WO2019148802A1 - 纳米钛光热制剂及其制备方法和应用 - Google Patents

纳米钛光热制剂及其制备方法和应用 Download PDF

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WO2019148802A1
WO2019148802A1 PCT/CN2018/100582 CN2018100582W WO2019148802A1 WO 2019148802 A1 WO2019148802 A1 WO 2019148802A1 CN 2018100582 W CN2018100582 W CN 2018100582W WO 2019148802 A1 WO2019148802 A1 WO 2019148802A1
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titanium
photothermal
nanosheet
dimensional
dimensional titanium
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张晗
陈世优
谢中建
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Shenzhen University
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    • 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
    • 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
    • A61K33/00—Medicinal preparations containing inorganic active ingredients
    • 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
    • A61K9/0002—Galenical forms characterised by the drug release technique; Application systems commanded by energy
    • A61K9/0009—Galenical forms characterised by the drug release technique; Application systems commanded by energy involving or responsive to electricity, magnetism or acoustic waves; Galenical aspects of sonophoresis, iontophoresis, electroporation or electroosmosis
    • 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
    • A61K9/0012—Galenical forms characterised by the site of application
    • A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
    • A61K9/0024—Solid, semi-solid or solidifying implants, which are implanted or injected in body tissue
    • 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
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents

Definitions

  • the invention relates to the field of biomedical nano materials, in particular to a nano titanium photothermal preparation, a preparation method and application thereof.
  • Near-infrared phototherapy based on nanomaterials is a new type of potential therapy, especially near-infrared light-responsive photothermal therapy, which solves the problem that light waves are easily absorbed, easily scattered, and have nucleic acids in short-wavelength phototherapy such as ultraviolet light and visible light. Toxicity, as well as poor tissue penetration.
  • the near-infrared light has a wavelength of about 700-1000 nm and has the advantages of low absorption and high penetration (more than 1 cm).
  • conventional photothermal therapy materials mainly include gold nanoparticles, carbon nanotubes, graphene and the like.
  • existing photothermal materials often fail to balance photothermal conversion efficiency and biocompatibility.
  • the present invention provides a nano titanium photothermal preparation with high photothermal conversion efficiency, good biocompatibility, and safety and non-toxicity.
  • a first aspect of the invention provides a nanotitanium photothermal formulation comprising a two-dimensional titanium nanosheet and a biocompatible material coated on the surface of the two-dimensional titanium nanosheet.
  • the mass ratio of the two-dimensional titanium nanosheet and the biocompatible material is 1:1-10.
  • the mass ratio of the two-dimensional titanium nanosheet to the biocompatible material is 1:1.
  • the two-dimensional titanium nanosheet has a thickness of 1-50 nm.
  • the two-dimensional titanium nanosheet has a thickness of 3-5 nm.
  • the two-dimensional titanium nanosheet has a length to width dimension of 10-50 nm.
  • the two-dimensional titanium nanosheet has a length to width dimension of 30-40 nm.
  • the biocompatible material comprises hyaluronic acid, dextran and derivatives thereof, chitosan and derivatives thereof, pectin, carboxymethyl cellulose, albumin, liposome, cell membrane, polyethylene One or more of pyrrolidone, polylactic acid-glycolic acid copolymer, polyethyleneimine, polyacrylic acid, and polyethylene glycol and derivatives thereof.
  • the biocompatible material is polyethylene glycol and a derivative thereof, and the polyethylene glycol and the derivative thereof have a molecular weight of 200-20000.
  • biocompatible material is adsorbed on the surface of the two-dimensional titanium nanosheet by electrostatic action.
  • the nanotitanium photothermal preparation further comprises a targeting material attached to the two-dimensional titanium nanosheet by chemical bonding or attached to the biocompatible material.
  • the targeting material is folic acid
  • the folic acid is attached to the polyethylene glycol and its derivative via an amide bond.
  • the nano titanium photothermal preparation provided by the first aspect of the invention has high photothermal conversion efficiency, good biocompatibility, safety and non-toxicity, and can be used for photothermal therapy including tumors.
  • a second aspect of the present invention provides a method for preparing a nano titanium photothermal preparation, comprising:
  • a biocompatible material is provided, and the two-dimensional titanium nanosheet and the biocompatible material are mixed and stirred at 10-30 ° C for 3-6 h to obtain a nano titanium photothermal preparation.
  • the preparation method of the nano titanium photothermal preparation specifically comprises: dissolving the biocompatible material in an appropriate amount of the first solvent to obtain a biocompatible material solution, and dispersing the two-dimensional titanium nanosheet in an appropriate amount.
  • a two-dimensional titanium nanosheet dispersion is obtained, and the biocompatible material solution is mixed with the two-dimensional titanium nanosheet dispersion at 10-30 ° C, and stirred for 3-6 hours to obtain nano titanium.
  • the stirring rate is from 100 rpm to 700 rpm.
  • the method for liquid phase stripping specifically comprises the following operations:
  • the titanium raw material is added to the solvent, and the probe is ultrasonicated for 8-15 hours in an ice bath environment; after the ultrasonication of the probe is finished, the water bath ultrasonic wave is continued, the ultrasonic time of the water bath is 3-10 h, and the temperature of the water bath is maintained. 5-15 ° C; after ultrasonication, centrifugation and drying to obtain two-dimensional titanium nanosheets.
  • the ultrasonic power of the probe is 200-250 W
  • the ultrasonic power of the water bath is 300-380 W.
  • the centrifugation operation comprises: firstly using a centrifugal force of 1800-2200 g, centrifuging for 20-35 min, taking the supernatant; then, the supernatant is continuously centrifuged by using a centrifugal force of 10000-13000 g to obtain a precipitate which is two-dimensional titanium. Nanosheets.
  • the second aspect of the invention provides a preparation method of the nano titanium photothermal preparation, the preparation method is simple and easy to operate, and the prepared nano titanium photothermal preparation has high light-heat conversion efficiency and good biocompatibility.
  • a third aspect of the present invention provides the use of the nano-titanium photothermal preparation as described above for the preparation of a photothermotherapy drug.
  • the nano titanium photothermal preparation provided by the invention has high photothermal conversion efficiency, good biocompatibility, safety and non-toxicity, and can be used for photothermal therapy including tumors;
  • the preparation method of the nano titanium photothermal preparation provided by the invention is simple and easy to operate.
  • Example 1 is a transmission electron micrograph of a two-dimensional titanium nanosheet prepared in Example 1;
  • Example 2 is an atomic force micrograph of a two-dimensional titanium nanosheet prepared in Example 1;
  • Example 3 is an absorption spectrum diagram of a liquid phase stripping process of a two-dimensional titanium nanosheet in Example 1;
  • Figure 4 is a photograph of a two-dimensional titanium nanosheet aqueous dispersion at different concentrations
  • Figure 6 is an extinction coefficient of a two-dimensional titanium nanosheet aqueous dispersion
  • Figure 8 is a photothermal conversion efficiency of a two-dimensional titanium nanosheet aqueous dispersion
  • Figure 9 is a graph showing the results of cytotoxicity measurement of two-dimensional titanium nanosheets.
  • Figure 10 is the effect of two-dimensional titanium nanosheets on body weight of mice
  • Figure 11 is a graph showing the effect of two-dimensional titanium nanosheets on tissue damage in mice.
  • Figure 13 is a graph showing the killing ability of a 50 ppm concentration of PEG2000-coated two-dimensional titanium nanosheets at different illumination times
  • Figure 14 is a graph showing changes in tumor site temperature in mouse tumor photothermal therapy
  • Figure 15 is a graph showing changes in tumor site temperature during phototherapy of mouse tumor
  • Figure 16 shows the volume change of tumors in tumor-bearing mice after photothermal therapy.
  • titanium nanosheet refers to elemental titanium.
  • a first aspect of the invention provides a nanotitanium photothermal formulation comprising a two-dimensional titanium nanosheet and a biocompatible material coated on the surface of the two-dimensional titanium nanosheet.
  • the two-dimensional titanium nanosheet has a thickness of 1-50 nm.
  • the two-dimensional titanium nanosheet has a thickness of 3-5 nm.
  • the two-dimensional titanium nanosheet has a thickness of 5-10 nm.
  • the two-dimensional titanium nanosheet has a thickness of 10-50 nm.
  • the two-dimensional titanium nanosheet has a thickness of 1 nm, 5 nm, 10 nm, 15 nm, 20 nm, 25 nm, 30 nm, 35 nm, 40 nm, 45 nm or 50 nm.
  • the two-dimensional titanium nanosheet has a length to width dimension of 10 to 50 nm.
  • the two-dimensional titanium nanosheet has a length to width dimension of 30-40 nm.
  • the two-dimensional titanium nanosheet has a length to width dimension of 10-30 nm.
  • the two-dimensional titanium nanosheet has a length to width dimension of 10 nm, 15 nm, 20 nm, 25 nm, 30 nm, 35 nm, 40 nm, 45 nm or 50 nm.
  • the mass ratio of the two-dimensional titanium nanosheet to the biocompatible material is 1:1-10.
  • the mass ratio of the two-dimensional titanium nanosheet to the biocompatible material is 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7. , 1:8, 1:9 or 1:10.
  • the two-dimensional titanium nanosheet has absorption from a visible region to a near-infrared region.
  • the two-dimensional titanium nanosheet has a light absorption wavelength ranging from 200 to 2000 nm.
  • the photothermal conversion efficiency of the two-dimensional titanium nanosheet is greater than or equal to 70%.
  • the two-dimensional titanium nanosheet provided by the first aspect of the invention has the advantages of environmental friendliness, biocompatibility, strong absorption of full spectrum and high photothermal conversion efficiency, and has excellent photothermal performance.
  • the biocompatible material comprises hyaluronic acid, dextran and derivatives thereof, chitosan and derivatives thereof, pectin, carboxymethyl cellulose, albumin, liposome, One or more of a cell membrane, polyvinylpyrrolidone, polylactic acid-glycolic acid copolymer, polyethyleneimine, polyacrylic acid, and polyethylene glycol and derivatives thereof. Further optionally, the biocompatible material comprises at least one of polyethylene glycol and derivatives thereof, polylactic acid-glycolic acid copolymer, albumin, liposomes, and cell membranes.
  • the biocompatible material comprises polyethylene glycol and derivatives thereof, the polyethylene glycol and its derivatives having a molecular weight between 200 and 20,000.
  • the end of the polyethylene glycol may be modified by an amino group.
  • the biocompatible material is adsorbed on the surface of the two-dimensional titanium nanosheet by electrostatic interaction.
  • the nanotitanium photothermal preparation further comprises a targeting material attached to the two-dimensional titanium nanosheet by chemical bonding or attached to the biocompatible material.
  • the targeting material is folic acid, and the folic acid is attached to the polyethylene glycol via an amide bond.
  • the nano titanium photothermal preparation may be dispersed in physiological saline, phosphate buffer or deionized water for subsequent application.
  • the nano titanium photothermal preparation provided by the first aspect of the invention has high photothermal conversion efficiency, good biocompatibility, safety and non-toxicity, and can be used for photothermal treatment including tumors.
  • a second aspect of the present invention provides a method for preparing a nano titanium photothermal preparation, comprising:
  • a biocompatible material is provided, and the two-dimensional titanium nanosheet and the biocompatible material are mixed and stirred at 10-30 ° C for 3-6 h to obtain a nano titanium photothermal preparation.
  • the two-dimensional titanium nanosheets and the biocompatible material are mixed at 25 ° C for 3-6 h.
  • the specific preparation method of the nano titanium photothermal preparation comprises: dissolving the biocompatible material in an appropriate amount of the first solvent to obtain a biocompatible material solution, and dispersing the two-dimensional titanium nanosheet. Obtaining a two-dimensional titanium nanosheet dispersion in an appropriate amount of the second solvent, mixing the biocompatible material solution with the two-dimensional titanium nanosheet dispersion at 10-30 ° C, and stirring for 3-6 hours. Nano titanium photothermal preparation.
  • the first solvent is a solvent that can dissolve the biocompatible material, such as when the biocompatible material is polyethylene glycol, the first solvent can be selected as deionized water.
  • the second solvent comprises deionized water.
  • the agitation rate is from 100 rpm to 700 rpm.
  • a two-dimensional titanium nanosheet coated with a biocompatible material is obtained, thereby obtaining a nano titanium photothermal preparation.
  • the method for liquid phase stripping specifically includes the following operations:
  • the titanium raw material is added to the solvent, and the probe is ultrasonicated for 8-15 hours in an ice bath environment; after the ultrasonication of the probe is finished, the water bath ultrasonic wave is continued, the ultrasonic time of the water bath is 3-10 h, and the water bath temperature is maintained at 5-15. °C; after ultrasonication, centrifugation and drying to obtain two-dimensional titanium nanosheets.
  • the solvent comprises at least one of isopropanol, ethanol, water, and methylpyrrolidone (i.e., N-methylpyrrolidone, NMP).
  • methylpyrrolidone i.e., N-methylpyrrolidone, NMP
  • the titanium raw material is present in the solvent at a concentration of from 1 to 7 mg/mL.
  • the probe has an ultrasonic power of 200-250W. Further optionally, the ultrasonic power of the probe is 240W.
  • the probe is sonicated for 10 hours.
  • the probe ultrasound is non-continuous ultrasound
  • the ultrasonic on/off time is selected to be 2/4 s, ie, ultrasonic for 2 s, then the ultrasound probe is turned off for 4 s, ultrasound is continued for 2 s, and so on.
  • the water bath has an ultrasonic power of 300-380 W. Further optionally, the water bath ultrasonic power is 360W.
  • the time of the water bath ultrasound is 8 h.
  • the water bath temperature is maintained at 10 °C.
  • the centrifugation operation comprises: firstly using a centrifugal force of 1800-2200 g, centrifuging for 20-35 min, taking the supernatant; and then centrifuging the supernatant with a centrifugal force of 10000-13000 g.
  • the precipitate is obtained as a two-dimensional titanium nanosheet.
  • the supernatant is taken by centrifugal force of 2000 g, and the supernatant is taken for 30 min; then the supernatant is centrifuged with 12,000 g to obtain a precipitate, and the obtained precipitate is dried to obtain a two-dimensional titanium nanosheet.
  • the manner of drying is not limited, and may be, for example, vacuum drying.
  • the prior art generally employs a liquid phase lift-off method for stripping a two-dimensional layered material.
  • the present invention has succeeded in stripping two-dimensional non-layered metal materials by liquid phase stripping.
  • the second aspect of the invention provides a preparation method of the nano titanium photothermal preparation, the preparation method is simple and easy to operate, and the prepared nano titanium photothermal preparation has high light-heat conversion efficiency and good biocompatibility.
  • a third aspect of the present invention provides a use of the nano titanium photothermal preparation as described above for the preparation of a photothermotherapy drug.
  • a method for preparing a nano titanium photothermal preparation comprising:
  • 500 mg of titanium powder was added to 100 ml of isopropanol. Then select the probe ultrasound 240W, ultrasound for 10h. The ultrasound on/off time was chosen to be 2/4 s and ultrasound was performed in an ice bath environment. After the probe is ultrasonicated, it is then ultrasonically probed in a water bath. The water bath ultrasonic power is 360W. The ultrasound time was 8 h. The bath temperature was maintained at 10 °C.
  • the desired metal elemental titanium nanosheets are obtained by centrifugation. First, the centrifugal force of 2000g was used and centrifuged for 30 minutes. The supernatant was taken, and then the supernatant was centrifuged at 12000 g to obtain a precipitate. The precipitate was vacuum dried to obtain a two-dimensional titanium nanosheet.
  • FIG. 1 is an electron mirror topography of the two-dimensional titanium nanosheet prepared in the step (1). Its size is less than 50 nm.
  • Figure 2 shows an atomic force micrograph. As can be seen from the figure, the thickness of the two-dimensional titanium nanosheet is about 3 nm. Therefore, by observation by transmission electron microscopy and atomic force microscopy, it is possible to peel off the two-dimensional metal elemental titanium nanosheet by the liquid phase stripping method.
  • the absorption spectra of the same concentration of two-dimensional titanium nanosheets were isopropyl alcohol (IPA) and water stripped, respectively. It is apparent that the absorption spectrum of the two-dimensional titanium nanosheet stripped in IPA has a higher absorption value and a larger slope (i.e., the upper curve in Fig. 3a). This shows that the larger titanium particles can be sufficiently stripped into smaller titanium nanosheets in IPA. Further, the absorption values of different stripping times (referred to as water bath ultrasonic time) of the same concentration of metal elemental titanium were compared (as shown in Fig. 3b), and it was found that as the stripping time increased, the absorption spectrum was continuously increased and appeared. A saturated state.
  • IPA isopropyl alcohol
  • a method for preparing a nano titanium photothermal preparation comprising:
  • 500 mg of titanium powder was added to 100 ml of isopropanol. Then select the probe ultrasound 200W, ultrasound for 15h. The ultrasound on/off time was chosen to be 2/4 s and ultrasound was performed in an ice bath environment. After the probe is ultrasonicated, it is then ultrasonically probed in a water bath. The water bath ultrasonic power is 300W. The ultrasound time was 10 h. The bath temperature is maintained at 15 ° C;
  • the desired metal elemental titanium nanosheets are obtained by centrifugation. First, centrifugal force of 1800 g was used and centrifuged for 35 min. The supernatant was taken, and then the supernatant was centrifuged at 10000 g to obtain a precipitate, which was vacuum dried to obtain a two-dimensional titanium nanosheet.
  • a method for preparing a nano titanium photothermal preparation comprising:
  • 500 mg of titanium powder was added to 100 ml of isopropanol. Then select the probe ultrasound 250W, ultrasound for 8h. The ultrasound on/off time was chosen to be 2/4 s and ultrasound was performed in an ice bath environment. After the probe is ultrasonicated, it is then ultrasonically probed in a water bath. The water bath ultrasonic power is 380W. The ultrasound time was 3 h. The bath temperature is maintained at 5 ° C;
  • the desired metal elemental titanium nanosheets are obtained by centrifugation.
  • the centrifugal force of 2200 g was used and centrifuged for 20 min. The supernatant was taken, and then the supernatant was centrifuged at 13,000 g to obtain a precipitate, which was vacuum dried to obtain a two-dimensional titanium nanosheet.
  • Different concentrations of two-dimensional titanium nanosheet aqueous dispersion were prepared to measure absorption spectrum and photothermal performance.
  • the absorption spectrum was measured using an ultraviolet-spectrophotometer.
  • the photothermal experiment used a 808 nm laser.
  • a two-dimensional titanium nanosheet aqueous dispersion of 10, 25, 50 and 100 ppm was prepared separately (as shown in Figure 4).
  • the prepared aqueous dispersion was separately placed in a quartz cuvette and placed in an ultraviolet spectrophotometer card slot to measure the absorbance.
  • the absorption curves of different concentrations are shown in Figure 5. According to the absorption at 808 nm, the extinction coefficient of the two-dimensional titanium nanosheet was 20.8 Lg -1 cm -1 (as shown in Fig. 6).
  • the photothermal conversion efficiency (73.4%) of the two-dimensional titanium nanosheet of the present invention is the highest among all reported photothermal agents, higher than the conventional two-dimensional photothermal agent of gold nanoparticles (21%), including MoS 2 ( 24.4%) black phosphorus quantum dots (28.4%) Ti 3 C 2 nanosheets (30.6%) and tantalum quantum dots (45.5%), therefore, the photothermal conversion efficiency of two-dimensional titanium nanosheets is significantly higher than other currently studied Photothermal agent. Therefore, the two-dimensional titanium nanosheets produced by the present invention have good photothermal properties.
  • Two-dimensional titanium nanosheets of different masses were dispersed in a cell culture medium, and then co-incubated with different cells, and the viability of the cells was measured.
  • hepatocyte cancer cells SMMC-7721, melanoma cells B16, and macrophage J774A.1 were separately plated into 96-well plates, and after the cells were attached, they were prepared for experiments.
  • Two-dimensional titanium nanosheet dispersions of 0, 10, 25, 50, and 100 ppm were prepared in DMEM high-sugar medium, 100 ⁇ l of the dispersion was taken, and the medium in the 96-well plate was replaced. After incubation for 24 hours, CCK8 was used.
  • the kit measures the viability of the cells in each well, and each set consists of 3 parallel wells. As shown in Fig. 9, in various cells, as the concentration of the two-dimensional titanium nanosheet increased (from 0 to 100 ppm), there was no significant decrease in cell viability as compared with the experimental group without the nanosheet. This indicates that the two-dimensional titanium nanosheets have no obvious cytotoxicity.
  • Embodiments of the invention also tested the toxicity of two-dimensional titanium nanosheets in model animals.
  • Two-dimensional titanium nanosheets and PEG2000-coated two-dimensional titanium nanosheets were separately dispersed in physiological saline to obtain a dispersion of 100 ppm.
  • Six weeks old female Balb/c nude mice were given 100 ⁇ l of physiological saline (control), 100 ⁇ l of 100 ppm two-dimensional titanium nanosheets (represented as "titanium nanosheets”) dispersion and 100 ⁇ l by subcutaneous injection.
  • a 100 ppm PEG-coated two-dimensional titanium nanosheet (indicated by "polyethylene glycol-coated titanium nanosheets") was injected subcutaneously under the right forelimb of the mouse.
  • mice The body weight of the mice was measured on the 1st, 3rd, 5th, 7th, 9th, 11th, 13th, and 15th day after the injection, and the mice were sacrificed on the 15th day, and the main organs, heart, liver, spleen, lung and kidney were taken for H&E. Dyeing to see if the nanosheets caused damage to tissues and organs in mice. As shown in Figure 10, the two-dimensional titanium nanosheets do not affect the change in body weight; as shown in Figure 11, the two-dimensional titanium nanosheets do not cause damage to mouse tissues and organs.
  • the two-dimensional titanium nanosheets are not toxic to cancer cells, normal cells, and in vivo conditions under in vitro conditions, showing their biocompatibility and safety and non-toxic advantages.
  • SMMC-7721, B16 and J774A.1 cells were plated in 96-well plates. After adhering the cells, different concentrations of two-dimensional titanium nanosheets dispersed in the cell culture medium were incubated with the cells. The concentration of the two-dimensional titanium nanosheets was respectively 0, 5, 10, 20, 30, 50 ppm, 3 parallel holes were set for each experimental group. After incubation for 2 hours, the cells were irradiated with a 808 nm laser at a power of 1 W cm -2 for 10 minutes, and 24 hours after the irradiation, the viability of the cells was measured using a CCK8 kit. As shown in Figure 12, the photothermal effect of low-concentration (5ppm) two-dimensional titanium nanosheets has partial cell killing ability.
  • the killing ability is already very significant (up to 20% for cancer cells, for macrophages). Up to 80%), at a concentration above 30ppm, the photothermal effect of two-dimensional titanium nanosheets can almost completely kill cells. Moreover, the PEG2000 coated or uncoated two-dimensional titanium nanosheets have the same photothermal effect killing ability.
  • PEG2000-coated two-dimensional titanium nanosheets were co-incubated with SMMC-7721 cells at 50 ppm, followed by irradiation at 808 nm, 1 W cm -2 , at different light times (0, 2, The killing ability was measured at 5, 8, 10 minutes. As shown in Fig. 13, only 2 minutes of light was sufficient to completely kill all cells.
  • the photothermal effect of two-dimensional titanium nanosheets can be used for cell killing, and good cell killing effect can be achieved under low concentration and short time near-infrared light conditions, showing excellent photothermal conversion efficiency.
  • SMMC-7721 cells were subcutaneously injected into the skin of female Balb/c nude mice. After about 10 days, the tumor volume reached 100-200 mm 3 , and the liver cancer model was used as the target of two-dimensional titanium nanosheet photothermal therapy.
  • the PEG2000 coated two-dimensional titanium nanosheets were dispersed in phosphate buffer at a concentration of 100 ppm.
  • 100 ⁇ l of nanosheet dispersion was injected intratumorally, then the mice were anesthetized, and the tumor site containing titanium nanosheets was irradiated with a laser of 808 nm wavelength at a power of 1 Wcm -2 for 5 minutes to perform photothermal treatment on the tumor. treatment.
  • Figure 14 shows the change in temperature per minute of the tumor site recorded by an infrared detector
  • Figure 15 shows the temperature profile of the tumor site during treatment.
  • a photothermal preparation comprising a two-dimensional titanium nanosheet can be effectively used for photothermal treatment of tumors.

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Abstract

一种纳米钛光热制剂、其制备方法及其在制备光热治疗药物中的应用，该纳米钛光热制剂包括二维钛纳米片和包覆在二维钛纳米片表面的生物相容性材料，其制备方法包括：提供钛原料，采用液相剥离的方法对所述钛原料进行剥离，得到二维钛纳米片；提供生物相容性材料，将二维钛纳米片和生物相容性材料在10-30℃下混合3-6h，得到纳米钛光热制剂。

Description

纳米钛光热制剂及其制备方法和应用
本发明要求于2018年01月30日递交的申请号为201810089749.6，发明名称为“纳米钛光热制剂及其制备方法和应用”的在先申请的优先权，上述在先申请的内容以引入的方式并入本文本中。
技术领域
本发明涉及生物医用纳米材料领域，具体涉及一种纳米钛光热制剂及其制备方法和应用。
背景技术
目前，对于癌症的治疗，在传统的手术治疗、放射性治疗和化疗之外，又开发出了免疫疗法、细胞疗法等新型疗法。但是，目前的疗法虽然能够在一定程度上延长患者的生命，仍有其局限性，如副作用大、无法完全清除肿瘤细胞、无法治愈等等。因此，开发新的疗法仍然是癌症领域的热点。
基于纳米材料的近红外光疗是一种新型的，有潜力的疗法，特别是近红外光响应的光热治疗，解决了紫外光以及可见光等短波长光疗中光波易被吸收、易散射、具有核酸毒性，以及组织穿透力差的问题。近红外光波长大约在700-1000纳米之间，具有低吸收、高穿透性的优点(1厘米以上)。
目前常规的光热治疗材料主要有金纳米颗粒、碳纳米管以及石墨烯等等。但现有的光热材料往往无法兼顾光热转换效率以及生物相容性。
因此，亟需寻找一种光热转换效率高、生物相容性好、安全无毒的光热材料，用于包括肿瘤在内的光热治疗。
发明内容
为解决上述问题，本发明提供了一种光热转换效率高、生物相容性好，安全无毒的纳米钛光热制剂。
本发明第一方面提供了一种纳米钛光热制剂，包括二维钛纳米片和包覆在所述二维钛纳米片表面的生物相容性材料。
其中，所述二维钛纳米片和所述生物相容性材料的质量比为1∶1-10。
其中，所述二维钛纳米片和所述生物相容性材料的质量比为1∶1。
其中，所述二维钛纳米片的厚度为1-50nm。
其中，所述二维钛纳米片的厚度为3-5nm。
其中，所述二维钛纳米片的长宽尺寸为10-50nm。
其中，所述二维钛纳米片的长宽尺寸为30-40nm。
其中，所述生物相容性材料包括透明质酸、葡聚糖及其衍生物、壳聚糖及其衍生物、果胶、羧甲基纤维素、白蛋白、脂质体、细胞膜、聚乙烯吡咯烷酮、聚乳酸-羟基乙酸共聚物、聚乙烯亚胺、聚丙烯酸和聚乙二醇及其衍生物中的一种或多种。
其中，所述生物相容性材料为聚乙二醇及其衍生物，所述聚乙二醇及其衍生物的分子量为200-20000。
其中，所述聚乙二醇的末端由氨基修饰。
其中，所述生物相容性材料通过静电作用吸附在所述二维钛纳米片的表面。
其中，所述纳米钛光热制剂还包括靶向材料，所述靶向材料通过化学键连接在所述二维钛纳米片或连接在所述生物相容性材料上。
其中，所述靶向材料为叶酸，所述叶酸通过酰胺键连接在所述聚乙二醇及其衍生物上。
本发明第一方面提供的纳米钛光热制剂，光热转换效率较高、生物相容性良好，安全无毒，可用于包括肿瘤在内的光热治疗。
本发明第二方面提供了一种纳米钛光热制剂的制备方法，包括：
提供钛原料，采用液相剥离的方法对所述钛原料进行剥离，得到二维钛纳米片；
提供生物相容性材料，将所述二维钛纳米片和生物相容性材料在10-30℃ 下混合搅拌3-6h，得到纳米钛光热制剂。
其中，所述纳米钛光热制剂的制备方法具体包括：将生物相容性材料溶于适量的第一溶剂中后，得到生物相容性材料溶液，将所述二维钛纳米片分散于适量的第二溶剂中，得到二维钛纳米片分散液，将所述生物相容性材料溶液与所述二维钛纳米片分散液在10-30℃下混合后搅拌3-6h，得到纳米钛光热制剂。
其中，所述搅拌速率为100转/min-700转/min。
其中，所述液相剥离的方法具体包括以下操作：
将所述钛原料加入至溶剂中，在冰浴环境下采用探头超声8-15h；所述探头超声结束后，继续采用水浴超声，所述水浴超声时间为3-10h，所述水浴的温度保持5-15℃；超声后，进行离心和干燥得到二维钛纳米片。
其中，所述探头超声的功率为200-250W，所述水浴超声功率为300-380W。
其中，所述离心的操作包括：首先采用1800-2200g的离心力，离心20-35min，取上清液；然后将所述上清液采用10000-13000g的离心力继续离心，得到沉淀即为二维钛纳米片。
本发明第二方面提供了一种纳米钛光热制剂的制备方法，制备方法简单易操作，制得的纳米钛光热制剂光热转换效率高、生物相容性好。
本发明第三方面提供了如上述所述的纳米钛光热制剂在制备光热治疗药物中的应用。
综上，本发明有益效果包括以下几个方面：
1、本发明提供的纳米钛光热制剂，光热转换效率高、生物相容性好，安全无毒，可用于包括肿瘤在内的光热治疗；
2、本发明提供的纳米钛光热制剂的制备方法，方法简单易操作。
附图说明
图1为实施例1制得的二维钛纳米片的透射电镜图片；
图2为实施例1制得的二维钛纳米片的原子力显微图片；
图3为实施例1中二维钛纳米片的液相剥离过程的吸收光谱图；
图4为不同浓度的二维钛纳米片水分散液照片；
图5为不同浓度的二维钛纳米片水分散液的吸收光谱图；
图6为二维钛纳米片水分散液的消光系数；
图7为不同浓度的二维钛纳米片水分散液的升温曲线；
图8为二维钛纳米片水分散液的光热转换效率；
图9为二维钛纳米片的细胞毒性测定结果图；
图10为二维钛纳米片对小鼠体重的影响；
图11为二维钛纳米片对引起小鼠组织器官损伤影响图；
图12为不同浓度的二维钛纳米片对细胞的光热杀伤能力效果图；
图13为50ppm浓度的PEG2000包覆的二维钛纳米片在不同光照时间下对细胞的杀伤能力的测定；
图14为小鼠肿瘤光热治疗时肿瘤部位温度的变化；
图15为小鼠肿瘤光热治疗时肿瘤部位温度变化的曲线；
图16为荷瘤小鼠接受光热治疗后肿瘤的体积变化。
具体实施方式
以下所述是本发明的优选实施方式，应当指出，对于本技术领域的普通技术人员来说，在不脱离本发明原理的前提下，还可以做出若干改进和润饰，这些改进和润饰也视为本发明的保护范围。
本发明提到的“二维钛纳米片”、“钛纳米颗粒”或“钛”，除特殊说明，均指的是单质钛。
本发明第一方面提供了一种纳米钛光热制剂，包括二维钛纳米片和包覆在所述二维钛纳米片表面的生物相容性材料。
本发明实施方式中，所述二维钛纳米片的厚度为1-50nm。可选地，所述二维钛纳米片的厚度为3-5nm。可选地，所述二维钛纳米片的厚度为5-10nm。可选地，所述二维钛纳米片的厚度为10-50nm。进一步可选地，所述二维钛纳 米片的厚度为1nm、5nm、10nm、15nm、20nm、25nm、30nm、35nm、40nm、45nm或50nm。
本发明实施方式中，所述二维钛纳米片的长宽尺寸为10-50nm。可选地，所述二维钛纳米片的长宽尺寸为30-40nm。可选地，所述二维钛纳米片的长宽尺寸为10-30nm。进一步可选地，所述二维钛纳米片的长宽尺寸为10nm、15nm、20nm、25nm、30nm、35nm、40nm、45nm或50nm。
本发明实施方式中，所述二维钛纳米片和所述生物相容性材料的质量比为1∶1-10。可选地，所述二维钛纳米片和所述生物相容性材料的质量比为1∶1、1∶2、1∶3、1∶4、1∶5、1∶6、1∶7、1∶8、1∶9或1∶10。
本发明实施方式中，所述二维钛纳米片具有从可见光区到近红外光区的吸收。可选地，所述二维钛纳米片的光吸收波长范围为200-2000nm。
本发明实施方式中，所述二维钛纳米片的光热转换效率为大于或等于70％。
本发明第一方面提供的二维钛纳米片具有环境友好、生物兼容性、全光谱的强吸收和较高的光热转换效率等优点，具有优良的光热性能。
本发明实施方式中，所述生物相容性材料包括透明质酸、葡聚糖及其衍生物、壳聚糖及其衍生物、果胶、羧甲基纤维素、白蛋白、脂质体、细胞膜、聚乙烯吡咯烷酮、聚乳酸-羟基乙酸共聚物、聚乙烯亚胺、聚丙烯酸和聚乙二醇及其衍生物中的一种或多种。进一步可选地，所述生物相容性材料包括聚乙二醇及其衍生物、聚乳酸-羟基乙酸共聚物、白蛋白、脂质体和细胞膜中的至少一种。进一步可选地，所述生物相容性材料包括聚乙二醇及其衍生物，所述聚乙二醇及其衍生物的分子量在200-20000之间。可选地，所述聚乙二醇的末端可以由氨基修饰。可选地，所述生物相容性材料通过静电作用吸附在所述二维钛纳米片的表面。
本发明实施方式中，所述纳米钛光热制剂还包括靶向材料，所述靶向材料通过化学键连接在所述二维钛纳米片或连接在所述生物相容性材料上。可选地，所述靶向材料为叶酸，所述叶酸通过酰胺键连接在所述聚乙二醇上。
本发明实施方式中，所述纳米钛光热制剂可以分散在生理盐水、磷酸盐缓冲液或去离子水中用于后续应用。
本发明第一方面提供的纳米钛光热制剂，光热转换效率高、生物相容性好，安全无毒，可用于包括肿瘤在内的光热治疗。
本发明第二方面提供了一种纳米钛光热制剂的制备方法，包括：
提供钛原料，采用液相剥离的方法对所述钛原料进行剥离，得到二维钛纳米片；
提供生物相容性材料，将所述二维钛纳米片和所述生物相容性材料在10-30℃下混合搅拌3-6h，得到纳米钛光热制剂。
本发明实施方式中，将二维钛纳米片和生物相容性材料在25℃下混合3-6h。
本发明实施方式中，所述纳米钛光热制剂的具体制备方法包括：将生物相容性材料溶于适量的第一溶剂中后，得到生物相容性材料溶液，将二维钛纳米片分散于适量的第二溶剂中，得到二维钛纳米片分散液，将所述生物相容性材料溶液与所述二维钛纳米片分散液在10-30℃下混合后搅拌3-6h，得到纳米钛光热制剂。
可选地，所述第一溶剂为可以溶解生物相容性材料的溶剂，如当生物相容性材料为聚乙二醇时，第一溶剂可以选择去离子水。
可选地，所述第二溶剂包括去离子水。
本发明实施方式中，所述搅拌速率为100转/min-700转/min。
本发明实施方式中，搅拌后，将所得混合物离心干燥后，得到生物相容性材料包覆的二维钛纳米片，即得纳米钛光热制剂。
本发明实施方式中，所述液相剥离的方法具体包括以下操作：
将所述钛原料加入至溶剂中，在冰浴环境下采用探头超声8-15h；所述探头超声结束后，继续采用水浴超声，所述水浴超声时间为3-10h，水浴温度保持5-15℃；超声后，进行离心和干燥得到二维钛纳米片。
可选地，所述溶剂包括异丙醇、乙醇、水和甲基吡咯烷酮(即N-甲基吡 咯烷酮，NMP)中的至少一种。
可选地，所述钛原料在所述溶剂中的浓度为1-7mg/mL。
可选地，所述探头超声的功率为200-250W。进一步可选地，所述探头超声的功率为240W。
可选地，所述探头超声的时间为10h。
可选地，所述探头超声是非连续超声，选择超声开/关时间为2/4s，即先超声2s，然后关闭超声探头保持4s，在继续超声2s，以此类推。
可选地，所述水浴超声功率为300-380W。进一步可选地，所述水浴超声功率为360W。
可选地，所述水浴超声的时间为8h。
可选地，所述水浴温度保持10℃。
可选地，超声后，进行离心，所述离心的操作包括：首先采用1800-2200g的离心力，离心20-35min，取上清液；然后将所述上清液采用10000-13000g的离心力继续离心，得到沉淀即为二维钛纳米片。进一步可选地，首先采用2000g的离心力，离心30min，取上清液；然后将所述上清液采用12000g的离心力继续离心，得到沉淀，将所得沉淀干燥后即得二维钛纳米片。可选地，所述干燥的方式不限，例如可为真空干燥。
现有技术通常采用液相剥离法用来剥离二维层状材料。而本发明首次采用液相剥离法剥离二维非层状金属材料，并取得成功。
本发明第二方面提供了一种纳米钛光热制剂的制备方法，制备方法简单易操作，制得的纳米钛光热制剂光热转换效率较高、生物相容性良好。
本发明实施方式第三方面提供了一种如上述所述的纳米钛光热制剂在制备光热治疗药物中的应用。
实施例1：
一种纳米钛光热制剂的制备方法，包括：
(1)制备二维钛纳米片；
将500mg的钛粉加入100ml的异丙醇中。然后选择探头超声240W，超声10h。选择超声开/关时间为2/4s，并且是在冰浴环境下进行超声。探头超声完后，接着采用水浴超声。水浴超声功率为360W。超声时间为8h。水浴温度保持10℃。
超声过后采用离心的办法得到需要的金属单质钛纳米片。首先采用2000g的离心力，离心30min。取上清，然后将上清采用12000g继续离心，得到沉淀。该沉淀，真空干燥后即得二维钛纳米片。
(2)提供PEG2000溶液，将步骤(1)制得的二维钛纳米片分散在适量水中得到二维钛纳米片分散液，将二维钛纳米片分散液与PEG2000溶液混合，其中，二维钛纳米片与PEG2000的质量比为1∶1，在25℃下混合搅拌5h，离心干燥后，得到PEG2000包覆的二维钛纳米片即得纳米钛光热制剂。
如图1所示，图1为步骤(1)制得的二维钛纳米片的电镜形貌图。其尺寸小于50nm。图2显示的是原子力显微图片。由图可以看出，二维钛纳米片的厚度在3nm左右。因此通过透射电镜和原子力显微镜的观察，通过液相剥离法确实可以剥离出二维金属单质钛纳米片。
如图3a所示，分别为异丙醇(IPA)和水中剥离的，相同浓度的二维钛纳米片的吸收光谱。很明显，IPA中剥离的二维钛纳米片的吸收光谱具有更高的吸收值和更大的斜率(即图3a中上面的一条曲线)。这说明IPA中可以充分将比较大的钛颗粒剥离成较小的钛纳米片。进一步，比较了相同浓度的金属单质钛的不同剥离时间(指的是水浴超声时间)的吸收值(如图3b所示)，发现随着剥离时间的增加，吸收光谱在不断增加，而且会出现一个饱和的状态。
实施例2：
一种纳米钛光热制剂的制备方法，包括：
(1)制备二维钛纳米片；
将500mg的钛粉加入100ml的异丙醇中。然后选择探头超声200W，超声15h。选择超声开/关时间为2/4s，并且是在冰浴环境下进行超声。探头超声完 后，接着采用水浴超声。水浴超声功率为300W。超声时间为10h。水浴温度保持15℃；
超声过后采用离心的办法得到需要的金属单质钛纳米片。首先采用1800g的离心力，离心35min。取上清，然后将上清采用10000g继续离心，得到沉淀，真空干燥后即得二维钛纳米片。
(2)提供PEG2000溶液，将步骤(1)制得的二维钛纳米片分散在适量水中得到二维钛纳米片分散液，将二维钛纳米片分散液与PEG2000溶液混合，其中，二维钛纳米片与PEG2000的质量比为1∶10，在30℃下混合搅拌3h，离心干燥后，得到PEG2000包覆的二维钛纳米片即得纳米钛光热制剂。
实施例3：
一种纳米钛光热制剂的制备方法，包括：
(1)制备二维钛纳米片；
将500mg的钛粉加入100ml的异丙醇中。然后选择探头超声250W，超声8h。选择超声开/关时间为2/4s，并且是在冰浴环境下进行超声。探头超声完后，接着采用水浴超声。水浴超声功率为380W。超声时间为3h。水浴温度保持5℃；
超声过后采用离心的办法得到需要的金属单质钛纳米片。首先采用2200g的离心力，离心20min。取上清，然后将上清采用13000g继续离心，得到沉淀，真空干燥后即得二维钛纳米片。
(2)提供PEG2000溶液，将步骤(1)制得的二维钛纳米片分散在适量水中得到二维钛纳米片分散液，将二维钛纳米片分散液与PEG2000溶液混合，其中，二维钛纳米片与PEG2000的质量比为1∶5，在10℃下混合搅拌6h，离心干燥后，得到PEG2000包覆的二维钛纳米片即得纳米钛光热制剂。
效果实施例
(1)吸收光谱和光热性能的测试
配制不同浓度的二维钛纳米片水分散液测量吸收光谱和光热性能。吸收光 谱采用紫外-分光光度计测量。光热实验采用808nm激光。分别配制10，25，50和100ppm的二维钛纳米片水分散液(如图4所示)。将配制的水分散液分别装入石英比色皿中，放入紫外分光光度计卡槽中测量吸收度。不同浓度的吸收曲线如图5所示。根据808nm处的吸收可以得到二维钛纳米片的消光系数为20.8Lg -1cm -1(如图6所示)。该值高于黑磷(14.8Lg -1cm -1)。对于光热实验的测量，取1ml二维钛纳米片水分散液加入比色皿中，采用808nm激光进行照射，并同时用热电偶记录温度曲线。图7显示的是不同浓度的温度随激光照射时间的升温图。通过定量的计算，可以得到二维钛纳米片的光热转换效率为73.4％(如图8所示)。
本发明的二维钛纳米片的光热转换效率(73.4％)在所有报道的光热剂中最高，高于传统的金纳米颗粒(21％)新兴的二维光热剂，包括MoS 2(24.4％)黑磷量子点(28.4％)Ti 3C 2纳米片(30.6％)和碲量子点(45.5％)，因此，二维钛纳米片光热转换效率值明显高于其他当前正在研究的光热剂。因此，本发明制得的二维钛纳米片光热性能良好。
(2)二维钛纳米片的生物毒性测试
将不同质量的二维钛纳米片分散于细胞培养基，再与不同的细胞共孵育，再测定细胞的活力。首先将肝细胞癌细胞SMMC-7721、黑色素瘤细胞B16以及巨噬细胞J774A.1分别铺到96孔板中，待细胞贴壁后，准备用于实验。以DMEM高糖培养基分别配制0，10，25，50，100ppm浓度的二维钛纳米片分散液，取100μl分散液，置换前述96孔板中的培养基，在孵育24小时之后，使用CCK8试剂盒测定每个孔里面细胞的活力，每一组实验设置3个平行孔。如图9所示，在多种细胞中，随着二维钛纳米片浓度的提高(从0到100ppm)，和未加纳米片的实验组对比，其细胞活力并无明显的下降。这说明二维钛纳米片没有明显的细胞毒性。
本发明实施例也在模式动物中检测了二维钛纳米片的毒性。分别将二维钛纳米片和PEG2000包覆的二维钛纳米片分散于生理盐水，得到100ppm的分散液备用。取6周龄的雌性Balb/c裸鼠，通过皮下注射的方式将100μl的生理盐水 (对照)、100μl的100ppm二维钛纳米片(图中以“钛纳米片”表示)分散液和100μl的100ppm PEG包覆的二维钛纳米片(图中以“聚乙二醇包覆的钛纳米片”表示)分散液注射到小鼠右前肢下方的皮下。分别在注射后的第1，3，5，7，9，11，13，15天测定小鼠的体重，并且在第15天时将小鼠处死，取其主要脏器心肝脾肺肾，进行H&E染色，观察纳米片是否导致小鼠组织器官损伤。如图10所示，二维钛纳米片不会影响其体重的变化；如图11所示，二维钛纳米片也不会对小鼠组织器官造成损伤。
综上所述，二维钛纳米片在体外条件下对癌细胞、正常细胞，以及在体内条件下都没有毒性，显示了其生物相容性以及安全无毒的优点。
(3)二维钛纳米片光热效应的细胞杀伤能力测定
将SMMC-7721、B16以及J774A.1细胞分别铺于96孔板，细胞贴壁后，将不同浓度的分散于细胞培养基的二维钛纳米片与细胞孵育，二维钛纳米片浓度为分别0，5，10，20，30，50ppm，每个实验组设置3个平行孔。孵育2小时后，用808nm的激光，以1W cm -2的功率照射10分钟，照射之后24小时，用CCK8试剂盒测定细胞的活力。如图12所示，低浓度(5ppm)二维钛纳米片的光热效应就有部分的细胞杀伤能力，在10ppm条件下，杀伤能力已经非常显著(对于癌细胞可达到20％，对于巨噬细胞可达到80％)，在30ppm以上的浓度，二维钛纳米片的光热效应已经基本可以完全杀死细胞。而且PEG2000包覆的或者未包覆的二维钛纳米片其光热效应杀伤能力一致。
同样地，在50ppm的条件下，将PEG2000包覆的二维钛纳米片与SMMC-7721细胞共孵育，随后用808nm，1W cm -2的功率辐照，在不同的光照时间(0，2，5，8，10分钟)下测定其杀伤能力，如图13所示，仅仅2分钟的光照，就足以完全杀死所有细胞。
综上所述，二维钛纳米片的光热效应可用于细胞杀伤，并且在低浓度、短时间的近红外光条件下就能达到良好的细胞杀伤效果，显示了其优异的光热转换效率。
(4)二维钛纳米片光热效应的肿瘤光热治疗效果测定
将5×10 6SMMC-7721细胞皮下注射到雌性Balb/c裸鼠的皮下，大约10天之后，肿瘤体积达到100-200mm 3，以此肝癌模型作为二维钛纳米片光热治疗的对象。将PEG2000包覆的二维钛纳米片分散到磷酸盐缓冲液中，浓度为100ppm。肿瘤治疗时，瘤内注射100μl纳米片分散液，然后将小鼠麻醉，利用808纳米波长的激光，在1Wcm -2的功率下照射含有钛纳米片的肿瘤部位5分钟，以对肿瘤进行光热治疗。图14是用红外线探测仪记录了肿瘤部位每分钟温度的变化情况，图15则是治疗过程中肿瘤部位的温度变化曲线，这两个图片说明在进行光热治疗时，二维钛纳米片导致的温度升高非常快速，1分钟内可以升高20℃。而在大约1分钟后，温度缓慢上升逐渐保持稳定。随后，在治疗后的第1，3，5，7，9，11，13，15天分别测定肿瘤体积，如图16所示，可知，经过光热治疗后肿瘤大小迅速降低，在整个治疗的过程中肿瘤组织被杀伤，肿瘤生长得到抑制，取得了良好的治疗效果。
因此，包含二维钛纳米片的光热制剂可以有效地用于肿瘤的光热治疗。
以上所述实施例仅表达了本发明的几种实施方式，其描述较为具体和详细，但并不能因此而理解为对本发明专利范围的限制。应当指出的是，对于本领域的普通技术人员来说，在不脱离本发明构思的前提下，还可以做出若干变形和改进，这些都属于本发明的保护范围。因此，本发明专利的保护范围应以所附权利要求为准。

Claims (20)

  1. 一种纳米钛光热制剂，其中，包括二维钛纳米片和包覆在所述二维钛纳米片表面的生物相容性材料。
  2. 如权利要求1所述的纳米钛光热制剂，其中，所述二维钛纳米片和所述生物相容性材料的质量比为1∶1-10。
  3. 如权利要求2所述的纳米钛光热制剂，其中，所述二维钛纳米片和所述生物相容性材料的质量比为1∶1。
  4. 如权利要求1所述的纳米钛光热制剂，其中，所述二维钛纳米片的厚度为1-50nm。
  5. 如权利要求4所述的纳米钛光热制剂，其中，所述二维钛纳米片的厚度为3-5nm。
  6. 如权利要求1所述的纳米钛光热制剂，其中，所述二维钛纳米片的长宽尺寸为10-50nm。
  7. 如权利要求6所述的纳米钛光热制剂，其中，所述二维钛纳米片的长宽尺寸为30-40nm。
  8. 如权利要求1所述的纳米钛光热制剂，其中，所述生物相容性材料包括透明质酸、葡聚糖及其衍生物、壳聚糖及其衍生物、果胶、羧甲基纤维素、白蛋白、脂质体、细胞膜、聚乙烯吡咯烷酮、聚乳酸-羟基乙酸共聚物、聚乙烯亚胺、聚丙烯酸和聚乙二醇及其衍生物中的一种或多种。
  9. 如权利要求8所述的纳米钛光热制剂，其中，所述生物相容性材料为聚乙二醇及其衍生物，所述聚乙二醇及其衍生物的分子量为200-20000。
  10. 如权利要求8所述的纳米钛光热制剂，其中，所述聚乙二醇的末端由氨基修饰。
  11. 如权利要求1所述的纳米钛光热制剂，其中，所述生物相容性材料通过静电作用吸附在所述二维钛纳米片的表面。
  12. 如权利要求8所述的纳米钛光热制剂，其中，所述纳米钛光热制剂还 包括靶向材料，所述靶向材料通过化学键连接在所述二维钛纳米片或连接在所述生物相容性材料上。
  13. 如权利要求12所述的纳米钛光热制剂，其中，所述靶向材料为叶酸，所述叶酸通过酰胺键连接在所述聚乙二醇及其衍生物上。
  14. 一种纳米钛光热制剂的制备方法，其中，包括：
    提供钛原料，采用液相剥离的方法对所述钛原料进行剥离，得到二维钛纳米片；
    提供生物相容性材料，将所述二维钛纳米片和所述生物相容性材料在10-30℃下混合后搅拌3-6h，得到纳米钛光热制剂。
  15. 如权利要求14所述的纳米钛光热制剂的制备方法，其中，所述纳米钛光热制剂的制备方法具体包括：将生物相容性材料溶于适量的第一溶剂中后，得到生物相容性材料溶液，将所述二维钛纳米片分散于适量的第二溶剂中，得到二维钛纳米片分散液，将所述生物相容性材料溶液与所述二维钛纳米片分散液在10-30℃下混合后搅拌3-6h，得到纳米钛光热制剂。
  16. 如权利要求15所述的纳米钛光热制剂的制备方法，其中，所述搅拌速率为100转/min-700转/min。
  17. 如权利要求14所述的纳米钛光热制剂的制备方法，其中，所述液相剥离的方法具体包括以下操作：
    将所述钛原料加入至溶剂中，在冰浴环境下采用探头超声8-15h；所述探头超声结束后，继续采用水浴超声，所述水浴超声时间为3-10h，所述水浴的温度保持5-15℃；超声后，进行离心和干燥得到二维钛纳米片。
  18. 如权利要求17所述的纳米钛光热制剂的制备方法，其中，所述探头超声的功率为200-250W，所述水浴超声功率为300-380W。
  19. 如权利要求17所述的纳米钛光热制剂的制备方法，其中，所述离心的操作包括：首先采用1800-2200g的离心力，离心20-35min，取上清液；然后将所述上清液采用10000-13000g的离心力继续离心，得到沉淀即为二维钛纳米片。
  20. 如权利要求1-13中任一项所述的纳米钛光热制剂在制备光热治疗药物中的应用。
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