ZnWO4 / TiO2 composite material and application of preparation method thereof in sonodynamic induction of tumor apoptosis
By using ZnWO4/TiO2 composite materials as sound sensitivity agents, apoptosis of glioblastoma cells was induced in acoustic dynamic therapy, which solved the problem that traditional treatment methods were difficult to completely prevent glioblastoma recurrence, and achieved efficient and biosafety acoustic dynamic therapy effect.
Patent Information
- Application Number
- CN202411631578.7
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-11-15
- Publication Date
- 2025-02-28
AI Technical Summary
The existing methods for treating glioblastoma have limitations. Traditional surgical resection is difficult to completely prevent recurrence, and traditional therapies have side effects, so new non-invasive treatments need to be sought.
Using ZnWO4/TiO2 composite material as a sound sensitivity agent, tumor apoptosis is induced by ultrasonic radiation in acoustic dynamic therapy, and using the efficiency and biosafety of the composite material to provide a new therapeutic strategy.
ZnWO4/TiO2 compound sound sensitizer showed a damage rate of 21.65±6.14% on U251 glioma cells under 1MHz ultrasonic irradiation, which significantly improved the efficacy of sound dynamic therapy. The material was non-toxic at high concentrations and had good biosafety.
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Figure CN119528213A_ABST
Abstract
Description
Technical Field
[0001] The present invention belongs to the field of medical technology, and specifically relates to a ZnWO4 / TiO2 composite material and a preparation method thereof, and an application thereof in sonodynamically inducing tumor apoptosis. Background Art
[0002] Glioblastoma (GM) is the most common primary malignant tumor of the nervous system. It has diverse sites of disease occurrence, can destroy functional areas, and has higher disability and mortality rates than other brain diseases. It is one of the most intractable tumors in the world. Currently, the only treatment for this disease is surgical resection or combined radiotherapy and chemotherapy to delay the progression of the disease. Moreover, due to the high infiltrative and invasive nature of GM cells, traditional surgical resection cannot completely prevent recurrence. As an emerging non-invasive treatment method, sonodynamic therapy (SDT) is considered to be a strong competitor for the treatment of tumors because of its advantages such as high selectivity and few side effects. The present invention provides a new sonosensitizer for the treatment of brain gliomas by systematically studying the application of TiO2-based composites in SDT. Summary of the invention
[0003] The purpose of the present invention is to provide a preparation method of a ZnWO4 / TiO2 composite sonosensitizer and its application in sonodynamic therapy. The composite sonosensitizer is highly efficient and has high biosafety, provides a new strategy for the treatment of tumors, and has good application prospects.
[0004] The technical solution adopted by the present invention is: a ZnWO4 / TiO2 composite material, wherein the ZnWO4 / TiO2 composite material contains 1%-10% of ZnWO4 by mass percentage.
[0005] The above-mentioned preparation method of a ZnWO4 / TiO2 composite material is as follows: a certain mass of Zn(COOCH3)2·2H2O and TiO2 are weighed and ultrasonically dispersed in distilled water, and then Na2WO4·2H2O is weighed and dissolved in distilled water. Under magnetic stirring, the Na2WO4·2H2O solution is dropped into the TiO2 dispersion liquid, stirred at room temperature for 30 minutes, and then transferred to a reactor for hydrothermal reaction. The obtained reactant is filtered, washed, and dried. After drying, it is ground into powder and weighed for standby use.
[0006] In the above preparation method, the molar ratios of Zn(COOCH3)2·2H2O:TiO2:Na2WO4·2H2O are 1:150:1, 1:80:1, 1:50:1 and 1:25:1 respectively.
[0007] In the above-mentioned preparation method, the hydrothermal reaction conditions are: reaction temperature at 453K for 24 hours.
[0008] The drying conditions of the above preparation method are drying at a temperature of 353 K for 4 hours.
[0009] The above-mentioned ZnWO4 / TiO2 composite material is used for the application in sonodynamic induction of tumor apoptosis.
[0010] The above application method is as follows: ZnWO4 / TiO2 composite material is added to the U251 glioma cell line as a model of sonodynamically induced tumor apoptosis, and ultrasonic irradiation is performed.
[0011] In the above application, U251 glioma cells in the logarithmic growth phase were added with ZnWO4 / TiO2 composite materials diluted in liquid culture medium and incubated with the cells for 24 hours, followed by ultrasonic irradiation, and the cell activity was detected after further culturing for 24 hours.
[0012] In the above application, the concentration of the added ZnWO4 / TiO2 composite sonosensitizer was 0 μg / mL, 12.5 μg / mL, 25 μg / mL, 50 μg / mL, 100 μg / mL and 200 μg / mL.
[0013] In the above application, the ultrasonic irradiation time is 1 min and the ultrasonic frequency is 1.0 MHz.
[0014] The beneficial effects of the present invention are
[0015] The application of SDT in cancer treatment is increasing worldwide, but the invention of effective and non-toxic sonosensitizers is still the top priority of SDT research. The introduction of nanomaterials as nucleation sites for SDT significantly reduces the threshold for cavitation effect and improves the efficacy of SDT. The results of Example 5 show that the ZnWO4 / TiO2 composite sonosensitizer forms a heterojunction structure complex ZT-3. When the addition amount is 200μg / mL, after 1MHz US irradiation for 1min, the damage to the U251 glioma cell line reaches 21.65±6.14%, which has a good sonodynamic therapy effect. BRIEF DESCRIPTION OF THE DRAWINGS
[0016] Figure 1 XRD patterns of ZnWO4 / TiO2 composite materials prepared with different ZnWO4 mass ratios.
[0017] Figure 2 The SEM images of pure TiO2, pure ZnWO4 and ZT-3 composite materials are shown in Figure 1. (A) TiO2; (B) ZnWO4; (C) ZT-3.
[0018] Figure 3EDX images of ZnWO4 / TiO2 composite materials, where (A) is the SEM image of the test area, (B), (C), (D), (E), and (F) are the EDX images of W, O, Ti, C, and Zn elements in the ZnWO4 / TiO2 composite materials, respectively.
[0019] Figure 4 This is a comparison chart of the active oxygen generation effect of ZnWO4 / TiO2 composite materials.
[0020] Figure 5 This is a comparison chart of the SDT results of ZT-3, TiO2 and ZnWO4.
[0021] Figure 6 This is a comparison chart of SDT results of different concentrations of ZT-3. DETAILED DESCRIPTION
[0022] The present invention is described in detail below with reference to the embodiments.
[0023] Example 1 Preparation of ZnWO4 / TiO2 Composite Material
[0024] Accurately weigh Zn(COOCH3)2·2H2O and TiO2 and disperse them in 30mL of distilled water under ultrasonic action. Then weigh Na2WO4·2H2O and dissolve it in 30mL of distilled water. Under magnetic stirring, drop the Na2WO4·2H2O solution into the dispersion of Zn(COOCH3)2·2H2O and TiO2. Stir thoroughly for 30min, transfer the mixed solution into a 100mL polytetrafluoroethylene reactor, and react at 453K for 24h. After the reaction is completed, cool naturally to room temperature, filter, and wash repeatedly with distilled water and anhydrous ethanol. The obtained white product is dried at 353K for 4h, ground into powder with an agate mortar, and weighed for use. According to the different amounts of Zn(COOCH3)2·2H2O and Na2WO4·2H2O added, ZT-1, ZT-3, ZT-5, and ZT-10 are obtained respectively.
[0025] In Example 1, the added amounts (g) of Na2WO4·2H2O, Zn(COOCH3)2·2H2O and TiO2 are respectively ZT-1: 0.0191, 0.0182, 1.0069, ZT-3: 0.0514, 0.0345, 1.0064, ZT-5: 0.0976, 0.0582, 1.0057, and ZT-10: 0.1521, 0.1036, 1.0076.
[0026] Preparation of pure ZnWO4: Weigh 1.0975g of Zn(COOCH3)2·2H2O and 1.6491g of Na2WO4·2H2O and fully dissolve them. Then, mix and stir the two solutions for 30 minutes and transfer them to a 100mL polytetrafluoroethylene liner. Then put the liner into a reactor and fully react in a constant temperature drying oven at 453K for 24h. Finally, filter, wash and dry at 353K for 4h. After drying, grind into powder, which is ZnWO4.
[0027] Example 2 XRD analysis of pure phase TiO2, pure phase ZnWO4 and prepared ZT-1, ZT-3, ZT-5 and ZT-10 composite materials
[0028] like Figure 1 As shown, the results show that the peak positions of pure phase elements and composites are basically the same, and with the increase of composite ratio, the characteristic peak intensity of ZnWO4 increases, and the characteristic peak intensity of TiO2 weakens. ZT-1, ZT-3, ZT-5, and ZT-10 composite materials were successfully prepared.
[0029] Example 3 Scanning electron microscope (SEM) analysis of the morphology and microstructure of pure phase TiO2, pure phase ZnWO4 and ZT-3.
[0030] Use as Figure 2 As shown in (A), the prepared TiO2 is nano-spherical, evenly distributed, compact in structure, and has a particle size of about 25nm; Figure 2 (B) shows that the prepared ZnWO4 is in the form of nanoparticles and is evenly distributed; Figure 2 In (C), ZnWO4 is evenly distributed on the TiO2 flakes, and the whole is in the shape of nanospheres with uniform and compact particle size.
[0031] Example 4 EDX spectrum analysis of ZT-3 composite material
[0032] like Figure 3 As shown, W, O, Ti, and Zn elements are present in the prepared ZT-3 sample. The results show that the elements Zn, W, and Ti are evenly distributed in the material, and the distribution of Ti and O elements is more dense, indicating that the composite ZT-3 was successfully synthesized.
[0033] Example 5 Comparison of active oxygen generation effects of ZnWO4 / TiO2 composite materials
[0034] Under US irradiation, the sonosensitizer can react with the solution to generate a large amount of reactive oxygen. The ability to generate reactive oxygen can reflect the activity of the material, and this result can be used as a basis for screening the best material. The DPBF (1,3-diphenylisobenzofuran) probe can be oxidized by reactive oxygen, resulting in a decrease in its characteristic ultraviolet absorption peak at 411nm, and is widely used to detect the reactive oxygen generation activity of materials.
[0035] like Figure 4 Compared with pure phase TiO2 and pure phase ZnWO4, ZT-3 with a composite ratio of 3% of ZnWO4 has the most significant effect. Therefore, ZT-3 is used as an example in subsequent experiments.
[0036] Example 6 Application of ZT-3 in sonodynamic induction of tumor apoptosis
[0037] The U251 glioma cell line was used as a model for sonodynamic antitumor research, and the cell activity after ultrasound irradiation was used as an indicator to investigate the sonodynamic antitumor activity of the ZnWO4 / TiO2 composite sonosensitizer.
[0038] U251 glioma cells in the logarithmic growth phase were uniformly inoculated into a 96-well plate and cultured for 24 hours. Then, materials diluted with culture medium were added and incubated with the cells for 24 hours, followed by US irradiation for 1 minute with a US parameter of 1.0 MHz. The cells were then cultured for another 24 hours. The activity of surviving cells was detected by the MTT method, and the absorption at 490 nm was tested by an enzyme marker. Further processing was performed to obtain the therapeutic effect at the cellular level.
[0039] like Figure 5 , when the material concentration was 200μg / mL, after 1min of ultrasonic irradiation, the cell activity remained above 99%, indicating that the material was non-toxic even at a high concentration of 200μg / mL. The survival rate of cells treated with US alone after SDT was 72.52±3.44%. Under the good effect of the cell survival rate of the ZnWO4+US group reduced to 53.56±6.08% and the cell survival rate of the TiO2+US group reduced to 38.23±2.59%, the cell survival rate of ZT-3 as a composite material after SDT was only 21.65±6.14%. The results showed that when the drug concentration was 200μg / mL, the sonosensitizer showed significant cell sonotoxicity after US, among which the sonodynamic therapy effect of ZT-3 was significantly better than that of the US treatment groups of ZnWO4 and TiO2, and had a good therapeutic effect.
[0040] Example 7 Effects of different concentrations of ZT-3 on apoptosis of glioma cells U251
[0041] The pro-apoptotic effect of different concentrations of ZT-3 on glioma U251 cells was investigated. U251 cell lines were incubated with gradient concentrations for 24 hours and then irradiated with US. Cell activity was measured after another 24 hours of incubation.
[0042] like Figure 6 , the added concentration of ZT-3 was 12.5 μg·mL -1 , 25 μg·mL -1 , 50μg·mL -1 , 100 μg·mL -1 and 200 μg mL -1 When irradiated with US, the cell activity decreased to 49.08%, 43.84%, 39.60%, 28.60% and 21.65%. Compared with the control group and the -US group, +US irradiation had a good killing effect on U251 cancer cells, and the effect was enhanced with the increase of ZT-3 concentration.
[0043] (Using one-way ANOVA, *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001).
Claims
1. A ZnWO4 / TiO2 composite material, characterized in that: The ZnWO4 / TiO2 composite material contains 1%-10% of ZnWO4 by mass percentage.
2. The method for preparing a ZnWO4 / TiO2 composite material according to claim 1, characterized in that: Weigh a certain mass of Zn(COOCH3)2·2H2O and TiO2 and ultrasonically disperse them in distilled water. Then weigh Na2WO4·2H2O and dissolve it in distilled water. Under magnetic stirring, add the Na2WO4·2H2O solution dropwise into the TiO2 dispersion. After stirring at room temperature for 30 minutes, transfer the solution to a reactor for hydrothermal reaction. Filter the obtained reactants, wash and dry them. After drying, grind them into powder and weigh them for later use.
3. The preparation method according to claim 2, characterized in that: In molar ratios, Zn(COOCH3)2·2H2O:TiO2:Na2WO4·2H2O are 1:150:1, 1:80:1, 1:50:1 and 1:25:1 respectively.
4. The preparation method according to claim 2, characterized in that: The hydrothermal reaction conditions are 453 K for 24 h.
5. The preparation method according to claim 2, characterized in that: The drying conditions are: drying at 353K for 4 hours.
6. Use of the ZnWO4 / TiO2 composite material according to claim 1 in sonodynamically inducing tumor apoptosis.
7. The use according to claim 6, characterized in that: The method is as follows: ZnWO4 / TiO2 composite material is added to the U251 glioma cell line as a model of sonodynamically induced tumor apoptosis, and ultrasonic irradiation is performed.
8. The use according to claim 7, characterized in that: U251 glioma cells in the logarithmic growth phase were incubated with ZnWO4 / TiO2 composite materials diluted in liquid culture medium for 24 hours and then irradiated with ultrasound. Cell activity was detected after further culturing for 24 hours.
9. The use according to claim 8, characterized in that: The concentrations of the added ZnWO4 / TiO2 composite sonosensitizer were 0 μg / mL, 12.5 μg / mL, 25 μg / mL, 50 μg / mL, 100 μg / mL and 200 μg / mL.
10. The use according to claim 8, characterized in that: The ultrasonic irradiation time was 1 min, and the ultrasonic frequency was 1.0 MHz.