CN114177289B - Composite nano material for photodynamic and photothermal combined treatment and preparation method and application thereof - Google Patents
Composite nano material for photodynamic and photothermal combined treatment and preparation method and application thereof Download PDFInfo
- Publication number
- CN114177289B CN114177289B CN202111538603.3A CN202111538603A CN114177289B CN 114177289 B CN114177289 B CN 114177289B CN 202111538603 A CN202111538603 A CN 202111538603A CN 114177289 B CN114177289 B CN 114177289B
- Authority
- CN
- China
- Prior art keywords
- carbon nitride
- phase carbon
- graphite
- copper sulfide
- loaded
- 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
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K41/00—Medicinal preparations obtained by treating materials with wave energy or particle radiation ; Therapies using these preparations
- A61K41/0057—Photodynamic 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
-
- 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
- A61K33/24—Heavy metals; Compounds thereof
- A61K33/32—Manganese; Compounds thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K41/00—Medicinal preparations obtained by treating materials with wave energy or particle radiation ; Therapies using these preparations
- A61K41/0052—Thermotherapy; Hyperthermia; Magnetic induction; Induction heating therapy
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
- A61K9/50—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals
- A61K9/51—Nanocapsules; Nanoparticles
- A61K9/5107—Excipients; Inactive ingredients
- A61K9/513—Organic macromolecular compounds; Dendrimers
- A61K9/5146—Organic macromolecular compounds; Dendrimers obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyethylene glycol, polyamines, polyanhydrides
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
- A61K9/50—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals
- A61K9/51—Nanocapsules; Nanoparticles
- A61K9/5192—Processes
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y30/00—Nanotechnology for materials or surface science, e.g. nanocomposites
-
- 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
-
- C—CHEMISTRY; METALLURGY
- C01—INORGANIC CHEMISTRY
- C01B—NON-METALLIC ELEMENTS; COMPOUNDS THEREOF; METALLOIDS OR COMPOUNDS THEREOF NOT COVERED BY SUBCLASS C01C
- C01B21/00—Nitrogen; Compounds thereof
- C01B21/06—Binary compounds of nitrogen with metals, with silicon, or with boron, or with carbon, i.e. nitrides; Compounds of nitrogen with more than one metal, silicon or boron
- C01B21/0605—Binary compounds of nitrogen with carbon
-
- C—CHEMISTRY; METALLURGY
- C01—INORGANIC CHEMISTRY
- C01G—COMPOUNDS CONTAINING METALS NOT COVERED BY SUBCLASSES C01D OR C01F
- C01G3/00—Compounds of copper
- C01G3/12—Sulfides
-
- C—CHEMISTRY; METALLURGY
- C01—INORGANIC CHEMISTRY
- C01G—COMPOUNDS CONTAINING METALS NOT COVERED BY SUBCLASSES C01D OR C01F
- C01G45/00—Compounds of manganese
- C01G45/02—Oxides; Hydroxides
-
- C—CHEMISTRY; METALLURGY
- C01—INORGANIC CHEMISTRY
- C01P—INDEXING SCHEME RELATING TO STRUCTURAL AND PHYSICAL ASPECTS OF SOLID INORGANIC COMPOUNDS
- C01P2004/00—Particle morphology
- C01P2004/80—Particles consisting of a mixture of two or more inorganic phases
Landscapes
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Nanotechnology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Organic Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Epidemiology (AREA)
- Physics & Mathematics (AREA)
- Inorganic Chemistry (AREA)
- Crystallography & Structural Chemistry (AREA)
- Molecular Biology (AREA)
- Optics & Photonics (AREA)
- Condensed Matter Physics & Semiconductors (AREA)
- General Physics & Mathematics (AREA)
- Biomedical Technology (AREA)
- Manufacturing & Machinery (AREA)
- Biochemistry (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Composite Materials (AREA)
- Materials Engineering (AREA)
- Biophysics (AREA)
- Biotechnology (AREA)
- General Engineering & Computer Science (AREA)
- Medical Informatics (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
The invention relates to the technical field of medical materials, in particular to a composite nano material for photodynamic and photothermal combined treatment, and a preparation method and application thereof. The preparation method of the composite nano material comprises the following steps: dispersing graphite-phase carbon nitride in water, adding copper acetate, stirring, mixing the obtained solution with thiourea aqueous solution, stirring to obtain a suspension, and carrying out hydrothermal reaction on the suspension to obtain copper sulfide-loaded graphite-phase carbon nitride; dispersing graphite-phase carbon nitride loaded with copper sulfide in water, adding a potassium permanganate aqueous solution and a polyallylamine hydrochloride solution, mixing and stirring to obtain graphite-phase carbon nitride loaded with copper sulfide and manganese dioxide; and dispersing graphite-phase carbon nitride loaded with copper sulfide and manganese dioxide into F127 aqueous solution, and stirring overnight to obtain the composite nano material. The composite nano material has good photo-thermal conversion capability, active oxygen generation capability, glutathione consumption capability and the like, and is suitable for photodynamic and photo-thermal combined treatment.
Description
Technical Field
The application relates to the technical field of medical materials, in particular to a composite nano material for photodynamic and photothermal combined treatment, and a preparation method and application thereof.
Background
The traditional cancer treatment means mainly comprise surgical excision, chemotherapy, radiotherapy and the like, but the traditional cancer treatment means have the defects of low curative effect and large side effect, so the traditional cancer treatment means are more effective and have smaller toxic and side effect on the body of a patient, and become research hot spots.
Among these new cancer treatments, photodynamic therapy (Photodynamic Therapy, PDT for short) has received attention because of its potential for therapeutic effects and relatively little harm to patients. However, when applied to tumor treatment, the technology can limit the treatment efficiency of photodynamic therapy due to the existence of hypoxic tumor microenvironment, and even lead to infiltration and metastasis of tumors, so that improvement of the technology is needed.
Disclosure of Invention
The application discloses a composite nano material for photodynamic and photothermal combined treatment, a preparation method and application thereof, and aims to solve the defects of the existing photodynamic therapy in the aspects of cancer treatment efficiency and the like.
In a first aspect, the present application provides a method for preparing a composite nanomaterial for photodynamic and photothermal combination therapy, the method comprising the steps of:
Dispersing graphite-phase carbon nitride in water, adding copper acetate, stirring, mixing and stirring the obtained solution with thiourea aqueous solution to obtain a suspension, and carrying out hydrothermal reaction on the suspension to obtain copper sulfide-loaded graphite-phase carbon nitride;
dispersing the graphite-phase carbon nitride loaded with copper sulfide in water, adding a potassium permanganate aqueous solution and a polyallylamine hydrochloride solution, mixing and stirring to obtain the graphite-phase carbon nitride loaded with copper sulfide and manganese dioxide;
and dispersing the graphite-phase carbon nitride loaded with copper sulfide and manganese dioxide into F127 aqueous solution, and stirring overnight to obtain the composite nano material.
Further, the preparation method further comprises the step of preparing the graphite phase carbon nitride before the step of preparing the graphite phase carbon nitride loaded with copper sulfide, wherein the preparation step of the graphite phase carbon nitride is as follows: calcining melamine in air, and grinding to obtain the graphite-phase carbon nitride.
Further, the preparation steps of the graphite phase carbon nitride are as follows: under the air condition, heating melamine to 550 ℃ at a heating rate of 2.5 ℃/min, cooling to room temperature after heating for 4 hours, collecting yellow powder, washing with deionized water and drying;
The yellow powder was ball milled at 500rpm for 6 hours and the finely milled graphite phase carbon nitride was collected by filtration.
Further, in the step of preparing the graphite-phase carbon nitride loaded with copper sulfide, the copper sulfide accounts for 3-10% of the graphite-phase carbon nitride by mass, and the hydrothermal reaction condition is that the reaction is carried out for 20-30 hours at 140-160 ℃.
Further, the preparation method of the graphite phase carbon nitride loaded with copper sulfide comprises the following steps: dispersing 0.5g of graphite phase carbon nitride in 60mL of deionized water, reacting for 1 hour under ultrasonic conditions, adding 0.068g of copper acetate, and continuously stirring for 2 hours;
mixing the obtained solution with 40mL of the thiourea aqueous solution, and stirring for 5 hours under the magnetic force condition to obtain a suspension; wherein the thiourea aqueous solution contains 0.074g of thiourea;
placing the suspension in an autoclave lined with polytetrafluoroethylene, and reacting for 24 hours at 150 ℃;
and (3) centrifugally separating to obtain a precipitate, washing the precipitate for multiple times, and drying the precipitate at 80 ℃ to obtain the graphite phase carbon nitride loaded with copper sulfide.
Further, in the step of preparing the graphite-phase carbon nitride loaded with copper sulfide and manganese dioxide, the manganese dioxide accounts for 1-3% of the graphite-phase carbon nitride loaded with copper sulfide;
In the step of preparing the composite nanomaterial, the mass ratio of the graphite phase carbon nitride loaded with copper sulfide and manganese dioxide to F127 is 5:4.
Further, the preparation method of the graphite phase carbon nitride loaded with copper sulfide and manganese dioxide comprises the following steps: dispersing 0.15g of the graphite-phase carbon nitride loaded with copper sulfide in 60mL of deionized water at room temperature, dripping 10mL of potassium permanganate aqueous solution with the concentration of 0.5mg/mL, mixing with 6.4mL of cationic polyacrylamide aqueous solution with the concentration of 5mg/mL, stirring for 30min, washing, centrifuging and drying the obtained product to obtain the graphite-phase carbon nitride loaded with copper sulfide and manganese dioxide;
the preparation method of the composite nano material comprises the following steps: 50mg of the graphite phase carbon nitride loaded with copper sulfide and manganese dioxide is dispersed into 20mL of the F127 aqueous solution with the concentration of 2mg/mL, stirred overnight, centrifugally filtered, and the composite nano material is collected.
In a second aspect, the present application provides a composite nanomaterial for photodynamic and photothermal combination therapy, the composite nanomaterial being produced by the production method as described in the first aspect.
In a third aspect, the present application provides an application of the composite nanomaterial for photodynamic and photothermal combined treatment according to the second aspect in preparing a medicament for treating a tumor disease.
The application also provides an application of the composite nanomaterial for photodynamic and photothermal combined treatment in preparing a medicament for photodynamic and photothermal combined treatment of tumor diseases.
Compared with the prior art, the application has the following beneficial effects:
in the embodiment of the application, graphite-phase carbon nitride, copper acetate and thiourea are subjected to hydrothermal reaction to successfully obtain the composite material, namely the copper sulfide-loaded graphite-phase carbon nitride, which takes the graphite-phase carbon nitride as a carrier and copper sulfide nano particles as a photo-thermal conversion agent. On the basis, the graphite-phase carbon nitride loaded with copper sulfide reacts with potassium permanganate and cationic polyacrylamide hydrochloride, the potassium permanganate is reduced into manganese dioxide nano particles by utilizing the cationic polyacrylamide hydrochloride, and the manganese dioxide nano particles are successfully attached to the surface of the graphite-phase carbon nitride loaded with copper sulfide, so that the graphite-phase carbon nitride loaded with copper sulfide nano particles and manganese dioxide nano particles simultaneously is obtained. Finally, the graphite phase carbon nitride loaded with the copper sulfide nano particles and the manganese dioxide nano particles is successfully coated by using a surface active substance F127, so that the composite nano material for photodynamic and photothermal combined treatment is obtained.
The composite nanomaterial can be used for photodynamic and photothermal combination therapy. Wherein the graphite phase carbon nitride (i.e., g-C 3 N 4 ) Has the advantages of high active oxygen yield, good biocompatibility, no toxicity and the like, and can be used as the photosensitizer of the embodiment of the application. Copper sulfide (namely CuS) has a wider light absorption range, higher light-heat conversion efficiency and light stability, and can be used as a light-heat conversion agent. Compared with traditional photothermal agents such as gold nanoparticles, platinum and the like, the copper sulfide is nontoxic, lower in price, better in biological safety and more suitable for application in the field of biological medicine. Manganese dioxide (MnO) 2 ) The nanomaterial is an enzyme similar to catalase and can be used for combining H in tumor environment 2 O 2 Reaction at H + O production in the presence of 2 Improving anoxia in tumor environment and improving photodynamic therapy efficiency. In addition, manganese dioxide can reduce glutathione levels in tumor cells by consuming glutathione, thereby reducing active oxygen consumption and improving anticancer efficiency of photodynamic therapy. Polyether F127 as a hydrophilic polymer can remarkably improve the water dispersibility of the nano particles, so that the stability of the composite nano material is stronger.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present application, the drawings that are needed in the embodiments will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present application, and other drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.
FIG. 1 is a microstructure of the graphite phase carbon nitride, composite nanomaterial of example 1;
FIG. 2 is an XRD pattern of the composite nanomaterial of example 1 and an intermediate product of the synthesis process thereof;
FIG. 3 is an XPS spectrum of the graphite phase carbon nitride and composite nanomaterial of example 1;
FIG. 4 is an XPS spectrum of the composite nanomaterial of example 1 at Cu 2p, mn 2 p;
FIG. 5 is an ultraviolet-visible absorption spectrum of the composite nanomaterial of example 1 and an intermediate product of the synthesis process thereof;
FIG. 6 is an ultraviolet-visible absorption spectrum of DPBF blended with composite nanomaterial, graphite phase carbon nitride, respectively, of example 1;
FIG. 7 is a graph showing the photo-thermal conversion ability and photo-stability test results of the composite nanomaterial of example 1;
FIG. 8 is a graph showing glutathione consumption of the composite nanomaterial of example 1 and an intermediate product of the synthesis process thereof;
FIG. 9 is a graph of intracellular ROS production of the composite nanomaterial of example 1 and an intermediate product of the synthesis process;
FIG. 10 is a graph showing the results of oxygen generation of the composite nanomaterial of example 1 and an intermediate product of the synthesis process thereof;
FIG. 11 is a graph showing the results of darkroom toxicity analysis of the composite nanomaterial of example 1 and the intermediate products of its synthesis on HepG2 cells at various concentrations;
FIG. 12 is a graph showing the results of the viability of the composite nanomaterial of example 1 and the cells treated as intermediates in the synthesis process under different light conditions.
Detailed Description
The following description of the embodiments of the present invention will be made clearly and completely with reference to the accompanying drawings, in which it is apparent that the embodiments described are only some embodiments of the present invention, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
In the present invention, the terms "upper", "lower", "left", "right", "front", "rear", "top", "bottom", "inner", "outer", "middle", "vertical", "horizontal", "lateral", "longitudinal" and the like indicate an azimuth or a positional relationship based on that shown in the drawings. These terms are only used to better describe the present invention and its embodiments and are not intended to limit the scope of the indicated devices, elements or components to the particular orientations or to configure and operate in the particular orientations.
Also, some of the terms described above may be used to indicate other meanings in addition to orientation or positional relationships, for example, the term "upper" may also be used to indicate some sort of attachment or connection in some cases. The specific meaning of these terms in the present invention will be understood by those of ordinary skill in the art according to the specific circumstances.
Furthermore, the terms "mounted," "configured," "provided," "connected," and "connected" are to be construed broadly. For example, it may be a fixed connection, a removable connection, or a unitary construction; may be a mechanical connection, or an electrical connection; may be directly connected, or indirectly connected through intervening media, or may be in internal communication between two devices, elements, or components. The specific meaning of the above terms in the present invention can be understood by those of ordinary skill in the art according to the specific circumstances.
Furthermore, the terms "first," "second," and the like, are used primarily to distinguish between different devices, elements, or components (the particular species and configurations may be the same or different), and are not used to indicate or imply the relative importance and number of devices, elements, or components indicated. Unless otherwise indicated, the meaning of "a plurality" is two or more.
In addition, the reaction materials, reagents, and solvents used in the examples of the present application are commercially available.
The technical scheme of the application will be further described with reference to specific embodiments and drawings.
Conventional cancer treatment methods (such as surgical excision, chemotherapy and radiotherapy) generally have the disadvantages of low curative effect, large toxic and side effects on patients and the like, so researchers are always searching for new cancer treatment technologies to overcome the problems.
Photodynamic therapy and photothermal therapy are both emerging cancer treatments in the medical field. Among them, photodynamic therapy is a minimally invasive anticancer treatment method, which has the advantage of mild injury to patients, and it is realized that three conditions, a photosensitizer, a light source and tissue oxygen, are required to be satisfied, and when the photosensitizer is exposed to light of a certain wavelength, its energy is transferred to surrounding molecular oxygen to generate active oxygen (Reactive Oxygen Species, abbreviated as ROS), by which cancer cells are killed. However, there are certain limitations to this therapy: solid tumors typically present a hypoxic tumor microenvironment (Tumor Microenvironment, TME for short), which not only limits the therapeutic efficiency of photodynamic therapy, but also promotes tumor infiltration and metastasis. In addition, oxygen depletion in photodynamic therapy can further exacerbate tumor hypoxia, leading to failure of the therapy in the treatment of cancer.
Near infrared mediated phototherapy (Photothermal Therapy, PTT for short) is mainly used to kill tumor cells by using the local thermal effect of the photothermal conversion material. However, the application of photothermal therapy is limited due to the limited penetration depth of near infrared light and the thermal damage to normal cells and tissues caused by local overheating during the hyperthermia process.
In order to solve the problems of the emerging cancer treatment methods in the related art, so that the emerging cancer treatment methods are more efficiently suitable for cancer treatment, the toxic and side effect influence of the cancer treatment methods on patients is reduced as much as possible, the embodiment of the application provides a composite nanomaterial for photodynamic and photothermal combined treatment, and a preparation method and application thereof, and the composite nanomaterial can realize the cooperative treatment of the cancers by photodynamic therapy and photothermal therapy, avoid multiple administration and can more effectively inhibit the cancers.
In a first aspect, an embodiment of the present application provides a method for preparing a composite nanomaterial for photodynamic and photothermal combined therapy, including the steps of:
dispersing graphite-phase carbon nitride in water, adding copper acetate, stirring, mixing and stirring the obtained solution with thiourea aqueous solution to obtain a suspension, and carrying out hydrothermal reaction on the suspension to obtain copper sulfide-loaded graphite-phase carbon nitride;
Dispersing the graphite-phase carbon nitride loaded with copper sulfide in water, adding a potassium permanganate aqueous solution and a polyallylamine hydrochloride solution, mixing and stirring to obtain the graphite-phase carbon nitride loaded with copper sulfide and manganese dioxide;
and dispersing the graphite-phase carbon nitride loaded with copper sulfide and manganese dioxide into F127 aqueous solution, and stirring overnight to obtain the composite nano material.
In the embodiment of the application, graphite-phase carbon nitride, copper acetate and thiourea are subjected to hydrothermal reaction to successfully obtain a composite material with the graphite-phase carbon nitride as a carrier and copper sulfide nano particles as a photo-thermal conversion agent, namely the copper sulfide-loaded graphite-phase carbon nitride, which is abbreviated as CNs-CuS, wherein CNs is the graphite-phase carbon nitride and CuS is the copper sulfide. On the basis, CNs-CuS, potassium permanganate and cationic polyacrylate amine hydrochloride (PAH for short, with molecular formula of (C) 3 H 7 N) n xHCl), reducing potassium permanganate into manganese dioxide nano particles by utilizing a cationic polyelectrolyte PAH, and enabling the manganese dioxide nano particles to be successfully attached to the CNs-CuS surface to obtain graphite-phase carbon nitride which is simultaneously loaded with copper sulfide nano particles and manganese dioxide nano particles, wherein the graphite-phase carbon nitride is called CNs-CuS/MnO for short 2 . Finally, the graphite loaded with both copper sulfide nano particles and manganese dioxide nano particles is successfully coated by a surface active substance F127Phase carbon nitride to obtain composite nanometer material for photodynamic and photothermal combined treatment, F127@CNs-CuS/MnO for short 2 。
Wherein F127 is polyether F127, a commercial reagent, commercially available.
Further, the preparation method further comprises the step of preparing the graphite phase carbon nitride before the step of preparing the graphite phase carbon nitride loaded with copper sulfide, wherein the preparation step of the graphite phase carbon nitride is as follows: calcining melamine in air, and grinding to obtain the graphite-phase carbon nitride.
Further, the preparation steps of the graphite phase carbon nitride specifically include: under the air condition, melamine is heated for 4 hours at 550 ℃ and at the heating rate of 2.5 ℃, then cooled to room temperature, yellow powder is collected, washed by deionized water and dried; the yellow powder was milled at 500rpm for 6 hours and the ultrafine graphite phase carbon nitride was collected by filtration. The grinding is specifically to put the graphite phase carbon nitride obtained after calcination into a planetary ball mill rotating at 500rpm for 6 hours to obtain the graphite phase carbon nitride nano material with small particle size.
When the graphite-phase carbon nitride nano material is prepared, melamine is calcined in air to obtain flaky graphite-phase carbon nitride with larger particle size, and the graphite-phase carbon nitride is ground into ultrafine particles so as to reduce the size of the graphite-phase carbon nitride, so that the graphite-phase carbon nitride can pass through cell membranes more easily and meets the application requirements of biology.
Further, in the step of preparing the graphite-phase carbon nitride loaded with copper sulfide, the copper sulfide accounts for 3-10% of the graphite-phase carbon nitride by mass, and the hydrothermal reaction condition is that the reaction is carried out for 20-30 hours at 140-160 ℃.
It is understood that the copper sulfide content of 3 to 10% by mass of the graphite phase carbon nitride includes any value within this range, for example, 3%, 5%, 7% or 10% by mass of the graphite phase carbon nitride. Preferably, the copper sulfide accounts for 7% of the graphite phase carbon nitride by mass. The reaction temperature of the hydrothermal reaction is 140 to 160℃and any value within this temperature range is included, and the conditions of the hydrothermal reaction are 140℃to 150℃or 160 ℃.
Further, the step of preparing the graphite phase carbon nitride loaded with copper sulfide comprises the following steps: dispersing 0.5g of the graphite-phase carbon nitride nanomaterial in 60mL of deionized water, reacting for 1 hour under ultrasonic conditions, adding 0.068g of copper acetate, and continuously stirring for 2 hours;
Mixing the obtained solution with 40mL of the thiourea aqueous solution, and stirring for 5 hours under the magnetic force condition to obtain a suspension; wherein the thiourea aqueous solution contains 0.074g of thiourea;
placing the suspension in an autoclave lined with polytetrafluoroethylene, and reacting for 24 hours at 150 ℃;
and (3) centrifugally separating to obtain a precipitate, washing the precipitate for multiple times, and drying the precipitate at 80 ℃ to obtain the graphite phase carbon nitride loaded with copper sulfide.
When the mass of the generated copper sulfide is 7% of that of the graphite phase carbon nitride, the effect of the finally obtained composite nano material is optimal. In addition, in the step of synthesizing the graphite-phase carbon nitride loaded with the copper sulfide, experiments show that when the hydrothermal reaction temperature is lower than 140 ℃, the problems of insufficient crystallization and incomplete reaction can occur, so that the hydrothermal reaction temperature of the embodiment of the application is controlled at 140-160 ℃.
Further, in the step of preparing the graphite-phase carbon nitride loaded with copper sulfide and manganese dioxide, the manganese dioxide accounts for 1-3% of the graphite-phase carbon nitride loaded with copper sulfide; in the step of preparing the composite nanomaterial, the mass ratio of the graphite phase carbon nitride loaded with copper sulfide and manganese dioxide to F127 is 5:4.
Wherein the manganese dioxide comprises 1 to 3% by mass of the copper sulfide-loaded graphite phase carbon nitride and any value within the range, for example, 1%, 1.5%, 2% or 3% by mass of the copper sulfide-loaded graphite phase carbon nitride. Preferably, the manganese dioxide accounts for 2% of the graphite phase carbon nitride loaded with copper sulfide.
Further, the preparation of the graphite phase carbon nitride loaded with copper sulfide and manganese dioxide comprises the following steps: dispersing 0.15g of the graphite-phase carbon nitride loaded with copper sulfide in 60mL of deionized water at room temperature, dripping 10mL of potassium permanganate aqueous solution with the concentration of 0.5mg/mL, mixing with 6.4mL of cationic polyacrylamide aqueous solution with the concentration of 5mg/mL, stirring for 30min, washing, centrifuging and drying the obtained product to obtain the graphite-phase carbon nitride loaded with copper sulfide and manganese dioxide;
the preparation method of the composite nano material comprises the following steps: 50mg of the graphite phase carbon nitride loaded with copper sulfide and manganese dioxide is dispersed into 20mL of the F127 aqueous solution with the concentration of 2mg/mL, stirred overnight, centrifugally filtered, and the composite nano material is collected.
When the mass of the generated manganese dioxide accounts for about 2% of the mass of the graphite phase carbon nitride loaded with the copper sulfide, the effect of the finally obtained composite nano material is optimal. In addition, the inventor also adopts other methods for loading manganese dioxide, such as a reaction of manganese sulfate and potassium permanganate, to obtain manganese dioxide, but experimental results show that the manganese dioxide prepared by the method has too large particle size and cannot be effectively adhered to the surface of graphite-phase carbon nitride.
In a second aspect, the embodiment of the application also provides a composite nanomaterial for photodynamic and photothermal combined treatment prepared by the preparation method, wherein the composite nanomaterial is obtained by coating graphite-phase carbon nitride loaded with copper sulfide and manganese dioxide in F127, and is abbreviated as F127@CNs-CuS/MnO 2 。
The composite nano material prepared by the embodiment of the application can be used for photodynamic and photothermal combined therapy. Wherein the graphite phase carbon nitride (i.e., g-C 3 N 4 ) Has the advantages of high active oxygen yield, good biocompatibility, no toxicity and the like, and can be used as the photosensitizer of the embodiment of the application. Copper sulfide (namely CuS) has a wider light absorption range, higher photo-thermal conversion efficiency and light stability, and can be used as photo-thermal A conversion agent. Compared with traditional photothermal agents such as gold nanoparticles, platinum and the like, the copper sulfide is nontoxic, lower in price, better in biological safety and more suitable for application in the field of biological medicine. Manganese dioxide (MnO) 2 ) The nanomaterial is an enzyme similar to catalase and can be used for combining H in tumor environment 2 O 2 Reaction at H + O production in the presence of 2 Improving anoxia in tumor environment and improving photodynamic therapy efficiency. In addition, manganese dioxide can reduce glutathione levels in tumor cells by consuming glutathione, thereby reducing active oxygen consumption and improving anticancer efficiency of photodynamic therapy. Polyether F127 as a hydrophilic polymer can remarkably improve the water dispersibility of the nano particles, so that the stability of the composite nano material is stronger.
In a third aspect, an embodiment of the present application provides an application of the above-mentioned composite nanomaterial for photodynamic and photothermal combined therapy in preparing a medicament for treating a tumor disease, and in particular, an application of the above-mentioned composite nanomaterial for photodynamic and photothermal combined therapy in preparing a medicament for treating a tumor disease.
The embodiment of the application discovers that the composite nano material has the capabilities of photo-thermal conversion capability, photodynamic capability, self-production of active oxygen and the like after a large number of performance tests are carried out on the prepared composite nano material, so that the composite nano material can be applied to the photodynamic-photo-thermal combined treatment, and the synergistic promotion effect of a photodynamic treatment method and a photothermal treatment method in cancer treatment is realized. On the one hand, the photodynamic therapy effect can be promoted to be improved by the photothermal therapy, namely, the photothermal therapy can not only induce apoptosis of tumor cells by increasing the temperature of the tumor, but also enhance the endocytosis of the photosensitizer, thereby promoting the photodynamic therapy effect to be improved. On the other hand, the anticancer effect of the photothermal therapy can be promoted by the photodynamic therapy, namely, the active oxygen generated by the photodynamic therapy can reduce the expression of heat shock proteins in cancer cells and improve the photothermal effect, thereby promoting the anticancer effect of the photothermal therapy. Therefore, when the composite nanomaterial is used for photodynamic and photothermal combined treatment, the synergistic effect of the composite nanomaterial is higher than that of the composite nanomaterial by using any monotherapy.
In order to describe the technical scheme and the technical effect of the present application in more detail, the present application will be further described by more specific examples, related test results, and the like.
Example 1
The embodiment provides a composite nanomaterial for photodynamic and photothermal combined treatment, and the preparation method comprises the following steps:
preparing graphite phase carbon nitride: 5g of melamine was placed in a crucible with a lid, then in a muffle furnace, heated to 550 ℃ at a heating rate of 2.5 ℃/min, after 4 hours of heating, cooled to room temperature, the yellow powder was collected and washed with deionized water and dried. The yellow powder thus prepared was put into a planetary ball mill rotating at 500rpm and milled for 6 hours. After grinding, the product is filtered and collected, and is ultrafine graphite phase carbon nitride which is a flaky nano material, called CNs for short. Wherein the superfine graphite phase carbon nitride refers to graphite phase carbon nitride with the grain diameter less than or equal to 200 nm.
Preparing graphite phase carbon nitride loaded with copper sulfide: after dispersing 0.5g CNs in 60mL deionized water and sonicating for 1 hour, 0.068g copper acetate was added and stirring was continued for 2 hours. The resulting stirred solution was then mixed with 40mL of an aqueous thiourea solution and stirred under magnetic force for 5 hours to give a suspension (wherein the aqueous thiourea solution contained 0.074g of thiourea). The resulting suspension was placed in a polytetrafluoroethylene-lined autoclave and maintained at 150℃for 24 hours. And (3) centrifugally separating after the reaction to obtain a precipitate, washing the precipitate for multiple times, and drying the precipitate at 80 ℃ to obtain graphite-phase carbon nitride loaded with copper sulfide, which is called CNs-CuS for short.
Preparing graphite phase carbon nitride loaded with copper sulfide and manganese dioxide: dispersing 0.15g CNs-CuS in 60mL deionized water at room temperature, then dripping 10mL of potassium permanganate aqueous solution with the concentration of 0.5mg/mL into the deionized water, mixing with 6.4mL of polyallylamine hydrochloride solution with the concentration of 5mg/mL, stirring for 30min, washing, centrifuging and drying the obtained product to obtain graphite-phase carbon nitride loaded with copper sulfide and manganese dioxide, namely CNs-CuS/MnO 2 。
Preparing a composite nano material: 50mg CNs-CuS/MnO 2 Dispersing into 20mL F127 water solution with concentration of 2mg/mL, stirring overnight, centrifugally filtering, collecting the product, namely the composite nano material, namely F127@CNs-CuS/MnO 2 。
Example 2
This example differs from example 1 only in that in the step of preparing the graphite-phase carbon nitride loaded with copper sulfide, copper sulfide accounts for 3% by mass of the graphite-phase carbon nitride.
Example 3
This example differs from example 1 only in that in the step of preparing the graphite-phase carbon nitride loaded with copper sulfide, copper sulfide accounts for 5% by mass of the graphite-phase carbon nitride.
Example 4
This example differs from example 1 only in that in the step of preparing the graphite-phase carbon nitride loaded with copper sulfide, copper sulfide accounts for 10% by mass of the graphite-phase carbon nitride.
Example 5
This example differs from example 1 only in that in the step of preparing the graphite-phase carbon nitride loaded with copper sulfide and manganese dioxide, the mass percentage of manganese dioxide to the graphite-phase carbon nitride loaded with copper sulfide is 1%.
Example 6
This example differs from example 1 only in that in the step of preparing the graphite-phase carbon nitride loaded with copper sulfide and manganese dioxide, the mass percentage of manganese dioxide to the graphite-phase carbon nitride loaded with copper sulfide is 1.5%.
Example 7
This example differs from example 1 only in that in the step of preparing the graphite-phase carbon nitride loaded with copper sulfide and manganese dioxide, the mass percentage of manganese dioxide to the graphite-phase carbon nitride loaded with copper sulfide is 3%.
Comparative example
This comparative example differs from example 1 only in that manganese sulfate was used instead of polyallylamine hydrochloride in the step of preparing graphite phase carbon nitride loaded with copper sulfide and manganese dioxide to prepare manganese dioxide.
Structural characterization test
Taking example 1 as an example, characterization tests were performed on the product structure.
FIG. 1 uses a transmission electron microscope, a high resolution transmission electron microscope for CNs, F127@CNs-CuS/MnO 2 Microstructure diagram for microstructure observation. Wherein FIG. 1 (a) is a TEM transmission electron microscope image of graphite phase carbon nitride, FIG. 1 (b) and FIG. 1 (c) are F127@CNs-CuS/MnO at different ratios 2 FIG. 1 (d) is a TEM transmission electron micrograph of F127@CNs-CuS/MnO 2 HRTEM high resolution transmission electron microscopy.
As can be seen from fig. 1 (a) and 1 (c), the graphite phase carbon nitride nanoplatelets after ball milling have a relatively uniform size, and have a diameter of about 150nm. As can be seen from FIG. 1 (b), mnO 2 And CuS nano particles are attached to the surfaces of CNs, F127@CNs-CuS/MnO 2 The composite nanoparticles are highly dispersed. CNs, cuS, mnO is clearly seen in FIG. 1 (d) 2 Is a lattice stripe of the composite nanomaterial F127@CNs-CuS/MnO 2 Is successfully prepared, cuS and MnO 2 Indeed, successful loading on CNs. In FIG. 1 (d), g-C 3 N 4 Namely graphite phase carbon nitride.
FIG. 2 is a graph of F127@CNs-CuS/MnO 2 And XRD analysis of intermediate products of the synthesis process, successful synthesis of F127@CNs-CuS/MnO2 can be demonstrated by figure 2. In FIG. 2, CNs-MnO are arranged in sequence from bottom to top 2 、CNs-CuS/MnO 2 、F127@CNs-CuS/MnO 2 XRD diffraction patterns of CNs-CuS, it should be noted that the inventors of the present application have studied to load MnO only on CNs during the course of studying the composite nanomaterial of the embodiment of the present application 2 And the related properties thereof, CNs-MnO are also provided herein 2 Related test results of (C) and other tests will also refer to CNs-MnO hereinafter 2 Is a test result of (a). From fig. 2 the following can be concluded:
(1) All the synthesized materials have two obvious diffraction peaks at 13.3 degrees and 27.6 degrees, and the two diffraction peaks respectively belong to graphite phase carbon nitride g-C 3 N 4 (002) and (100) faces of (PDF # 87-1526), indicating all combinationsThe graphite phase carbon nitride is used as a carrier in the formed material.
(2) In CNs-MnO 2 、CNs-CuS/MnO 2 、F127@CNs-CuS/MnO 2 The distinct diffraction peaks appearing at 37.06℃and 67℃are the (311) and (400) planes of manganese dioxide (PDF#42-1169), respectively, indicating that the diffraction peaks are observed in CNs-MnO 2 、CNs-CuS/MnO 2 、F127@CNs-CuS/MnO 2 Is truly successful in loading MnO on CNs 2 。
(3) In CNs-CuS/MnO 2 、F127@CNs-CuS/MnO 2 In XRD patterns of CNs-CuS, the X-ray diffraction pattern of the CuS nano particles is consistent with the CuS standard pattern (JCPDS 06-0464), no impurity peak is observed, and the successful loading of the CuS on CNs is indicated.
(4) With CNs-CuS/MnO 2 In comparison, F127@CNs-CuS/MnO 2 The crystallinity of (c) is slightly reduced, possibly due to the encapsulated amorphous F127.
FIG. 3 is CNs, F127@CNs-CuS/MnO 2 The XPS spectrum diagram of (2) is that the composite nano material F127@CNs-CuS/MnO is positioned at the upper part 2 The XPS spectrum diagram of the carbon nitride CNs of the graphite phase is positioned below. As can be seen from FIG. 3, F127@CNs-CuS/MnO 2 Contains C, N, O, mn, S, cu element. Further referring to FIG. 4, FIG. 4 (a) and FIG. 4 (b) are composite nanomaterial F127@CNs-CuS/MnO, respectively 2 From FIG. 4 (a), it is clear that the two peaks at 930.4eV and 951.1eV correspond to Cu (II) 2p3/2 and Cu (II) 2p 1/2, respectively, and from FIG. 4 (b), the two peaks at about 639.5eV and 651.2eV correspond to Mn 2p3/2 and Mn 2p 1/2, respectively, and the spin energy resolved to 11.7eV, corresponding to a typical tetravalent Mn species. Thus, in combination with FIGS. 3 and 4, it can be further demonstrated that CuS and MnO are successfully loaded on CNs on the basis of FIGS. 1 and 2 2 And (3) nanoparticles.
In addition, FIG. 5 is F127@CNs-CuS/MnO 2 And the ultraviolet-visible absorption spectrum of the intermediate product of the synthesis process. As can be seen from FIG. 5, other synthetic products, except for pure CNs, exhibit full spectral absorption in the wavelength range of 200nm to 1200nm, such that a wider light absorption range and intensity are beneficial for enhancing the practice of the present applicationThe composite nanomaterial has photo-thermal conversion efficiency and photo-catalytic oxidation activity.
Composite nanomaterial property testing
Reactive oxygen species production capability assay
CNs and F127@CNs-CuS/MnO 2 For the test subjects, CNs and F127@CNs-CuS/MnO were detected using the chemical probe 1, 3-Dimethylbenzofuran (DPBF) 2 Reactive Oxygen Species (ROS) generated. Specifically, 400. Mu.g of CNs and F127@CNs-CuS/MnO, respectively 2 Dispersing into 4mL of absolute ethanol containing DPBF (20 μg) and irradiating with 410nm light (0.25 mW/cm) 2 ) And (5) irradiating. The absorption spectrum of DPBF at 412nm was recorded with an ultraviolet-visible spectrophotometer at each time point (0, 5, 10 and 15 minutes) and the results are shown in FIG. 6 (a) and FIG. 6 (b), and the results of the experiment indicate F127@CNs-CuS/MnO 2 Has higher active oxygen generating capability and good photodynamic property.
FIG. 6 (a) is a schematic diagram showing the reaction of F127@CNs-CuS/MnO 2 The ultraviolet-visible absorption spectrum (at 412nm wavelength) of the mixed DPBF, and fig. 6 (b) is the ultraviolet-visible absorption spectrum (at 412nm wavelength) of the DPBF mixed with CNs. As can be seen from fig. 6 (a) and 6 (b), the absorbance at 412nm of both sets of reaction solutions decreased to different extents over time, indicating that Reactive Oxygen Species (ROS) were indeed formed, resulting in a decrease in absorbance at 412nm of DPBF with increasing ROS due to oxidation of ROS that may be formed by DPBF. In particular FIG. 6 (a), which is F127@CNs-CuS/MnO 2 The fluorescence intensity is reduced to about 35% after 15min, which indicates F127@CNs-CuS/MnO 2 Generates more Reactive Oxygen Species (ROS) than CNs and has stronger photodynamic properties.
Photo-thermal conversion capability and photo-stability test
Composite nano material F127@CNs-CuS/MnO 2 And the intermediate product of the synthesis process are used as test objects, 1mL of CNs and CNs-MnO with different concentrations (0, 50, 100, 200 and 300 mu g/mL) are used as test objects 2 、CNs-CuS、CNs-CuS/MnO 2 And F127@CNs-CuS/MnO 2 The aqueous dispersion (i.e., an aqueous dispersion of these substances dispersed well in deionized water) was exposed to 808nm laser light (2W/cm 2 ) For 10min, and useThe thermal imaging camera records the temperature of the different samples at each time point. Deionized water was set as a negative control. To further verify the photostability, F127@CNs-CuS/MnO was used 2 An aqueous dispersion of composite nanomaterial (200. Mu.g/mL) was exposed to 808nm laser (2W/cm) 2 ) The next 10 minutes, then cooled to ambient temperature (15 min) while the laser was turned off, and the process was repeated four times. The results are shown in FIG. 7 (a) and FIG. 7 (b), and the results of this experiment indicate F127@CNs-CuS/MnO 2 Has high photo-thermal conversion capability and photo-stability, and can effectively inhibit cancer cells.
FIG. 7 (a) is a composite nanomaterial F127@CNs-CuS/MnO 2 And the temperature profile of the intermediate product of the synthesis thereof (808 nm, 2W/cm 2 10 min) of the power radiation of (a) and (b) of FIG. 7 is a composite nanomaterial F127@CNs-CuS/MnO 2 Is a light cycle temperature profile of (a). As can be seen from FIG. 7 (a), the wavelength is 2W/cm at 808nm 2 After 10min of power irradiation of CNs-CuS/MnO 2 、F127@CNs-CuS/MnO 2 The temperature of (a) was raised to 52.1 ℃ and 54.5 ℃ respectively, while the temperature of water (control) was raised by only 3.9 ℃. The above material can successfully inhibit cancer cells because cancer cells are effectively inhibited after several minutes when the temperature is raised to 40-60 ℃. Wherein, with CNs-CuS/MnO 2 Compared with the composite nanomaterial F127@CNs-CuS/MnO under the same condition 2 The temperature rise is faster and the temperature is higher, which shows that the photo-thermal conversion capability is stronger because the F127 outer layer improves CNs-CuS/MnO 2 Stability and dispersibility of nanoparticles.
As can be seen from FIG. 7 (b), in the four laser irradiation cycle test, F127@CNs-CuS/MnO 2 No obvious change appears in the photo-thermal effect of the nano-composite material, which indicates that the composite nano-material F127@CNs-CuS/MnO 2 Has good light stability.
Glutathione consumption Capacity test
Glutathione is an antioxidant on the surface of tumors, can react with active oxygen to influence the killing capacity of photodynamic therapy on tumor cells, so that the application hopes to consume certain glutathione through the synthesized composite nanomaterial, further reduce the influence of the glutathione on photodynamic therapy efficiency and improve the photodynamic therapy efficiency in cooperative therapy.
Composite nano material F127@CNs-CuS/MnO 2 And the intermediate product of the synthesis process is used as a test object, 15mmol/L Glutathione Solution (GSH), CNs and CNs-MnO are added 2 、CNs-CuS、CNs-CuS/MnO 2 、F127@CNs-CuS/MnO 2 The aqueous dispersions (each at a concentration of 200. Mu.g/mL) were mixed well and water was used as the control. Subsequently, the residual amount of GSH was determined using GSH/GGSH assay kit. As shown in FIG. 8, FIG. 8 is a composite nanomaterial F127@CNs-CuS/MnO 2 And glutathione consumption profiles of intermediates in the synthesis process thereof.
As can be seen from FIG. 8, CNs-MnO 2 、CNs-CuS/MnO 2 F127@CNs-CuS/MnO 2 The glutathione content in the samples loaded with manganese dioxide nano particles all shows a decreasing trend, which indicates that the sample is coated with F127@CNs-CuS/MnO 2 The nano manganese dioxide can play a role in consuming glutathione. Thus, when the composite nanomaterial of the embodiment of the application is used for photodynamic and photothermal combined treatment, the level of glutathione in tumor cells can be effectively reduced, so that the consumption of active oxygen by glutathione is reduced, and the anticancer efficiency of photodynamic treatment is improved.
In vitro cellular reactive oxygen species production level test
HepG2 cells were seeded into 6-well plates and cultured in DMEM medium for 24h. CNs, CNs-MnO, respectively 2 、CNs-CuS、CNs-CuS/MnO 2 And F127@CNs-CuS/MnO 2 Cells were treated (at a concentration of 200. Mu.g/mL). After 4h, 10. Mu.L of 2, 7-dichlorofluorescein diacetate (DCFH-DA) was added to each well and incubated with cells for 30min. The 6-well plate was then exposed to 410nm light for 1h. After washing 3 times with PBS, the photoluminescent intensity of the mixture was measured with a fluorescence spectrophotometer. The result is shown in FIG. 9, FIG. 9 is a composite nanomaterial F127@CNs-CuS/MnO 2 And intracellular ROS production profiles of intermediates in their synthesis.
Non-fluorescent DCFH-DA is cleaved into DCFH by esterase after passively entering cells, and is oxidized into green fluorescence strong dichloro fluorescein (DCF) by ROS in cells, and the fluorescence absorption intensity of DCF can be used for a tableThe amount of ROS in the cells is characterized. As can be seen from FIG. 9, CNs-MnO after light exposure 2 The highest ROS yield, followed by CNs-CuS-MnO 2 F127@CNs-CuS/MnO 2 。
O 2 Is detected by (a)
Monitoring of O using RDPP fluorescent probes 2 Is generated. First, CNs-MnO are added in a certain amount 2 、CNs-CuS、CNs-CuS/MnO 2 、F127@CNS-CuS/MnO 2 Dispersing in 2mL PBS (pH 6.5) to form suspension (each at 200. Mu.g/mL), and adding 50. Mu.L of RDPP (10 mmol/L) ethanol solution and 200. Mu.L of H, respectively 2 O 2 . Finally, the RDPP fluorescence intensity at each time point (0, 2, 4, 6, 8, 10, 12, 14, 16 minutes) was recorded with a fluorescence spectrophotometer. As shown in FIG. 10, FIG. 10 shows a composite nanomaterial F127@CNs-CuS/MnO 2 And an oxygen production result graph of an intermediate product of the synthesis process thereof.
As can be seen from FIG. 10, under the same conditions, CNs-MnO 2 Generated O 2 The maximum amount is F127@CNs-CuS/MnO 2 Indicating F127@CNs-CuS/MnO 2 Has higher oxygen generating capacity.
In vitro cytotoxicity experiments
Determination of CNs, CNs-MnO Using Standard CCK-8 assay kit 2 、CNs-CuS、CNs-CuS/MnO 2 And F127@CNs-CuS/MnO 2 Cytotoxicity of nanoparticles on HepG2 cells included cytotoxicity tests under no light and specified light conditions, respectively.
For no-light testing, hepG2 cells (2×105/well) were inoculated into 96-well plates, and after bottom attachment, the cells were incubated with different concentrations (0, 50, 100, 200, 300. Mu.g/mL) of CNs, CNs-MnO in the absence of irradiation 2 、CNs-CuS、CNs-CuS/MnO 2 And F127@CNs-CuS/MnO 2 Incubate for 24 hours. Cell-free medium containing the same concentration samples was used as a control group. Then, the cells were washed 3 times with PBS.
For the illumination condition test, hepG2 cells and CNs with different concentrations and CNs-MnO 2 、CNs-CuS、CNs-CuS/MnO 2 、F127@CNs-CuS/MnO 2 Incubation 12After hours, the substrates were exposed to light having a wavelength of 410nm (0.25W/cm 2 ，60min)、808nm(2W/cm 2 ，2min)、404nm(0.25W/cm 2 ，60min)+808nm(2W/cm 2 2 min), 808nm (2W/cm) 2 ，2min)+404nm(0.25W/cm 2 60 min). After 12 hours, cell viability was determined using the CCK-8 assay kit. Independent replicates were run for each condition and the results are expressed as mean standard deviation. Cell viability was expressed as mean standard deviation. The difference of the one-way analysis of variance comparison is statistically significant (p<0.05). The results are shown in fig. 11 and 12.
FIG. 11 is a graph showing different concentrations of CNs, CNs-CuS/MnO 2 、F127@CNs-CuS/MnO 2 Results of darkroom toxicity analysis on HepG2 cells. As shown in FIG. 11, CNs-CuS/MnO 2 F127@CNs-CuS/MnO 2 Cytotoxicity in the absence of irradiation was negligible, indicating that the biocompatible sample was a potential biophototherapeutic agent. In particular F127@CNs-CuS/MnO 2 The treated cells still maintain a higher viability (86.6%) at a concentration of 200. Mu.g/mL, and the viability is slightly higher than CNs-CuS/MnO at the same concentration 2 F127 proved to be able to improve the biocompatibility of the composite nanoparticle. Sample concentration and incubation time were optimized using the CCK-8 method, and cytotoxicity detection results in all experiments were characterized by a concentration of 200. Mu.g/mL.
FIG. 12 shows the viability of cells treated with different materials (at a concentration of 200. Mu.g/mL) under different light conditions, by which the in vitro anticancer efficiency of the composite nanomaterial of the present application was evaluated. From fig. 12, it can be seen that the decrease in cell viability was more pronounced after PDT and PTT co-treatment than with single PDT or PTT. In particular F127@CNs-CuS/MnO 2 The treated cells were at 808nm (2W/cm) 2 ，2min)+410nm(0.25W/cm 2 60 min) and the cell inactivation rate under the combined illumination reaches 88.6%, which shows that the curative effect of the synergistic treatment is obviously better than that of the monotherapy.
Furthermore, in the case of co-therapy, 808nm (2W/cm 2 ，2min)+404nm(0.25W/cm 2 The 60min group showed a specific 410nm (0.25W/cm) 2 ，60min)+808nm(2W/cm 2 2 min) better cell inactivation efficiency, which may be due to endocytosis of the composite nanomaterial caused by the photothermal effect of PTT, and a significant increase in oxygen content, resulting in more singlet oxygen production.
It should be noted that CNs-MnO in PDT groups alone under the same conditions 2 The highest inactivation efficiency. There are two possible reasons: first, nano manganese dioxide and intracellular endogenous H 2 O 2 Reaction to produce O 2 ，O 2 Additional oxygen is provided for PDT. Second, manganese dioxide nanoparticles react with antioxidant GSH and are overexpressed in cancer cells.
Results of this test and the in vitro cellular reactive oxygen species level test, O, described above 2 The test results of (2) are consistent, and also describe O again 2 The deep influence of photodynamic reaction and the application of the composite nanomaterial in the embodiment of the application to photodynamic and photothermal combined treatment are illustrated, and the effect is much better than that of single photodynamic or photothermal treatment.
The above-mentioned embodiment of the present application discloses a composite nanomaterial for photodynamic and photothermal combined therapy, and a preparation method, an application and a detailed description thereof, wherein specific examples are applied to illustrate principles and embodiments of the present application, and the above-mentioned embodiment is only used to help understand the electronic device and core ideas thereof; meanwhile, as those skilled in the art will vary in the specific embodiments and application scope according to the idea of the present application, the present disclosure should not be construed as limiting the present application in summary.
Claims (9)
1. A preparation method of a composite nanomaterial for photodynamic and photothermal combined treatment, characterized in that the preparation method comprises the following steps:
dispersing graphite-phase carbon nitride in water, adding copper acetate, stirring, mixing and stirring the obtained solution with thiourea aqueous solution to obtain a suspension, and carrying out hydrothermal reaction on the suspension to obtain copper sulfide-loaded graphite-phase carbon nitride; the copper sulfide accounts for 3-10% of the graphite phase carbon nitride by mass;
dispersing the graphite-phase carbon nitride loaded with copper sulfide in water, adding a potassium permanganate aqueous solution and a cationic polyacrylamide acid aqueous solution, mixing and stirring to obtain the graphite-phase carbon nitride loaded with copper sulfide and manganese dioxide; the manganese dioxide accounts for 1-3% of the graphite phase carbon nitride loaded with the copper sulfide;
dispersing the graphite-phase carbon nitride loaded with copper sulfide and manganese dioxide into F127 aqueous solution, and stirring overnight to obtain the composite nanomaterial;
the preparation method further comprises the step of preparing the graphite phase carbon nitride loaded with copper sulfide before the step of preparing the graphite phase carbon nitride, wherein the preparation step of the graphite phase carbon nitride is as follows: calcining melamine in air, and grinding to obtain the graphite-phase carbon nitride.
2. The method of claim 1, wherein the graphite phase carbon nitride is prepared by the steps of: under the air condition, heating melamine to 550 ℃ at a heating rate of 2.5 ℃/min, cooling to room temperature after heating for 4 hours, collecting yellow powder, washing with deionized water and drying;
the yellow powder was ball milled at 500rpm for 6 hours and the finely milled graphite phase carbon nitride was collected by filtration.
3. The method according to claim 1, wherein in the step of producing the copper sulfide-supported graphite phase carbon nitride, the hydrothermal reaction is carried out at 140 to 160 ℃ for 20 to 30 hours.
4. A method according to claim 3, wherein the step of preparing the copper sulfide-loaded graphite phase carbon nitride comprises: dispersing 0.5g of graphite phase carbon nitride in 60mL of deionized water, reacting for 1 hour under ultrasonic conditions, adding 0.068g of copper acetate, and continuously stirring for 2 hours;
mixing the obtained solution with 40mL of the thiourea aqueous solution, and stirring for 5 hours under the magnetic force condition to obtain a suspension; wherein the thiourea aqueous solution contains 0.074g of thiourea;
placing the suspension in an autoclave lined with polytetrafluoroethylene, and reacting for 24 hours at 150 ℃;
And (3) centrifugally separating to obtain a precipitate, washing the precipitate for multiple times, and drying the precipitate at 80 ℃ to obtain the graphite phase carbon nitride loaded with copper sulfide.
5. The method according to claim 1, wherein in the step of preparing the composite nanomaterial, the mass ratio of the graphite phase carbon nitride loaded with copper sulfide and manganese dioxide to F127 is 5:4.
6. The method according to claim 5, wherein the step of preparing the graphite-phase carbon nitride loaded with copper sulfide and manganese dioxide comprises: dispersing 0.15g of the graphite-phase carbon nitride loaded with copper sulfide in 60mL of deionized water at room temperature, dripping 10mL of potassium permanganate aqueous solution with the concentration of 0.5mg/mL, mixing with 6.4mL of cationic polyacrylamide aqueous solution with the concentration of 5mg/mL, stirring for 30min, washing, centrifuging and drying the obtained product to obtain the graphite-phase carbon nitride loaded with copper sulfide and manganese dioxide;
the preparation method of the composite nano material comprises the following steps: 50mg of the graphite phase carbon nitride loaded with copper sulfide and manganese dioxide is dispersed into 20mL of the F127 aqueous solution with the concentration of 2mg/mL, stirred overnight, centrifugally filtered, and the composite nano material is collected.
7. A composite nanomaterial for photodynamic and photothermal combination therapy, characterized in that the composite nanomaterial is produced by the production method according to any one of claims 1 to 6.
8. Use of the composite nanomaterial for photodynamic and photothermal combination therapy as claimed in claim 7 in the preparation of a medicament for treating a neoplastic disease.
9. Use of the composite nanomaterial for photodynamic and photothermal combination therapy as claimed in claim 7 in the preparation of a medicament for photodynamic and photothermal combination therapy of a neoplastic disease.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111538603.3A CN114177289B (en) | 2021-12-15 | 2021-12-15 | Composite nano material for photodynamic and photothermal combined treatment and preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111538603.3A CN114177289B (en) | 2021-12-15 | 2021-12-15 | Composite nano material for photodynamic and photothermal combined treatment and preparation method and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN114177289A CN114177289A (en) | 2022-03-15 |
| CN114177289B true CN114177289B (en) | 2023-10-20 |
Family
ID=80605210
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202111538603.3A Active CN114177289B (en) | 2021-12-15 | 2021-12-15 | Composite nano material for photodynamic and photothermal combined treatment and preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114177289B (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116019923A (en) * | 2023-02-20 | 2023-04-28 | 武汉科技大学 | Preparation method and application of copper sulfide nano-drug coated with tetravalent platinum prodrug |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108786874A (en) * | 2017-04-26 | 2018-11-13 | 中国科学院上海硅酸盐研究所 | Load the graphite phase carbon nitride nanometer sheet material and its preparation method and application of manganese dioxide |
| CN109966489A (en) * | 2017-12-28 | 2019-07-05 | 南京邮电大学 | A kind of nanocomposite and the preparation method and application thereof with light power and photo-thermal combination therapy function |
| CN110982521A (en) * | 2019-12-31 | 2020-04-10 | 山西医科大学 | Graphite phase carbon nitride quantum dot composite and synthetic method and biological application thereof |
| CN111202720A (en) * | 2020-01-30 | 2020-05-29 | 上海工程技术大学 | Copper sulfide/silicon dioxide/manganese dioxide nano composite particle, preparation method and application |
| CN111250138A (en) * | 2020-02-28 | 2020-06-09 | 韶关学院 | Porous nano flaky graphite phase carbon nitride and preparation method and application thereof |
| CN113219016A (en) * | 2021-05-25 | 2021-08-06 | 济南大学 | Preparation method of electrochemical immunosensor based on uracil modified graphite-like phase carbon nitride |
-
2021
- 2021-12-15 CN CN202111538603.3A patent/CN114177289B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108786874A (en) * | 2017-04-26 | 2018-11-13 | 中国科学院上海硅酸盐研究所 | Load the graphite phase carbon nitride nanometer sheet material and its preparation method and application of manganese dioxide |
| CN109966489A (en) * | 2017-12-28 | 2019-07-05 | 南京邮电大学 | A kind of nanocomposite and the preparation method and application thereof with light power and photo-thermal combination therapy function |
| CN110982521A (en) * | 2019-12-31 | 2020-04-10 | 山西医科大学 | Graphite phase carbon nitride quantum dot composite and synthetic method and biological application thereof |
| CN111202720A (en) * | 2020-01-30 | 2020-05-29 | 上海工程技术大学 | Copper sulfide/silicon dioxide/manganese dioxide nano composite particle, preparation method and application |
| CN111250138A (en) * | 2020-02-28 | 2020-06-09 | 韶关学院 | Porous nano flaky graphite phase carbon nitride and preparation method and application thereof |
| CN113219016A (en) * | 2021-05-25 | 2021-08-06 | 济南大学 | Preparation method of electrochemical immunosensor based on uracil modified graphite-like phase carbon nitride |
Non-Patent Citations (4)
| Title |
|---|
| Enguo Ju等.Copper(II)–Graphitic Carbon Nitride Triggered Synergy: Improved ROS Generation and Reduced Glutathione Levels for Enhanced Photodynamic Therapy.《Angew. Chem.》.2016,第55卷第11467-11471页. * |
| Zhenhe Xu等.In situ growth of CuS nanoparticles on g-C3N4 nanosheets for H2 production and the degradation of organic pollutant under visible-light irradiation.《RSC Adv.》.2019,第9卷第25638-25646页. * |
| 方杰等.g-C3N4@C-TiO2 纳米颗粒增强可见光驱动的体外光动力灭活HL60 细胞的试验研究.《激光生物学报》.2021,第30卷(第1期),第43-51页. * |
| 谭诗杨等.g-C3N4/MnO2 复合光催化剂的制备及可见光下降解四环素性能.《功能材料》.2020,第51卷(第6期),第06164-06168页和第06214页. * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN114177289A (en) | 2022-03-15 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105031671B (en) | Based on Prussian blue synergic nano diagnosis and treatment agent of intelligent pH triggering MRI monitoring insoluble drug releases and preparation method thereof | |
| CN113398285B (en) | Preparation method of bimetal nano enzyme composite material with anti-tumor effect | |
| CN110038128B (en) | Au @ Se @ Pt-Ce6 nano composite probe and preparation method thereof | |
| CN112480925B (en) | Near-infrared two-region luminous long-afterglow nanoprobe based on X-ray excitation, preparation method and application thereof in living body imaging analysis | |
| US20220160892A1 (en) | Single nir irradiation triggered upconversion nano system for synergistic photodynamic and photothermal cancer therapy | |
| CN113773667A (en) | Organic small-molecule near-infrared two-region fluorescent dye and preparation method and application thereof | |
| CN114177289B (en) | Composite nano material for photodynamic and photothermal combined treatment and preparation method and application thereof | |
| CN113975411A (en) | Preparation method of near-infrared light response up-conversion mesoporous tin dioxide diagnosis and treatment nanocapsule | |
| CN115845086A (en) | Photo-thermal Fenton-like reaction artificial nano enzyme and preparation method and application thereof | |
| Li et al. | Multifunctional nanoplatform based on g-C3N4, loaded with MnO2 and CuS nanoparticals for oxygen self-generation photodynamic/photothermal synergistic therapy | |
| CN112891536B (en) | Bismuth-based nano heterojunction material and preparation method and application thereof | |
| CN110623940A (en) | Selenium/silicon dioxide/gold nano composite particle and preparation method and application thereof | |
| Yang et al. | Mesoporous semiconductors combined with up-conversion nanoparticles for enhanced photodynamic therapy under near infrared light | |
| CN117482231A (en) | Porous copper-manganese bimetallic nano material applied to tumor immunotherapy and preparation method thereof | |
| CN109550050B (en) | Melanin-loaded molybdenum dioxide drug-loaded compound and preparation and application thereof | |
| CN112336858A (en) | Bismuth-manganese-based composite particle and preparation method and application thereof | |
| CN114477108B (en) | Preparation method of near-infrared light-responsive manganese ion-doped vanadium diselenide nanosheet | |
| Uribe-Robles et al. | TiO2 hollow nanospheres functionalized with folic acid and ZnPc for targeted photodynamic therapy in glioblastoma cancer | |
| CN115006523A (en) | Preparation method of near-infrared two-region responsive nanoparticles | |
| Liang et al. | Facile synthesis of phycocyanin/polydopamine hierarchical nanocomposites for synergizing PTT/PDT against cancer | |
| CN112891532B (en) | Nano photo-thermal conversion material and preparation method and application thereof | |
| CN110240170B (en) | Yolk-eggshell type UCNP @ MgSiO3Process for preparing nanoparticles | |
| CN112156186B (en) | Infrared dye-sensitized carbon nitride ternary heterojunction nano material and preparation method and application thereof | |
| CN108904472A (en) | The modified photosensitizer nanocomposite of graphene and its application | |
| CN117084981B (en) | Preparation method and application of iron ion synergistic artemisinin-quercetin synergistic treatment nano system |
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 |