CN112316140B - Preparation method of Bi-DMSNs @ PCM multifunctional nano composite material - Google Patents

Preparation method of Bi-DMSNs @ PCM multifunctional nano composite material Download PDF

Info

Publication number
CN112316140B
CN112316140B CN202011246485.4A CN202011246485A CN112316140B CN 112316140 B CN112316140 B CN 112316140B CN 202011246485 A CN202011246485 A CN 202011246485A CN 112316140 B CN112316140 B CN 112316140B
Authority
CN
China
Prior art keywords
dmsns
pcm
solution
preparing
temperature
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
Application number
CN202011246485.4A
Other languages
Chinese (zh)
Other versions
CN112316140A (en
Inventor
贺飞
杜亚倩
贾培培
杨飘萍
盖世丽
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Harbin Engineering University
Original Assignee
Harbin Engineering University
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by Harbin Engineering University filed Critical Harbin Engineering University
Priority to CN202011246485.4A priority Critical patent/CN112316140B/en
Publication of CN112316140A publication Critical patent/CN112316140A/en
Application granted granted Critical
Publication of CN112316140B publication Critical patent/CN112316140B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Images

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
    • A61K41/0052—Thermotherapy; Hyperthermia; Magnetic induction; Induction heating therapy
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K49/00—Preparations for testing in vivo
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K49/00—Preparations for testing in vivo
    • A61K49/04—X-ray contrast preparations
    • 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
    • A61K9/5115—Inorganic compounds
    • 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
    • A61K9/5138—Organic macromolecular compounds; Dendrimers obtained by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyvinyl pyrrolidone, poly(meth)acrylates
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents

Abstract

A preparation method of Bi-DMSNs @ PCM multifunctional nano composite material relates to a preparation method of photo-thermal material. The invention aims to solve the problems that the existing Bi nano particles are easy to oxidize and have poor stability, and the existing technology synthesizes the Bi nano particles by a simple method, but the problem that an organic solvent harmful to human bodies is inevitably used. The preparation method comprises the following steps: 1. preparing dendritic mesoporous silica; 2. preparing the Bi-DMSNs @ PCM nano composite material. The invention is used for preparing the Bi-DMSNs @ PCM multifunctional nano composite material.

Description

Preparation method of Bi-DMSNs @ PCM multifunctional nano composite material
Technical Field
The invention relates to a preparation method of a photo-thermal material.
Background
Currently, near-infrared laser-induced photothermal therapy has characteristics of high efficiency, small invasiveness, high selectivity and the like, is intensively studied in preclinical and clinical studies of cancer treatment, and is expected to convert near-infrared laser into heat by a photothermal agent to thermally ablate surrounding cancer cells. In addition, the near infrared region II (1000 nm to 1350 nm) has attracted much attention in recent years because of the deeper penetration depth of tissues, compared with the near infrared region I. Therefore, there is an urgent need to search for a photothermal agent having excellent properties in the near-infrared region II. The nano-photothermal agents can be mainly classified into four types: the first is the use of rare earth elements as radiosensitizers; the second type is nanoparticles based on the tungsten (W) element, such as ultra-small WO 3-x Nanodots and WS 2 Quantum dots; the third type is Au derived nano structure (Au nano particle, au @ Pt nanocrystalline, PB @ Au nuclear satellite nano particle and Au @ FeS nuclear shell nano particle); the last type is a bismuth (Bi) -based nanoparticle comprising a semimetallic Bi nanoparticle, bi 2 S 3 Nanodot, bi 2 Se 3 Nanodot, cu 3 BiS 3 Ternary semiconductor nanorod, mnSe @ Bi 2 Se 3 Core-shell nanoparticles and FeSe 2 Modified Bi 2 Se 3 Nanoplatelets, all of which have both imaging and therapeutic capabilities. It is needless to say that nanomaterials with a single component have the advantage of being easy to synthesize, making their use very valuable. Therefore, the ideal therapeutic agent for photothermal therapy should be a single-component nanoparticle. However, pure Bi nanomaterials are easily oxidized in air, thereby losing photo-thermal properties. For example, bi nanotubes are stored in saturated NaBH 4 In the middle, oxidation is avoided.
Bi has very high atomic number, has the unique property of good X-ray attenuation in a plurality of non-radioactive elements, is nontoxic and cheap green metal compared with other heavy metal elements, and has wide prospect when being used for synthesizing a photo-thermal reagent. In addition, bi is also one of the most biocompatible heavy metal elements, and a compound thereof (such as colloidal bismuth nitrite) is used as a clinical drug for treating gastrointestinal diseases in daily life. In the case of Bi colloidal nanomaterials, they can be prepared by reductive methods, solvothermal methods, microwave synthesis or metal-organic precursors. The reduction method, the solvothermal method and the microwave method are simpler, but the experimental process is not easy to control, and the morphology of the Bi nanoparticles is usually poor. The laser ablation method can effectively avoid using toxic raw materials, but the process consumes energy seriously. The prior art synthesizes bismuth nano-particles by a simple method, but organic solvents harmful to human bodies, such as 2-dimethylimidazole and n-dodecyl mercaptan, are inevitably used. Therefore, it is necessary to develop an ultra-simple strategy for synthesizing high-quality pure metal Bi nanoparticles to further expand the CT imaging prospect, and the light absorption thereof can be expanded to the near-infrared region, so as to better perform photothermal imaging and photothermal therapy.
Disclosure of Invention
The invention provides a preparation method of a Bi-DMSNs @ PCM multifunctional nano composite material, aiming at solving the problems that the existing Bi nano particles are easy to oxidize and have poor stability, and the existing technology synthesizes the bismuth nano particles by a simple method, but the problem that an organic solvent harmful to human bodies is inevitably used.
A preparation method of Bi-DMSNs @ PCM multifunctional nano composite material is carried out according to the following steps:
1. preparing the dendritic mesoporous silica:
adding triethanolamine into water, stirring for 0.4-0.6 h in an oil bath with a magnetic stirring speed of 90-150 rpm and a temperature of 70-90 ℃ to obtain a solution A, adding cetyl trimethyl ammonium bromide and sodium salicylate into the solution A, stirring for 0.5-1.5 h under the conditions of a magnetic stirring speed of 90-150 rpm and a temperature of 70-90 ℃ to obtain a solution B, adding a mixture of tetraethoxysilane and 1, 2-bis (triethoxysilyl) ethane into the solution B, stirring for 11-13 h under the conditions of a magnetic stirring speed of 150-200 rpm and a temperature of 70-90 ℃, centrifuging, washing, extracting, standing and drying to obtain the dendritic mesoporous silica;
the mass ratio of the triethanolamine to the water is 1g (355-375) mL; the mass ratio of the triethanolamine to the hexadecyl trimethyl ammonium bromide is 1 (4-6); the mass ratio of the triethanolamine to the sodium salicylate is 1 (2-3); the volume ratio of the mass of the triethanolamine to the volume of the tetraethoxysilane is 1g (55-65) mL; the volume ratio of the mass of the triethanolamine to the 1, 2-bis (triethoxysilyl) ethane is 1g (20-25) mL;
2. preparing a Bi-DMSNs @ PCM nano composite material:
dissolving polyvinylpyrrolidone and bismuth nitrate in a mixture of glycerol and ethanol, heating to fully melt bismuth nitrate at the heating temperature of 60-70 ℃ to obtain a solution C, adding dendritic mesoporous silica into the solution C, stirring for 0.5-2 min at the stirring speed of 150-200 rpm to obtain a solution D, adding sodium borohydride into the solution D, stirring for 1-3 h at the stirring speed of 150-200 rpm to obtain a solution E, adding tetradecanol into the solution E, stirring for 3-6 h at the stirring speed of 150-200 rpm, centrifuging, washing, and drying to obtain the Bi-DMSNs @ PCM multifunctional nanocomposite;
the mass ratio of the polyvinylpyrrolidone to the bismuth nitrate is 1 (0.2-0.4); the volume ratio of the mass of the polyvinylpyrrolidone to the mixture of the glycerol and the ethanol is 1g (40-60) mL; the volume ratio of the glycerol to the ethanol in the mixture of the glycerol and the ethanol is 1 (0.4-0.6); the mass ratio of the polyvinylpyrrolidone to the dendritic mesoporous silica is 1 (0.15-0.35); the mass ratio of the polyvinylpyrrolidone to the sodium borohydride is 1 (0.1-0.25); the mass ratio of the polyvinylpyrrolidone to the tetradecanol is 1 (0.01-0.15).
The beneficial effects of the invention are:
in order to avoid the oxidation of Bi nanoparticles and better improve the stability of PVP-Bi nanoparticles, dendritic Mesoporous Silica Nanoparticles (DMSNs) are used as carriers, have unique center-radial hole structures and are very promising biomedical nano materials, and compared with conventional mesoporous silica nanoparticles, the Dendritic Mesoporous Silica Nanoparticles (DMSNs) prepared by the method have the advantages of large pore diameter and large surface area, and the pore diameter of the prepared Dendritic Mesoporous Silica Nanoparticles (DMSNs) can reach 15-20 nm. PVP-Bi directly grows at the in-situ position of the dendritic mesoporous silica nano particles (DMSNs), then tetradecanol (PCM) is introduced, a layer of tetradecanol (PCM) is coated outside the PVP-Bi nano dots, and the Bi-DMSNs @ PCM composite nano material is synthesized. The melting point of the PCM is 38 ℃, when the nanocomposite is irradiated by a laser, the PCM on the outermost layer of the material is melted, and then the Bi nanodots are released, so that the characteristic that the Bi nanodots are oxidized can be effectively prevented, and the stability of the composite is improved.
The method adopts an in-situ growth method, PVP-Bi directly grows at the in-situ position of Dendritic Mesoporous Silica Nanoparticles (DMSNs), and then tetradecanol (PCM) is introduced, wherein in the PVP-Bi synthesis process, a proper amount of DMSNs is added before sodium borohydride is added, then sodium borohydride is added, so that Bi single substances directly grow in the holes of the DMSNs, and then the PCM is added, and the PCM is successfully coated outside. If the PVP-Bi nanodots are directly coated with the PCM on the basis of synthesizing the PVP-Bi nanodots, the added PCM coats a layer outside a large number of PVP-Bi nanodots instead of coating outside a single nanosphere; if PVP-Bi nanodots and DMSNs particles are synthesized respectively, the two solutions are connected together by stirring through the action of electrostatic adsorption, and then the two solutions are coated outside the Bi-DMSNs through a method of directly adding PCM, but the final result is that Bi does not grow in the pores of the DMSNs, and the PCM is not coated outside the Bi-DMSNs. The PVP-Bi nanodots are synthesized by a chemical reduction method, dendritic silicon Dioxide (DMSNs) is synthesized to serve as a carrier, and a layer of tetradecanol (PCM) is coated outside the Bi-DMSNs to prevent Bi from being oxidized, so that the characteristic that PVP-Bi quantum dots are easily oxidized is avoided, and the stability of the material can be well maintained. According to the process flow method, the nano-sized Bi-DMSNs @ PCM composite material can be obtained, the characteristic that Bi is easy to oxidize is solved, and a brand-new solution and process flow are provided for preventing Bi from being oxidized.
The Bi-DMSNs @ PCM nano material prepared by the invention has good stability and oxidation resistance and higher photothermal conversion efficiency (the photothermal conversion efficiency can reach 32.3%), and the aqueous solution prepared by the prepared Bi-DMSNs @ PCM multifunctional nano composite material is very stable, so that the Bi nano points are prevented from being easily oxidized, and an optimized, quick and low-cost selection is provided for preparing a novel photothermal material.
The Bi-DMSNs @ PCM nano material prepared by the invention is 0 mu g/mL -1 ～500μg·mL -1 The survival rate of the L929 cells is more than 85 percent in the whole concentration range, and the Bi-DMSNs @ PCM has no obvious toxic or side effect on the L929 cells, has very small toxicity on normal cells and has good biocompatibility. Bi-DMSNs @ PCM has the best anti-tumor effect than PVP-Bi, stronger photothermal capacity and better treatment effect, and the power of a laser is 0.5W-cm -2 And the lowest survival rate of the HeLa cells and the 4T1 cells can be reduced to below 10 percent under the irradiation of 10 min.
The method is quick, simple and convenient, low in required temperature, good in safety performance, free of special reaction equipment, environment-friendly and applicable to industrialization.
In conclusion, the process is simple and easy to implement, the repeatability is good, the whole reaction system is environment-friendly and safe, and the proposed process route has good application prospect and practical value. The prepared novel nano composite material is a potential tumor treatment reagent which can be used for photothermal treatment and has CT and photothermal imaging effects.
The invention is used for a preparation method of a Bi-DMSNs @ PCM multifunctional nano composite material.
Drawings
FIG. 1 is a TEM image with a PVP-Bi nanodots prepared in comparative experiment one, b Bi @ PCM of coated PCM prepared in comparative experiment two, c DMSNs prepared in example one step one, d Bi-DMSNs coated PCM prepared in comparative experiment three, e Bi-DMSNs @ PCM prepared in example one at 100nm magnification, f single Bi-DMSNs @ PCM prepared in example one at 50nm magnification;
FIG. 2 is the distribution diagram of Bi-DMSNs @ PCM prepared in the first embodiment, a is HAADF diagram, b is C element, C is O element, d is Si element, e is Bi element, f is element composite diagram;
FIG. 3 is a diagram of the UV-VIS absorption spectra of Bi-DMSNs @ PCM and PVP-Bi nanodots, 1 is the Bi-DMSNs @ PCM prepared in the first example, and 2 is the PVP-Bi nanodots prepared in the first comparative experiment;
FIG. 4 is Zeta potential diagram, a is PVP-Bi nanodots prepared in the first comparative experiment, b is DMSNs prepared in the first step of the example, c is PCM, d is Bi-DMSNs @ PCM prepared in the first step of the example;
FIG. 5 is a photograph of an aqueous solution of Bi-DMSNs @ PCM before and after standing;
FIG. 6 is a diagram showing the UV-VIS absorption spectra before and after the Bi-DMSNs @ PCM aqueous solution is allowed to stand, wherein a is before standing and b is after standing for 14 days;
FIG. 7 is a photograph of an aqueous solution of Bi-DMSNs @ PCM before and after heating;
FIG. 8 is a graph showing the change of the ultraviolet-visible absorption curve of the aqueous solution of Bi-DMSNs @ PCM with the heating time, wherein a is unheated, b is heated 60min, c is heated 120min, and d is heated 180min;
FIG. 9 shows Bi-DMSNs @ PCM aqueous solutions of different concentrations at 1064nm and 0.5W cm -2 The temperature rise profile within 10min of laser irradiation of (1) is 0. Mu.g.mL -1 And 2 is 50. Mu.g.mL -1 And 3 is 100. Mu.g.mL -1 And 4 is 200. Mu.g.mL -1 5 is 500. Mu.g.mL -1 ；
FIG. 10 shows Bi-DMSNs @ PCM aqueous solutions of different concentrations at 1064nm and 0.5W cm -2 Irradiating for 10min, and then carrying out temperature change versus concentration curve chart;
FIG. 11 is an infrared thermal imaging plot of PVP-Bi nanodot aqueous solution and Bi-DMSNs @ PCM aqueous solution under 1064nm laser irradiation, a being PVP-Bi nanodot aqueous solution and irradiated with 1064nm laser, b being Bi-DMSNs @ PCM aqueous solution and irradiated with 1064nm laser;
FIG. 12 shows 1064nm and 0.5W cm -2 The temperature rise and cooling curve diagram of the solution after laser irradiation, 1 is Bi-DMSNs @ PCM aqueous solution, 2 is PVP-Bi aqueous solution, and 3 is water;
fig. 13 is a natural logarithm graph of the cooling time and the driving temperature in fig. 12, c is a natural logarithm curve of the cooling time and the driving temperature corresponding to curve 2, and d is a natural logarithm curve of the cooling time and the driving temperature corresponding to curve 1;
FIG. 14 is a graph showing the photo-thermal cycling curves after repeated irradiation of an aqueous Bi-DMSNs @ PCM solution;
FIG. 15 is a graph showing the comparison of cell viability after 24h of Bi-DMSNs @ PCM and L929 cell culture at different concentrations;
FIG. 16 is a graph showing comparison of cell viability after HeLa cells were cultured under different conditions, a is the addition of Bi-DMSNs @ PCM prepared in example one and irradiation of the cells with a 1064nm laser, b is the addition of PVP-Bi nanodots prepared in comparative experiment one and irradiation of the cells with a 1064nm laser, c is the addition of Bi-DMSNs @ PCM prepared in example one, and d is the addition of pure medium and treatment with a 1064nm laser;
FIG. 17 is a graph showing comparison of cell viability after culturing 4T1 cells under different conditions, wherein a is the addition of Bi-DMSNs @ PCM prepared in example one and irradiating the cells with a 1064nm laser, b is the addition of PVP-Bi nanodots prepared in comparative experiment one and irradiating the cells with a 1064nm laser, c is the addition of Bi-DMSNs @ PCM prepared in example one, and d is the addition of pure medium and irradiating with a 1064nm laser;
FIG. 18 is a confocal laser microscopy image of HeLa cells after staining with calcein and propidium iodide, a being laser treatment with 1064nm only, b being treatment with addition of Bi-DMSNs @ PCM prepared in example one, c being addition of PVP-Bi nanodots prepared in comparative experiment one and irradiating the cells with 1064nm laser, d being addition of Bi-DMSNs @ PCM prepared in example one and irradiating the cells with 1064nm laser;
FIG. 19 is confocal laser microscopy images after staining with JC-1, (1) PVP-Bi plus 1064nm laser treated 4T1 cells prepared in comparative experiment one, (2) Bi-DMSNs @ PCM plus 1064nm laser treated 4T1 cells prepared in example one.
Detailed Description
The first embodiment is as follows: the embodiment of the invention relates to a preparation method of a Bi-DMSNs @ PCM multifunctional nano composite material, which comprises the following steps:
1. preparing dendritic mesoporous silica:
adding triethanolamine into water, stirring for 0.4-0.6 h in an oil bath with a magnetic stirring speed of 90-150 rpm and a temperature of 70-90 ℃ to obtain a solution A, adding cetyl trimethyl ammonium bromide and sodium salicylate into the solution A, stirring for 0.5-1.5 h under the conditions of the magnetic stirring speed of 90-150 rpm and the temperature of 70-90 ℃ to obtain a solution B, adding a mixture of tetraethoxysilane and 1, 2-bis (triethoxysilyl) ethane into the solution B, stirring for 11-13 h under the conditions of the magnetic stirring speed of 150-200 rpm and the temperature of 70-90 ℃, centrifuging, washing, extracting, standing and drying to obtain the dendritic mesoporous silica;
the mass ratio of the triethanolamine to the water is 1g (355-375) mL; the mass ratio of the triethanolamine to the hexadecyl trimethyl ammonium bromide is 1 (4-6); the mass ratio of the triethanolamine to the sodium salicylate is 1 (2-3); the volume ratio of the mass of the triethanolamine to the volume of the tetraethoxysilane is 1g (55-65) mL; the volume ratio of the mass of the triethanolamine to the 1, 2-bis (triethoxysilyl) ethane is 1g (20-25) mL;
2. preparing Bi-DMSNs @ PCM nano composite material:
dissolving polyvinylpyrrolidone and bismuth nitrate in a mixture of glycerol and ethanol, heating to fully melt bismuth nitrate at the heating temperature of 60-70 ℃ to obtain a solution C, adding dendritic mesoporous silica into the solution C, stirring for 0.5-2 min at the stirring speed of 150-200 rpm to obtain a solution D, adding sodium borohydride into the solution D, stirring for 1-3 h at the stirring speed of 150-200 rpm to obtain a solution E, adding tetradecanol into the solution E, stirring for 3-6 h at the stirring speed of 150-200 rpm, centrifuging, washing, and drying to obtain the Bi-DMSNs @ PCM multifunctional nanocomposite;
the mass ratio of the polyvinylpyrrolidone to the bismuth nitrate is 1 (0.2-0.4); the volume ratio of the mass of the polyvinylpyrrolidone to the mixture of the glycerol and the ethanol is 1g (40-60) mL; the volume ratio of the glycerol to the ethanol in the mixture of the glycerol and the ethanol is 1 (0.4-0.6); the mass ratio of the polyvinylpyrrolidone to the dendritic mesoporous silica is 1 (0.15-0.35); the mass ratio of the polyvinylpyrrolidone to the sodium borohydride is 1 (0.1-0.25); the mass ratio of the polyvinylpyrrolidone to the tetradecanol is 1 (0.01-0.15).
The washing purpose in the step one is to remove residual reactants; the placement is for the purpose of removing the template.
The beneficial effects of this embodiment are:
in order to avoid the oxidation of the PVP-Bi nanoparticles and better improve the stability of the PVP-Bi nanoparticles, the Dendritic Mesoporous Silica Nanoparticles (DMSNs) are used as carriers, the Dendritic Mesoporous Silica Nanoparticles (DMSNs) have unique center-radial hole structures and are very promising biomedical nano materials, the pore diameter is large compared with that of the conventional mesoporous silica nanoparticles, the surface area is large, and the pore diameter of the Dendritic Mesoporous Silica Nanoparticles (DMSNs) prepared by the embodiment can reach 15-20 nm. PVP-Bi directly grows at the in-situ position of the dendritic mesoporous silica nano particles (DMSNs), then tetradecanol (PCM) is introduced, a layer of tetradecanol (PCM) is coated outside the PVP-Bi nano points, and the Bi-DMSNs @ PCM composite nano material is synthesized. The melting point of the PCM is 38 ℃, when the nanocomposite is irradiated by a laser, the PCM on the outermost layer of the material is melted, and then the Bi nanodots are released, so that the characteristic that the Bi nanodots are oxidized can be effectively prevented, and the stability of the composite is improved.
In the embodiment, an in-situ growth method is adopted, PVP-Bi directly grows at the in-situ position of Dendritic Mesoporous Silica Nanoparticles (DMSNs), and then tetradecanol (PCM) is introduced. If the PVP-Bi nanodots are directly coated with the PCM outside on the basis of synthesizing the PVP-Bi nanodots, the added PCM coats a layer outside a large amount of PVP-Bi nanodots instead of coating the single nanospheres; if PVP-Bi nanodots and DMSNs particles are synthesized respectively, the PVP-Bi nanodots and the DMSNs particles are connected together by stirring through the electrostatic adsorption effect, and then the PVP-Bi nanodots and the DMSNs particles are coated outside the Bi-DMSNs through a method of directly adding PCM, but finally, bi does not grow in pores of the DMSNs, and the PCM does not coat outside the Bi-DMSNs. In the embodiment, the PVP-Bi nano dots are synthesized by a chemical reduction method, dendritic silicon Dioxide (DMSNs) is synthesized to serve as a carrier, and a layer of tetradecanol (PCM) is coated outside the Bi-DMSNs to prevent the PVP-Bi from being oxidized, so that the characteristic that PVP-Bi quantum dots are easily oxidized is avoided, and the stability of the material can be well kept. According to the method for preparing the nano-sized Bi-DMSNs @ PCM composite material, the characteristic that Bi is easy to oxidize is solved, and a brand-new solution and process flow are provided for preventing PVP-Bi from being oxidized.
The Bi-DMSNs @ PCM nano material prepared by the embodiment has good stability, oxidation resistance and higher photo-thermal conversion efficiency (the photo-thermal conversion efficiency can reach 32.3%), and the aqueous solution prepared from the prepared Bi-DMSNs @ PCM multifunctional nano composite material is very stable, so that the Bi nano points are prevented from being easily oxidized, and an optimized, quick and low-cost selection is provided for preparing a novel photo-thermal material.
The Bi-DMSNs @ PCM nano material prepared by the embodiment is 0 mu g/mL -1 ～500μg·mL -1 The survival rate of the L929 cells is more than 85 percent in the whole concentration range, and the Bi-DMSNs @ PCM has no obvious toxic or side effect on the L929 cells, has very small toxicity on normal cells and has good biocompatibility. Bi-DMSNs @ PCM has the best anti-tumor effect than PVP-Bi, stronger photothermal capacity and better treatment effect, and the power of a laser is 0.5W-cm -2 And the minimum survival rate of the HeLa cells and the 4T1 cells can be reduced to below 10 percent under the irradiation of 10 min.
The method of the embodiment is rapid, simple and convenient, requires low temperature, has good safety performance, does not need special reaction equipment, is environment-friendly, and can be used for industrialization.
In conclusion, the method has the advantages of simple and easy process, good repeatability, environmental protection and safety of the whole reaction system, and the provided process route has good application prospect and practical value. The prepared novel nano composite material is a potential tumor treatment reagent which can be used for photothermal treatment and has CT and photothermal imaging effects.
The second embodiment is as follows: the first difference between the present embodiment and the specific embodiment is: the centrifugation in the step one is high-speed centrifugation collection under the condition that the centrifugation speed is 10000 rpm-12000 rpm. The rest is the same as the first embodiment.
The third concrete implementation mode: this embodiment is different from the first or second embodiment in that: the washing in the step one is washing with ethanol for 3 times. The other is the same as in the first or second embodiment.
The fourth concrete implementation mode: the difference between this embodiment and one of the first to third embodiments is: the extraction in the step one is carried out by using a mixed solution of hydrochloric acid and methanol as an extracting agent at the temperature of 100-120 ℃; the mixed solution of hydrochloric acid and methanol is obtained by mixing 36-38% of hydrochloric acid and 99.5-100% of methanol according to the volume ratio of 1 (8-10). The rest is the same as the first or second embodiment.
The fifth concrete implementation mode is as follows: the difference between this embodiment and one of the first to fourth embodiments is: the first step is to place the mixture for 5 to 7 hours at the temperature of between 50 and 70 ℃. The rest is the same as the first to fourth embodiments.
The sixth specific implementation mode: the present embodiment is different from one or more of the first to fifth embodiments in that: in the step one, the extraction and the placement are repeated for 3 to 4 times. The rest is the same as the first to fifth embodiments.
The seventh embodiment: the difference between this embodiment and one of the first to sixth embodiments is: the drying in the step one is carried out in a vacuum drying oven at the temperature of 50-70 ℃ for overnight. The others are the same as the first to sixth embodiments.
The specific implementation mode is eight: the present embodiment differs from one of the first to seventh embodiments in that: and the centrifugation in the step two is centrifugal collection under the condition that the centrifugation speed is 12000 rpm-14000 rpm. The rest is the same as the first to seventh embodiments.
The specific implementation method nine: the present embodiment differs from the first to eighth embodiments in that: the washing in the second step is carried out according to the following steps: (1) washing with absolute ethyl alcohol and deionized water in sequence; (2) and (3) repeating the step (2) for three times. The other points are the same as those in the first to eighth embodiments.
The detailed implementation mode is ten: the present embodiment differs from one of the first to ninth embodiments in that: the drying in the second step is drying in a vacuum drying oven at the temperature of 50-70 ℃. The other points are the same as those in the first to ninth embodiments.
The following examples were used to demonstrate the beneficial effects of the present invention:
the first embodiment is as follows:
a preparation method of Bi-DMSNs @ PCM multifunctional nano composite material is carried out according to the following steps:
1. preparing dendritic mesoporous silica:
adding 0.068g of triethanolamine into 25mL of water, stirring for 0.5h in an oil bath with a magnetic stirring speed of 100rpm and a temperature of 80 ℃ to obtain a solution A, adding 0.38g of hexadecyl trimethyl ammonium bromide and 0.168g of sodium salicylate into the solution A, stirring for 1h under the conditions of a magnetic stirring speed of 110rpm and a temperature of 80 ℃ to obtain a solution B, adding a mixture of 4mL of tetraethoxysilane and 1.6mL of 1, 2-bis (triethoxysilyl) ethane into the solution B, stirring for 12h under the conditions of a magnetic stirring speed of 180rpm and a temperature of 80 ℃, centrifuging, washing, extracting, standing and drying to obtain the dendritic mesoporous silica;
2. preparing a Bi-DMSNs @ PCM nano composite material:
dissolving 0.3g of polyvinylpyrrolidone and 0.1g of bismuth nitrate in a mixture of glycerol and ethanol, heating to the temperature of 75 ℃ until the bismuth nitrate is fully melted to obtain a solution C, adding 0.08g of dendritic mesoporous silica into the solution C, stirring for 1min at the stirring speed of 180rpm to obtain a solution D, adding 0.05g of sodium borohydride into the solution D, stirring for 2h at the stirring speed of 180rpm to obtain a solution E, adding 0.02g of tetradecanol into the solution E, stirring for 5h at the stirring speed of 180rpm, centrifuging, washing, and drying to obtain the Bi-DMSNs @ PCM multifunctional nano composite material;
the mixture of the glycerol and the ethanol is formed by mixing 10mL of the glycerol and 5mL of the ethanol.
The centrifugation in the step one is high-speed centrifugation and collection under the condition that the centrifugation speed is 11000 rpm.
The washing in the step one is washing with ethanol for 3 times.
The extraction in the step one is carried out by using a mixed solution of hydrochloric acid and methanol as an extracting agent at the temperature of 110 ℃.
The mixed solution of hydrochloric acid and methanol is obtained by mixing 37% by mass of hydrochloric acid and 99.9% by mass of methanol according to a volume ratio of 1.
The first step is to place the mixture for 6 hours at the temperature of 60 ℃.
In step one, extraction and standing are repeated for 3 times.
The drying in the step one is carried out in a vacuum drying oven at the temperature of 60 ℃ overnight.
And the centrifugation in the second step is centrifugal collection under the condition that the centrifugation speed is 12000 rpm.
The washing in the second step is carried out according to the following steps: (1) washing with absolute ethyl alcohol and deionized water in sequence; (2) and (3) repeating the step (2) for three times.
And the drying in the step two is drying in a vacuum drying oven at the temperature of 60 ℃.
The dendritic mesoporous silica obtained in the step one is named as DMSNs; the Bi-DMSNs @ PCM multifunctional nano composite material prepared in the embodiment I is named as Bi-DMSNs @ PCM.
Comparison experiment one: dissolving 0.3g of polyvinylpyrrolidone and 0.1g of bismuth nitrate in a mixture of glycerol and ethanol, heating at 75 ℃ until the bismuth nitrate is fully melted, then adding 0.05g of sodium borohydride, stirring until no bubbles are generated and no heat is released, and after the reaction is finished, centrifuging, washing and drying to obtain the PVP-Bi nanodots; the mixture of the glycerol and the ethanol is formed by mixing 10mL of the glycerol and 5mL of the ethanol. The centrifugation, washing and drying are the same as the second step of the embodiment.
Comparative experiment two: dissolving 0.3g of polyvinylpyrrolidone and 0.1g of bismuth nitrate in a mixture of glycerol and ethanol, heating to the temperature of 75 ℃ until the bismuth nitrate is fully melted, then adding 0.05g of sodium borohydride, stirring until no bubbles are generated and no heat is released, adding 0.02g of tetradecanol, stirring for 5 hours at the stirring speed of 180rpm, centrifuging, washing and drying to obtain the Bi @ PCM coated with the PCM.
A third comparative experiment: and (3) stirring the PVP-Bi nano-dots prepared in the first comparative experiment and the DMSNs prepared in the first step of the embodiment for 30min at the stirring speed of 200rpm, adding 0.02g of tetradecanol, stirring for 5h at the stirring speed of 180rpm, centrifuging, washing and drying to obtain the Bi-DMSNs coated with the PCM.
FIG. 1 is a TEM image, a is PVP-Bi nanodots prepared in the first comparative experiment, b is Bi @ PCM coated with PCM prepared in the second comparative experiment, c is DMSNs prepared in the first step of the example, d is PCM coated with Bi-DMSNs prepared in the third comparative experiment, e is Bi-DMSNs @ PCM prepared in the first example at 100nm rate, f is single Bi-DMSNs @ PCM prepared in the first example at 50nm rate; a is a spherical shape with regular appearance, uniform grain diameter and obvious lattice fringes; b is a TEM image of Bi @ PCM directly coating PCM outside the PVP-Bi nanodots on the basis of synthesizing the PVP-Bi nanodots, and the added PCM coats a layer outside a large amount of PVP-Bi nanodots instead of coating the single nanospheres; c is synthesized Dendritic Mesoporous Silica (DMSNs), the appearance is uniform, the center-radial hole structure is unique, the grain diameter of the dendritic mesoporous silica is about 210nm, d is that PVP-Bi nanodots and DMSNs particles are respectively synthesized, then the two solutions are connected together by stirring under the action of electrostatic adsorption, and then the solutions are coated outside the Bi-DMSNs by directly adding PCM, the expected effect can be found not to be realized, bi does not grow in the holes of the DMSNs, and the PCM is not coated outside the Bi-DMSNs; e-f adopts an in-situ growth method, PVP-Bi directly grows at the in-situ position of the DMSNs, and then Bi-DMSNs @ PCM is introduced into the PCM, so that the expected effect is achieved, and the PCM is successfully coated outside.
FIG. 2 is the distribution diagram of elements of Bi-DMSNs @ PCM prepared in the first example, wherein a is HAADF diagram, b is C element, C is O element, d is Si element, e is Bi element, and f is element composite diagram; the single element distribution and superposition of the four elements of C, O, bi and Si can be seen, and the successful synthesis of Bi-DMSNs @ PCM can be seen.
FIG. 3 is a diagram of the UV-VIS absorption spectra of Bi-DMSNs @ PCM and PVP-Bi nanodots, 1 is the Bi-DMSNs @ PCM prepared in the first example, and 2 is the PVP-Bi nanodots prepared in the first comparative experiment; as can be seen from the figure, the nano-dots have wide absorption from ultraviolet to near infrared regions, which indicates that the nano-dots have good dispersibility in water, and the absorption of the Bi-DMSNs @ PCM in the near infrared region is better than that of PVP-Bi, which indicates that the Bi-DMSNs @ PCM improves the absorption in the near infrared region.
FIG. 4 is a Zeta potential diagram, a is PVP-Bi nanodots prepared in comparative experiment one, b is DMSNs prepared in example one step one, c is PCM, d is Bi-DMSNs @ PCM prepared in example one; as can be seen from the figure, PVP-Bi with positive charge is adhered to DMSNs with negative charge through electrostatic adsorption, and positive charge displayed by Bi-DMSNs @ PCM can analyze that PCM is successfully coated outside the PVP-Bi nanodots.
The Bi-DMSNs @ PCM prepared in example one was formulated to a concentration of 100. Mu.g.mL -1 Then, the aqueous solution of Bi-DMSNs @ PCM is allowed to stand at an ambient temperature of 25 ℃. FIG. 5 is a photograph of an aqueous solution of Bi-DMSNs @ PCM before and after standing; as can be seen from the figure, the aqueous solution of Bi-DMSNs @ PCM was black before it was allowed to stand at 25 ℃ and was still black after it was allowed to stand at 25 ℃ for 14 days. FIG. 6 is the UV-visible absorption spectra before and after the Bi-DMSNs @ PCM aqueous solution is left standing, wherein a is before the standing and b is after the standing for 14 days; as can be seen from the figure, the absorption value of Bi-DMSNs @ PCM before and after standing does not change, which indicates that the material has good stability and is not oxidized.
The Bi-DMSNs @ PCM prepared in example one was formulated to a concentration of 100. Mu.g.mL -1 In aqueous Bi-DMSNs @ PCM solution, then strip at a temperature of 80 DEG CHeating under the condition of workpiece. FIG. 7 is a photograph of an aqueous solution of Bi-DMSNs @ PCM before and after heating; it can be seen that the solution turns white after heating the aqueous solution of Bi-DMSNs @ PCM for 180min, whereas the existing PVP-Bi is immediately oxidized under heating, thus greatly slowing down the rate of oxidation of Bi compared to PVP-Bi. FIG. 8 is a graph showing the change of the ultraviolet-visible absorption curve of the aqueous solution of Bi-DMSNs @ PCM with the heating time, wherein a is unheated, b is heated 60min, c is heated 120min, and d is heated 180min; it was found that the absorption value of Bi-DMSNs @ PCM decreased after a certain period of time, indicating that Bi was released and oxidized due to the melting of PCM after heating Bi-DMSNs @ PCM for a short period of time at a certain temperature, but the time required for the oxidation of Bi-DMSNs @ PCM was longer and the material whitening rate was slower compared to PVP-Bi, thereby satisfying the stability of the material during the treatment.
The Bi-DMSNs @ PCM prepared in the first example is prepared into aqueous solutions with different concentrations, and the concentrations of the aqueous solutions of the Bi-DMSNs @ PCM are 0 mug. ML -1 、50μg·mL -1 、100μg·mL -1 、200μg·mL -1 And 500. Mu.g.mL -1 Irradiated with a laser at the same power (0.5W cm) –2 1064 nm) were measured every 10s during the irradiation with an infrared thermography. FIG. 9 shows Bi-DMSNs @ PCM aqueous solutions of different concentrations at 1064nm and 0.5W cm -2 The temperature rise profile within 10min of laser irradiation of (1) is 0. Mu.g.mL -1 2 is 50. Mu.g/mL -1 And 3 is 100. Mu.g/mL -1 And 4 is 200. Mu.g.mL -1 5 is 500. Mu.g/mL -1 (ii) a FIG. 10 shows Bi-DMSNs @ PCM aqueous solutions of different concentrations at 1064nm and 0.5W cm -2 Irradiating for 10min, and then carrying out temperature change-concentration curve graph; as can be seen from the figure, the curve of the concentration of Bi-DMSNs @ PCM and the temperature change is almost linear, and the temperature change within 10min can reach 30 ℃, so that the temperature is greatly changed. The concentration dependence of the thermal effect of Bi-DMSNs @ PCM is demonstrated by FIGS. 9 and 10. These results show that Bi-DMSNs @ PCM has good photothermal effect.
Determination of Bi-DMSNs @ PCM photo-thermal stability: the concentration was 1000. Mu.g.mL -1 In the aqueous solution of Bi-DMSNs @ PCM and having a concentration of 1000. Mu.g.mL -1 PVP-Bi nanodot water of (1)The solutions were placed in sample bottles using a 0.5 W.cm meter –2 The 1064nm laser is used for irradiation until the temperature does not rise any more, then the laser is switched off, and the temperature change of the sample and the surrounding environment is recorded by an infrared thermal imaging instrument. FIG. 11 is the infrared thermal imaging graph of PVP-Bi nanodot aqueous solution and Bi-DMSNs @ PCM aqueous solution under the irradiation of 1064nm laser, a is PVP-Bi nanodot aqueous solution and irradiated by 1064nm laser, b is Bi-DMSNs @ PCM aqueous solution and irradiated by 1064nm laser; it can be seen that the temperature rise speed of the sample Bi-DMSNs @ PCM under the irradiation of a 1064nm laser is faster, and the temperature rise speed is higher in the same time, which indicates that the photo-thermal performance of the Bi-DMSNs @ PCM under the irradiation of the 1064nm laser is better.
Measurement of Bi-DMSNs @ PCM photothermal conversion efficiency: the concentration was 1000. Mu.g.mL -1 In an aqueous Bi-DMSNs @ PCM solution and having a concentration of 1000. Mu.g.mL -1 The PVP-Bi aqueous solution (2) was placed in a sample bottle using a 0.5 W.cm –2 The 1064nm laser is used for irradiation until the temperature stabilizes and does not rise any more. And then closing the laser for natural cooling, measuring the temperature once every 10s by using an infrared thermal imager in the natural cooling process, and naturally cooling the solution to the ambient temperature.
FIG. 12 shows 1064nm and 0.5W cm -2 The temperature rise and cooling curve diagram of the solution after laser irradiation, 1 is Bi-DMSNs @ PCM aqueous solution, 2 is PVP-Bi aqueous solution, and 3 is water; fig. 13 is a natural logarithm graph of the cooling time and the driving temperature in fig. 12, c is a natural logarithm curve of the cooling time and the driving temperature corresponding to curve 2, and d is a natural logarithm curve of the cooling time and the driving temperature corresponding to curve 1; photothermal conversion efficiency (η T) of Bi-DMSNs @ PCM was determined as previously reported. The photothermal conversion efficiency of the Bi-DMSNs @ PCM can be calculated by an energy conservation law method, and the photothermal conversion efficiency eta T of the PVP-Bi is calculated to be 32.3%, which indicates that the Bi-DMSNs @ PCM can be used in the field of photothermal therapy.
Determination of Bi-DMSNs @ PCM photo-thermal stability: the concentration of the mixed solution is 1000 mug. Multidot.mL -1 The aqueous solution of Bi-DMSNs @ PCM (E) was put in a sample bottle using a 0.5 W.cm –2 Irradiating with 1064nm laser until the temperature is not raised, turning off the laser, and recording the sample and the surrounding environment with infrared thermal imaging instrumentWhen the temperature of the Bi-DMSNs @ PCM aqueous solution is not reduced any more, the laser is turned on again, the sample is irradiated by the same power until the temperature is not increased any more, then the laser is turned off, and the process is repeated for four times to record the temperature change. FIG. 14 is a graph showing the photo-thermal cycling curves after repeated irradiation of an aqueous Bi-DMSNs @ PCM solution; after the irradiation is repeated for four times and the solution is naturally cooled, the whole process lasts 70min, the rising temperature of the Bi-DMSNs @ PCM solution is basically consistent, the temperature rising capability of the sample is not changed, and the Bi-DMSNs @ PCM has good photo-thermal stability and is very suitable for long-term and repeated irradiation treatment.
And (3) detecting biocompatibility: first, well-grown L929 cells were plated in 96-well plates at appropriate densities, approximately 200. Mu.L per well, at 5% CO 2 And culturing at 37 deg.C for 24 hr to make the cells grow adherently. Then, the Bi-DMSNs @ PCM prepared in example one was dissolved in 100. Mu.L of the medium to prepare different concentrations, and the concentrations of the Bi-DMSNs @ PCM solutions were 0. Mu.g/mL, 15.625. Mu.g/mL, 31.25. Mu.g/mL, 62.5. Mu.g/mL, 125. Mu.g/mL, 250. Mu.g/mL and 500. Mu.g/mL, respectively, and then added to each well separately to continue the culture for 24 hours. Wherein, pure culture medium was added as a control group. Thereafter, 20. Mu.L of a stock MTT solution at a concentration of 5mg/mL was added to each well seeded with L929 cells, and then the culture of the cells was continued for 4 hours according to the previous environmental conditions. After 4h, 150 mu L of dimethyl sulfoxide solution is added into each hole, and after 10min, the absorbance of the suspension in each hole at 490nm is detected by a microplate reader, so that the survival rate of the cells with different concentrations of the sample is calculated.
FIG. 15 is a graph showing the comparison of cell viability after 24h of Bi-DMSNs @ PCM and L929 cell culture at different concentrations; the survival rate of the cells does not change obviously with the increase of the sample concentration, and is between 0 and 500 mu g.mL -1 The survival rate of all cells is more than 85 percent in the whole concentration range, which indicates that the Bi-DMSNs @ PCM has no obvious toxic or side effect on L929 cells, has very small toxicity on normal cells and simultaneously has good biocompatibility, and indicates that the synthesized nano composite material can be applied to the field of biomedicine;
and (3) detecting cytotoxicity:
testing of Bi-DMSNs @ PCM on HeLa cells by MTT methodAnd 4T1 cell toxicity. First, 4T1 cells grown in good condition at appropriate density were seeded in 96-well plates at 200. Mu.L per well in 5% CO 2 And incubating for 24h at 37 ℃ to ensure that the cells grow adherently. The cytotoxicity experiment process of the 4T1 cell is similar to the biocompatibility of the L929 cell, and specifically, the PVP-Bi nanodot prepared in the first comparative experiment and the Bi-DMSNs @ PCM prepared in the first example are respectively dissolved in 100 mu L of culture medium to prepare different concentrations, and the concentrations of the PVP-Bi nanodot solution and the Bi-DMSNs @ PCM solution are respectively 15.625 mu g/mL -1 、31.25μg·mL -1 、62.5μg·mL -1 、125μg·mL -1 、250μg·mL -1 And 500. Mu.g.mL -1 Then added separately to each well at 37 ℃ with 5% CO 2 Incubate for 4h under standard conditions. The 4T1 cells were then treated as follows: pure culture medium is treated by 1064nm laser, bi-DMSNs @ PCM prepared in the first example is treated, PVP-Bi nanodots prepared in the first comparative experiment are added, 1064nm laser is used for irradiating cells, bi-DMSNs @ PCM is added, 1064nm laser is used for irradiating the cells, the irradiation time is 10min, and the laser power is 0.5W-cm -2 Wherein the pure medium group is set as a control group. Then, the reaction was continued at 37 ℃ and 5% CO 2 4T1 cells were cultured for 48h under standard conditions. Next, the cells were treated with MTT solution, cultured for 4 hours, and then the supernatant was removed. And respectively adding 150 mu L of dimethyl sulfoxide into each hole, and detecting the absorbance of the suspension in each hole at 490nm by using a microplate reader after 10min, so as to calculate the survival rate of the 4T1 cells with different concentrations of the sample, wherein the cytotoxicity experimental process of the HeLa cells is the same as that of the 4T1 cells.
FIG. 16 is a graph showing comparison of cell viability after HeLa cells were cultured under different conditions, a is the addition of Bi-DMSNs @ PCM prepared in example one and irradiation of the cells with a 1064nm laser, b is the addition of PVP-Bi nanodots prepared in comparative experiment one and irradiation of the cells with a 1064nm laser, c is the addition of Bi-DMSNs @ PCM prepared in example one, and d is the addition of pure medium and treatment with a 1064nm laser; MTT results show that the survival rate of HeLa cells in a group which is added with pure culture medium and irradiated by 1064nm laser is more than 80%, which shows that the cells can not be effectively killed only by irradiation of 1064nm near-infrared laser, while the lowest survival rate of the cells is only 8% by adding Bi-DMSNs @ PCM and irradiating the cells by 1064nm laser.
FIG. 17 is a graph showing comparison of cell viability after culturing 4T1 cells under different conditions, wherein a is the addition of Bi-DMSNs @ PCM prepared in example one and irradiating the cells with a 1064nm laser, b is the addition of PVP-Bi nanodots prepared in comparative experiment one and irradiating the cells with a 1064nm laser, c is the addition of Bi-DMSNs @ PCM prepared in example one, and d is the addition of pure medium and irradiating with a 1064nm laser; the MTT results show that the survival rate of the 4T1 cells in the group which is added with the pure culture medium and irradiated by the 1064nm laser is more than 80 percent, which shows that the cells can not be effectively killed only by irradiating the cells by the 1064nm near-infrared laser. Therefore, treatment with Bi-DMSNs @ PCM and exhibiting the strongest tumor inhibition with 1064nm light irradiation showed the lowest survival rate of only 10%.
Cell staining experiment: heLa cells of appropriate cell density were seeded in 6-well plates, 2mL per well, at 5% CO 2 And incubating for 24h at 37 ℃ to ensure that the cells grow adherently. The 6-well cells were randomly divided into four groups, the first group treated with 1064nm laser only, the second group treated with addition of only Bi-DMSNs @ PCM prepared in example one, the third group added PVP-Bi nanodots prepared in comparative experiment one and irradiated with 1064nm laser, and the fourth group added Bi-DMSNs @ PCM prepared in example one and irradiated with 1064nm laser. The PVP-Bi nanodots prepared in the first comparative experiment and the Bi-DMSNs @ PCM prepared in the first example were dissolved in the medium respectively, and 1mL of the solution was added at a concentration of 500. Mu.g.mL -1 PVP-Bi solution of (1 mL) and a concentration of 500. Mu.g.mL -1 The Bi-DMSNs @ PCM solutions of (A) were added to the corresponding wells, respectively, at 37 ℃ and 5% CO 2 Respectively culturing for a period of time under standard conditions until the cells engulf materials, then respectively irradiating a first group, a third group and a fourth group with 1064nm, then continuously culturing for a period of time, then washing with PBS three times for washing out samples which are not engulfed, respectively adding a certain amount of calcein (AM) and Propidium Iodide (PI) into a 6-well plate so as to dye the cells, then respectively using 1mL of 2.5% glutaraldehyde, standing for 10min, and then observing the cell staining condition by using a laser confocal microscope.
FIG. 18 is a confocal laser microscopy image of HeLa cells after staining with calcein and propidium iodide, a being laser treatment with 1064nm only, b being treatment with addition of Bi-DMSNs @ PCM prepared in example one, c being addition of PVP-Bi nanodots prepared in comparative experiment one and irradiating the cells with 1064nm laser, d being addition of Bi-DMSNs @ PCM prepared in example one and irradiating the cells with 1064nm laser; it was found that when HeLa cells were irradiated with only 1064nm near-infrared light, there were almost no red cells, indicating that the cells were substantially alive, indicating that irradiation of the cells with only 1064nm near-infrared laser light was not effective in killing them. For the cells treated by PVP-Bi and irradiated by 1064nm near-infrared laser, a part of the cells are found to be red and a part of the cells are green, which proves that PVP-Bi has a certain inhibiting effect on tumor cells under the irradiation of 1064nm laser. After the material is cultured with Bi-DMSNs @ PCM and irradiated by 1064nm near infrared light, almost all red cells in a picture are red due to a photo-thermal effect, and the number of the red cells is more than that of the red cells when the cells are treated by PVP-Bi and 1064nm laser, which shows that the material Bi-DMSNs @ PCM has stronger photo-thermal capability and better treatment effect under the irradiation of 1064nm laser.
FIG. 19 is confocal laser microscopy images after staining with JC-1, (1) PVP-Bi plus 1064nm laser treated 4T1 cells prepared in comparative experiment one, (2) Bi-DMSNs @ PCM plus 1064nm laser treated 4T1 cells prepared in example one. The group treated with Bi-DMSNs @ PCM plus 1064nm laser fluoresced more strongly in green, showing the highest fluorescence ratio of green to red, indicating that a large number of mitochondria were destroyed. The results show that the anti-tumor effect is best when Bi-DMSNs @ PCM is used compared with PVP-Bi.

Claims (10)

1. A preparation method of Bi-DMSNs @ PCM multifunctional nano composite material is characterized by comprising the following steps:
1. preparing dendritic mesoporous silica:
adding triethanolamine into water, stirring for 0.4-0.6 h in an oil bath with a magnetic stirring speed of 90-150 rpm and a temperature of 70-90 ℃ to obtain a solution A, adding cetyl trimethyl ammonium bromide and sodium salicylate into the solution A, stirring for 0.5-1.5 h under the conditions of a magnetic stirring speed of 90-150 rpm and a temperature of 70-90 ℃ to obtain a solution B, adding a mixture of tetraethoxysilane and 1, 2-bis (triethoxysilyl) ethane into the solution B, stirring for 11-13 h under the conditions of a magnetic stirring speed of 150-200 rpm and a temperature of 70-90 ℃, centrifuging, washing, extracting, standing and drying to obtain the dendritic mesoporous silica;
the mass ratio of the triethanolamine to the water is 1g (355-375) mL; the mass ratio of the triethanolamine to the hexadecyl trimethyl ammonium bromide is 1 (4-6); the mass ratio of the triethanolamine to the sodium salicylate is 1 (2-3); the volume ratio of the mass of the triethanolamine to the volume of the tetraethoxysilane is 1g (55-65) mL; the volume ratio of the mass of the triethanolamine to the 1, 2-bis (triethoxysilyl) ethane is 1g (20-25) mL;
2. preparing a Bi-DMSNs @ PCM nano composite material:
dissolving polyvinylpyrrolidone and bismuth nitrate in a mixture of glycerol and ethanol, heating to fully melt bismuth nitrate at the heating temperature of 60-70 ℃ to obtain a solution C, adding dendritic mesoporous silica into the solution C, stirring for 0.5-2 min at the stirring speed of 150-200 rpm to obtain a solution D, adding sodium borohydride into the solution D, stirring for 1-3 h at the stirring speed of 150-200 rpm to obtain a solution E, adding tetradecanol into the solution E, stirring for 3-6 h at the stirring speed of 150-200 rpm, centrifuging, washing, and drying to obtain the Bi-DMSNs @ PCM multifunctional nanocomposite;
the mass ratio of the polyvinylpyrrolidone to the bismuth nitrate is 1 (0.2-0.4); the volume ratio of the mass of the polyvinylpyrrolidone to the mixture of the glycerol and the ethanol is 1g (40-60) mL; the volume ratio of the glycerol to the ethanol in the mixture of the glycerol and the ethanol is 1 (0.4-0.6); the mass ratio of the polyvinylpyrrolidone to the dendritic mesoporous silica is 1 (0.15-0.35); the mass ratio of the polyvinylpyrrolidone to the sodium borohydride is 1 (0.1-0.25); the mass ratio of the polyvinylpyrrolidone to the tetradecanol is 1 (0.01-0.15).
2. The method for preparing the Bi-DMSNs @ PCM multifunctional nanocomposite material according to claim 1, wherein the centrifugation in the step one is high-speed centrifugation collection under the condition that the centrifugation speed is 10000 rpm-12000 rpm.
3. The method for preparing the Bi-DMSNs @ PCM multifunctional nanocomposite material according to claim 1, wherein the washing in the step one is 3 times of washing with ethanol.
4. The method for preparing the Bi-dmsns @ pcm multifunctional nanocomposite as claimed in claim 1, wherein the extraction in the step one is performed by using a mixed solution of hydrochloric acid and methanol as an extractant at a temperature of 100 ℃ to 120 ℃; the mixed solution of hydrochloric acid and methanol is obtained by mixing 36-38% of hydrochloric acid and 99.5-100% of methanol according to the volume ratio of 1 (8-10).
5. The method for preparing the Bi-dmsns @ pcm multifunctional nanocomposite as claimed in claim 1, wherein the placing in the step one is for 5h to 7h at a temperature of 50 ℃ to 70 ℃.
6. The method for preparing the Bi-dmsns @ pcm multifunctional nanocomposite as claimed in claim 1, wherein the extraction and the standing are repeated 3 to 4 times in the step one.
7. The method for preparing the Bi-dmsns @ pcm multifunctional nanocomposite as claimed in claim 1, wherein the drying in the step one is carried out overnight in a vacuum drying oven at a temperature of 50 ℃ to 70 ℃.
8. The method for preparing the Bi-dmsns @ pcm multifunctional nanocomposite as claimed in claim 1, wherein the centrifugation in the step two is performed at a centrifugation speed of 12000rpm to 14000 rpm.
9. The preparation method of the Bi-dmsns @ pcm multifunctional nanocomposite material according to claim 1, wherein the washing in the second step is specifically performed according to the following steps: (1) washing with absolute ethyl alcohol and deionized water in sequence; (2) and (3) repeating the step (1) for three times.
10. The method for preparing the Bi-dmsns @ pcm multifunctional nanocomposite as claimed in claim 1, wherein the drying in the step two is drying in a vacuum drying oven at a temperature of 50 ℃ to 70 ℃.
CN202011246485.4A 2020-11-10 2020-11-10 Preparation method of Bi-DMSNs @ PCM multifunctional nano composite material Active CN112316140B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN202011246485.4A CN112316140B (en) 2020-11-10 2020-11-10 Preparation method of Bi-DMSNs @ PCM multifunctional nano composite material

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN202011246485.4A CN112316140B (en) 2020-11-10 2020-11-10 Preparation method of Bi-DMSNs @ PCM multifunctional nano composite material

Publications (2)

Publication Number Publication Date
CN112316140A CN112316140A (en) 2021-02-05
CN112316140B true CN112316140B (en) 2022-10-28

Family

ID=74317362

Family Applications (1)

Application Number Title Priority Date Filing Date
CN202011246485.4A Active CN112316140B (en) 2020-11-10 2020-11-10 Preparation method of Bi-DMSNs @ PCM multifunctional nano composite material

Country Status (1)

Country Link
CN (1) CN112316140B (en)

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN115531534B (en) * 2021-06-29 2024-02-09 国家纳米科学中心 Nanometer diagnosis and treatment agent, preparation method and application

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2013031767A1 (en) * 2011-08-29 2013-03-07 地方独立行政法人東京都立産業技術研究センター Process for producing particles held in porous silica, porous silica, and particles held in porous silica
CN106362149A (en) * 2016-09-18 2017-02-01 天津大学 Door control type medicine composition integrating cancer imaging and phototherapy and preparation method
CN110484236A (en) * 2019-07-29 2019-11-22 浙江大学 The preparation method of bismuth quanta point material is grown in a kind of mesoporous silica nano-particle

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2013031767A1 (en) * 2011-08-29 2013-03-07 地方独立行政法人東京都立産業技術研究センター Process for producing particles held in porous silica, porous silica, and particles held in porous silica
CN106362149A (en) * 2016-09-18 2017-02-01 天津大学 Door control type medicine composition integrating cancer imaging and phototherapy and preparation method
CN110484236A (en) * 2019-07-29 2019-11-22 浙江大学 The preparation method of bismuth quanta point material is grown in a kind of mesoporous silica nano-particle

Non-Patent Citations (5)

* Cited by examiner, † Cited by third party
Title
"Bismuth embedded silica nanoparticles loaded with autophagy suppressant to promote photothermal therapy";Tong Chen等;《Biomaterials》;20190809;第221卷;第1-9页 *
"Enhanced stability and catalytic activity of bismuth nanoparticles by modified with porous silica";Kejun Chen等;《Journal of Physics and Chemistry of Solids》;20170526;第110卷;第9-14页 *
"On-Demand Triggered Chemodynamic Therapy by NIR-II Light on Oxidation-Prevented Bismuth Nanodots";Lu Yang等;《ACS Appl. Mater. Interfaces》;20220504(第14期);第21787-21799页 *
"基于 Bi 的纳米复合材料的制备及肿瘤诊疗性能研究";贾培培;《哈尔滨工程大学硕士学位论文》;20210422;第1-70页 *
氨基功能化介孔二氧化硅纳米药物传递载体的制备与性能研究;赵松峰等;《中国药学杂志》;20180822(第16期);全文 *

Also Published As

Publication number Publication date
CN112316140A (en) 2021-02-05

Similar Documents

Publication Publication Date Title
Zhu et al. Red carbon dots: Optical property regulations and applications
Yu et al. Black hollow silicon oxide nanoparticles as highly efficient photothermal agents in the second near-infrared window for in vivo cancer therapy
Jiang et al. Bifunctional Fe3O4–Ag heterodimer nanoparticles for two‐photon fluorescence imaging and magnetic manipulation
CN107184978B (en) Copper sulfide @ mesoporous silica nanocomposite and preparation method and application thereof
Yan et al. Recent advances in the rational design of copper chalcogenide to enhance the photothermal conversion efficiency for the photothermal ablation of cancer cells
Xu et al. High-yield synthesis of gold bipyramids for in vivo CT imaging and photothermal cancer therapy with enhanced thermal stability
Song et al. A novel photothermal nanocrystals of Cu 7 S 4 hollow structure for efficient ablation of cancer cells
CN106075443B (en) A kind of gold cladding copper selenide nanoparticles and the preparation method and application thereof
CN112316140B (en) Preparation method of Bi-DMSNs @ PCM multifunctional nano composite material
CN114346253B (en) Preparation method of silver nanocluster-palladium composite nanosheets, product and application thereof
CN110408377B (en) Rare earth doped NaCeF4Near-infrared fluorescent nano probe and preparation method and biological application thereof
Fang et al. Zeolitic imidazole framework coated Au nanorods for enhanced photothermal therapy and stability
CN111388448A (en) Nano particles for photothermal treatment of tumors and preparation and application thereof
CN107986318B (en) Water-soluble CuS nanocrystal and synthesis method and application thereof
Du et al. Engineering Cu 2− x S-conjugated upconverting nanocomposites for NIR-II light-induced enhanced chemodynamic/photothermal therapy of cancer
CN110623940A (en) Selenium/silicon dioxide/gold nano composite particle and preparation method and application thereof
CN112891536B (en) Bismuth-based nano heterojunction material and preparation method and application thereof
CN107952071B (en) Preparation method of molybdenum disulfide quantum dot-loaded periodic mesoporous organosilicon nano drug-loaded compound
CN109589408A (en) A kind of meter of shape liquid metal nanoparticle and its synthetic method and application
Xu et al. Restriction of molecular motion to a higher level: Towards bright AIE dots for biomedical applications
CN108888778B (en) Mesoporous silica nano composite material based on near-infrared Aza-Bodipy composite, and preparation method and application thereof
CN114767883B (en) Preparation method and application of nano composite material based on Au-PbS heterostructure
CN106902352B (en) The controllable method for preparing of the selenium category compound nano piece of copper for tumor thermal therapy
CN106913871B (en) Platinum-copper nanocrystalline used as tumor photothermal ablation diagnosis and treatment agent and preparation method thereof
CN114177289A (en) Composite nano material for photodynamic and photothermal combined treatment and preparation method and application thereof

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