CN115381945B - A kind of Mn-In2S3/InOOH nanoparticles and preparation method and application - Google Patents

Abstract

The invention discloses a Mn-In ₂ S ₃ /InOOH nanoparticle and its preparation method and application. The preparation method includes: adding MnCl ₂ ·4H ₂ O and In(NO ₃ ) ₃ solution to the Na ₂ S solution, Then adjust the pH with dilute nitric acid solution to obtain a precursor solution; add the precursor solution to a hydrothermal reactor for reaction, and centrifuge and wash after the reaction to obtain Mn-In ₂ S ₃ /InOOH nanoparticles. Through two methods, heterojunction and metal ion doping, the nanoparticles greatly improve the efficiency of sonosensitizers and show strong sonodynamic activity, which can effectively solve the problem of the small number of sonosensitizers. Poor structural design and low acoustic and dynamic efficiency.

Description

A kind of Mn-In2S3/InOOH nanoparticles and preparation method and application

Technical field

The invention belongs to the field of biological nanomaterials, and in particular relates to Mn-In ₂ S ₃ /InOOH nanoparticles and preparation methods and applications.

Background technique

Malignant tumors remain one of the most lethal diseases threatening human health. In the past few decades, ROS-mediated tumor treatment has been widely used in the development of tumor treatment agents due to its minimal invasiveness, spatiotemporal controllability, targeting, and activation at specific locations. . Among them, ultrasonic dynamic therapy (SDT), as a new non-invasive treatment method, has shown great potential in tumor treatment. As a typical non-invasive radiation source, ultrasound has low tissue attenuation efficiency and can penetrate deeper body tissues without obvious energy loss. In addition, ultrasound can control the activation of sonosensitizers in space and time, thereby ensuring The biosafety of SDT in treatment was confirmed. SDT induces bubble bursting through the cavitation effect of ultrasound. The high temperature and high pressure generated during the bubble bursting process can induce phenomena such as sonoluminescence and pyrolysis, thereby activating the sonosensitizer accumulated specifically at the tumor site to produce ROS and cause oxidation inside the tumor cells. Stress kills cells and causes tumor ablation. In the process of SDT, the presence of sonosensitizer and its efficiency play a crucial role. Research on organic and inorganic sonosensitizers has increased significantly over the past few decades. Traditional organic sonosensitizers (such as chlorophyll derivatives, ATX-70, porphyrins, etc.) have the characteristics of hydrophobicity, low bioavailability, and high skin sensitivity, which greatly limits their application. In comparison, inorganic sonosensitizers have shown broad application prospects in the biomedical field due to their unique physical and chemical properties, inherent biological effects, high stability, and low photosensitivity. Among them, due to the sonoluminescence properties of ultrasound, various semiconductor materials are increasingly being studied for use in ultrasonic power therapy. Under ultrasonic irradiation, semiconductor materials can absorb the energy of ultrasonic waves to cause the transition of electrons in the valence band, generating activated electrons and holes, thereby generating redox reactions with reactants in the environment to generate reactive oxygen species. The recombination of electrons and holes is a reaction we do not want to occur in this process. In order to promote the separation of electrons and holes and improve quantum yield, the design of heterojunctions and the doping of metal ions are reasonable and effective ways.

Therefore, in the process of realizing the present invention, the inventor found that there are at least the following problems in the prior art: fewer types of sonosensitizers, poor structural design of the sonosensitizers, and low sonodynamic efficiency.

Contents of the invention

The purpose of the present invention is to provide a kind of Mn-In ₂ S ₃ /InOOH nanoparticles, preparation methods and applications in view of the shortcomings of the existing technology, so as to solve the problem of few types of sonosensitizers and the structural design of sonosensitizers in related technologies. Poor performance and low acoustic power efficiency.

The object of the present invention is achieved through the following technical solutions: a preparation method of Mn-In ₂ S ₃ /InOOH nanoparticles, including:

Add MnCl ₂ ·4H ₂ O and In(NO ₃ ) ₃ to water, and disperse with ultrasonic to obtain Mn and In source solutions;

Add Na ₂ S to water and disperse it ultrasonically to obtain a Na ₂ S solution;

Add the Mn and In source solutions dropwise to the Na ₂ S solution to form a uniform light yellow solution;

Add the dilute nitric acid solution dropwise to the above light yellow solution, adjust the pH to below 3, and obtain the Mn-In ₂ S ₃ /InOOH precursor solution that has not yet formed stable particles;

The Mn-In ₂ S ₃ /InOOH precursor solution that has not yet formed stable particles is transferred to a hydrothermal reactor for reaction. After the reaction, it is centrifuged and washed to obtain a Mn-In ₂ S ₃ /InOOH solution.

Preferably, in the Mn and In source solution, the concentration of MnCl ₂ ·4H ₂ O is 0-3.5mg/ml, and the concentration of In(NO ₃ ) ₃ is 12-24mM.

Preferably, the concentration of Na ₂ S in the Na ₂ S solution is 30-60 mM.

Preferably, the Mn and In source precursor solutions are added dropwise to the Na ₂ S precursor solution at a dropping speed of 0.5-2 ml/min.

Preferably, the pH is adjusted to 2.6-3.

Preferably, the reaction temperature of the reaction in the hydrothermal reactor is 180°C, and the reaction time is 12-24 hours.

Preferably, the Mn-In ₂ S ₃ /InOOH nanoparticles have a particle size of 10-30 nm, irregular shape, and good dispersion stability.

The invention also provides Mn-In ₂ S ₃ /InOOH nanoparticles prepared by the preparation method.

The invention also provides an application of the Mn-In ₂ S ₃ /InOOH nanoparticles in preparing sonodynamic therapy preparations.

Beneficial effects of the present invention: The present invention combines In ₂ S ₃ and InOOH to form a heterojunction through a one-step hydrothermal method, and in this process, the metal ion Mn is doped into the composite material, which is beneficial to the sonodynamic process of the material. It absorbs ultrasound and promotes the separation of electron holes, which greatly improves the efficiency of sonodynamic force and achieves a good tumor inhibition effect.

The present invention synthesizes Mn-In ₂ S ₃ /InOOH nanoparticles through a one-step hydrothermal method. In the Mn-In ₂ S ₃ /InOOH nanoparticles in the present invention, the formation of heterojunction promotes the absorption of ultrasonic waves by the composite material. Under ultrasonic irradiation, both In ₂ S ₃ and InOOH can absorb ultrasonic waves to realize the transition of electrons from the valence band to the conduction band. Due to the formation of the heterojunction and the different Fermi levels of In ₂ S ₃ and InOOH, according to the lowest energy The principle is that the electrons in the conduction band of In ₂ S ₃ will migrate to the conduction band of InOOH; the holes in the valence band of InOOH will migrate to the valence band of In ₂ S ₃ , thereby promoting the separation of electrons and holes and improving the quantum yield. In addition, the doping of metal ions Mn can form a defect energy level near the conduction band of In ₂ S ₃ and InOOH, thereby capturing the activated electrons during their transition to the valence band, thereby further reducing the recombination of electron holes. , promoting the separation of electrons and holes. The activated electrons can undergo a reduction reaction with surface-adsorbed O ₂ to produce toxic O ₂ · ^- and ¹ O ₂ , thereby killing tumor cells. Such a design verifies the feasibility of Mn-In ₂ S ₃ /InOOH as a sonosensitizer, and provides guidance for the application of heterojunction and metal ion doping in the field of tumor treatment. In the present invention, the Mn-doped In ₂ S ₃ /InOOH heterostructure is synthesized through a one-step hydrothermal method, thereby realizing an efficient sonodynamic treatment method. So far, the field has not developed a sonodynamic treatment method based on Mn-In ₂ S ₃ /InOOH nanoparticles. The present invention fills this gap. The preparation method of the invention is simple, has good dispersion stability and has great application prospects.

Description of the drawings

The accompanying drawings, which are incorporated in and constitute a part of this specification, illustrate embodiments consistent with the application and together with the description, serve to explain the principles of the application.

Figure 1 is a transmission electron microscope image of In ₂ S ₃ /InOOH nanoparticles with different Mn doping concentrations in the present invention. (a) is an undoped image, (b) is a 1% doped image, and (c) is a 5% doping image. doping map;

Figure 2 is the XRD pattern of In ₂ S ₃ /InOOH nanoparticles with different Mn doping concentrations in the present invention;

Figure 3 is the degradation diagram of DPBF of In ₂ S ₃ /InOOH nanoparticles with different Mn doping concentrations in the present invention after different ultrasonic irradiation times. (a) is the Control (DPBF itself) diagram, (b) is the undoped Doping images, (c) is a 1% doping image, (d) is a 5% doping image;

Figure 4 is a diagram showing the degradation percentage of DPBF by different materials in the present invention;

Figure 5 is a solid diffuse reflection spectrum diagram of different materials in the present invention, (a) is a solid diffuse reflection spectrum diagram, (b) is a solid diffuse reflection conversion curve diagram;

Figure 6 is an AC impedance spectrum diagram of different materials in the present invention;

Figure 7 is a graph of cytotoxicity of different materials after culture for 24 hours in the present invention;

Figure 8 is a flow cytometry apoptosis detection chart under different groups in the present invention;

Figure 9 is a graph of ROS levels in different groups in the present invention. (a) is a ROS fluorescence photo graph, and (b) is a flow cytometer ROS level detection graph.

Detailed ways

Exemplary embodiments will be described in detail herein, examples of which are illustrated in the accompanying drawings. When the following description refers to the drawings, the same numbers in different drawings refer to the same or similar elements unless otherwise indicated. The implementations described in the following exemplary embodiments do not represent all implementations consistent with this application. Rather, they are merely examples of apparatus and methods consistent with aspects of the application as detailed in the appended claims.

The terminology used in this application is for the purpose of describing particular embodiments only and is not intended to be limiting of the application. As used in this application and the appended claims, the singular forms "a," "the" and "the" are intended to include the plural forms as well, unless the context clearly dictates otherwise. It will also be understood that the term "and/or" as used herein refers to and includes any and all possible combinations of one or more of the associated listed items.

The present invention will be further described below in conjunction with the accompanying drawings and specific examples.

It should be understood that the following examples are only used to further illustrate the present invention and cannot be understood as limiting the protection scope of the present invention. Some non-essential improvements and adjustments made by those skilled in the art based on the above content of the present invention all belong to the present invention. protected range. The specific process parameters of the following examples are only an example of the appropriate range, that is, those skilled in the art can make selections within the appropriate range through the description herein, and are not limited to the specific numerical values of the examples below.

Example 1

A method for preparing Mn-In ₂ S ₃ /InOOH nanoparticles may include the following steps:

Step (1), add MnCl ₂ 4H ₂ O and In(NO ₃ ) ₃ to ultrapure water, and disperse with ultrasonic to obtain Mn and In source solutions;

Specifically, weigh 3.5 mg of MnCl ₂ ·4H ₂ O and 72 mg of In(NO ₃ ) ₃ (24mM), dissolve them in 10 ml of ultrapure water, and disperse them with an ultrasonic cleaner for 10 minutes to obtain a uniform, clear and transparent solution.

Step (2), add Na ₂ S to ultrapure water and disperse it ultrasonically to obtain a Na ₂ S solution;

Specifically, 144 mg of Na ₂ S was weighed, dissolved in 10 ml of ultrapure water, and dispersed with an ultrasonic cleaner for 10 minutes to obtain a uniform, clear and transparent solution.

Step (3), add the Mn and In source solution dropwise to the Na ₂ S solution to form a uniform light yellow solution;

Specifically, the homogeneous transparent clear solution obtained by mixing the Mn and In sources was slowly dropped into the Na ₂ S solution at a dropping speed of 2 ml/min, and stirred evenly to form a light yellow solution.

Step (4), add the dilute nitric acid solution dropwise to the above light yellow solution, adjust the pH to below 3, and obtain the Mn-In ₂ S ₃ /InOOH precursor solution;

Specifically, dilute nitric acid (10% wt concentrated nitric acid) was slowly added dropwise to the above mixed solution, and a pH meter was used to detect the real-time pH change of the solution. Adjust the pH of the solution to 2.67, and then continue stirring at room temperature for 10 minutes to obtain a Mn-In ₂ S ₃ /InOOH precursor solution.

Step (5), transfer the Mn-In ₂ S ₃ /InOOH precursor solution to the hydrothermal reactor for reaction, and centrifuge and wash after the reaction to obtain the Mn-In ₂ S ₃ /InOOH solution;

Specifically, the obtained solution was transferred to a hydrothermal reaction kettle with a volume of 50 ml and sealed. The reaction kettle was placed in a 180°C oven to react for 12 hours. It was naturally cooled to room temperature and then centrifuged and washed 3-4 times. After washing, Mn-In was obtained. ₂ S ₃ /InOOH nanoparticles (1-MISO). For solid-liquid separation after the reaction, high-speed centrifugation at 12,000 rpm was used for 8 minutes. The described washing method is to use ultrapure water for washing, and the washed product is redispersed in ultrapure water.

Example 2

A method for preparing Mn-In ₂ S ₃ /InOOH nanoparticles may include the following steps:

Step (1), add MnCl ₂ 4H ₂ O and In(NO ₃ ) ₃ to ultrapure water, and disperse with ultrasonic to obtain Mn and In source solutions;

Specifically, weigh 17.5 mg of MnCl ₂ ·4H ₂ O and 72 mg of In(NO ₃ ) ₃ (24mM), dissolve them in 10 ml of ultrapure water, and disperse them with an ultrasonic cleaner for 10 minutes to obtain a uniform, clear and transparent solution.

Step (2), add Na ₂ S to ultrapure water and disperse it ultrasonically to obtain a Na ₂ S solution;

Specifically, 144 mg of Na ₂ S was weighed, dissolved in 10 ml of ultrapure water, and dispersed with an ultrasonic cleaner for 10 minutes to obtain a uniform, clear and transparent solution.

Step (3), add the Mn and In source solutions dropwise to the Na ₂ S precursor solution to form a uniform light yellow solution;

Specifically, the homogeneous transparent clear solution obtained by mixing the Mn and In sources was slowly dropped into the Na ₂ S solution at a dropping speed of 2 ml/min, and stirred evenly to form a light yellow solution.

Step (4), add the dilute nitric acid solution dropwise to the above light yellow solution, adjust the pH to below 3, and obtain the Mn-In ₂ S ₃ /InOOH precursor solution;

Specifically, dilute nitric acid (10% wt concentrated nitric acid) was slowly added dropwise to the above mixed solution, and a pH meter was used to detect the real-time pH change of the solution. Adjust the pH of the solution to 2.67, and then continue stirring at room temperature for 10 minutes to obtain a Mn-In ₂ S ₃ /InOOH precursor solution.

Step (5), transfer the Mn-In ₂ S ₃ /InOOH precursor solution to the hydrothermal reactor for reaction, and centrifuge and wash after the reaction to obtain the Mn-In ₂ S ₃ /InOOH solution;

Specifically, the obtained solution was transferred to a hydrothermal reaction kettle with a volume of 50 ml and sealed. The reaction kettle was placed in a 180°C oven for reaction for 12 hours. It was naturally cooled to room temperature and then centrifuged and washed 3-4 times. After washing, Mn-In was obtained. ₂ S ₃ /InOOH nanoparticles (5-MISO). For solid-liquid separation after the reaction, high-speed centrifugation at 12,000 rpm was used for 8 minutes. The described washing method is to use ultrapure water for washing, and the washed product is redispersed in ultrapure water.

Example 3

A method for preparing In ₂ S ₃ /InOOH nanoparticles may include the following steps:

Step (1), add In(NO ₃ ) ₃ to ultrapure water and disperse it ultrasonically to obtain an In source solution;

Specifically, weigh 72 mg of In(NO ₃ ) ₃ (24 mM), dissolve it in 10 ml of ultrapure water, and disperse it with an ultrasonic cleaner for 10 minutes to obtain a uniform, clear and transparent solution.

Step (2), add Na ₂ S to ultrapure water and disperse it ultrasonically to obtain a Na ₂ S solution;

Specifically, 144 mg of Na ₂ S was weighed, dissolved in 10 ml of ultrapure water, and dispersed with an ultrasonic cleaner for 10 minutes to obtain a uniform, clear and transparent solution.

Step (3), add the In source solution dropwise to the Na ₂ S precursor solution to form a uniform light yellow solution;

Specifically, the uniform transparent clear solution after mixing the In source was slowly dropped into the Na ₂ S solution at a dropping speed of 2 ml/min, and stirred evenly to form a light yellow solution.

Step (4), add the dilute nitric acid solution dropwise to the above light yellow solution, adjust the pH to below 3, and obtain the In ₂ S ₃ /InOOH precursor solution;

Specifically, dilute nitric acid (10% wt concentrated nitric acid) was slowly added dropwise to the above mixed solution, and a pH meter was used to detect the real-time pH change of the solution. Adjust the pH of the solution to 2.67, and then continue stirring at room temperature for 10 minutes to obtain an In ₂ S ₃ /InOOH precursor solution.

Step (5), transfer the In ₂ S ₃ /InOOH precursor solution to the hydrothermal reactor for reaction, and centrifuge and wash after the reaction to obtain the In ₂ S ₃ /InOOH solution;

Specifically, the obtained solution was transferred to a hydrothermal reaction kettle with a volume of 50 ml and sealed. The reaction kettle was placed in an oven at 180°C for 12 hours, cooled naturally to room temperature, and then centrifuged and washed 3-4 times. After washing, In ₂ S was obtained. ₃ /InOOH nanoparticles (0-MISO). For solid-liquid separation after the reaction, high-speed centrifugation at 12,000 rpm was used for 8 minutes. The described washing method is to use ultrapure water for washing, and the washed product is redispersed in ultrapure water.

Example 4

The difference between this example and Example 1 is that 36mg of In(NO ₃ ) ₃ (12mM) was added to step (1) to obtain the Mn and In source solution; 72mg of Na ₂ S (30mM) was added to step (2). A Na2S solution was obtained.

Example 5

The difference between this embodiment and Example 1 is that the dropping speed in step (3) is 0.5 ml/min.

Example 6

The difference between this embodiment and Example 1 is that in step (4), the dilute nitric acid solution is added dropwise to the above light yellow solution, and the pH is adjusted to 3.0 to obtain an In ₂ S ₃ /InOOH precursor solution.

Example 7

The difference between this example and Example 1 is that in the hydrothermal reaction in step (5), the reaction time is 24 hours.

Mn-In ₂ S ₃ /InOOH nanoparticles were prepared by a one-step hydrothermal method. Pictures (a), (b), and (c) in Figure 1 are transmission electron microscope pictures of In ₂ S ₃ /InOOH nanoparticles with different Mn doping concentrations. It can be seen that different doping concentrations do not Change the morphology of the nanoparticles, the size is about 20nm, and the shape is irregular.

Figure 2 shows the XRD patterns of In2S3/InOOH nanoparticles doped with different Mn concentrations. It can be seen that In ₂ S ₃ /InOOH nanoparticles were successfully synthesized, and their crystal phases are β-In ₂ S ₃ (JCPDS 25-0390) and InOOH phase (JCPDS 17-0549) with a tetragonal structure, and the crystallization condition is good; so JCPDS 25-0390 is the PDF card number. In addition, it can be seen that the doping of Mn affects the ratio of In ₂ S ₃ and InOOH in the composite material, with less Mn doping (1-MISO) relative to undoped nanoparticles (0-MISO) , Na _0.6 MnO ₂ phase appears (JCPDS 69-0060), the proportion of In ₂ S ₃ in more Mn doping (5-MISO) decreases, and an obvious peak of In(OH) ₃ phase appears at 22.26° (JCPDS 01-85-1338).

Example 8

This embodiment provides an application of Mn-In ₂ S ₃ /InOOH nanoparticles prepared in the above embodiment in sonodynamic therapy of tumors.

The application includes: under ultrasonic irradiation, both In ₂ S ₃ and InOOH can absorb ultrasonic waves (US) to realize the transition of electrons from the valence band to the conduction band. Due to the formation of heterojunction and the different costs of In ₂ S ₃ and InOOH meter energy level, according to the principle of minimum energy, electrons in the conduction band of In ₂ S ₃ will migrate to the conduction band of InOOH; holes in the valence band of InOOH will migrate to the valence band of In ₂ S ₃ , thus promoting The separation of electrons and holes increases the quantum yield. In addition, the doping of metal ions Mn can form a defect energy level near the conduction band of In ₂ S ₃ and InOOH, thereby capturing the activated electrons during their transition to the valence band, thereby further reducing the recombination of electron holes. , promoting the separation of electrons and holes. The activated electrons can undergo a reduction reaction with surface-adsorbed O ₂ to produce toxic O ₂ · ^- and ¹ O ₂ , thereby killing tumor cells.

The following examples characterize the performance of the MISO nanoparticles in Examples 1, 2, and 3.

Example 9

The typical ROS detection probe DPBF was used to detect the ROS generation ability of MISO nanoparticles under ultrasonic irradiation. Reactive oxygen species (ROS, including O ₂ · ^- , ¹ O _{2 ,} etc.) generated during the sonodynamic process can degrade DPBF and reduce the characteristic absorption peak of DPBF at 415 nm. 100 μl of DPBF (2mM) was added to 3 ml of MISO hydroalcoholic mixed solution (50 μg/ml), and then the mixture was exposed to ultrasound (1.0MHZ, 1.0W/cm ² ) in the dark, and UV-visible spectroscopy was used to detect differences. Absorbance change under irradiation time (every two minutes).

Figure 3 shows the sonodynamic degradation curves of DPBF by In ₂ S ₃ /InOOH nanoparticles with different Mn doping concentrations. It can be seen that MISO can effectively degrade DPBF under ultrasonic irradiation (US), which demonstrates the sonodynamic properties of MISO. Figure 4 shows the degradation percentage of DPBF by different materials. It can be found that the degradation percentages of Control, 0-MISO, 1-MISO, and 5-MISO within 10 minutes are 16.68%, 53.94%, 100%, and 96.51% respectively. Among them, 1-MISO shows the strongest sonodynamic efficiency. This shows that the heterojunction of In ₂ S ₃ /InOOH promotes ultrasonic power; Mn doping is beneficial to improving the sonodynamic efficiency of In ₂ S ₃ /InOOH. But too much doping will reduce its activity.

Example 10

In order to scientifically study the reasons for the different properties of MISO nanoparticles, this study used solid diffuse reflectance spectroscopy to calculate the band gap of MISO with different Mn doping concentrations.

(a) in Figure 5 shows the solid diffuse reflectance spectrum of MISO, and (b) shows the conversion curve calculated based on Figure 5(a) and the Tauc equation. It can be seen from Figure 5(a) that Mn doping is beneficial to improving the material’s absorption of ultrasonic waves. The band gaps of different materials can be calculated from Figure 5(b). It can be found that the band gaps of 0-MISO, 1-MISO, and 5-MISO are 3.72eV, 2.677eV, and 3.2398eV respectively. The band gap of 0-MISO confirms that the formation of heterojunction is conducive to the generation of electron holes. The effect of Mn doping on the band gap means that metal ions act as defect energy levels. However, increasing the doping concentration cannot continuously reduce the band gap. At the same time, this study also used electrochemical impedance spectroscopy (EIS) to evaluate the charge transfer capability and carrier separation efficiency of MISO. Figure 6 shows that the EIS Nyquist plot of Mn-doped nanoparticles shows a smaller semicircle than 0-MISO, indicating that its charge transfer resistance becomes smaller. Mn doping can induce higher electron-hole separation efficiency, further confirming that Metal ion Mn doping acts as a defect energy level that can capture transition electrons and promote the separation of electrons and holes. Therefore, it can be speculated that the mechanism is: first, InOOH has a band gap of 3.75eV, and In ₂ S ₃ has a band gap of 2.12 eV. Both semiconductors can cause the separation of electrons and holes while absorbing ultrasonic energy, The compatibility of the two can cause the transition of their respective electrons from the valence band to the conduction band when they absorb ultrasonic energy. Due to the difference in the Fermi level of In ₂ S ₃ and InOOH and the principle of minimum energy, In ₂ S ₃ conducts The electrons in the band migrate to the conduction band of InOOH, and the holes in the valence band of InOOH migrate to the valence band of In ₂ S ₃ . On the other hand, the doping of Mn causes a defect energy level near the conduction band of In2S3 and InOOH, which can capture electrons during the electron-hole recombination process. The synergistic effect of heterojunction and doping reduces the recombination of electrons and holes in the material and improves the quantum yield. In addition, we expect that the redistributed activated electrons can drive the reduction of dissolved oxygen molecules present in the environment, producing toxic O ₂ · ^- , ¹ O ₂ .

Example 11

This experiment illustrates the application of the material in tumor treatment through its killing effect at the cellular level. The cells used were mouse breast cancer cells (4T1), and the material used was 1-MISO. 4T1 cells were co-cultured with 50 μg/ml 1-MISO, and the cell survival rate after 24 h of culture with or without ultrasonic irradiation was measured. As shown in Figure 7, ultrasound itself and the material itself have no killing effect on cells. However, under the combined action of materials and ultrasound, the survival rate of 4T1 cells is significantly reduced, showing noteworthy sonodynamic efficiency.

This experiment also used the principle of double staining of Annexin V-fluorescein isothiocyanate (Annexin-FTIC) and PI to further verify the sonodynamic effect of the material using flow cytometry. As shown in Figure 8, most cells were killed by MISO+US treatment, indicating the high ultrasound toxicity displayed by ultrasound-assisted MISO on cells. It itself has very good biocompatibility.

The ability of the material to generate ROS in cells was verified through the oxidation-sensitive probe 2’,7’-dichlorofluorescein diacetate (DCFH-DA). This probe can be oxidized into dichlorofluorescein (DCF) in the presence of ROS and exhibits green fluorescence. Figure 9(a) shows the fluorescence images of cells after being treated by different groups. It can be found that cells treated with MISO+US show enhanced fluorescence, which proves that MISO can produce ultrasound-induced ROS. However, US itself and MISO itself did not show obvious DCF fluorescence. Figure 9(b) uses flow cytometry to quantitatively measure the ROS levels in cells after treatment in different groups, further confirming the above conclusion.

Claims (5)

1. A method for preparing Mn-In2S3/InOOH nanoparticles, which is characterized by comprising: Weigh 3.5mg of MnCl2·4H2O and 72mg, 24mM of In(NO3)3, dissolve them in 10ml of ultrapure water, and disperse them with an ultrasonic cleaner for 10 minutes to obtain a uniform, clear and transparent solution; Weigh 144 mg of Na2S, dissolve it in 10 ml of ultrapure water, and disperse it with an ultrasonic cleaner for 10 minutes to obtain a uniform, clear and transparent solution; Slowly add the homogeneous transparent clear solution after mixing the Mn and In sources into the Na2S solution at a dropping speed of 2ml/min, stir evenly to form a light yellow solution; Slowly add 10%wt concentrated nitric acid dropwise into the above mixed solution, and use a pH meter to detect the real-time pH change of the solution; adjust the pH of the solution to 2.67, and then continue stirring at room temperature for 10 minutes to obtain the Mn-In2S3/InOOH precursor solution; Transfer the obtained solution to a hydrothermal reaction kettle with a volume of 50 ml and seal it. Place the reaction kettle in a 180°C oven to react for 12 hours. After natural cooling to room temperature, centrifuge and wash 3-4 times. After washing, Mn-In2S3/InOOH is obtained. Nanoparticles; the solid-liquid separation after the reaction is completed uses high-speed centrifugation at 12,000 rpm for 8 minutes; the washing method is to use ultrapure water for washing, and the washed products are redispersed in ultrapure water. 2. A method for preparing Mn-In2S3/InOOH nanoparticles, which is characterized by including: Weigh 17.5mg of MnCl2·4H2O and 72mg, 24mM of In(NO3)3, dissolve them in 10ml of ultrapure water, and disperse them with an ultrasonic cleaner for 10 minutes to obtain a uniform, clear and transparent solution; Weigh 144 mg of Na2S, dissolve it in 10 ml of ultrapure water, and disperse it with an ultrasonic cleaner for 10 minutes to obtain a uniform, clear and transparent solution; Slowly add the homogeneous transparent clear solution after mixing the Mn and In sources into the Na2S solution at a dropping speed of 2ml/min, stir evenly to form a light yellow solution; Slowly add 10%wt concentrated nitric acid dropwise into the above mixed solution, and use a pH meter to detect the real-time pH change of the solution; adjust the pH of the solution to 2.67, and then continue stirring at room temperature for 10 minutes to obtain the Mn-In2S3/InOOH precursor solution; Transfer the obtained solution to a hydrothermal reaction kettle with a volume of 50 ml and seal it. Place the reaction kettle in a 180°C oven for reaction for 12 hours. After natural cooling to room temperature, centrifuge and wash 3-4 times. After washing, Mn-In2S3/InOOH is obtained. Nanoparticles; the solid-liquid separation after the reaction is completed uses high-speed centrifugation at 12,000 rpm for 8 minutes; the washing method is to use ultrapure water for washing, and the washed products are redispersed in ultrapure water. 3. The preparation method of Mn-In2S3/InOOH nanoparticles as claimed in claim 1 or 2, characterized in that the Mn-In2S3/InOOH nanoparticles have a particle size of 20nm and an irregular shape. 4. A Mn-In2S3/InOOH nanoparticle prepared by the preparation method described in any one of claims 1-3. 5. The use of Mn-In2S3/InOOH nanoparticles as claimed in claim 4 in the preparation of sonodynamic therapy preparations.