D-histidine prepared in example 1.
Fig. 6 is an ultraviolet-visible light absorption spectrum of the chiral two-dimensional sulfur nanosheet prepared in example 1.
Fig. 7 is a circular dichroism plot of chiral two-dimensional sulfur nanoplates prepared in example 1.
Fig. 8 is a 7-day stability plot for chiral two-dimensional sulfur nanosheets and pure two-dimensional sulfur nanosheets prepared in example 1.
Fig. 9 is a transmission electron microscope image of renaturation of the chiral two-dimensional sulfur nanosheets prepared in example 1 after lyophilization.
Fig. 10 is a graph of the antimicrobial performance of chiral two-dimensional sulfur nanoplates and sublimed sulfur powder (S), pure two-dimensional sulfur nanoplates (S-NSs), and L-His @ S-NSs prepared by L-histidine (L-His) prepared in example 1 against methicillin-resistant staphylococcus aureus (MRSA) and escherichia coli (e.
Fig. 11 is a fluorescence microscope image of the chiral two-dimensional sulfur nanosheets and sublimed sulfur powder (S) prepared in example 1, pure two-dimensional sulfur nanosheets (S-NSs), and L-His @ S-NSs prepared by L-histidine (L-His), after treatment of methicillin-resistant staphylococcus aureus (MRSA) and escherichia coli (e.
FIG. 12 is a graph showing the growth of methicillin-resistant Staphylococcus aureus treated with the chiral two-dimensional sulfur nanosheets prepared in example 1, Phosphate Buffered Saline (PBS), sublimed sulfur powder (S), pure two-dimensional sulfur nanosheets (S-NSs), and D-His, respectively.
Fig. 13 is a graph of biocompatibility of chiral two-dimensional sulfur nanoplates at different concentrations.
FIG. 14 is a graph of the biocompatibility of sulfur powder (S), pure two-dimensional sulfur nanoplates (S-NSs), L-His @ S-NSs, and D-His @ S-NSs at the same concentrations.
Detailed Description
In order to make the aforementioned objects, features and advantages of the present invention more comprehensible, specific embodiments thereof will be described in detail with reference to the following examples. The following is merely exemplary and illustrative of the inventive concept and various modifications, additions and substitutions of similar embodiments may be made to the described embodiments by those skilled in the art without departing from the inventive concept or exceeding the scope of the claims defined thereby.
Example 1
This example prepares chiral sulfur nanoplates as follows:
(1) according to the reported preparation method of the two-dimensional sulfur nanosheet, 0.5g of sublimed sulfur powder and 5mg of Bovine Serum Albumin (BSA) are weighed, placed in 50mL of deionized water, subjected to ultrasonic treatment for 72 hours (the ultrasonic power is 425W), then centrifuged at 5000rpm for 30min, and the supernatant is collected to obtain the aqueous solution of the pure two-dimensional sulfur nanosheet. The concentration of S in the solution was calculated by the standard curve by ICP (inductively coupled plasma spectroscopy) test.
(2) According to the mass ratio of the chiral amino acid to the pure two-dimensional sulfur nanosheets being 100: adding D-histidine (D-His) into a water solution of a pure two-dimensional sulfur nano-sheet, ultrasonically dispersing uniformly (the power of ultrasonic is 300W, the time is 30min), and then stirring and reacting at room temperature for 12 h: and (3) carrying out ultrafiltration centrifugal separation on the obtained product (the centrifugal rotation speed is 2500rpm, and the centrifugal time is 30min), and washing the obtained product by using deionized water to obtain the chiral two-dimensional sulfur nanosheet.
Fig. 2 is a transmission electron microscope image of the chiral two-dimensional sulfur nanosheet obtained in the present embodiment, and the characterization method thereof is as follows: and dropwise adding the aqueous dispersion of the chiral sulfur nanosheets onto a transmission electron microscope copper mesh, airing, and observing in the transmission electron microscope. The diameter of the nano-sheet is 200-300 nm.
Fig. 3 is an atomic force microscope image of the chiral sulfur nanosheet obtained in the present embodiment, and the characterization method thereof is: dropwise adding the water dispersion of the chiral sulfur nanosheets onto a mica sheet, airing, and placing into an atomic force microscope for observation. The thickness of the nano-sheet is 4-6 nm.
Fig. 4 is a Zeta potential diagram of the chiral two-dimensional sulfur nanosheet obtained in this embodiment, and its characterization method is as follows: 680 μ L of an aqueous dispersion of chiral two-dimensional sulfur nanoplates was added to a Zeta potential sample cell and tested using a NanoZS90 malvern particle sizer. It can be seen from the figure that: the Zeta potential value of the pure two-dimensional sulfur nanosheet is-11.2 mV, and the Zeta potential value is reduced to-33.8 mV after modification by the chiral amino acid molecules, so that the results show that the amino acid molecules are successfully loaded on the surface of the two-dimensional sulfur nanosheet, and the reduction of the Zeta potential value is probably caused by negative charge of a carboxyl dissociation proton band of the amino acid molecules.
Fig. 5 is a raman spectrum comparison graph of the chiral two-dimensional sulfur nanosheet, the pure two-dimensional sulfur nanosheet and D-histidine obtained in the present embodiment, and the characterization method thereof is as follows: and (3) dripping the chiral two-dimensional sulfur nanosheet, the pure two-dimensional sulfur nanosheet or the water dispersion of D-histidine on a quartz plate, airing, and testing by using a microscopic confocal laser Raman spectrometer. The figure shows that the chiral two-dimensional sulfur nanosheet shows an obvious Raman peak which is completely consistent with the Raman peaks appearing on the pure two-dimensional sulfur nanosheet and the chiral amino acid, and the fact that D-His is successfully modified on the two-dimensional sulfur nanosheet is proved, and the prepared chiral two-dimensional sulfur nanosheet has a unit cell structure similar to that of the two-dimensional sulfur nanosheet.
Fig. 6 is an ultraviolet-visible light absorption spectrum of the chiral two-dimensional sulfur nanosheet obtained in the present embodiment, and the characterization method thereof is as follows: preparing water dispersions (5 mu g/mL, 10 mu g/mL, 15 mu g/mL, 20 mu g/mL and 25 mu g/mL) of chiral two-dimensional sulfur nano-sheets with different concentrations, and testing ultraviolet-visible absorption spectrograms of the water dispersions. As can be seen from the figure, the ultraviolet-visible light absorption spectrum of the prepared chiral two-dimensional sulfur nanosheet clearly shows full-band absorption at 200-600 nm, the absorption is concentration-dependent, and the absorption value is increased along with the increase of the concentration.
Fig. 7 is a circular dichroism chart of the chiral two-dimensional sulfur nanosheet obtained in the present example, and the characterization method thereof is as follows: and (3) adding 2mL of the aqueous dispersion of the chiral two-dimensional sulfur nanosheet into a quartz cuvette, and testing the chirality of the cuvette by using a circular dichrograph. From the figure, the nanosheet shows an obvious CD absorption band between 200 nm and 250nm, and the corresponding L-His @ S-NSs prepared by L-histidine (L-His) according to the same method in the embodiment 1 shows an obvious CD absorption band with opposite sign and similar amplitude between 200 nm and 250nm, and the pure two-dimensional sulfur nanosheet does not have a meaningful CD signal, so that the chiral two-dimensional sulfur nanosheet prepared by the embodiment has obvious chiral characteristics.
Fig. 8 is a 7-day stability chart of the chiral two-dimensional sulfur nanosheet and the pure two-dimensional sulfur nanosheet obtained in the present embodiment, and the characterization method thereof is as follows: the two-dimensional sulfur nanosheet or the aqueous dispersion of the chiral two-dimensional sulfur nanosheet is stored in a refrigerator at 4 ℃, and the size of the aqueous dispersion is tested by using a laser particle sizer every other day.
Fig. 9 is a transmission electron microscope image of renaturation of the chiral two-dimensional sulfur nanosheet obtained in the present embodiment after lyophilization, and the characterization method thereof is as follows: the chiral two-dimensional sulfur nanosheets obtained in the example were lyophilized into powder by a lyophilizer and then redispersed in deionized water. The figure shows that the stability of the chiral sulfur nanosheet is good, and the morphology of the chiral sulfur nanosheet cannot be changed by freeze-drying.
To verify the selective antibacterial performance of the chiral two-dimensional sulfur nanosheet obtained in this example, the following tests were performed: mu.L of LB broth containing methicillin-resistant Staphylococcus aureus (MRSA) with a colony density of 1X 107CFU/mL) and 180 mu L and 150 mu g/mL of chiral two-dimensional sulfur nanosheet (D-His @ S-NSs) prepared in the embodiment are mixed, incubated at 37 ℃ for 4h, and 100 mu L of bacterial liquid is sucked and evenly coated on an agar culture plateAnd incubating at 37 ℃ for 48h, and judging the growth condition of the bacteria according to the colony number on an agar culture dish. For comparison, sublimed sulfur powder (S), pure two-dimensional sulfur nanosheets (S-NSs), and L-His @ S-NSs prepared by the same method with L-histidine (L-His) were also tested in the same manner in this example. A sample to which the above-mentioned material was not added was used as a control group. To verify the selective killing effect of the chiral two-dimensional sulfur nanosheet obtained in this example on gram-positive bacteria and gram-negative bacteria, the following tests were performed: the antibacterial performance of the chiral two-dimensional sulfur nanosheet, the sublimed sulfur powder (S), the pure two-dimensional sulfur nanosheet (S-NSs) and the L-His @ S-NSs on escherichia coli (E.coli) is tested according to the same method, and a sample without the materials is used as a control group. As shown in FIG. 10, it can be seen that the growth of the D-His @ S-NSs treated MRSA was significantly inhibited, and bacterial colonies from other groups of dishes showed incomplete killing of the material to varying degrees. Coli, the bacterial colonies showed normal growth of the bacteria. The results show that the chiral two-dimensional sulfur nanosheet obtained in the example has antibacterial activity for selectively killing gram-positive bacteria (such as MRSA).
To further prove the antibacterial performance of the chiral two-dimensional sulfur nanosheet obtained in this example, the following tests were performed: centrifuging and washing LB broth culture medium containing methicillin-resistant staphylococcus aureus (MRSA) with normal saline, wherein the centrifugation speed is 6000rpm, the centrifugation time is 5min, washing is carried out for 3-5 times, and the normal saline is used for fixing the volume until the colony density of MRSA is 2 multiplied by 109CFU/mL, 20 μ L of bacterial liquid was mixed with 180 μ L and 150 μ g/mL of the chiral two-dimensional sulfur nanosheet (D-His @ S-NSs) prepared in this example, incubated at 37 ℃ for 4h, subjected to bacterial death and survival staining of syto9/PI, and the death and survival of the bacteria were judged by observing the emitted fluorescence through a fluorescence microscope. For comparison, sublimed sulfur powder (S), pure two-dimensional sulfur nanosheets (S-NSs), and L-His @ S-NSs prepared by the same method with L-histidine (L-His) were also tested in the same manner in this example. A sample to which the above-mentioned material was not added was used as a control group. To further verify the selective killing effect of the chiral two-dimensional sulfur nanosheet obtained in this example on gram-positive bacteria and gram-negative bacteria, the following tests were performed: testing according to the same methodBacterial death and survival staining of Escherichia coli (E.coli) by chiral two-dimensional sulfur nanosheets, sublimed sulfur powder (S), pure two-dimensional sulfur nanosheets (S-NSs) and L-His @ S-NSs prepared by L-histidine (L-His) according to the same method are disclosed. The results are shown in fig. 11, and it can be seen that the MRSA treated by D-His @ S-NSs shows strong red fluorescence, indicating that the group of bacteria is killed, other control groups all show a large amount of green fluorescence, and the e.coli treated by the chiral two-dimensional sulfur nanosheet also shows a large amount of green fluorescence, and the results are consistent with the phenomenon of bacteria plating experiment. Further shows that the chiral two-dimensional sulfur nano-plate obtained in the embodiment has antibacterial activity for selectively killing gram-positive bacteria (such as MRSA).
To prove the antibacterial performance of the chiral two-dimensional sulfur nanosheet obtained in this example again, the following tests were performed: mu.L of LB broth containing methicillin-resistant Staphylococcus aureus (MRSA) with a colony density of 1X 107CFU/mL) and 180. mu.L of 150. mu.g/mL of chiral two-dimensional sulfur nanosheet (D-His @ S-NSs) prepared in the example, incubating at 37 ℃ for 4h, measuring the light absorption value at OD600nm by using an enzyme-labeling instrument, and measuring once every 1h to evaluate the growth state of the bacteria. For comparison, the present example also tested Phosphate Buffered Saline (PBS), sublimed sulfur powder (S), pure two-dimensional sulfur nanosheets (S-NSs), and D-His in the same manner, and the results are shown in fig. 12, and it can be seen that the growth of the bacteria was inhibited from the first 1h by the MRSA treated with D-His @ S-NSs, and the bacteria did not grow and reproduce significantly with the passage of time; the absorbance of bacteria increased over time for all other controls, indicating incomplete killing.
Fig. 13 is a biocompatibility map of chiral sulfur nanoplates at different concentrations, characterized by: diluting the D-His @ S-NSs aqueous dispersion to 2 mu g/mL, 50 mu g/mL, 75 mu g/mL, 100 mu g/mL and 150 mu g/mL, respectively mixing 0.2mL of the above solutions with 0.2mL of treated blood (500 mu L of fresh blood, adding 4.5mL of physiological saline, centrifuging for 5-8 times, wherein the centrifuging speed is 3000rpm and the centrifuging time is 10min, removing the supernatant after the supernatant is clear and transparent, diluting to 5mL with physiological saline), mixing the mixture with 0.6mL of physiological saline, incubating at 37 ℃ for 4h, centrifuging at 3000rpm for 10min, sucking the supernatant, measuring the absorbance at OD541nm, and calculating the hemolysis rate. From the figure, the hemolysis rate of the chiral two-dimensional sulfur nano-sheet with different concentrations is lower than 5%, which indicates that the biocompatibility of the material is good.
FIG. 14 is a comparison graph of hemolysis rates calculated by the above method using 150. mu.g/mL sublimed sulfur powder (S), two-dimensional sulfur nano-sheet (S-NSs), L-His @ S-NSs and D-His @ S-NSs, respectively, from which it can be seen that the hemolysis rates of each group of materials are all lower than 5%, indicating that elemental sulfur has good biosafety.
Example 2
In this example, chiral two-dimensional sulfur nanosheets were prepared as follows:
(1) according to the reported preparation method of the two-dimensional sulfur nanosheet, 0.5g of sublimed sulfur powder and 5mg of polyethylene glycol (PEG) are weighed, placed in 50mL of deionized water, subjected to ultrasonic treatment for 72 hours (the ultrasonic power is 425W), then centrifuged at 5000rpm for 30min, and the supernatant is collected to obtain the aqueous solution of the pure two-dimensional sulfur nanosheet. The concentration of S in the solution was calculated by the standard curve by ICP (inductively coupled plasma spectroscopy) test.
(2) According to the mass ratio of the chiral amino acid to the pure two-dimensional sulfur nanosheets being 50: adding D-histidine (D-His) into a water solution of a pure two-dimensional sulfur nano-sheet, ultrasonically dispersing uniformly (the power of ultrasonic is 300W, the time is 30min), and then stirring and reacting at room temperature for 12 h: and (3) carrying out ultrafiltration centrifugal separation on the obtained product (the centrifugal rotation speed is 2500rpm, and the centrifugal time is 30min), and washing the obtained product by using deionized water to obtain the chiral two-dimensional sulfur nanosheet.
Through characterization and testing, the chiral two-dimensional sulfur nanosheet obtained in the embodiment has a diameter of 200-300 nm and a thickness of 3-5 nm, and has good selective antibacterial activity on gram-positive bacteria.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents and improvements made within the spirit and principle of the present invention are intended to be included within the scope of the present invention.
Claims (8)
1. A chiral two-dimensional sulfur nanoplatelet for selective resistance to gram-positive bacteria, characterized by: the chiral two-dimensional sulfur nanosheet takes a two-dimensional sulfur nanosheet as a core, and chiral amino acid molecules are modified on the surface of the two-dimensional sulfur nanosheet.
2. Chiral two-dimensional sulfur nanoplatelets according to claim 1 characterized in that: the thickness of the chiral two-dimensional sulfur nanosheet is 3-6 nm, and the diameter of the chiral two-dimensional sulfur nanosheet is 200-300 nm.
3. Chiral two-dimensional sulfur nanoplatelets according to claim 1 characterized in that: the chiral amino acid molecule is dextrorotatory amino acid.
4. A method for preparing chiral two-dimensional sulfur nanosheets of any one of claims 1-3, wherein:
adding chiral amino acid into the aqueous solution of the pure two-dimensional sulfur nanosheets, ultrasonically dispersing the chiral amino acid uniformly, and then stirring the mixture at room temperature for 12-24 h: and carrying out ultrafiltration centrifugal separation on the obtained product to obtain the chiral two-dimensional sulfur nanosheet.
5. The method of manufacturing according to claim 4, characterized in that: the mass ratio of the chiral amino acid to the pure two-dimensional sulfur nanosheets is 10-100: 1.
6. the method of claim 4, wherein: the power of the ultrasonic wave is 300W, and the time is 0.5-1 h.
7. The method of claim 4, wherein: the rotation speed of the centrifugation is 2500rpm, and the centrifugation time is 15-30 min.
8. Use of a chiral two-dimensional sulfur nanosheet as defined in any one of claims 1 to 3, wherein: is used for preparing selective antibacterial agent for gram-positive bacteria.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111168521.4A CN114681481B (en) | 2021-09-30 | 2021-09-30 | Chiral two-dimensional sulfur nano-sheet for selectively resisting gram-positive bacteria and preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111168521.4A CN114681481B (en) | 2021-09-30 | 2021-09-30 | Chiral two-dimensional sulfur nano-sheet for selectively resisting gram-positive bacteria and preparation method and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN114681481A true CN114681481A (en) | 2022-07-01 |
| CN114681481B CN114681481B (en) | 2023-05-02 |
Family
ID=82135448
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202111168521.4A Active CN114681481B (en) | 2021-09-30 | 2021-09-30 | Chiral two-dimensional sulfur nano-sheet for selectively resisting gram-positive bacteria and preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114681481B (en) |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH06125970A (en) * | 1992-09-02 | 1994-05-10 | Fuji Shirishia Kagaku Kk | Antibacterial material having selectivity to gram-positive bacteria and selective elimination method thereof |
| WO2013187846A1 (en) * | 2012-06-15 | 2013-12-19 | Institut "Jožef Stefan" | Functionalized hydroxyapatite/gold composites as "green" materials with antibacterial activity and the process for preparing and use thereof |
| CN104706665A (en) * | 2015-03-26 | 2015-06-17 | 中国科学院长春应用化学研究所 | Method for inhibiting aggregation of Abeta by using WS2 nanosheet and method for de-aggregating formed Abeta fiber aggregate |
| CN108042565A (en) * | 2017-11-02 | 2018-05-18 | 深圳先进技术研究院 | A kind of black phosphorus nano material with antibiotic effect and preparation method thereof |
| US20210289791A1 (en) * | 2018-07-30 | 2021-09-23 | Arizona Board Of Regents On Behalf Of Arizona State University | Biopolymer-coated two-dimensional transition metal chalcogenides having potent antimicrobial activity |
-
2021
- 2021-09-30 CN CN202111168521.4A patent/CN114681481B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH06125970A (en) * | 1992-09-02 | 1994-05-10 | Fuji Shirishia Kagaku Kk | Antibacterial material having selectivity to gram-positive bacteria and selective elimination method thereof |
| WO2013187846A1 (en) * | 2012-06-15 | 2013-12-19 | Institut "Jožef Stefan" | Functionalized hydroxyapatite/gold composites as "green" materials with antibacterial activity and the process for preparing and use thereof |
| CN104706665A (en) * | 2015-03-26 | 2015-06-17 | 中国科学院长春应用化学研究所 | Method for inhibiting aggregation of Abeta by using WS2 nanosheet and method for de-aggregating formed Abeta fiber aggregate |
| CN108042565A (en) * | 2017-11-02 | 2018-05-18 | 深圳先进技术研究院 | A kind of black phosphorus nano material with antibiotic effect and preparation method thereof |
| US20210289791A1 (en) * | 2018-07-30 | 2021-09-23 | Arizona Board Of Regents On Behalf Of Arizona State University | Biopolymer-coated two-dimensional transition metal chalcogenides having potent antimicrobial activity |
Also Published As
| Publication number | Publication date |
|---|---|
| CN114681481B (en) | 2023-05-02 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Zhang et al. | Synthesis of lanthanum doped carbon dots for detection of mercury ion, multi-color imaging of cells and tissue, and bacteriostasis | |
| Wu et al. | Photothermally triggered nitric oxide nanogenerator targeting type IV pili for precise therapy of bacterial infections | |
| Seifi et al. | Anti-pathogenic activity of graphene nanomaterials: A review | |
| Abbas et al. | Cytotoxicity and antimicrobial efficiency of selenium nanoparticles biosynthesized by Spirulina platensis | |
| Suner et al. | Versatile fluorescent carbon dots from citric acid and cysteine with antimicrobial, anti-biofilm, antioxidant, and AChE enzyme inhibition capabilities | |
| Neethu et al. | Surface functionalization of central venous catheter with mycofabricated silver nanoparticles and its antibiofilm activity on multidrug resistant Acinetobacter baumannii | |
| CN108042565B (en) | Black phosphorus nano material with antibacterial effect and preparation method thereof | |
| Wang et al. | Antibacterial fluorescent nano-sized lanthanum-doped carbon quantum dot embedded polyvinyl alcohol for accelerated wound healing | |
| Le et al. | Novel silver nanoparticles: synthesis, properties and applications | |
| Abishad et al. | Green synthesized silver nanoparticles using Lactobacillus acidophilus as an antioxidant, antimicrobial, and antibiofilm agent against multi-drug resistant enteroaggregative Escherichia coli | |
| Luo et al. | An NIF-doped ZIF-8 hybrid membrane for continuous antimicrobial treatment | |
| Du et al. | Size-dependent antibacterial of carbon dots by selective absorption and differential oxidative stress of bacteria | |
| CN113548657B (en) | Natural plant impatiens balsamina carbon dots and preparation method and application thereof | |
| Tong et al. | Daptomycin and AgNP co-loaded rGO nanocomposites for specific treatment of Gram-positive bacterial infection in vitro and in vivo | |
| Erci et al. | Antimicrobial and antibiofilm activity of green synthesized silver nanoparticles by using aqueous leaf extract of Thymus serpyllum | |
| Sekar et al. | Green nanotechnology in cadmium sulphide nanoparticles and understanding its toxicity and antimicrobial properties | |
| Zhang et al. | A series of carboxymethyl cellulose-based antimicrobial peptide mimics were synthesized for antimicrobial applications | |
| Huang et al. | Rapid synthesis of bismuth-organic frameworks as selective antimicrobial materials against microbial biofilms | |
| Edan et al. | Effect of silver nanoparticles synthesized by pulsed laser ablation in liquid on the hematological, hepatic, and renal functions of albino rats | |
| Krishnaraj et al. | Green synthesis of Ag and Au NPs decorated rGO nanocomposite for high impedimetric electrochemical sensor as well as enhanced antimicrobial performance against foodborne pathogens | |
| Wang et al. | Silver nanoparticles modified with polygonatum sibiricum polysaccharide improve biocompatibility and infected wound bacteriostasis | |
| CN114470149A (en) | Antibacterial solution with size-controllable protein-based nanoparticles | |
| Gao et al. | A rational design of copper–selenium nanoclusters that cures sepsis by consuming endogenous H 2 S to trigger photothermal therapy and ROS burst | |
| CN114681481B (en) | Chiral two-dimensional sulfur nano-sheet for selectively resisting gram-positive bacteria and preparation method and application thereof | |
| Tilwani et al. | Preparation, physicochemical characterization, and in vitro biological properties of selenium nanoparticle synthesized from exopolysaccharide of enterococcus faecium MC-5 |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |