Preparation method and application of nanoparticles for ultrasonic treatment of osteosarcoma
Through the preparation method of modified tin dioxide nanoparticles, the problem of low utilization rate of sound sensitizers in acoustic dynamic treatment osteosarcoma is solved, targeted tumor delivery and multiple strikes are achieved, and efficient and non-invasive osteosarcoma treatment plan is provided.
Patent Information
- Application Number
- CN202510417117.8
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-04-03
- Publication Date
- 2025-07-04
AI Technical Summary
The current sound-sensitive agent utilization rate of acoustic dynamics in the treatment of osteosarcoma is low, resulting in poor treatment effect. Traditional treatment methods such as surgical resection and chemotherapy have side effects, making it difficult to achieve accurate and efficient targeted tumor delivery and non-invasive treatment.
Using tin dioxide nanoparticles as carriers, by introducing oxygen vacancies and modifying with folic acid, the acoustic dynamic performance and biocompatibility are enhanced. The prepared nanoparticles can mediate the tumor-targeted delivery of therapeutic agents, and use the tips of octahedral nanoparticles to penetrate the cell membrane to achieve multiple blows.
Efficient and non-invasive osteosarcoma treatment has been achieved. By combining acoustic dynamic therapy and physical damage under low-dose ultrasound radiation, it enhances the treatment effect and reduces side effects, providing a new strategy for the accurate and efficient treatment of clinical osteosarcoma.
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Figure CN120242038A_ABST
Abstract
Description
Technical Field
[0001] The present invention belongs to the technical field of biomedicine, and specifically relates to a preparation method and application of nanoparticles for ultrasonic treatment of osteosarcoma. Background Art
[0002] Osteosarcoma is the most common type of primary bone malignancy, which develops from mesenchymal cell lines. Most cases occur in children and young people aged 10 to 30 years, with the peak incidence during puberty. The main clinical manifestations include fractures, osteolytic lesions, synovial joint cavity effusion, and peripheral nerve compression, etc. Its prevalence accounts for about 0.2% of all malignancies. Currently, the main treatment strategies for osteosarcoma in clinical practice mainly include surgical resection, radiotherapy, and chemotherapy. Surgical resection often causes extensive damage to surrounding normal tissues and cells, and residual tumor cells are prone to cause tumor recurrence and distant metastasis. Chemotherapeutic drugs (such as cisplatin, doxorubicin, ifosfamide) lack selectivity and sensitivity to tumor cells, and their side effects of organ damage seriously weaken the therapeutic effect of osteosarcoma. In addition, most bone tumors are insensitive to radiation, and the risk of radiotherapy in areas such as the spine is relatively high. Therefore, developing a safe and efficient new method for osteosarcoma treatment, inducing specific cell death in tumor lesions, delaying tumor metastasis and recurrence, prolonging the survival time of patients, and reducing treatment side effects is a frontier topic in scientific research.
[0003] Sonodynamic therapy (SDT) is a treatment method that uses low-frequency (<3 MHz) and low-intensity (<3 W / cm2) ultrasound (US) to excite a photosensitizer to generate reactive oxygen species (ROS) to damage tumor cells, and has characteristics such as the unique high tissue penetration ability of ultrasound and excellent biosafety. First, the US used in SDT is a mechanical wave that can penetrate tissues up to several centimeters and reach deep tumor sites. Second, by controlling the US conditions, deep tumor tissues can be accurately located, and different combinations of US frequencies and intensities can bring excellent treatment effects, providing new ideas and directions for cancer treatment. In recent years, the number of reports related to SDT has been increasing year by year. Especially in cancer treatment, SDT has been used to treat various cancer types such as breast cancer, melanoma, glioma, pancreatic cancer, gastric cancer, and lung cancer. The US Food and Drug Administration (FDA) approved SDT for clinical treatment experiments of recurrent glioblastoma and other malignant gliomas through the fast track in 2022. As of now, multiple sonodynamic clinical treatment projects including the treatment of atherosclerosis and glioma have been approved by the FDA.
[0004] However, the utilization rate of sonosensitizers greatly restricts the efficacy of SDT. Sonosensitizers mainly include two categories: organic sonosensitizers and inorganic sonosensitizers. Organic sonosensitizers include porphyrin, porphyrin derivatives, and some chemotherapeutic drugs such as doxorubicin (DOX) and curcumin. However, these organic sonosensitizers have poor water solubility, low accumulation in tumor tissues, and low bioavailability. Inorganic sonosensitizers include titanium dioxide (TiO2) and manganese dioxide (MnO2), etc. Compared with organic sonosensitizers, these inorganic sonosensitizers exhibit more excellent performance in the field of SDT. For example, these inorganic sonosensitizers have unique semiconductor properties and can generate more ROS by triggering the generation of electrons (e - ) and holes (h + ). At the same time, the size and morphology of these inorganic materials are easier to control, which helps their further application in tumor targeting or drug delivery. Therefore, developing multifunctional inorganic sonosensitizer nanomaterials is an excellent strategy to improve sonodynamic therapy efficacy. Summary of the Invention
[0005] The technical problem to be solved by the present invention is to provide a preparation method of nanoparticles for ultrasonic treatment of osteosarcoma in view of the deficiencies of the above-mentioned prior art. The present invention uses tin dioxide with sonosensitive properties as a carrier, modifies its outer surface by introducing oxygen vacancies to enhance its sonodynamic performance, and performs surface modification with folic acid to improve its biocompatibility. The prepared nanoparticles can mediate the tumor-targeted delivery of therapeutic agents. At the same time, the tip of the octahedral nanoparticles endows the nanoparticles with stronger ability to penetrate cell membranes, and has the functions of efficiently integrating tumor-targeted delivery, physical killing, and sonosensitivity. It can perform multiple strikes on osteosarcoma to achieve the purpose of highly efficient and non-invasive treatment of osteosarcoma, and can achieve the purpose of enhancing efficacy and reducing toxicity in the treatment of osteosarcoma by combining sonodynamic therapy and physical damage under low-dose ultrasonic irradiation, providing new methods and strategies for the precise, efficient, and combined treatment of clinical osteosarcoma.
[0006] To solve the above technical problems, the technical solution adopted by the present invention is: a preparation method of nanoparticles for ultrasonic treatment of osteosarcoma, characterized by including the following steps:
[0007] Step 1: Add stannous chloride dihydrate and polyvinylpyrrolidone to an ethanol aqueous solution in sequence to obtain a reaction system. Stir the reaction system for 0.3 h to 1 h, stop the reaction, then add concentrated hydrochloric acid and transfer it to an autoclave, and keep it at a constant temperature of 100 °C to 200 °C for 5 h to 15 h. Subsequently, centrifuge, and wash the precipitate with water and ethanol to obtain tin dioxide nanoparticles;
[0008] Step 2: Vacuum-dry the tin dioxide nanoparticles obtained in Step 1, and then place them in a tube furnace and calcine them at 200 °C to 600 °C for 2 h to 5 h to obtain oxygen vacancy-modified tin dioxide nanoparticles;
[0009] Step 3: Disperse the oxygen vacancy-modified tin dioxide nanoparticles described in Step 2 in the folic acid-polyethylene glycol-thiol solution, stir vigorously at 4°C for 12 h to 24 h, then centrifuge and wash with deionized water to obtain the nanoparticles for ultrasonic treatment of osteosarcoma.
[0010] The method for preparing the nanoparticles for ultrasonic treatment of osteosarcoma as described above is characterized in that the particle size of the tin dioxide nanoparticles in Step 1 is 150 nm to 250 nm.
[0011] The method for preparing the nanoparticles for ultrasonic treatment of osteosarcoma as described above is characterized in that the concentration of stannous chloride dihydrate in the reaction system in Step 1 is 0.005 M to 0.2 M.
[0012] The method for preparing the nanoparticles for ultrasonic treatment of osteosarcoma as described above is characterized in that the concentration of polyvinylpyrrolidone in the reaction system in Step 1 is 0.05 g / mL to 0.3 g / mL.
[0013] The method for preparing the nanoparticles for ultrasonic treatment of osteosarcoma as described above is characterized in that the volume ratio of ethanol to water in the ethanol aqueous solution in Step 1 is 1:(1 to 4).
[0014] The method for preparing the nanoparticles for ultrasonic treatment of osteosarcoma as described above is characterized in that the volume of the concentrated hydrochloric acid in Step 1 is 9% to 17% of the volume of the ethanol aqueous solution.
[0015] The method for preparing the nanoparticles for ultrasonic treatment of osteosarcoma as described above is characterized in that the concentration of the folic acid-polyethylene glycol-thiol solution in Step 3 is 50 μg / mL to 150 μg / mL.
[0016] The method for preparing the nanoparticles for ultrasonic treatment of osteosarcoma as described above is characterized in that the mass ratio of the oxygen vacancy-modified tin dioxide nanoparticles to folic acid-polyethylene glycol-thiol in Step 3 is 1:(1 to 10).
[0017] Furthermore, the present invention provides a kind of nanoparticles for ultrasonic treatment of osteosarcoma prepared by the above preparation method.
[0018] Even further, the present invention provides an application of the above nanoparticles in the preparation of medicaments for treating osteosarcoma.
[0019] The present invention has the following advantages compared with the prior art:
[0020] 1. The nanoparticles for ultrasonic treatment of osteosarcoma according to the present invention have good biocompatibility and stability, and do not cause adverse reactions in vivo and in vitro; they can be actively targeted and enriched in the tumor site; the exposed octahedral tips can easily penetrate the cell membrane, improving the sonodynamic therapeutic effect.
[0021] 2. The present invention uses tin dioxide with sonosensitizing properties as a carrier, modifies its outer surface by introducing oxygen vacancies to enhance its sonodynamic properties, and performs surface modification with folic acid to improve its biocompatibility. The prepared nanoparticles can mediate the tumor-targeted delivery of therapeutic agents. At the same time, the tips of the octahedral nanoparticles endow the nanoparticles with a stronger ability to penetrate the cell membrane, integrating multiple functions of efficient tumor-targeted delivery, physical killing, and sonosensitization. It can perform multiple strikes on osteosarcoma, achieving the purpose of efficient and non-invasive treatment of osteosarcoma. By combining sonodynamic therapy and physical damage under low-dose ultrasonic irradiation, it can achieve the purpose of enhancing efficacy and reducing toxicity in the treatment of osteosarcoma, providing new methods and strategies for the precise, efficient, and combined treatment of clinical osteosarcoma.
[0022] The following further describes the technical solutions of the present invention in detail with reference to the drawings and embodiments. Description of the Drawings
[0023] Figure 1 Transmission electron microscope photograph of SnO 2-x -FA nanoparticles synthesized in Example 1 of the present invention.
[0024] Figure 2 Scanning electron microscope photograph of SnO 2-x -FA nanoparticles synthesized in Example 1 of the present invention.
[0025] Figure 3 X-ray diffraction pattern of SnO 2-x and SnO 2-x -FA nanoparticles synthesized in Example 1 of the present invention and the X-ray diffraction pattern of standard SnO2 (No. 41-1445).
[0026] Figure 4 Oxygen levels generated by SnO2, SnO 2-x and SnO 2-x -FA nanoparticles synthesized in Example 6 of the present invention at different times under ultrasonic action. 1 O2 level.
[0027] Figure 5 Cytotoxic effects of different concentrations of SnO 2-x -FA nanoparticles synthesized in Example 6 of the present invention and their ultrasonic irradiation on HOS cells.
[0028] Figure 6The SnO synthesized in Example 6 of the present invention 2-x -FA nanoparticles' effect on the cell viability of NIH-3T3 cells. Detailed implementation manners
[0029] The present invention will be specifically described below through examples, which are only used for further illustration of the present invention and cannot be construed as limiting the protection scope of the present invention. For the experimental methods without specific conditions noted in the examples, they are generally carried out according to the conventional conditions and the conditions described in the manuals, or according to the conditions recommended by the manufacturers; for the equipment, materials, reagents, etc., if not otherwise specified, they can be obtained from commercial channels.
[0030] Example 1
[0031] This example provides SnO 2-x -FA nanoparticles for ultrasonic treatment of osteosarcoma, and its preparation method includes:
[0032] Step 1: Add stannous chloride dihydrate and 0.5 g of polyvinylpyrrolidone to 8 mL of an ethanol aqueous solution (volume ratio of ethanol to water is 1:1) in sequence to obtain a reaction system, and the concentration of stannous chloride dihydrate in the reaction system is 0.005 M; stir the reaction system for 0.6 h, stop stirring, then add 1.2 mL of concentrated hydrochloric acid (concentration 36% - 38%) and transfer it to a polytetrafluoroethylene reaction kettle, keep it at a constant temperature of 200 °C for 12 h, cool the reaction kettle to room temperature, collect the product by centrifugation, and wash the precipitate once with water and once with ethanol to obtain tin dioxide nanoparticles;
[0033] Step 2: Vacuum-dry the tin dioxide nanoparticles obtained in Step 1 at 60 °C, then place them in a tubular furnace and calcine at 200 °C for 5 h to obtain oxygen vacancy-modified tin dioxide nanoparticles, hereinafter referred to as SnO 2-x ;
[0034] Step 3: Disperse 1 mg of the SnO 2-x obtained in Step 2 in 10 mL of a folic acid-polyethylene glycol-thiol solution with a concentration of 100 μg / mL, stir vigorously at 4 °C for 20 h, then centrifuge and wash 3 times with ultrapure water to obtain nanoparticles SnO 2-x -FA for ultrasonic treatment of osteosarcoma.
[0035] Performance evaluation
[0036] Figure 1 is the transmission electron microscope photo of the SnO 2-x -FA nanoparticles in Example 1. Figure 2 is the scanning electron microscope photo of the SnO 2-x -FA nanoparticles in Example 1. According to Figure 1 and Figure 2 it can be seen that SnO2-x The particle size of the -FA nanoparticles is 150 - 250 nm.
[0037] Figure 3 For SnO in Example 1 2-x and SnO 2-x The X-ray diffraction patterns of the -FA nanoparticles and the X-ray diffraction pattern of standard SnO2 (No. 41-1445). The testing method includes: using XRD to conduct crystal structure test and analysis on the freeze-dried sample powder. According to Figure 3 It can be seen that the stronger characteristic diffraction peaks in the SnO 2-x nanoparticles match the diffraction peaks of No. 41-1445 in the XRD standard card (JCPDS), confirming the successful preparation of the SnO 2-x nanoparticles. Meanwhile, after being modified with folic acid, the characteristic peaks of the SnO 2-x nanoparticles are still maintained.
[0038] Example 2
[0039] This example provides a kind of SnO 2-x -FA nanoparticles for ultrasonic treatment of osteosarcoma, and its preparation method includes:
[0040] Step 1: Add stannous chloride dihydrate and 1.2 g of polyvinylpyrrolidone into 4 mL of ethanol aqueous solution (the volume ratio of ethanol to water is 1:2) in sequence to obtain a reaction system, and the concentration of stannous chloride dihydrate in the reaction system is 0.055 M; stir the reaction system for 0.5 h, stop stirring, then add 0.4 mL of concentrated hydrochloric acid (concentration 36% - 38%), and transfer it into a polytetrafluoroethylene reaction kettle, keep it at a constant temperature of 180 °C for 15 h, cool the reaction kettle to room temperature, collect the product by centrifugation, and wash the precipitate with water and ethanol once each to obtain tin dioxide nanoparticles;
[0041] Step 2: Vacuum-dry the tin dioxide nanoparticles obtained in Step 1 at 60 °C, then place them in a tube furnace and calcine them at 350 °C for 4 h to obtain oxygen vacancy-modified tin dioxide nanoparticles, hereinafter referred to as SnO 2-x ;
[0042] Step 3: Disperse 1 mg of the SnO 2-x obtained in Step 2 in 70 mL of folic acid-polyethylene glycol-thiol solution with a concentration of 100 μg / mL, stir vigorously at 4 °C for 12 h, then centrifuge, and wash with ultrapure water 3 times to obtain the nanoparticles SnO 2-x -FA for ultrasonic treatment of osteosarcoma.
[0043] The physical and chemical properties of the SnO 2-x -FA nanoparticles prepared in this example are basically the same as those in Example 1.
[0044] Example 3
[0045] This example provides SnO 2-x -FA nanoparticles for ultrasonic treatment of osteosarcoma, and its preparation method includes:
[0046] Step 1: Add stannous chloride dihydrate and 1.5 g of polyvinylpyrrolidone into 12 mL of ethanol aqueous solution (volume ratio of ethanol to water is 1:4) in sequence to obtain a reaction system, and the concentration of stannous chloride dihydrate in the reaction system is 0.2 M; stir the reaction system for 0.3 h, stop stirring, then add 2 mL of concentrated hydrochloric acid (concentration 36% - 38%) and transfer it into a polytetrafluoroethylene reaction kettle, keep it at a constant temperature of 200 °C for 8 h, cool the reaction kettle to room temperature, collect the product by centrifugation, and wash the precipitate with water and ethanol once each to obtain tin dioxide nanoparticles;
[0047] Step 2: Vacuum-dry the tin dioxide nanoparticles obtained in Step 1 at 60 °C, then place them in a tubular furnace and calcine at 350 °C for 4 h to obtain oxygen vacancy-modified tin dioxide nanoparticles, hereinafter referred to as SnO 2-x ;
[0048] Step 3: Disperse 1 mg of the SnO 2-x obtained in Step 2 in 100 mL of folic acid-polyethylene glycol-thiol solution with a concentration of 100 μg / mL, stir vigorously at 4 °C for 24 h, then centrifuge and wash with ultrapure water 3 times to obtain nanoparticles SnO 2-x -FA for ultrasonic treatment of osteosarcoma.
[0049] The physicochemical properties of the SnO 2-x -FA nanoparticles prepared in this example are basically the same as those in Example 1.
[0050] Example 4
[0051] This example provides SnO 2-x -FA nanoparticles for ultrasonic treatment of osteosarcoma, and its preparation method includes:
[0052] Step 1: Add stannous chloride dihydrate and 2 g of polyvinylpyrrolidone into 20 mL of ethanol aqueous solution (volume ratio of ethanol to water is 1:3) in sequence to obtain a reaction system, and the concentration of stannous chloride dihydrate in the reaction system is 0.15 M; stir the reaction system for 1 h, stop stirring, then add 1.8 mL of concentrated hydrochloric acid (concentration 36% - 38%) and transfer it into a polytetrafluoroethylene reaction kettle, keep it at a constant temperature of 100 °C for 10 h, cool the reaction kettle to room temperature, collect the product by centrifugation, and wash the precipitate with water and ethanol once each to obtain tin dioxide nanoparticles;
[0053] Step 2: Vacuum dry the tin dioxide nanoparticles described in Step 1 at 60°C, then place them in a tube furnace and calcine at 600°C for 2 h to obtain oxygen vacancy-modified tin dioxide nanoparticles, hereinafter referred to as SnO 2-x ;
[0054] Step 3: Disperse 1 mg of the SnO 2-x described in Step 2 in 45 mL of a folic acid-polyethylene glycol-thiol solution with a concentration of 50 μg / mL, vigorously stir at 4°C for 24 h, then centrifuge and wash 3 times with ultrapure water to obtain the nanoparticles SnO 2-x -FA for ultrasonic treatment of osteosarcoma.
[0055] The physicochemical properties of the SnO 2-x -FA nanoparticles prepared in this example are basically the same as those in Example 1.
[0056] Example 5
[0057] This example provides SnO 2-x -FA nanoparticles for ultrasonic treatment of osteosarcoma, and its preparation method includes:
[0058] Step 1: Add stannous chloride dihydrate and 0.8 g of polyvinylpyrrolidone to 10 mL of an ethanol aqueous solution (volume ratio of ethanol to water is 1:3) in sequence to obtain a reaction system, and the concentration of stannous chloride dihydrate in the reaction system is 0.1 M; stir the reaction system for 0.7 h, stop stirring, then add 0.9 mL of concentrated hydrochloric acid (concentration 36% - 38%) and transfer it to a polytetrafluoroethylene reaction kettle, keep it at a constant temperature of 130°C for 5 h, cool the reaction kettle to room temperature, collect the product by centrifugation, and wash the precipitate once with water and once with ethanol to obtain tin dioxide nanoparticles;
[0059] Step 2: Vacuum dry the tin dioxide nanoparticles described in Step 1 at 60°C, then place them in a tube furnace and calcine at 400°C for 3.5 h to obtain oxygen vacancy-modified tin dioxide nanoparticles, hereinafter referred to as SnO 2-x ;
[0060] Step 3: Disperse 1 mg of the SnO 2-x described in Step 2 in 30 mL of a folic acid-polyethylene glycol-thiol solution with a concentration of 150 μg / mL, vigorously stir at 4°C for 18 h, then centrifuge and wash 3 times with ultrapure water to obtain the nanoparticles SnO 2-x -FA for ultrasonic treatment of osteosarcoma.
[0061] The physicochemical properties of the SnO 2-x -FA nanoparticles prepared in this example are basically the same as those in Example 1.
[0062] Example 6
[0063] This embodiment provides SnO 2-x -FA nanoparticles for ultrasonic treatment of osteosarcoma, and the preparation method thereof includes:
[0064] Step 1: Add stannous chloride dihydrate and 1 g of polyvinylpyrrolidone into 15 mL of ethanol aqueous solution (volume ratio of ethanol to water is 1:3) in sequence to obtain a reaction system, and the concentration of stannous chloride dihydrate in the reaction system is 0.18 M; stir the reaction system for 0.5 h, stop stirring, then add 1.4 mL of concentrated hydrochloric acid (concentration 36% - 38%), and transfer it into a polytetrafluoroethylene reaction kettle, keep it at a constant temperature of 170 °C for 12 h, cool the reaction kettle to room temperature, collect the product by centrifugation, and wash the precipitate with water and ethanol once each to obtain tin dioxide nanoparticles;
[0065] Step 2: Vacuum-dry the tin dioxide nanoparticles obtained in Step 1 at 60 °C, then place them in a tube furnace, calcine at 300 °C for 4 h to obtain oxygen vacancy-modified tin dioxide nanoparticles, hereinafter referred to as SnO 2-x ;
[0066] Step 3: Disperse 1 mg of the SnO 2-x obtained in Step 2 in 60 mL of folic acid-polyethylene glycol-thiol solution with a concentration of 100 μg / mL, stir vigorously at 4 °C for 24 h, then centrifuge, and wash with ultrapure water 3 times to obtain nanoparticles SnO 2-x -FA for ultrasonic treatment of osteosarcoma.
[0067] The physical and chemical properties of the SnO 2-x -FA nanoparticles prepared in this embodiment are basically the same as those in Example 1.
[0068] Performance evaluation
[0069] Figure 4 For the O2 levels generated by the SnO2, SnO 2-x and SnO 2-x -FA nanoparticles synthesized in Example 6 at different times under ultrasonic irradiation. The test method includes: using the ultraviolet probe 1,3-diphenylisobenzofuran (DPBF) to detect the O2 level generated by SnO 1 -FA under ultrasonic irradiation. Add 60 μL of DMSO solution containing DPBF (1 mg / mL) into 1 mL of SnO2, SnO 2-x and SnO 1 -FA solutions with a concentration of 100 μg / mL respectively. Perform ultrasonic irradiation on each sample solution for different times (0, 1, 2, 3, 4, 5 min), and the ultrasonic parameters are set to 1 W / cm 2-x 2-x 2 , 5 min, 1 MHz, 50% duty cycle. After sonication, the absorbance of each sample at 425 nm was detected by ultraviolet-visible spectroscopy, and the results are as Figure 4 shown. As can be seen from the figure, under ultrasonic irradiation, the absorbance value of SnO 2-x -FA nanoparticles at 425 nm decreased more significantly, indicating that more 1 O2 was generated by the nanoparticles, suggesting that the nanocomposite SnO 2-x -FA has strong sonosensitizing performance.
[0070] Figure 5 For the SnO 2-x -FA nanoparticles with different concentrations synthesized in Example 6 and their killing effect on HOS cells under ultrasonic irradiation. The test method includes: inoculating HOS cells onto a 96-well plate and allowing them to grow for 24 h, then adding SnO 2-x -FA nanoparticles with different concentrations and incubating for 4 h; subjecting the ultrasonic group to ultrasonic treatment with ultrasonic conditions of 1 W / cm 2 , 5 min, 1 MHz, 50% duty cycle, and continuing to culture for 24 h; discarding the culture medium, adding 100 μL of a 1 mg / mL solution of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide to each well and continuing to culture for 4 h; discarding the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide solution, adding 150 μL of dimethyl sulfoxide to each well, and detecting the absorbance value (OD) at 490 nm with an enzyme-linked immunosorbent assay reader, and calculating the cell survival rate. The survival rate = (OD value of the experimental group - OD value of the blank control group) / (OD value of the control group - OD value of the blank control group) × 100%. The results are as Figure 5 shown. As can be seen from the figure, simple ultrasonic irradiation does not affect cell growth, and the cell survival rate can reach 85.5%. Under ultrasonic irradiation with the same parameters, the cell survival rate gradually decreases with the increase in the concentration of SnO 2-x -FA; when the concentration of SnO 2-x -FA is 100 μg / mL, the cell survival rate drops to 34.5%; when the concentration of SnO 2-x -FA increases to 200 μg / mL, the cell survival rate is only 17.6%. The above experiments show that the sonodynamic effect of SnO 2-x -FA can effectively kill HOS cells.
[0071] Figure 6 For the effect of the SnO 2-x -FA nanoparticles synthesized in Example 6 on the cell viability of mouse fibroblast NIH-3T3 cells. The test method includes: inoculating NIH-3T3 onto a 96-well plate and allowing them to grow for 24 h, then adding SnO 2-x-FA nanoparticles and continue culturing for 24 h, 48 h, and 72 h; discard the culture medium, add 100 μL of 1 mg / mL tetramethyl azoazolium salt solution to each well, and continue culturing for 4 h; discard the tetramethyl azoazolium salt solution, add 150 μL of dimethyl sulfoxide to each well, measure the absorbance value at 490 nm with an enzyme-linked immunosorbent assay (ELISA) reader, calculate the cell survival rate, and the survival rate = (OD value of the experimental group - OD value of the blank control group) / (OD value of the control group - OD value of the blank control group) × 100%. The results are as Figure 6 shown. As can be seen from the figure, SnO 2-x -FA has no obvious effect on cell viability in the concentration range of 0 - 400 μg / mL, indicating that SnO 2-x -FA has good biocompatibility at the cellular level.
[0072] In the present invention, tin dioxide with sonosensitizing properties is used as a carrier, its outer surface is modified by introducing oxygen vacancies to enhance its sonodynamic performance, and it is surface-modified with folic acid to improve its biocompatibility. The prepared nanoparticles can mediate the tumor-targeted delivery of therapeutic agents. At the same time, the tip of the octahedral nanoparticles endows the nanoparticles with a stronger ability to penetrate cell membranes, integrating the functions of efficient tumor-targeted delivery, physical killing, and sonosensitivity. It can perform multiple strikes on osteosarcoma, achieving the purpose of highly non-invasive treatment of osteosarcoma. By combining sonodynamic therapy and physical damage under low-dose ultrasound irradiation, it can achieve the purpose of enhancing efficacy and reducing toxicity in the treatment of osteosarcoma, providing new methods and strategies for the precise, efficient, and combined treatment of clinical osteosarcoma.
[0073] The above are only the preferred embodiments of the present invention, and do not limit the present invention in any way. Any simple modifications, changes, and equivalent structural changes made to the above embodiments according to the technical essence of the invention still fall within the protection scope of the technical solution of the present invention.
Claims
1. A preparation method of nanoparticles for ultrasonic treatment of osteosarcoma, characterized in that, It includes the following steps: Step 1: Add stannous chloride dihydrate and polyvinylpyrrolidone into an ethanol aqueous solution in sequence to obtain a reaction system. Stir the reaction system for 0.3 h to 1 h, stop the reaction, then add concentrated hydrochloric acid and transfer it into an autoclave. Keep it at a constant temperature of 100°C to 200°C for 5 h to 15 h, then centrifuge. Wash the precipitate with water and ethanol to obtain tin dioxide nanoparticles. Step 2: Vacuum-dry the tin dioxide nanoparticles obtained in Step 1, then place them in a tube furnace and calcine them at 200°C to 600°C for 2 h to 5 h to obtain oxygen vacancy-modified tin dioxide nanoparticles. Step 3: Disperse the oxygen vacancy-modified tin dioxide nanoparticles obtained in Step 2 in a folic acid-polyethylene glycol-thiol solution, stir vigorously at 4°C for 12 h to 24 h, then centrifuge and wash to obtain nanoparticles for ultrasonic treatment of osteosarcoma.
2. The preparation method of the nanoparticles for ultrasonic treatment of osteosarcoma according to claim 1, wherein, The particle size of the tin dioxide nanoparticles described in Step 1 is 150 nm to 250 nm.
3. The preparation method of the nanoparticles for ultrasonic treatment of osteosarcoma according to claim 1, wherein, The concentration of stannous chloride dihydrate in the reaction system described in Step 1 is 0.005 M to 0.2 M.
4. The preparation method of the nanoparticles for ultrasonic treatment of osteosarcoma according to claim 1, wherein, The concentration of polyvinylpyrrolidone in the reaction system described in Step 1 is 0.05 g / mL to 0.3 g / mL.
5. The preparation method of the nanoparticles for ultrasonic treatment of osteosarcoma according to claim 1, characterized in that, In the ethanol aqueous solution described in Step 1, the volume ratio of ethanol to water is 1: (1～4)。 6. The preparation method of the nanoparticles for ultrasonic treatment of osteosarcoma according to claim 1, characterized in that, The volume of the concentrated hydrochloric acid described in Step 1 is 9% to 17% of the volume of the ethanol aqueous solution.
7. The preparation method of the nanoparticles for ultrasonic treatment of osteosarcoma according to claim 1, characterized in that, The concentration of the folic acid-polyethylene glycol-thiol solution described in Step 3 is 50 μg / mL to 150 μg / mL.
8. The preparation method of the nanoparticles for ultrasonic treatment of osteosarcoma according to claim 1, characterized in that, The mass ratio of the oxygen vacancy-modified tin dioxide nanoparticles to the folic acid-polyethylene glycol-thiol described in Step 3 is 1:(1 to 10).
9. A nanoparticle for ultrasonic treatment of osteosarcoma prepared by the preparation method according to any one of claims 1 to 8.
10. An application of the nanoparticle according to claim 9 in the preparation of a medicament for treating osteosarcoma.