CN115252776B - Preparation of up-conversion-metal phenolic network composite nano material and application thereof in tumor treatment - Google Patents

Preparation of up-conversion-metal phenolic network composite nano material and application thereof in tumor treatment Download PDF

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CN115252776B
CN115252776B CN202210583052.0A CN202210583052A CN115252776B CN 115252776 B CN115252776 B CN 115252776B CN 202210583052 A CN202210583052 A CN 202210583052A CN 115252776 B CN115252776 B CN 115252776B
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刘金亮
秦腾
魏芹
陈晨
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University of Shanghai for Science and Technology
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Abstract

The invention relates to the field of biomedical nano materials, and discloses a preparation method of an up-conversion-metal phenolic network composite nano material. The method comprises the following steps: (1) Synthesizing rare earth up-conversion nano particles by adopting a thermal decomposition method; (2) Coating a layer of mesoporous silica on the surface of UCNPs to make the mesoporous silica water-soluble, and loading photosensitizer and chemotherapeutic drug DOX to UCNPs@mSiO 2 In the duct; (3) UCNPs@mSiO 2 And contains TA and Fe 3+ Is mixed with the aqueous solution of (2) to induce UCNPs@mSiO 2 And (3) rapidly forming the surface MPN film to obtain the nanocomposite. The invention also discloses a preparation method of the composite material and application of the composite material in tumor treatment. MPN films have pH response properties and photothermal properties for PTT treatment; MPN is gradually decomposed to realize DOX controllable release, and is used for chemotherapy; the MC540 generates ROS in response to green emissions for PDT.

Description

Preparation of up-conversion-metal phenolic network composite nano material and application thereof in tumor treatment
Technical Field
The invention relates to the field of biological nano material medicine, in particular to an up-conversion-metal phenolic network composite nano material which is used for synergizing PTT/PDT/chemotherapy in tumor treatment.
Background
Due to the complexity of the tumor microenvironment, the therapeutic effects of monotherapy are often unsatisfactory. Thus, cancer treatment has been shifted from monotherapy to combination therapy to achieve the optimal therapeutic effect of synergistic treatment. Chemotherapy relies on chemotherapeutic agents (e.g., doxorubicin (DOX), paclitaxel, etc.) to kill cancer cells. However, these drugs can be absorbed by cancer cells, and also can be absorbed by normal cells, which has a certain adverse effect on normal cells, and in addition, the drug premature ejaculation can be encountered in the process of delivering the drug to the tumor site, so that the problem of insufficient drug concentration at the cancer site and the like can be caused, so that the development of a controllable drug release system has guiding significance in the field of cancer treatment.
Photodynamic therapy (PDT) is a non-invasive, low-toxicity and low-side-effect treatment, with good prospects in cancer treatment. Photodynamic therapy is independent of the three conditions of photosensitizer, excitation light source and oxygen. Currently, more photosensitizer molecules are used to absorb light mainly in the ultraviolet or visible region, but due to the limited penetration depth of short wavelength light, there is a limit to tumor treatment at deep tissues.
Therefore, the development of a novel material which can realize controllable drug release and solve the problem of tissue penetration depth of an excitation light source in PDT is a problem which needs to be solved by researchers in the field.
Disclosure of Invention
The invention aims at the defects existing in the prior art, and provides a preparation method of the up-conversion-metal phenolic network composite nano material, which is simple and easy to operate, and the prepared material has uniform morphology; and the composite material synthesized after modification treatment can meet the requirements of clinical diagnosis and treatment integration, and a PTT/PDT/chemotherapy combined treatment platform under imaging guidance is constructed under near infrared light irradiation.
In order to achieve the aim of the invention, the invention adopts the following technical scheme:
the preparation method of the up-conversion-metal phenolic network-based composite nanomaterial is characterized by comprising the following steps of:
(1) Preparation of NaYF 4 Yb, er and Nd nano particles are used as the inner cores: by "thermal decomposition" method, in the presence of 6 mL oleic acid
(OA) and 15 mL Octadecene (ODE) in 100 mL three-neck flasks were charged with different proportions of Y (CH) 3 CO 2 ) 3 ，Yb(CH 3 CO 2 ) 3 ，Er(CH 3 CO 2 ) 3 And Nd (CH) 3 CO 2 ) 3 (1 mmol, Y: yb: er: nd=80.5%: 18%:0.5%: 1%). Placing the mixture on a heating sleeve, then heating to 160 ℃ and stirring at uniform speed (rpm is approximately equal to 700)Preserving heat for five minutes until the acetate powder is completely dissolved to form a rare earth-oleic acid complex, and obtaining a pale yellow clear and transparent solution at the moment; and then the heating is turned off, and the solution is cooled to below 60 ℃. 0.1 g sodium hydroxide and 0.15 g ammonium fluoride are weighed and dissolved in 5 mL methanol solution, ultrasonic dissolution is carried out, then the mixed solution is slowly added into a three-neck flask drop by drop, the temperature is raised to 110 ℃ for reaction for about 30 minutes, so that methanol is evaporated, then a vacuum system is utilized to vacuumize the environment in the three-neck flask for about 30 minutes, then the pumping and ventilating are carried out for 3 times, each time for 30 seconds, the temperature of reactants is raised to 300 ℃ in an argon environment, and the temperature is kept at 1 h. After the reaction was completed, the solution was cooled to room temperature, and the product was precipitated with absolute ethanol (NaYF 4 Yb, er, nd up-conversion nano-particles), washing with cyclohexane and ethanol respectively for multiple times to obtain uniform up-conversion nano-materials, and storing in cyclohexane;
(2) Preparation of NaYF 4 :Yb,Er@NaYF 4 Core-shell up-conversion nanoparticles: weighing metal-acetate Y (CH) 3 CO 2 ) 3 And Nd (CH) 3 CO 2 ) 3 (1 mmol, y: nd=80%: 20%) was added to the flask, and 12 mL OA and 30 mL ODE were added, and after heating to 160 ℃, stirring was carried out at a constant speed, and the mixture was kept for 10 min to completely dissolve the mixture; then cooling to below 80deg.C (cyclohexane boiling point), adding 5 mL above NaYF 4 Heating Yb, er and Nd cyclohexane solution to 150 ℃ to remove cyclohexane solvent until no liquid flow down and no bubble generation of the solution exist at the bottle mouth, and cooling to below 60 ℃; 0.0674 g NaOH and 0.0374 g NH were taken 4 F, adding the mixture into a 5 mL methanol solvent, slowly adding the mixture into a reaction solution dropwise after ultrasonic dissolution, heating to 110 ℃, and removing methanol until no fog and no bubbles exist; then vacuuming for 30 min, exchanging argon for 3 times, heating to 300 ℃ and reacting for 1 h. Purification steps such as NaYF 4 The preparation method comprises the steps of preparing Yb, er and Nd nano particles. Finally purified NaYF 4 :Yb,Er,Nd@NaYF 4 Nd (core-shell) nanoparticles are dispersed in 15 mL cyclohexane;
(3) Water-soluble mesoporous silica coated NaYF 4 :Yb,Er,Nd@NaYF 4 Preparation of Nd nano-particles: first, 0 is weighed.1 g CTAB in a beaker, adding 20 mL ultrapure water, heating to 70 ℃ and strongly stirring to clarify the solution; the heating was turned off and 5 mL of NaYF was added at room temperature 4 :Yb,Er,Nd@NaYF 4 Nd (15, mg) cyclohexane solution is stirred overnight until the solution is clarified again; after overnight, the solvent was reduced by evaporation of water, then the solution was made up to 20 mL, the above solution was transferred to a 250 mL three-neck flask, then 40 mL ultrapure water, 6 mL absolute ethanol and 300 μl (2 mol/L) NaOH aqueous solution were added, the mixture was placed under an oil bath at 70 ℃ for 30 min of condensation reflux (speed of rotation is maximized), 400 μl TEOS was slowly added dropwise with a 200 μl pipette, after stirring reaction 1 h, cooled to room temperature, centrifuged to obtain precipitate, and the precipitate was washed 3 times with absolute ethanol to remove free silicon, thereby obtaining mesoporous silica coated UCNPs. To remove CTAB template, a mesoporous structure was obtained. The precipitate was dissolved in 40 mL acid ethanol solution (ph=1.4 to 1.5) and reacted under reflux at 60 ℃ with stirring for 1 h. Centrifuging to obtain a precipitate, washing the precipitate with absolute ethyl alcohol for 2-3 times, dispersing the final product in 20 mL absolute ethyl alcohol, and repeating the reflux process for 2-3 times to fully remove CTAB and reduce biotoxicity;
（4）UCNPs@mSiO 2 -preparation of MC540/DOX nanocomposite: an aqueous solution of 0.2 mg/mL DOX and an aqueous solution of 0.2 mg/mL MC540 were prepared, and UCNPs@mSiO was then added 2 Centrifuging the aqueous solution to obtain solid precipitate, and collecting 4 mg UCNPs@mSiO 2 Soaking in 2 mL of mixed solution of DOX and MC540, firstly placing in an ultrasonic machine for ultrasonic treatment for 30 min, and then rapidly stirring on a magnetic stirring heating sleeve under the environment of room temperature and light shielding for 24 h. After physical adsorption is completed, removing supernatant liquid by centrifugation to leave sediment, dissolving the sediment with ultrapure water, centrifuging again to leave sediment, washing for three times, and removing free DOX and MC540;
（5）UCNPs@mSiO 2 preparation of @ MPN-MC540/DOX nanocomposite: 40. Mu.L TA and 40. Mu.L FeCl were stirred vigorously 3 (24 mM) solution was added to UCNPs@mSiO of 2 mL 2 -MC540/DOX in aqueous solution. Finally, collecting UCNPs@mSiO by centrifugal separation and washing with deionized water for three times 2 @MPN-MC540/DOX nanocomposite.
The step (1) comprises the following specific steps:
(1) Y (CH) in different proportions was added to a 100 mL three-neck flask containing 6 mL Oleic Acid (OA) and 15 mL Octadecene (ODE) 3 CO 2 ) 3 ，Yb(CH 3 CO 2 ) 3 ，Er(CH 3 CO 2 ) 3 And Nd (CH) 3 CO 2 ) 3 (1 mmol, Y: yb: er: nd=80.5%: 18%:0.5%: 1%). Placing the mixture on a heating sleeve, then heating to 160 ℃, stirring at a constant speed, and preserving heat for five minutes until acetate powder is completely dissolved to form a rare earth-oleic acid complex, wherein the rare earth-oleic acid complex is light yellow clear and transparent solution;
(2) And then the heating is turned off, and the solution is cooled to below 60 ℃. 0.1 g sodium hydroxide and 0.15 g ammonium fluoride are weighed and dissolved in 5 mL methanol solution, ultrasonic dissolution is carried out, then the mixed solution is slowly added into a three-neck flask drop by drop, the temperature is raised to 110 ℃ for reaction for about 30 minutes, so that methanol is evaporated, then a vacuum system is utilized to vacuumize the environment in the three-neck flask for about 30 minutes, then the pumping and ventilating are carried out for 3 times, each time for 30 seconds, the temperature of reactants is raised to 300 ℃ in an argon environment, and the temperature is kept at 1 h. After the reaction was completed, the solution was cooled to room temperature, and the product was precipitated with absolute ethanol (NaYF 4 Yb, er, nd up-conversion nano-particles), washing with cyclohexane and ethanol respectively for multiple times to obtain uniform up-conversion nano-materials, and storing in cyclohexane.
The rotating speed of the heating sleeve in the step (1) is 700 rpm.
And (3) in the step (2), reacting for 1 hour at a temperature of 300 ℃ in an argon environment.
The step (3) comprises the following specific steps:
(1) Firstly, weighing 0.1 g of CTAB in a beaker, adding 20 mL ultrapure water, heating to 70 ℃ and clarifying the solution under strong stirring; the heating was turned off and 5 mL of NaYF was added at room temperature 4 :Yb,Er,Nd@NaYF 4 Nd (15, mg) cyclohexane solution is stirred overnight until the solution is clarified again;
(2) After overnight, evaporating water to reduce the solvent, then adding water to 20 mL, transferring the solution into a 250 mL three-neck flask, adding 40 mL ultrapure water, 6 mL absolute ethyl alcohol and 300 mu L (2 mol/L) NaOH aqueous solution, placing under an oil bath kettle, condensing and refluxing at 70 ℃ for 30 min (the rotation speed is maximum), slowly adding 400 mu L of TEOS dropwise by using a 200 mu L pipetting gun, stirring for reacting 1 h, cooling to room temperature, centrifuging to obtain precipitate, washing 3 times by absolute ethyl alcohol, and removing free silicon to obtain mesoporous silica coated UCNPs;
(3) To remove CTAB template, a mesoporous structure was obtained. The precipitate was dissolved in 40 mL acid ethanol solution (ph=1.4 to 1.5) and reacted under reflux at 60 ℃ with stirring for 1 h. And (3) centrifuging to obtain a precipitate, washing the precipitate with absolute ethyl alcohol for 2-3 times, dispersing the final product in 20 mL absolute ethyl alcohol, and repeating the reflux process for 2-3 times to fully remove CTAB and reduce biotoxicity.
The temperature of heating and stirring in the step (3) is 70 ℃ and the rotating speed is 1250 rpm.
The NaYF is added in the step (3) 4 :Yb,Er,Nd@NaYF 4 The Nd cyclohexane solution was about 15. 15 mg, and after the overnight solution was clarified, the next experiment was performed.
In the step (3), the CTAB template agent is removed by refluxing in an acidic ethanol solution with the pH value of 1.4-1.5, and the process can be repeated for 2-3 times to fully remove CTAB and reduce biotoxicity.
The step (4) comprises the following specific steps:
(1) An aqueous solution of 0.2 mg/mL DOX and an aqueous solution of 0.2 mg/mL MC540 were prepared, and UCNPs@mSiO was then added 2 Centrifuging the aqueous solution to obtain solid precipitate, and collecting 4 mg UCNPs@mSiO 2 Soaking in 2 mL of mixed solution of DOX and MC540, firstly placing in an ultrasonic machine for ultrasonic treatment for 30 min, and then rapidly stirring on a magnetic stirring heating sleeve for 24 h at room temperature in a light-proof environment;
(2) After physical adsorption was completed, the supernatant was removed by centrifugation to leave a precipitate, then ultrapure water was dissolved, again centrifuged to leave a precipitate, and washed three times to remove free DOX and MC540.
In the step (5), 40. Mu.L of TA and 40. Mu.L of FeCl are stirred vigorously 3 (24 mM) solution was added to UCNPs@mSiO of 2 mL 2 -MC540/DOX in aqueous solution. FinallyCollecting UCNPs@mSiO by centrifugal separation and washing with deionized water three times 2 @MPN-MC540/DOX nanocomposite.
The preparation method of the up-conversion-metal phenolic network composite nano material is characterized in that the up-conversion-metal phenolic network composite nano material is synthesized by adopting a thermal decomposition method, has uniform size, good biocompatibility and excellent photo-thermal performance and pH response performance.
The preparation method of the up-conversion-metal phenolic network composite nano material is characterized in that the composite material realizes the tumor combined treatment of PTT/PDT/chemotherapy through in vitro cell treatment experiments and in vivo tumor treatment experiments of mice, and has higher combined treatment effect.
The invention has the advantages that:
(1) The preparation method of the up-conversion-metal phenolic network composite nano material provided by the invention is mainly characterized in that UCNPs are synthesized by adopting a thermal decomposition method. The synthesis process is simple and convenient to operate. The composite nano material has high and stable repetition rate in the preparation process. The invention can realize stable and uniform nanoscale structure and good biocompatibility.
(2) The up-conversion-metal phenolic network composite nano material provided by the invention is mainly characterized in that a layer of mesoporous silica is coated on the surface of UCNPs, so that the UCNPs become water-soluble and are used for subsequent biological experiments. The mesoporous structure of silicon dioxide is utilized to load photosensitizer MC540 and chemotherapeutic medicine DOX to UCNPs@mSiO 2 In the pore canal. Finally, UCNPs@mSiO is prepared by adopting a simple assembly method 2 And contains TA and Fe 3+ The strong adhesion of TA and complexation of metal ions induces UCNPs@mSiO 2 And (3) rapidly forming the surface MPN film.
(3) The up-conversion-metal phenolic network composite nano material provided by the invention has the key point that as MPN has dual performances (photo-thermal performance and pH response performance), UCNPs@mSiO is irradiated by 808 nm laser 2 The @ MPN-MC540/DOX shows good photo-thermal performance, the photo-thermal conversion efficiency is up to 35.13%, and photo-thermal treatment (PTT) can be realized; meanwhile, under the weak acid condition of tumor microenvironment, MPNCan be gradually decomposed to realize the controllable release of DOX, and is used for chemotherapy, and in addition, the influence of photo-heat on DOX release is explored, and the result shows that the photo-heat can promote the release of 5 percent of medicines; while photosensitizer MC540 responds 808 nm to excite 545 nm emission peak of UCNPs, so as to realize photodynamic therapy, and the synergistic effect can achieve the aim of killing tumor cells.
(4) The up-conversion-metal phenolic network composite nano material provided by the invention is mainly characterized in that results in anticancer experiments of Hela cells and tumor-bearing mice show that UCNPs@mSiO 2 The @ MPN-MC540/DOX nanocomposite is synergistic with PTT/PDT/chemotherapy under the excitation of 808 nm, has remarkably increased cancer cell mortality compared with single PTT treatment or PDT/chemotherapy (PTT/chemotherapy) synergistic treatment, and has good inhibition effect on tumor growth. The present invention will be described in further detail with reference to the accompanying drawings and detailed description.
Drawings
FIG. 1 is a schematic diagram of the preparation of nanomaterial N in example 1 2 Adsorption/desorption isotherms and pore size distribution plots.
FIG. 2 is an electron microscope image of the nanomaterial prepared in example 1.
FIG. 3 is an ultraviolet absorption diagram of the nanomaterial prepared in example 1.
Fig. 4 is a photo-thermal graph of the nanomaterial prepared in example 2.
FIG. 5 is a graph showing the time-dependent release of DOX in PBS buffers at different pH values (7.4 and 6.0,5.0) for the preparation of the nanomaterial of example 3.
Fig. 6 is a graph showing ROS release curves of the nanomaterial prepared in example 4 under different pH environments (ph=7.4, ph=6.0, and ph=5.0)
FIG. 7 is a graph showing in vitro cytotoxicity and in vitro therapeutic effects of the nanomaterial prepared in example 5.
FIG. 8 is a graph showing the change in mouse weight and the graph showing the growth of tumor in the experiment of inhibiting tumor in Hela cell tumor-bearing mice by preparing the nanomaterial of example 6.
Detailed Description
Referring to fig. 1 to 8, the following detailed description of the technical solution of the present invention is given by way of examples and drawings.
Example 1
The preparation method of the up-conversion-metal phenolic network composite nano material provided by the embodiment comprises the following steps:
(1) Preparation of NaYF 4 Yb, er and Nd nano particles are used as the inner cores: by "thermal decomposition" method, in the presence of 6 mL oleic acid
(OA) and 15 mL Octadecene (ODE) in 100 mL three-neck flasks were charged with different proportions of Y (CH) 3 CO 2 ) 3 ，Yb(CH 3 CO 2 ) 3 ，Er(CH 3 CO 2 ) 3 And Nd (CH) 3 CO 2 ) 3 (1 mmol, Y: yb: er: nd=80.5%: 18%:0.5%: 1%). Placing the mixture on a heating sleeve, then heating to 160 ℃, stirring at a constant speed (rpm is approximately equal to 700), and preserving the heat for five minutes until acetate powder is completely dissolved to form a rare earth-oleic acid complex, wherein the rare earth-oleic acid complex is light yellow clear and transparent solution; and then the heating is turned off, and the solution is cooled to below 60 ℃. 0.1 g sodium hydroxide and 0.15 g ammonium fluoride are weighed and dissolved in 5 mL methanol solution, ultrasonic dissolution is carried out, then the mixed solution is slowly added into a three-neck flask drop by drop, the temperature is raised to 110 ℃ for reaction for about 30 minutes, so that methanol is evaporated, then a vacuum system is utilized to vacuumize the environment in the three-neck flask for about 30 minutes, then the pumping and ventilating are carried out for 3 times, each time for 30 seconds, the temperature of reactants is raised to 300 ℃ in an argon environment, and the temperature is kept at 1 h. After the reaction was completed, the solution was cooled to room temperature, and the product was precipitated with absolute ethanol (NaYF 4 Yb, er, nd up-conversion nano-particles), washing with cyclohexane and ethanol respectively for multiple times to obtain uniform up-conversion nano-materials, and storing in cyclohexane;
(2) Preparation of NaYF 4 :Yb,Er@NaYF 4 Core-shell up-conversion nanoparticles: weighing metal-acetate Y (CH) 3 CO 2 ) 3 And Nd (CH) 3 CO 2 ) 3 (1 mmol, y: nd=80%: 20%) was added to the flask, and 12 mL OA and 30 mL ODE were added, and after heating to 160 ℃, stirring was carried out at a constant speed, and the mixture was kept for 10 min to completely dissolve the mixture; then cooling to below 80deg.C (cyclohexane boiling point), adding 5 mL above NaYF 4 Heating Yb, er and Nd cyclohexane solution to 150 ℃ to remove cyclohexane solvent until no liquid flow down and no bubble generation of the solution exist at the bottle mouth, and cooling to below 60 ℃; 0.0674 g NaOH and 0.0374 g NH were taken 4 F, adding the mixture into a 5 mL methanol solvent, slowly adding the mixture into a reaction solution dropwise after ultrasonic dissolution, heating to 110 ℃, and removing methanol until no fog and no bubbles exist; then vacuuming for 30 min, exchanging argon for 3 times, heating to 300 ℃ and reacting for 1 h. Purification steps such as NaYF 4 The preparation method comprises the steps of preparing Yb, er and Nd nano particles. Finally purified NaYF 4 :Yb,Er,Nd@NaYF 4 Nd (core-shell) nanoparticles are dispersed in 15 mL cyclohexane;
(3) Water-soluble mesoporous silica coated NaYF 4 :Yb,Er,Nd@NaYF 4 Preparation of Nd nano-particles: firstly, weighing 0.1 g of CTAB in a beaker, adding 20 mL ultrapure water, heating to 70 ℃ and clarifying the solution under strong stirring; the heating was turned off and 5 mL of NaYF was added at room temperature 4 :Yb,Er,Nd@NaYF 4 Nd (15, mg) cyclohexane solution is stirred overnight until the solution is clarified again; after overnight, the solvent was reduced by evaporation of water, then the solution was made up to 20 mL, the above solution was transferred to a 250 mL three-neck flask, then 40 mL ultrapure water, 6 mL absolute ethanol and 300 μl (2 mol/L) NaOH aqueous solution were added, the mixture was placed under an oil bath at 70 ℃ for 30 min of condensation reflux (speed of rotation is maximized), 400 μl TEOS was slowly added dropwise with a 200 μl pipette, after stirring reaction 1 h, cooled to room temperature, centrifuged to obtain precipitate, and the precipitate was washed 3 times with absolute ethanol to remove free silicon, thereby obtaining mesoporous silica coated UCNPs. To remove CTAB template, a mesoporous structure was obtained. The precipitate was dissolved in 40 mL acid ethanol solution (ph=1.4 to 1.5) and reacted under reflux at 60 ℃ with stirring for 1 h. Centrifuging to obtain a precipitate, washing the precipitate with absolute ethyl alcohol for 2-3 times, dispersing the final product in 20 mL absolute ethyl alcohol, and repeating the reflux process for 2-3 times to fully remove CTAB and reduce biotoxicity;
（4）UCNPs@mSiO 2 -preparation of MC540/DOX nanocomposite: an aqueous solution of 0.2 mg/mL DOX and an aqueous solution of 0.2 mg/mL MC540 were prepared, and UCNPs@mSiO was then added 2 Centrifuging the aqueous solution to obtain solid precipitate, and collecting 4 mg UCNPs@mSiO 2 Soaking in 2 mLThe mixed solution of DOX and MC540 is firstly placed in an ultrasonic machine for ultrasonic treatment for 30 min, and then is rapidly stirred on a magnetic stirring heating sleeve for 24 h under the environment of room temperature and light shielding. After physical adsorption is completed, removing supernatant liquid by centrifugation to leave sediment, dissolving the sediment with ultrapure water, centrifuging again to leave sediment, washing for three times, and removing free DOX and MC540;
（5）UCNPs@mSiO 2 preparation of @ MPN-MC540/DOX nanocomposite: 40. Mu.L TA and 40. Mu.L FeCl were stirred vigorously 3 (24 mM) solution was added to UCNPs@mSiO of 2 mL 2 -MC540/DOX in aqueous solution. Finally, collecting UCNPs@mSiO by centrifugal separation and washing with deionized water for three times 2 @MPN-MC540/DOX nanocomposite.
The step (1) comprises the following specific steps:
(1) Y (CH) in different proportions was added to a 100 mL three-neck flask containing 6 mL Oleic Acid (OA) and 15 mL Octadecene (ODE) 3 CO 2 ) 3 ，Yb(CH 3 CO 2 ) 3 ，Er(CH 3 CO 2 ) 3 And Nd (CH) 3 CO 2 ) 3 (1 mmol, Y: yb: er: nd=80.5%: 18%:0.5%: 1%). Placing the mixture on a heating sleeve, then heating to 160 ℃, stirring at a constant speed, and preserving heat for five minutes until acetate powder is completely dissolved to form a rare earth-oleic acid complex, wherein the rare earth-oleic acid complex is light yellow clear and transparent solution;
(2) And then the heating is turned off, and the solution is cooled to below 60 ℃. 0.1 g sodium hydroxide and 0.15 g ammonium fluoride are weighed and dissolved in 5 mL methanol solution, ultrasonic dissolution is carried out, then the mixed solution is slowly added into a three-neck flask drop by drop, the temperature is raised to 110 ℃ for reaction for about 30 minutes, so that methanol is evaporated, then a vacuum system is utilized to vacuumize the environment in the three-neck flask for about 30 minutes, then the pumping and ventilating are carried out for 3 times, each time for 30 seconds, the temperature of reactants is raised to 300 ℃ in an argon environment, and the temperature is kept at 1 h. After the reaction was completed, the solution was cooled to room temperature, and the product was precipitated with absolute ethanol (NaYF 4 Yb, er, nd up-conversion nano-particles), washing with cyclohexane and ethanol respectively for multiple times to obtain uniform up-conversion nano-materialThe material was then stored in cyclohexane.
The rotating speed of the heating sleeve in the step (1) is 700 rpm.
And (3) in the step (2), reacting for 1 hour at a temperature of 300 ℃ in an argon environment.
The step (3) comprises the following specific steps:
(1) Firstly, weighing 0.1 g of CTAB in a beaker, adding 20 mL ultrapure water, heating to 70 ℃ and clarifying the solution under strong stirring; the heating was turned off and 5 mL of NaYF was added at room temperature 4 :Yb,Er,Nd@NaYF 4 Nd (15, mg) cyclohexane solution is stirred overnight until the solution is clarified again;
(2) After overnight, evaporating water to reduce the solvent, then adding water to 20 mL, transferring the solution into a 250 mL three-neck flask, adding 40 mL ultrapure water, 6 mL absolute ethyl alcohol and 300 mu L (2 mol/L) NaOH aqueous solution, placing under an oil bath kettle, condensing and refluxing at 70 ℃ for 30 min (the rotation speed is maximum), slowly adding 400 mu L of TEOS dropwise by using a 200 mu L pipetting gun, stirring for reacting 1 h, cooling to room temperature, centrifuging to obtain precipitate, washing 3 times by absolute ethyl alcohol, and removing free silicon to obtain mesoporous silica coated UCNPs;
(3) To remove CTAB template, a mesoporous structure was obtained. The precipitate was dissolved in 40 mL acid ethanol solution (ph=1.4 to 1.5) and reacted under reflux at 60 ℃ with stirring for 1 h. And (3) centrifuging to obtain a precipitate, washing the precipitate with absolute ethyl alcohol for 2-3 times, dispersing the final product in 20 mL absolute ethyl alcohol, and repeating the reflux process for 2-3 times to fully remove CTAB and reduce biotoxicity.
The temperature of heating and stirring in the step (3) is 70 ℃ and the rotating speed is 1250 rpm.
The NaYF is added in the step (3) 4 :Yb,Er,Nd@NaYF 4 The Nd cyclohexane solution was about 15. 15 mg, and after the overnight solution was clarified, the next experiment was performed.
In the step (3), the CTAB template agent is removed by refluxing in an acidic ethanol solution with the pH value of 1.4-1.5, and the process can be repeated for 2-3 times to fully remove CTAB and reduce biotoxicity.
The step (4) comprises the following specific steps:
(1) An aqueous solution of 0.2 mg/mL DOX and an aqueous solution of 0.2 mg/mL MC540 were prepared, and UCNPs@mSiO was then added 2 Centrifuging the aqueous solution to obtain solid precipitate, and collecting 4 mg UCNPs@mSiO 2 Soaking in 2 mL of mixed solution of DOX and MC540, firstly placing in an ultrasonic machine for ultrasonic treatment for 30 min, and then rapidly stirring on a magnetic stirring heating sleeve for 24 h at room temperature in a light-proof environment;
(2) After physical adsorption was completed, the supernatant was removed by centrifugation to leave a precipitate, then ultrapure water was dissolved, again centrifuged to leave a precipitate, and washed three times to remove free DOX and MC540.
In the step (5), 40. Mu.L of TA and 40. Mu.L of FeCl are stirred vigorously 3 (24 mM) solution was added to UCNPs@mSiO of 2 mL 2 -MC540/DOX in aqueous solution. Finally, collecting UCNPs@mSiO by centrifugal separation and washing with deionized water for three times 2 @MPN-MC540/DOX nanocomposite.
The preparation method of the up-conversion-metal phenolic network composite nano material is characterized in that the up-conversion-metal phenolic network composite nano material is synthesized by adopting a thermal decomposition method, has uniform size, good biocompatibility and excellent photo-thermal performance and pH response performance.
The preparation method of the up-conversion-metal phenolic network composite nano material is characterized in that the composite material realizes the tumor combined treatment of PTT/PDT/chemotherapy through in vitro cell treatment experiments and in vivo tumor treatment experiments of mice, and has higher combined treatment effect.
The up-conversion-metal phenolic network composite nano material prepared by the method is synthesized by adopting a thermal decomposition method.
FIG. 1 is a graph showing pore diameter analysis of the mesoporous nanomaterial prepared in example 1. They were tested by nitrogen adsorption-desorption. As can be seen from the nitrogen adsorption-desorption isothermal curve, the mesoporous nanomaterial exhibits a typical type IV isothermal line, and the pore diameter of the synthesized mesoporous nanomaterial is about 2.2 and nm according to the nitrogen adsorption-desorption isothermal curve by using a Brunauer-Emmett-Teller (BET) method, and basically accords with the regulation of mesoporous size (2-50)nm); the specific surface area of the mesoporous nano particles is 979 m 2 /g。
FIG. 2 is a TEM photograph of the up-conversion-metal phenolic network composite nanomaterial prepared in example 1, and it is observed that MPN coated UCNPs@mSiO 2 Has good monodispersity and average particle diameter of about 101 nm, and has great significance for circulation in organisms.
Fig. 3 is an ultraviolet absorption spectrum of the up-conversion-metal phenolic network composite nanomaterial prepared in example 1. From the figure, UCNPs@mSiO 2 The @ MPN has a broad absorption peak at 560 nm due to Fe 3+ Formation of the TA complex is accompanied by charge transfer from tannic acid to iron ions.
Example 2
The preparation method and application of the up-conversion-metal phenolic network composite nanomaterial provided in this embodiment are basically the same as those in embodiment 1, and are different in that:
the up-conversion-metal phenolic network composite nanomaterial prepared in example 1 is specifically applied to a method for photo-thermal experiments, and comprises the following steps:
(1) Preparing the nano materials with the nano particle concentration of 0.1,0.2,0.4,0.8 mg/mL by using ultrapure water respectively;
(2) 1 mL each was placed in a cuvette and irradiated with a 808 nm laser at 2W/cm 2 Illumination is performed at a power density of (2);
(3) The temperature rise of materials with different concentrations along with the irradiation time is detected and monitored by a micro thermocouple.
Fig. 4 is a photo-thermal graph of the nanomaterial prepared in example 2. The observation shows that the temperature of the sample gradually rises along with the increase of the concentration of the sample, and the prepared nano material has good photo-thermal effect and high photo-stability.
Example 3
The preparation method and application of the up-conversion-metal phenolic network composite nanomaterial provided by the embodiment are basically the same as those of the embodiment 1-2, and are different in that:
the nanometer material loaded with the anticancer drug DOX is further used as a therapeutic agent and is applied to a method of chemotherapy, and the method comprises the following steps:
(1)2 mg UCNP@mSiO 2 adding into 2 mL doxorubicin hydrochloride solution (0.2 mg/mL), stirring in the dark, and adsorbing for 24 hr to obtain UCNPs@mSiO 2 The DOX product is separated by centrifugation and washed with water;
(2) 2 mLUCNPs@mSiO 2 The @ MPN-DOX (1 mg/mL) solution was placed in a dialysis bag with a molecular weight cut-off of 8000 Da, and the two ends were fastened with a thin wire to prevent leakage of fluid. Immersing the dialysis bag in a beaker containing 10 mL Phosphate Buffer (PBS) with different pH values (pH 7.4, pH 6.0 and pH 5.0) without or with NIR laser irradiation (2W/cm) 2 ) Slowly stirring under the condition;
(3) The dialysate was aspirated and collected at various time points, and then an equal amount of fresh buffer solution was rapidly added as a supplement. The time-dependent release of DOX under laser irradiation was determined by UV-visible absorption spectroscopy at 480 nm in PBS buffer solutions of different pH.
FIG. 5 is a graph showing the time-dependent release of DOX in PBS buffers at different pH values (7.4 and 6.0,5.0) for the upconverter-metal phenolic network composite nanomaterial prepared in example 3. It was observed that the loading of DOX was 10.03 wt% and that the release profile of DOX was pH dependent and the release rate increased under acidic conditions. In addition, under the irradiation of near infrared light, photo-thermal promotion is to 5% DOX release, which is that photo-thermal effect accelerates DOX molecular Brownian motion and promotes drug permeation.
Example 4
The preparation method and application of the up-conversion-metal phenolic network composite nanomaterial provided by the embodiment are basically the same as those of the embodiments 1-3, and are different in that:
the method specifically relates to an in-vitro photodynamic therapy method of a MC540 loaded nano diagnosis and treatment agent composite material, which comprises the following steps:
(1) Taking 2 mL UCNPs@mSiO 2 MC540 aqueous solution in cuvette, 10. Mu.L of DPBF (10 mM) ethanol solution was added. In a dark environment, a 808 nm laser (1W/cm 2 ) Irradiating for 30 min, measuring absorption spectrum of the solution every 5 min, and recordingAbsorbance values at the maximum absorption wavelength (417 nm). The solution can be evenly irradiated by continuously stirring during the irradiation process. The control groups were UCNPs@mSiO without laser irradiation 2 -MC540 in water and pure water to exclude the effect of natural degradation of laser and DPBF.
(2) In addition, UCNPs@mSiO is made 2 Experiment of ROS release in different pH Environment (pH=7.4, pH=6.0 and pH=5.0) with MPN-MC540/DOX, briefly, UCNPs@mSiO synthesized first 2 The @ MPN-MC540/DOX solution was left for a period of time in solutions of different pH values (ph=7.4, ph=6.0 and ph=5.0) until the ROS release experiment was performed after MPN decomposition, as detailed above.
FIG. 6 is UCNPs@mSiO prepared in example 4 2 Release profile of ROS at different pH values (ph=7.4, ph=6.0 and ph=5.0) for MPN-MC540/DOX. From the figure, ucnps@msio in solutions with ph=5.0 and ph=6.0 2 The @ MPN-MC540/DOX has a molecular structure equal to UCNPs @ mSiO 2 MC540 similarity 1 O 2 The production capacity can prove UCNPs@mSiO 2 The @ MPN-MC540/DOX has great potential for PDT in tumor microenvironments. But at ph=7.4 ucnps@msio 2 @MPN-MC540/DOX production 1 O 2 This may be due to the outer MPN film absorbing 808 nm light, converting the light into heat, and achieving PTT.
Example 5
The preparation method and application of the up-conversion-metal phenolic network composite nanomaterial provided by the embodiment are basically the same as those of the embodiments 1 to 4, and are different in that:
the nanometer diagnosis and treatment agent composite material loaded with DOX and MC540 is applied to in-vitro toxicity assessment, and comprises the following steps:
(1) Cervical cancer HeLa cells were cultured at 1.5X10 5 Density of individual cells/well were seeded in 96-well plates, incubated with DMEM containing 10% FBS and 1% penicillin/streptomycin, 5% CO at 37 °c 2 Culturing in a concentration incubator for 24 h;
(2) The old medium was aspirated with a pipette, 100. Mu.L of fresh medium containing different concentrations of material (0, 25, 50, 100. Mu.g/mL) was added, and the mixture was placed in an incubator for incubation 24 h.
(3) The mixed culture solution containing the nanomaterial was aspirated with a pipette and washed 3-4 times with PBS to remove free material, then fresh medium was added and the cells were placed in 808 nm (2W/cm) 2 ) Irradiating with laser for 5 min. After the irradiation was completed, the cells were replaced in a constant temperature incubator to culture 24. 24 h. After time, quantitative CCK-8 reagent was added and absorbance per well was measured using a microplate reader. The control group was set to control other variables in agreement without 808 and nm laser irradiation.
Fig. 7 shows that the nano material prepared in example 5 has no obvious toxicity to cells without adding the drug DOX, which indicates that the material has no obvious toxicity to cells and has good biocompatibility. When the DOX-containing nanocomposite is added, cell viability decreases significantly as the concentration of the nanocomposite increases. Notably, irradiation with 808 nm laser will trigger the sample material UCNPs@mSiO for PTT treatment 2 Sample material UCNPs@mSiO for@MPN, PTT/PDT treatment 2 Sample material UCNPs@mSiO for treatment of MPN-MC540, PTT/chemotherapy 2 Sample material UCNPs@mSiO combining MPN-DOX and PTT/PDT/chemotherapy 2 @ MPN-MC540/DOX. After 808 and nm laser irradiation for 5 minutes, the single PTT treatment effect is not obvious, and the survival rate of the cells can still reach about 60% even when the concentration reaches 100 mug/mL; in addition, when Hela cells and 100 mug/mLUCNPs@mSiO 2 Cell viability of @ MPN-MC540/DOX incubation with 100 μg/mL UCNPs @ mSiO 2 @MPN-MC540 or UCNPs@mSiO 2 The viability of cells was reduced to less than 10% after incubation with MPN-DOX, which fully demonstrates that PDT/PTT/chemotherapy combination therapy has better efficacy in inhibiting cancer cell growth than PTT or PTT/PDT (PTT/chemotherapy) alone.
Example 6
The preparation method and application of the up-conversion-metal phenolic network composite nanomaterial provided by the embodiment are basically the same as those of the embodiments 1 to 5, and are different in that:
the method for applying the nanometer diagnosis and treatment agent composite material loaded with DOX and MC540 to in-vivo tumor photothermal/photodynamic/chemotherapy treatment comprises the following steps of:
(1) 5X 10 subcutaneous injections were administered to the right axilla of female BALB/c mice 6 HeLa cells, until tumor volume grew to 100 mm 3 Left and right;
(2) Hela tumor-bearing mice were randomly divided into five groups: (1) PBS+808 nm, (2) UCNPs@mSiO 2 @MPN (10 mg/mL,100 [ mu ] L) +808 nm laser, (3) UCNPs@mSiO 2 @MPN-MC540 (10 mg/mL,100 μL) +808 nm laser, (4) UCNPs@mSiO 2 @MPN-DOX (10 mg/mL,100 μL) +808 nm laser, (5) UCNPs@mSiO 2 @ MPN-MC540/DOX (10 mg/mL,100 μl) +808 nm laser. After 1 hour of material injection, the tumor site of the mice was exposed to 808 nm laser at a power density of 2.5W/cm 2 Irradiation was performed for 15 minutes, but irradiation was continued after a pause of 3 minutes every 5 minutes in order to avoid overheating leading to burning of the skin tissue of the mice. The treatment was performed every other day. Mice body weight and tumor volume changes were recorded daily during treatment.
FIG. 8 example 6 preparation of a graph of tumor growth in an experiment of nanomaterial inhibition against tumors of Hela cell tumor-bearing mice. The body weight of the group of mice did not change much from the initial body weight, indicating that the material did not have a significant effect on the health status of the mice. We also recorded the change in tumor size for each group: (1) The volume of the group of tumors has obvious increasing trend, which indicates that the laser and the non-added materials have no inhibition effect on the growth of the tumors; (2) The group is a photothermal treatment group, and the tumor can be seen to be reduced; (3) The group is a photothermal treatment group and a photodynamic treatment group, and the tumor growth volume can be seen to be slow; (4) The group is a photo-thermal and drug combined treatment group, and the tumor is obviously smaller than the volume of the group (2); (5) The group is a photo-thermal, photodynamic and drug combined treatment group, and the tumor is obviously much smaller than the other groups, which indicates UCNPs@mSiO 2 The @ MPN-MC540/DOX composite nanomaterial has a good combined tumor treatment effect. The tumor photographs of the dissected mice in each group are consistent with the results, and the inhibition effect on tumors is more intuitively reflected, wherein the tumor volumes of the groups (3), (4) and (5) are obviously higher than those of the first two groupsSmall, wherein the combined treatment (5) group volume is minimal.
The method provided by the invention synthesizes NaYF 4 :Yb,Er,Nd@NaYF 4 Nd up-conversion nano particles, and coating a silicon dioxide layer to form UCNPs@mSiO 2 Improving the hydrophilicity and making the material loading capacity, and utilizing UCNPs@mSiO 2 Mesoporous loading DOX and MC540 to form UCNPs@mSiO 2 MC540/DOX nano composite material, finally coating a complex (MPN) formed by Fe-TA on the outermost layer, and finally forming UCNPs@mSiO 2 @MPN-MC540/DOX nanocomposite.
The invention focuses on utilizing the photo-thermal property and pH response property of MPN, UCNPs@mSiO under 808 nm laser irradiation 2 The @ MPN-MC540/DOX shows good photo-thermal performance, and photo-thermal treatment is realized; meanwhile, under the weak acid condition of tumor microenvironment, the MPN film is gradually decomposed, so that the controllable release of DOX is realized, and the DOX is used for chemotherapy; while photosensitizer MC540 excites the 545 nm emission peak of UCNPs in response to 808 nm, effecting photodynamic therapy. The synergistic effect can achieve the aim of killing tumor cells. UCNPs@mSiO 2 Under the excitation of 808 nm, the nano composite material of MPN-MC540/DOX has obviously raised cancer cell death rate and obvious tumor growth inhibiting effect compared with single PTT or PDT/PTT (PTT/chemotherapy) synergistic treatment effect.
The present invention is not limited to the above embodiments, and the method of using other similar nanocomposite materials obtained by the same or similar method is not limited to the above embodiments, and specific numerical values, different organic molecules for further improving water solubility and functionalization, etc. are specifically selected within the range of values of each component described in each embodiment of the present invention, and all embodiments of the present invention are not listed one by one.
The foregoing description of the exemplary embodiments of the invention is not intended to limit the invention to the precise form disclosed, and any modifications, equivalents, and alternatives falling within the spirit and principles of the invention are intended to be included within the scope of the invention.

Claims (5)

1. The preparation method of the up-conversion-metal phenolic network composite nano material is characterized by comprising the following steps of:
(1) Preparation of NaYF 4 Yb, er and Nd nano particles are used as the inner cores: by "thermal decomposition" method, in the presence of 6 mL oleic acid
And 15 mL octadecene in a 100 mL three neck flask was charged with 1mmol Y: yb: er: nd=80.5%: 18%:0.5%:1% of Y (CH) 3 CO 2 ) 3 ，Yb(CH 3 CO 2 ) 3 ，Er(CH 3 CO 2 ) 3 And Nd (CH) 3 CO 2 ) 3 The method comprises the steps of carrying out a first treatment on the surface of the Placing the mixture on a heating sleeve, then heating to 160 ℃, stirring at a constant speed, and preserving heat for five minutes until acetate powder is completely dissolved to form a rare earth-oleic acid complex, wherein the rare earth-oleic acid complex is light yellow clear and transparent solution; then heating is closed, and the solution is cooled to below 60 ℃; weighing 0.1 g sodium hydroxide and 0.15 g ammonium fluoride, dissolving in 5 mL methanol solution, performing ultrasonic dissolution, then slowly adding the mixed solution into a three-neck flask drop by drop, heating to 110 ℃ for reaction for 30 minutes, so that methanol is evaporated, then vacuumizing the environment in the three-neck flask for 30 minutes by using a vacuum system, exchanging and ventilating for 3 times each for 30 seconds, heating the reactant to 300 ℃ in an argon environment, and preserving heat for 1 h; after the reaction was completed, the solution was cooled to room temperature, and the product, naYF, was precipitated with absolute ethanol 4 The up-conversion nano-particles of Yb, er and Nd are washed for a plurality of times by cyclohexane and ethanol respectively, and finally the up-conversion nano-material with uniform particles is obtained and then stored in the cyclohexane;
(2) Preparation of NaYF 4 :Yb,Er@NaYF 4 Core-shell up-conversion nanoparticles: weigh 1mmol, Y: nd=80%: 20% of metal-acetate Y (CH) 3 CO 2 ) 3 And Nd (CH) 3 CO 2 ) 3 Adding 12 mL of OA and 30 mL of ODE into a flask, heating to 160 ℃, stirring at a constant speed, and keeping for 10 min to completely dissolve; then cooling to below 80deg.C, adding 5 mL above NaYF 4 Heating Yb, er and Nd cyclohexane solution to 150 ℃ to remove cyclohexane solvent until no liquid flow down and no bubble generation of the solution exist at the bottle mouth, and cooling to below 60 ℃; take 0.0674g NaOH and 0.0374 g NH 4 F, adding the mixture into a 5 mL methanol solvent, slowly adding the mixture into a reaction solution dropwise after ultrasonic dissolution, heating to 110 ℃, and removing methanol until no fog and no bubbles exist; then vacuumizing for 30 min, exchanging argon for 3 times, heating to 300 ℃ and reacting for 1 h; purification step and step (1) the NaYF 4 The preparation process of Yb, er and Nd nano particles is the same; finally purified NaYF 4 :Yb,Er,Nd@NaYF 4 Nd nanoparticles are dispersed in 15 mL cyclohexane;
(3) Water-soluble mesoporous silica coated NaYF 4 :Yb,Er,Nd@NaYF 4 Preparation of Nd nano-particles: firstly, weighing 0.1 g of CTAB in a beaker, adding 20 mL ultrapure water, heating to 70 ℃ and clarifying the solution under strong stirring; the heating was turned off and 5 mL of NaYF was added at room temperature 4 :Yb,Er,Nd@NaYF 4 Stirring Nd cyclohexane solution overnight until the solution is clarified again; after overnight, evaporating water to reduce the solvent, then supplementing water to 20 mL, transferring the solution into a 250 mL three-neck flask, adding 40 mL ultrapure water, 6 mL absolute ethyl alcohol and 300 mu L of 2 mol/L NaOH aqueous solution, placing under an oil bath kettle, condensing and refluxing for 30 min at 70 ℃, slowly adding 400 mu L of TEOS dropwise by using a 200 mu L pipetting gun, stirring for reacting 1 h, cooling to room temperature, centrifuging to obtain precipitate, washing 3 times by absolute ethyl alcohol, removing free silicon, and obtaining mesoporous silica coated UCNPs; to remove CTAB template agent, obtaining mesoporous structure; dissolving the precipitate in 40 mL acid ethanol solution with pH=1.4-1.5, and carrying out reflux stirring reaction at 60 ℃ for 1 h; centrifuging to obtain a precipitate, washing the precipitate with absolute ethyl alcohol for 2-3 times, dispersing the final product in 20 mL absolute ethyl alcohol, and repeating the reflux process for 2-3 times to fully remove CTAB and reduce biotoxicity;
（4）UCNPs@mSiO 2 -preparation of MC540/DOX nanocomposite: an aqueous solution of 0.2 mg/mL DOX and an aqueous solution of 0.2 mg/mL MC540 were prepared, and UCNPs@mSiO was then added 2 Centrifuging the aqueous solution to obtain solid precipitate, and collecting 4 mg UCNPs@mSiO 2 Soaking in 2 mL of mixed solution of DOX and MC540, firstly placing in an ultrasonic machine for ultrasonic treatment for 30 min, and then rapidly stirring on a magnetic stirring heating sleeve for 24 h at room temperature in a light-proof environment; after physical adsorption is completed, the supernatant is removed by centrifugationLeaving the precipitate, dissolving with ultrapure water, centrifuging again to leave the precipitate, washing for three times, and removing free DOX and MC540;
（5）UCNPs@mSiO 2 preparation of @ MPN-MC540/DOX nanocomposite: 40. Mu.L of TA and FeCl at a concentration of 24 mmol/L were stirred vigorously 3 40. Mu.L of solution was added to UCNPs@mSiO of 2 mL 2 -MC540/DOX in aqueous solution; finally, centrifugal separation and washing with deionized water are carried out for three times, and UCNPs@mSiO is obtained by collection 2 @MPN-MC540/DOX nanocomposite.
2. The method for preparing up-conversion-metal phenolic network composite nano-material according to claim 1, wherein the rotational speed of the heating sleeve used in the step (1) is 700 rpm.
3. The method for preparing up-conversion-metal phenolic network composite nano-material according to claim 1, wherein the heating and stirring temperature in the step (3) is 70 ℃ and the rotating speed is 1250 rpm.
4. An up-conversion-metal phenolic network composite nano material prepared by the method of any one of claims 1 to 3, which is characterized in that the up-conversion-metal phenolic network composite nano material is synthesized by a thermal decomposition method, has uniform size, good biocompatibility, and excellent photo-thermal performance and pH response performance.
5. The up-conversion-metal phenolic network composite nano material according to claim 4, wherein the up-conversion-metal phenolic network composite nano material is proved by an in vitro cell therapy experiment and a mouse in vivo tumor therapy experiment to realize the tumor combined therapy of PTT/PDT/chemotherapy, and has higher combined therapy effect.
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Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108192590A (en) * 2017-12-10 2018-06-22 湖北工业大学 A kind of preparation method of the poly- azobenzene multifunctional nanoparticle based on rare earth up-conversion
CN112245407A (en) * 2020-11-25 2021-01-22 山东大学 Preparation of targeting nano vaccine based on metal-polyphenol network structure and product thereof
CN112315941A (en) * 2020-11-18 2021-02-05 海南大学 Preparation method of nano vaccine with pH and reduction double sensitivity and obtained product
CN112484851A (en) * 2021-01-06 2021-03-12 福州大学 Perovskite lanthanide series composite nano material, preparation method thereof and application of perovskite lanthanide series composite nano material in broadband photoelectric detector
CN113181361A (en) * 2021-04-25 2021-07-30 上海大学 Phase-change nano composite material with bubbles promoting drug release, preparation method and application
CN113230401A (en) * 2021-04-26 2021-08-10 上海大学 Core-shell up-conversion MOFs photosensitive composite material, preparation method and application thereof
CN113975411A (en) * 2021-11-01 2022-01-28 哈尔滨工程大学 Preparation method of near-infrared light response up-conversion mesoporous tin dioxide diagnosis and treatment nanocapsule

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US11439150B2 (en) * 2020-11-02 2022-09-13 The Administrators Of The Tulane Educational Fund Methods and compositions for treating algae using metal phenolic networks

Patent Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108192590A (en) * 2017-12-10 2018-06-22 湖北工业大学 A kind of preparation method of the poly- azobenzene multifunctional nanoparticle based on rare earth up-conversion
CN112315941A (en) * 2020-11-18 2021-02-05 海南大学 Preparation method of nano vaccine with pH and reduction double sensitivity and obtained product
CN112245407A (en) * 2020-11-25 2021-01-22 山东大学 Preparation of targeting nano vaccine based on metal-polyphenol network structure and product thereof
CN112484851A (en) * 2021-01-06 2021-03-12 福州大学 Perovskite lanthanide series composite nano material, preparation method thereof and application of perovskite lanthanide series composite nano material in broadband photoelectric detector
CN113181361A (en) * 2021-04-25 2021-07-30 上海大学 Phase-change nano composite material with bubbles promoting drug release, preparation method and application
CN113230401A (en) * 2021-04-26 2021-08-10 上海大学 Core-shell up-conversion MOFs photosensitive composite material, preparation method and application thereof
CN113975411A (en) * 2021-11-01 2022-01-28 哈尔滨工程大学 Preparation method of near-infrared light response up-conversion mesoporous tin dioxide diagnosis and treatment nanocapsule

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
上转换介孔二氧化硅多功能肝癌诊疗纳米复合体系的构建;杨涵等;生物医学工程研究(01);6-11 *

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