CN114681481B - Chiral two-dimensional sulfur nano-sheet for selectively resisting gram-positive bacteria and preparation method and application thereof - Google Patents
Chiral two-dimensional sulfur nano-sheet for selectively resisting gram-positive bacteria and preparation method and application thereof Download PDFInfo
- Publication number
- CN114681481B CN114681481B CN202111168521.4A CN202111168521A CN114681481B CN 114681481 B CN114681481 B CN 114681481B CN 202111168521 A CN202111168521 A CN 202111168521A CN 114681481 B CN114681481 B CN 114681481B
- Authority
- CN
- China
- Prior art keywords
- chiral
- sheet
- dimensional
- dimensional sulfur
- nano
- 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.)
- Active
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)
- Life Sciences & Earth Sciences (AREA)
- Pharmacology & Pharmacy (AREA)
- Medicinal Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Nanotechnology (AREA)
- Epidemiology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Crystallography & Structural Chemistry (AREA)
- Medical Informatics (AREA)
- Condensed Matter Physics & Semiconductors (AREA)
- Molecular Biology (AREA)
- Communicable Diseases (AREA)
- Inorganic Chemistry (AREA)
- Biophysics (AREA)
- Biotechnology (AREA)
- General Engineering & Computer Science (AREA)
- General Physics & Mathematics (AREA)
- Manufacturing & Machinery (AREA)
- Physics & Mathematics (AREA)
- Oncology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Organic Chemistry (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
The invention discloses a chiral two-dimensional sulfur nano-sheet for selectively resisting gram-positive bacteria, a preparation method and application thereof. The chiral two-dimensional sulfur nano-sheet 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 nano sheet for selectively resisting gram-positive bacterial infection, and a preparation method and application thereof.
Background
The long term and overuse of antibiotics has led to greater resistance of bacteria, and multi-drug resistant bacterial infections have become one of the most urgent public health threats worldwide, and the development of new antimicrobial agents is urgently needed to solve this problem. Nanomaterials have unique chemical and physical properties, such as extremely high specific surface area and abundant surface active sites, which have been widely explored for combating multiple resistant bacteria. Heretofore, various metal nanoparticles (e.g., gold, silver, and copper) have been successfully synthesized for use as strong antimicrobial agents. However, cost effectiveness and unknown biosafety limit their further clinical use. The biological inertia of the noble metal itself makes the noble metal have degradation problem, and long-term retention has a certain biological safety risk. In addition, the antibacterial activity of such noble metal nanoparticles is mostly broad spectrum, killing almost all bacteria including beneficial bacteria. However, some non-pathogenic bacteria present in the body play an important role in the health of the body, and inaccuracy in broad-spectrum therapy may lead to death of beneficial bacteria, which in turn affect physiological function. Therefore, designing a nano-antimicrobial agent that can selectively kill specific bacteria has important clinical value.
Sulfur (S) is a nonmetallic oxygen species and has an atomic number of 16. Elemental sulfur is one of the most widely used elements in the biomedical field. The sulfur element is important for human body, because it is a constituent part of various amino acids such as methionine, cysteine, cystine, homocysteine, taurine, etc., and is an absolute dietary element. Has strong medicinal background in history, is widely used for treating various skin disease infections, rashes, sterilization, agricultural fungi killing and other aspects in the biomedical industry, and is also an antidote for acute contact with 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. Although various material designs are made based on sulfur elements so far, a series of sulfur nano particles are synthesized and applied to the field of biological medicine, the nano materials still lack effective targeting, and precise selective sterilization cannot be realized. Meanwhile, no research has been made on antibacterial applications involving two-dimensional sulfur nanoplatelets. Compared with the conventional nano material, the two-dimensional sulfur nano sheet has higher specific surface area and more active sites. Therefore, the antibacterial application of the two-dimensional sulfur nano-sheet is explored, the specific bacterial characteristic is targeted and killed by chiral modification, and the medical problem of selectively killing drug-resistant bacteria at present is hopeful to be overcome.
Disclosure of Invention
In order to solve the problems of high cost, difficult metabolism, difficult guarantee of biological safety and difficult specific killing of bacteria caused by the abuse of antibiotics and the traditional noble metal nano particles, the invention constructs the chiral two-dimensional sulfur nano sheet for selectively killing the gram-positive bacteria and the preparation method thereof, and applies the chiral two-dimensional sulfur nano sheet to targeted inhibition of gram-positive bacterial infection.
The invention adopts the following technical scheme to solve the technical problems:
the invention firstly discloses a chiral two-dimensional sulfur nano-sheet for selectively resisting gram-positive bacteria, which is characterized in that: the chiral two-dimensional sulfur nano-sheet takes a two-dimensional sulfur nano-sheet as a core, and chiral amino acid molecules are modified on the surface of the two-dimensional sulfur nano-sheet. The thickness of the chiral two-dimensional sulfur nano-sheet is 3-6 nm, and the diameter is 200-300 nm.
Further, the chiral amino acid molecule is a dextrorotatory amino acid, such as D-histidine, D-glutamic acid, and the like.
The preparation method of the chiral two-dimensional sulfur nano-sheet comprises the following steps: adding chiral amino acid into the water solution of the pure two-dimensional sulfur nano-sheet, uniformly dispersing by ultrasonic, and then stirring and reacting for 12-24 hours at room temperature: and carrying out ultrafiltration and centrifugal separation on the obtained product to obtain the chiral two-dimensional sulfur nano-sheet.
Further, the mass ratio of the chiral amino acid to the pure two-dimensional sulfur nano-sheet is 10-100: 1.
further, the power of the ultrasonic wave is 300W and the time is 0.5-1 h.
Further, the rotational speed of the centrifugation is 2500rpm, and the centrifugation time is 15-30 min.
The chiral two-dimensional sulfur nano-sheet has the function of targeted killing of gram-positive bacteria, no obvious toxicity to mammalian cells, good biological safety, can be used for preparing selective antibacterial agents against gram-positive bacteria,
the chiral two-dimensional sulfur nano-sheet is used for targeted identification of bacteria and selective killing of gram-positive bacteria, and has the implementation mechanism that: there is a significant difference between gram-positive bacteria and gram-negative bacterial envelopes, the outer membrane surface of gram-negative bacteria has specialized lipopolysaccharide, the surface layer of LPS contributes to the strict permeability characteristics of the outer membrane and is resistant to permeation by many compounds including sulfur and the like. In addition, during the growth and reproduction of bacteria, D-amino acids are taken up by transpeptidase, which participates in the synthesis of peptidoglycan, whereas L-amino acids are usually taken up by mammalian cells. Thus, the D-amino acid modified two-dimensional sulfur nanoplatelets can be selectively enriched on the surface of gram-positive bacteria. When the sulfur nano-sheets are attached to the surfaces of bacterial cells, on one hand, the strong interaction of the sulfur nano-sheets (with negative charges) and biological target molecules such as enzymes, proteins and the like existing on the surfaces of the cells forms pits, and then the membrane integrity, membrane potential and depolarization are changed, the content leaks, and finally the bacterial membrane is dissolved or the bacteria die; on the other hand, the ultrathin property of the two-dimensional sulfur nano-sheet can cause irreversible mechanical damage to microorganism cells, thereby killing bacteria.
The beneficial effects of the invention are as follows:
1. the chiral two-dimensional sulfur nano-sheet has good stability, biocompatibility and biodegradability. The sulfur nanomaterial undergoes partial reduction at its surface and partial deprotonation to form sulfide species, such as inorganic sulfide (S) 2- ) And polysulfide (S) X 2- ) Anions are metabolized into inorganic sulfides or organic sulfides when the sulfur nano-sheets contact the skin, and D-type amino acid molecules on the surfaces are not absorbed by cells, so that the safety of clinical application is improved; meanwhile, the specific uptake of the bacteria to the D-type amino acid realizes the specific targeted enrichment of the material to the bacteria.
2. The chiral two-dimensional sulfur nano-sheet is used for selectively resisting gram-positive bacteria infection, so that the antibacterial property of the traditional sulfur powder is remarkably improved, and the material consumption is remarkably reduced.
3. The chiral two-dimensional sulfur nano-sheet has the advantages of simple preparation process, mild conditions, possibility of large-scale production and potential of industrial and practical application.
4. The material used in the invention has good biocompatibility, no direct or indirect toxic action to human body, and no potential toxicity.
5. The two-dimensional nano-sheet has good dispersibility and stability, and is beneficial to clinical use.
Drawings
FIG. 1 is a schematic diagram of the synthesis of the present invention.
FIG. 2 is a transmission electron microscope image of the chiral two-dimensional sulfur nanoplatelets prepared in example 1.
FIG. 3 is an atomic force microscope image of chiral two-dimensional sulfur nanoplatelets prepared in example 1.
FIG. 4 is a Zeta potential diagram of chiral two-dimensional sulfur nanoplatelets prepared in example 1.
FIG. 5 is a Raman spectrum of chiral two-dimensional sulfur nanoplatelets and pure two-dimensional sulfur nanoplatelets and D-histidine prepared in example 1.
FIG. 6 is an ultraviolet-visible light absorption spectrum of the chiral two-dimensional sulfur nanoplatelets prepared in example 1.
FIG. 7 is a circular dichroism spectrum of chiral two-dimensional sulfur nanoplatelets prepared in example 1.
FIG. 8 is a 7-day stability graph of chiral two-dimensional sulfur nanoplatelets and pure two-dimensional sulfur nanoplatelets prepared in example 1.
Fig. 9 is a transmission electron microscope image of renaturation after lyophilization of the chiral two-dimensional sulfur nanoplatelets prepared in example 1.
FIG. 10 is a graph showing antibacterial properties of chiral two-dimensional sulfur nanoplatelets and sublimated sulfur powder (S), pure two-dimensional sulfur nanoplatelets (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.coli).
FIG. 11 is a fluorescence microscopy image of the chiral two-dimensional sulfur nanoplatelets and sublimated sulfur powder (S), pure two-dimensional sulfur nanoplatelets (S-NSs), and L-His@S-NSs prepared by L-histidine (L-His) prepared in example 1 after treatment with methicillin-resistant Staphylococcus aureus (MRSA) and Escherichia coli (E.coli).
FIG. 12 is a graph showing the growth of the chiral two-dimensional sulfur nanoplatelets prepared in example 1 and Phosphate Buffer (PBS), sublimed sulfur powder (S), pure two-dimensional sulfur nanoplatelets (S-NSs), D-His, respectively treated methicillin-resistant Staphylococcus aureus.
FIG. 13 is a graph of biocompatibility of chiral two-dimensional sulfur nanoplatelets at different concentrations.
FIG. 14 is a graph of biocompatibility of sulfur powder (S), pure two-dimensional sulfur nanoplatelets (S-NSs), L-His@S-NSs, and D-His@S-NSs at the same concentration.
Detailed Description
In order that the above objects, features and advantages of the invention will be readily understood, a more particular description of the invention will be rendered by reference to specific embodiments thereof which are illustrated in the appended drawings. The following is merely illustrative and explanatory of the principles of the invention, as various modifications and additions may be made to the specific embodiments described, or similar thereto, by those skilled in the art, without departing from the principles of the invention or beyond the scope of the appended claims.
Example 1
The chiral sulfur nanoplatelets were prepared as follows:
(1) Referring to the reported preparation method of the two-dimensional sulfur nano-sheet, 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 (ultrasonic power is 425W), and centrifuged at 5000rpm for 30 minutes, and the supernatant is collected to obtain the pure two-dimensional sulfur nano-sheet aqueous solution. The concentration of S in the solution was calculated by ICP (inductively coupled plasma spectroscopy) testing, standard curve.
(2) According to the mass ratio of chiral amino acid to pure two-dimensional sulfur nano-sheet of 100:1, adding dextro-histidine (D-His) into an aqueous solution of a pure two-dimensional sulfur nano sheet, uniformly dispersing by ultrasonic waves (the power of ultrasonic waves is 300W and the time is 30 min), and then stirring and reacting for 12h at room temperature: and carrying out ultrafiltration and centrifugal separation (the centrifugal speed is 2500rpm, the centrifugal time is 30 min) on the obtained product, and washing the obtained product with deionized water to obtain the chiral two-dimensional sulfur nano-sheet.
Fig. 2 is a transmission electron microscope image of the chiral two-dimensional sulfur nano-sheet obtained in this example, and the characterization method is as follows: dropping the aqueous dispersion of the chiral sulfur nano-sheet onto a transmission electron microscope copper mesh, airing, and putting into a transmission electron microscope for observation. The nanoplatelets can be seen to have diameters of 200 to 300nm.
FIG. 3 is an atomic force microscope image of chiral sulfur nanoplatelets obtained in this example, characterized by: dropping the aqueous dispersion of the chiral sulfur nano-sheet onto the mica sheet, airing, and observing by an atomic force microscope. The thickness of the nanoplatelets can be seen from the figure to be 4-6 nm.
Fig. 4 is a Zeta potential diagram of the chiral two-dimensional sulfur nano-sheet obtained in this example, and the characterization method is as follows: 680. Mu.L of the aqueous dispersion of chiral two-dimensional sulfur nanoplatelets was added to a Zeta potential sample cell and tested using a NanoZS90 Markov particle sizer. From the figures it can be seen that: the Zeta potential value of the pure two-dimensional sulfur nano-sheet is-11.2 mV, and after chiral amino acid molecule modification, the Zeta potential value is reduced to-33.8 mV, and the result shows that the amino acid molecule is successfully loaded on the surface of the two-dimensional sulfur nano-sheet, and the reduction of the Zeta potential value is probably caused by that carboxyl groups of the amino acid molecule dissociate from a proton to be negatively charged.
FIG. 5 is a comparison chart of Raman spectra of chiral two-dimensional sulfur nanoplatelets, pure two-dimensional sulfur nanoplatelets and D-histidine obtained in the present example, and the characterization method is as follows: and (3) dripping the chiral two-dimensional sulfur nano-sheet, the pure two-dimensional sulfur nano-sheet or the D-histidine water dispersion liquid on a quartz sheet, airing, and testing by using a microscopic confocal laser Raman spectrometer. From the figure, the chiral two-dimensional sulfur nano-sheet can be seen to show obvious Raman peaks, which are completely consistent with positions of Raman peaks of pure two-dimensional sulfur nano-sheet and chiral amino acid, and the fact that D-His is successfully modified to the two-dimensional sulfur nano-sheet is confirmed, and the prepared chiral two-dimensional sulfur nano-sheet has a unit cell structure similar to that of the two-dimensional sulfur nano-sheet.
FIG. 6 is an ultraviolet-visible light absorption spectrum of the chiral two-dimensional sulfur nanoplatelets obtained in this example, and is characterized by the following steps: aqueous dispersions of chiral two-dimensional sulfur nanoplatelets of different concentrations (5 μg/mL,10 μg/mL,15 μg/mL,20 μg/mL,25 μg/mL) were prepared and tested for UV-visible absorption spectra. As can be seen from the graph, the ultraviolet-visible light absorption spectrum of the prepared chiral two-dimensional sulfur nano-sheet clearly shows full-band absorption at 200-600 nm, the absorption is concentration-dependent, and the absorption value of the chiral two-dimensional sulfur nano-sheet is increased along with the increase of the concentration.
Fig. 7 is a circular dichroism spectrum of the chiral two-dimensional sulfur nano-sheet obtained in this example, and the characterization method is as follows: 2mL of aqueous dispersion of chiral two-dimensional sulfur nano-sheets is added into a quartz cuvette, and the chirality is tested by using a circular dichroscope. From the figure, it can be seen that the nano-sheet shows a distinct CD absorption band between 200 and 250nm, while the corresponding L-His@S-NSs prepared by the L-histidine (L-His) in the same way as in example 1 shows a distinct CD absorption band with opposite sign and similar amplitude between 200 and 250nm, and the pure two-dimensional sulfur nano-sheet does not show a significant CD signal, which proves that the chiral two-dimensional sulfur nano-sheet prepared in the example has a remarkable chiral characteristic.
FIG. 8 is a 7-day stability chart of chiral two-dimensional sulfur nanoplatelets and pure two-dimensional sulfur nanoplatelets obtained in this example, characterized by: the two-dimensional sulfur nano-sheet or the aqueous dispersion of the chiral two-dimensional sulfur nano-sheet is stored in a refrigerator at the temperature of 4 ℃, the size of the aqueous dispersion is tested by using a laser particle analyzer every other day, and the prepared chiral two-dimensional sulfur nano-sheet has good stability and basically unchanged size under the condition of water dispersion.
Fig. 9 is a transmission electron microscope image of renaturation of the chiral two-dimensional sulfur nano-sheet obtained in the present example after lyophilization, and the characterization method is as follows: the chiral two-dimensional sulfur nanoplatelets obtained in this example were lyophilized into powder by a lyophilizer and then redispersed in deionized water. The figure shows that the chiral sulfur nano-sheet has good stability, and the appearance of the chiral sulfur nano-sheet is not changed by freeze drying.
To verify the selective antibacterial properties of the chiral two-dimensional sulfur nanoplatelets obtained in this example, the following test was performed: mu.L of LB broth medium containing methicillin-resistant Staphylococcus aureus (MRSA) (colony density of MRSA in the medium was 1X 10) 7 CFU/mL) and 180 mu L of the chiral two-dimensional sulfur nano-sheet (D-His@S-NSs) prepared in the embodiment are mixed, incubated for 4 hours at 37 ℃, 100 mu L of bacterial liquid is absorbed and uniformly coated on an agar culture plate, incubated for 48 hours at 37 ℃, and the growth condition of bacteria is judged through the colony number on an agar culture dish. For comparison, this example also tested sublimated sulfur powder (S), pure two-dimensional sulfur nanoplatelets (S-NSs) and L-His@S-NSs prepared by L-histidine (L-His) in the same manner. And samples without the above materials were used as control groups. To verify the selective killing effect of the chiral two-dimensional sulfur nanoplatelets obtained in this example on gram-positive bacteria and gram-negative bacteria, the following test was performed: the antibacterial properties of chiral two-dimensional sulfur nanoplatelets, sublimed sulfur powder (S), pure two-dimensional sulfur nanoplatelets (S-NSs) and L-His@S-NSs against E.coli (E.coli) were tested in the same manner, and samples without the above materials were used as control groups. As a result, as shown in FIG. 10, it can be seen that the D-His@S-NSs treated MRSA growth was significantly inhibited, and other groups of petri dish bacterial colonies showed incomplete killing of the material to varying degrees. The petri dish bacterial colonies of all material groups treated e.coli showed normal growth of bacteria. The results show that the implementationThe chiral two-dimensional sulfur nano-sheet obtained by the method has antibacterial activity for selectively killing gram-positive bacteria (such as MRSA).
To further demonstrate the antimicrobial properties of the chiral two-dimensional sulfur nanoplatelets obtained in this example, the following test was performed: centrifuging and washing LB broth medium containing methicillin-resistant staphylococcus aureus (MRSA) with physiological saline at 6000rpm for 5min for 3-5 times until colony density of MRSA is 2×10 9 CFU/mL, 20 mu L of bacterial liquid is mixed with 180 mu L of chiral two-dimensional sulfur nano-sheets (D-His@S-NSs) prepared in the embodiment, incubation is carried out at 37 ℃ for 4 hours, bacterial dead-alive staining of syto9/PI is carried out, and the dead-alive of bacteria is judged by observing fluorescence emitted by a fluorescence microscope. For comparison, this example also tested sublimated sulfur powder (S), pure two-dimensional sulfur nanoplatelets (S-NSs) and L-His@S-NSs prepared by L-histidine (L-His) in the same manner. And samples without the above materials were used as control groups. To further verify the selective killing effect of the chiral two-dimensional sulfur nanoplatelets obtained in this example on gram-positive bacteria and gram-negative bacteria, the following test was performed: chiral two-dimensional sulfur nanoplatelets, sublimed sulfur powder (S), pure two-dimensional sulfur nanoplatelets (S-NSs) and bacterial dead-living staining of E.coli (E.coli) by L-His@S-NSs prepared by the same method as L-histidine (L-His) were tested in the same manner. The results are shown in fig. 11, and it can be seen that MRSA treated with D-his@s-NSs showed intense red fluorescence, indicating that the group of bacteria was killed, the other control group showed a large amount of green fluorescence, and the e.coll treated with chiral two-dimensional sulfur nanoplatelets also showed a large amount of green fluorescence, consistent with the bacterial plating experimental phenomenon. Further shows that the chiral two-dimensional sulfur nano-sheet obtained in the example has the antibacterial activity of selectively killing gram-positive bacteria (such as MRSA).
To demonstrate again the antibacterial properties of the chiral two-dimensional sulfur nanoplatelets obtained in this example, the following test was performed: mu.L of LB broth medium containing methicillin-resistant Staphylococcus aureus (MRSA) (colony density of MRSA in the medium was 1X 10) 7 CFU/mL) and 180. Mu.L, 150. Mu.g/mL of chiral two-dimensional sulfur nanoplatelets (D-H) prepared in this exampleis@S-NSs), incubated at 37℃for 4 hours, absorbance at OD600nm was measured using a microplate reader, once every 1 hour, and the growth state of the bacteria was evaluated. For comparison, in this example, phosphate Buffer (PBS), sublimed sulfur powder (S), pure two-dimensional sulfur nanoplatelets (S-NSs), and D-His were also tested in the same manner, and as shown in FIG. 12, it can be seen that MRSA treated with D-His@S-NSs was inhibited from the initial 1h of bacterial growth, and no significant bacterial growth and proliferation occurred over time; bacteria of other control groups all increased in absorbance over time, indicating incomplete killing of bacteria.
FIG. 13 is a graph of the biocompatibility of chiral sulfur nanoplatelets at different concentrations, characterized by: the D-His@S-NSs aqueous dispersion is diluted to 2 mug/mL, 50 mug/mL, 75 mug/mL, 100 mug/mL and 150 mug/mL, 0.2mL of the above solutions with different concentrations are respectively washed with 0.2mL of treated blood (500 mug fresh blood is added with 4.5mL of physiological saline for 5-8 times, the centrifugal speed is 3000rpm, the centrifugal time is 10min, the supernatant is discarded after the supernatant is clear and transparent, the physiological saline is used for constant volume to 5 mL) and 0.6mL of physiological saline are mixed for incubation for 4h at 37 ℃, then the supernatant is centrifuged at 3000rpm for 10min, the absorbance at OD541nm is measured by sucking the supernatant, and the hemolysis rate is calculated. From the graph, the hemolysis rate of chiral two-dimensional sulfur nano-sheets with different concentrations is lower than 5%, which indicates that the biocompatibility of the material is good.
FIG. 14 is a graph showing comparison of hemolysis rates calculated by the above method using 150. Mu.g/mL of sublimed sulfur powder (S), two-dimensional sulfur nanoplatelets (S-NSs), L-His@S-NSs and D-His@S-NSs, respectively, and it can be seen from the graph that the hemolysis rates of the materials of each group are lower than 5%, indicating that the sulfur element has good biosafety.
Example 2
The chiral two-dimensional sulfur nanoplatelets were prepared as follows:
(1) Referring to the reported preparation method of the two-dimensional sulfur nano-sheet, 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 (ultrasonic power is 425W), and centrifuged at 5000rpm for 30 minutes, and the supernatant is collected to obtain the pure two-dimensional sulfur nano-sheet aqueous solution. The concentration of S in the solution was calculated by ICP (inductively coupled plasma spectroscopy) testing, standard curve.
(2) The mass ratio of the chiral amino acid to the pure two-dimensional sulfur nano-sheet is 50:1, adding D-histidine (D-His) into an aqueous solution of a pure two-dimensional sulfur nano sheet, uniformly dispersing by ultrasonic waves (the power of ultrasonic waves is 300W and the time is 30 min), and then stirring and reacting for 12h at room temperature: and carrying out ultrafiltration and centrifugal separation (the centrifugal speed is 2500rpm, the centrifugal time is 30 min) on the obtained product, and washing the obtained product with deionized water to obtain the chiral two-dimensional sulfur nano-sheet.
The chiral two-dimensional sulfur nano-sheet obtained by the embodiment has the diameter of 200-300 nm and the thickness of 3-5 nm, and has good selective antibacterial activity on gram-positive bacteria.
The foregoing description of the preferred embodiments of the invention is not intended to be limiting, but rather is intended to cover all modifications, equivalents, and alternatives falling within the spirit and principles of the invention.
Claims (5)
1. A chiral two-dimensional sulfur nanoplatelet for selectively combating gram positive bacteria, characterized in that: the chiral two-dimensional sulfur nano-sheet takes a two-dimensional sulfur nano-sheet as a core, and chiral amino acid molecules are modified on the surface of the two-dimensional sulfur nano-sheet; the chiral amino acid molecule is dextrorotatory amino acid;
the preparation method of the chiral two-dimensional sulfur nano-sheet comprises the following steps: adding chiral amino acid into the water solution of the pure two-dimensional sulfur nano-sheet, uniformly dispersing by ultrasonic, and then stirring and reacting for 12-24 hours at room temperature: performing ultrafiltration and centrifugal separation on the obtained product to obtain chiral two-dimensional sulfur nano-sheets; wherein, the mass ratio of the chiral amino acid to the pure two-dimensional sulfur nano-sheet is 10-100: 1.
2. the chiral two-dimensional sulfur nanoplatelets of claim 1, wherein: the thickness of the chiral two-dimensional sulfur nano-sheet is 3-6 nm, and the diameter is 200-300 nm.
3. The chiral two-dimensional sulfur nano-sheet according to claim 1, wherein the power of the ultrasonic wave is 300W and the time is 0.5-1 h.
4. The chiral two-dimensional sulfur nanoplatelets of claim 1, wherein: the rotational speed of the centrifugation is 2500rpm, and the centrifugation time is 15-30 min.
5. Use of the chiral two-dimensional sulfur nanoplatelets of any of claims 1 to 4, characterized in that: a selective antimicrobial agent for the preparation of 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 CN114681481A (en) | 2022-07-01 |
| CN114681481B true 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 (4)
| 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 |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 (4)
| 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 |
Also Published As
| Publication number | Publication date |
|---|---|
| CN114681481A (en) | 2022-07-01 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Vijayakumar et al. | Biosynthesis, characterisation and anti-bacterial effect of plant-mediated silver nanoparticles using Artemisia nilagirica | |
| Zhang et al. | Synergetic antibacterial effects of silver nanoparticles@ aloe vera prepared via a green method | |
| Logpriya et al. | Preparation and characterization of ascorbic acid-mediated chitosan–copper oxide nanocomposite for anti-microbial, sporicidal and biofilm-inhibitory activity | |
| Öztürk | Intracellular and extracellular green synthesis of silver nanoparticles using Desmodesmus sp.: Their antibacterial and antifungal effects | |
| Hungund et al. | Comparative evaluation of antibacterial activity of silver nanoparticles biosynthesized using fruit juices | |
| Rajgovind et al. | Pterocarpus marsupium derived phyto-synthesis of copper oxide nanoparticles and their antimicrobial activities | |
| WO2020028391A1 (en) | Biopolymer-coated two-dimensional transition metal chalcogenides having potent antimicrobial activity | |
| WO2009063508A2 (en) | Nanoparticle composition and process thereof | |
| CN109845761B (en) | Application of lysozyme two-dimensional nano-film as antibacterial material | |
| Saleh et al. | Bio-synthesis of silver nanoparticles from bacteria Klebsiella pneumonia: Their characterization and antibacterial studies | |
| Du et al. | Size-dependent antibacterial of carbon dots by selective absorption and differential oxidative stress of bacteria | |
| Fatih et al. | Antimicrobial and antibiofilm activity of green synthesized silver nanoparticles by using aqueous leaf extract of Thymus serpyllum | |
| Sundara Selvam et al. | Cadmium oxide-zinc oxide nanocomposites synthesized using waste eggshell membrane and its in-vitro assessments of the antimicrobial activities and minimum inhibitory concentration | |
| Parveen et al. | Emphasized mechanistic antimicrobial study of biofunctionalized silver nanoparticles on model Proteus mirabilis | |
| Anand et al. | Carbon dots for bacterial detection and antibacterial applications-a mini review | |
| Bhatia et al. | Duchsnea indica plant extract mediated synthesis of copper oxide nanomaterials for antimicrobial activity and free-radical scavenging assay | |
| 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 | |
| Uzair et al. | Targeting microbial biofilms: by Arctium lappa l. synthesised biocompatible CeO2‐NPs encapsulated in nano‐chitosan | |
| Sivakamavalli et al. | Garcinia cambogia assisted synthesis of Zno nanoparticles coupled with chitosan for antibacterial, antibiofilm, cytotoxic, anticancer and ecotoxicity assessment | |
| Abalaka et al. | The antibacterial efficacy of gold nanoparticles derived from Gomphrena celosioides and Prunus amygdalus (Almond) leaves on selected bacterial pathogens | |
| Shnawa et al. | Antimicrobial activity of plant-extract-mediated synthesis of Silver-Zinc Oxide nanocomposites and their acaricidal efficacy on Hyalomma marginatum ticks | |
| CN114681481B (en) | Chiral two-dimensional sulfur nano-sheet for selectively resisting gram-positive bacteria and preparation method and application thereof | |
| Shah et al. | Histopathological and hematological investigations of mice model inoculated with nickel oxide nanoparticles and bacterial pathogens: In-vitro and in-vivo antibacterial studies | |
| Nguyen et al. | Enhancement of antibacterial activity by a copper (II) and zinc (II) in chelation with ethylenediaminetetra-acetic acid and urea complex | |
| Pham et al. | Facile ultrasound-assisted green synthesis of nio/chitosan nanocomposite from mangosteen peel extracts as antibacterial agents |
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 |