BiVO4-at-TiO2 nano-composite sound-sensitive agent, preparation method thereof and application of BiVO4-at-TiO2 nano-composite sound-sensitive agent in sonodynamic antitumor
By developing BiVO4@TiO2 nanocomposite sound sensitivity agent, the problems of high risks and side effects of traditional glioma treatment methods have been solved, and efficient and biosafety acoustic dynamic treatment effects have been achieved.
Patent Information
- Application Number
- CN202510369719.0
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-03-27
- Publication Date
- 2025-05-16
AI Technical Summary
Traditional glioma treatment methods such as surgery, radiotherapy, chemotherapy, etc. have high risks and serious side effects, and it is difficult to completely prevent glioma recurrence.
A BiVO4@TiO2 nanocomposite sound sensitizer was developed, and BiVO4 and TiO2 nanomaterials were combined through the preparation method to form a composite sound sensitizer with high efficiency acoustic dynamic response and biosafety.
This compound sound sensitizer significantly reduces cell viability under ultrasound irradiation, has good acoustic dynamic treatment effect, selective killing of glioma cells, and reduces side effects.
Smart Images
Figure CN120000788A_ABST
Abstract
Description
Technical Field
[0001] The invention belongs to the technical field of medical materials, and specifically relates to a BiVO4@TiO2 nanocomposite sonosensitizer and a preparation method thereof and application thereof in sonodynamic anti-tumor treatment. Background Art
[0002] Glioma (GM) is the most common primary malignant brain tumor with poor prognosis, high recurrence rate, median survival of 15 months and 5-year survival rate of <5%. The most commonly used treatment method is surgical resection combined with chemotherapy or radiotherapy. Since GM cells are highly infiltrative and invasive, traditional surgical resection cannot completely prevent the recurrence of GM, and traditional treatment methods such as surgery, radiotherapy, chemotherapy, etc. are often accompanied by high risks and serious side effects. 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 method for cancer treatment 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 BiVO4@TiO2 nanocomposite sonosensitizer and its preparation method and application in sonodynamic anti-tumor. 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:
[0005] A BiVO4@TiO2 nanocomposite sonosensitizer, wherein the BiVO4@TiO2 nanocomposite sonosensitizer contains 40% TiO2 by mass.
[0006] The preparation method of the above-mentioned BiVO4@TiO2 nanocomposite sonosensitizer comprises the following steps: dissolving bismuth nitrate pentahydrate in 50 mL of HNO3 solution with a mass concentration of 10%, then adding citric acid thereto, stirring until colorless, to obtain solution A; dissolving ammonium metavanadate in 50 mL of deionized water, stirring at 80°C until the solution is a clear lemon yellow, then adding citric acid thereto, to obtain a brown solution, which is solution B; adding solution A dropwise into solution B to form a dark blue mixture, then adding TiO2, adjusting the pH value to 6.5 with ammonia water, forming a gel under continuous stirring at 100°C, drying, and finally calcining, and grinding to obtain the BiVO4@TiO2 nanocomposite sonosensitizer.
[0007] Furthermore, in the above preparation method, the amount of bismuth nitrate pentahydrate added is 4.85 g, the amount of ammonium metavanadate added is 1.17 g, the amount of citric acid added twice is 4.2 g, and the amount of TiO2 added is 1.25 g.
[0008] Furthermore, in the above preparation method, the speed of adding solution A to solution B is 20-30 mL / min.
[0009] Furthermore, in the above preparation method, the drying condition is 100° C. for 12 h.
[0010] Furthermore, in the above preparation method, the calcination condition is calcination at 500° C. for 2 hours.
[0011] The application of the above-mentioned BiVO4@TiO2 nanocomposite sonosensitizer in sonodynamic anti-tumor therapy.
[0012] Furthermore, in the above application, the anti-tumor effect is the inhibition of human glioma cells.
[0013] Furthermore, the above application method is as follows: human glioma cells in the logarithmic growth phase after 24 hours of culture are added with BiVO4@TiO2 composite sonosensitizer diluted with liquid culture medium, incubated for 24 hours, then irradiated with ultrasound, and cultured for another 24 hours.
[0014] Furthermore, in the above application, the ultrasonic irradiation time is 1 min and the ultrasonic frequency is 1.0 MHz.
[0015] Furthermore, in the above application, the concentration of the BiVO4@TiO2 composite sonosensitizer is 200 μg / mL.
[0016] The beneficial effects of the present invention are:
[0017] 1. High cavitation threshold and limited treatment area are the main limitations of SDT, while the introduction of nanomaterials as nucleation sites in the treatment medium significantly reduces the threshold for cavitation. - will be transferred to the CB of TiO2, and the h generated by the VB of TiO2 + will be transferred to the VB of BiVO4, which can promote the generation of e - -h + The EVB of BiVO4 and TiO2 are both higher than the standard reaction potential of ·OH / H2O. Therefore, the surface h + and H2O and OH - The reaction generates OH.
[0018] 2. The BiVO4@TiO2 composite sonosensitizer formed a heterojunction structure complex BiVO4@TiO2. When the addition amount was 200μg / mL, after 1MHz US irradiation for 1min, the viability of the U251 glioma cell line decreased from 73.12%±0.4% to 29.12±2.84%, which had a good sonodynamic therapy effect. BRIEF DESCRIPTION OF THE DRAWINGS
[0019] Figure 1 XRD patterns of BiVO4@TiO2, TiO2 and BiVO4.
[0020] Figure 2 This is the SEM image of BiVO4.
[0021] Figure 3 This is the SEM image of TiO2.
[0022] Figure 4 This is the SEM image of BiVO4@TiO2.
[0023] Figure 5 Comparison of cell viability with and without ultrasonic irradiation at different concentrations of BiVO4@TiO2.
[0024] Figure 6 Comparison of cell viability of 200 μg / mL BiVO4@TiO2, TiO2 and BiVO4 with / without ultrasonic irradiation (one-way ANOVA, *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001).
[0025] Figure 7 This is the AO / EB experimental results of BiVO4@TiO2. DETAILED DESCRIPTION
[0026] Example 1
[0027] (I) Preparation of BiVO4@TiO2 nanocomposite sonosensitizer
[0028] 4.85g Bi(NO3)3·5H2O and 1.17g NH4VO3 were accurately weighed, and Bi(NO3)3·5H2O was dissolved in 50mL HNO3 solution (mass concentration was 10%), and 4.2g citric acid was added and stirred until colorless to obtain solution A for standby use; NH4VO3 was dissolved in 50mL deionized water, and magnetic stirring was performed at 80°C until the solution was clear lemon yellow, and 4.2g citric acid was added to form a brown solution, which was solution B. Solution A was added dropwise to solution B at a rate of 25mL / min to form a dark blue mixture, and 1.25g TiO2 was added and the pH was adjusted to 6.5 with ammonia water, and a gel was formed under continuous stirring at 100°C, and then dried at 100°C overnight, calcined at 500°C for 2h, and ground to obtain BiVO4@TiO2 nanocomposite sonosensitizer.
[0029] (II) Preparation of pure phase BiVO4
[0030] Accurately weigh 4.85g Bi(NO3)3·5H2O and 1.17g NH4VO3, dissolve Bi(NO3)3·5H2O in 50mL HNO3 solution (mass concentration is 10%), add 4.2g citric acid and stir until colorless to obtain solution A for standby use; dissolve NH4VO3 in 50mL deionized water, stir magnetically at 80℃ until the solution is clear lemon yellow, add 4.2g citric acid to form a brown solution, which is solution B. Solution A is added dropwise to solution B at a rate of 25mL / min to form a dark blue mixture, adjust the pH to 6.5 with ammonia water, form a gel under continuous stirring at 100℃, dry at 100℃ overnight, calcine at 500℃ for 2h, grind, and obtain BiVO4.
[0031] (iii) Pure phase TiO2 is purchased industrial grade TiO2.
[0032] (IV) XRD analysis of pure BiVO4, pure TiO2 and BiVO4@TiO2 nanocomposite sonosensitizers
[0033] like Figure 1 As shown, the peak positions of the pure phase single substance are basically consistent with those of BiVO4@TiO2, and the peak positions at 19°, 25.3°, 28.7°, etc. all show characteristic peaks corresponding to the pure phase single substance, proving the successful preparation of the material.
[0034] (V) Scanning electron microscopy (SEM) analysis of the morphology and microstructure of BiVO4, TiO2 and BiVO4@TiO2
[0035] like Figure 2 , 3 4 are SEM images of BiVO4, TiO2 and BiVO4@TiO2, respectively. Figure 2 BiVO4 presents a nanosheet structure. Figure 3 The TiO2 in the sample is in the form of nano-spheres. Figure 4 There are some attached spherical structures on the BiVO4@TiO2 nanosheets, indicating the successful composite of TiO2.
[0036] Example 2 Application of BiVO4@TiO2 in sonodynamic therapy
[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 BiVO4@TiO2 composite sonosensitizer.
[0038] U251 glioma cells in the logarithmic growth phase were uniformly inoculated into a 96-well plate. After culturing for 24 hours, BiVO4, TiO2 and BiVO4@TiO2 (concentrations of 0, 12.5, 25, 50, 100 and 200 μg / mL, respectively) diluted in liquid culture medium were added and incubated with the cells for 24 hours. The cells were irradiated with US for 1 minute with a US parameter of 1.0 MHz and 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 a microplate reader. Further processing was performed to obtain the therapeutic effect at the cellular level.
[0039] like Figure 5 When the addition amount of the composite material BiVO4@TiO2 was 0μg / mL, that is, the Control group, only the effect of ultrasound on the survival rate of U251 was examined. After 60s of ultrasonic irradiation, the cell viability reached 80.64%±1.22%, indicating that simple ultrasonic irradiation has a certain killing effect on U251 cells, but the effect is limited.
[0040] like Figure 5 When the concentration of the composite material BiVO4@TiO2 was 12.5μg / mL and 25μg / mL, the cell viability was 77.47%±5.42% and 79.33%±0.2% respectively after 60s of ultrasonic irradiation. This shows that when the concentration of the sonosensitizer is low, its effect is not much different from that of simple ultrasound, and ultrasonic irradiation plays the main role.
[0041] like Figure 5 When the concentration of the composite material BiVO4@TiO2 was increased again to 50 and 100 μg / mL, the cell viability decreased to 63.72%±7.53% and 55.92%±6.06% respectively after 60s of ultrasonic irradiation. This indicates that after the concentration of the material was increased, the material was excited by ultrasound and the reactive oxygen produced killed the cancer cells.
[0042] like Figure 5Finally, when the concentration reached 200 μg / mL, the cell viability dropped to 29.11% ± 2.84%. It is worth mentioning that with the increase of concentration, the material also showed microcytotoxicity, and the cell viability dropped to 73.12% ± 0.41%. It can be concluded that the composite material BiVO4@TiO2 has good sonodynamic activity and has microcytotoxicity when it exceeds the half lethal concentration.
[0043] Figure 6 After determining the concentration of the composite material BiVO4@TiO2 to be 200μg / mL, it was verified whether the U251 killing effect of the composite material BiVO4@TiO2 was better than that of the pure phase element at this concentration. The results showed that after 60s of ultrasonic irradiation, the cell viability of pure phase TiO2 dropped to 80.64±1.22%, that of pure phase BiVO4 was 63.00%±6.49%, and that of the composite material BiVO4@TiO2 was 43.21%±2.67%, which was significantly lower than that of pure phase TiO2. Figure 5 The results proved to be consistent, the material has good sonodynamic activity, exceeds the half lethal concentration and also has slight cytotoxicity. On the basis of pure phase TiO2, the introduction of BiVO4 semiconductor material enhances the performance of the material.
[0044] Example 3 AO / EB experiment of BiVO4@TiO2 to verify its sonodynamic activity
[0045] Figure 7 This is an AO / EB staining experiment of cells. Acridine orange (AO) can penetrate cells with intact cell membranes and embed into nuclear DNA, causing them to emit bright green fluorescence. Ethidium bromide (EB) can only penetrate cells with damaged cell membranes and embed into nuclear DNA, emitting orange-red fluorescence. Apoptotic cells show enhanced staining, brighter fluorescence, and uniform round or condensed or lumpy structures. Non-apoptotic cell nuclei show structural characteristics with varying depths of fluorescence. The two are very different in morphology and are easy to distinguish. Under a fluorescence microscope, four cell morphologies can be seen: living cells (VN), with green nuclear chromatin and normal structure; early apoptotic cells (VA), with green nuclear chromatin and condensed or beaded; non-apoptotic dead cells (NVN), with orange-red nuclear chromatin and normal structure; late apoptotic cells (NVA), with orange-red nuclear chromatin and condensed or beaded.
[0046] U251 cells in the logarithmic growth phase were irradiated with ultrasound for 60 seconds and then incubated for one day for AO / EB staining. The results showed that the composite material BiVO4@TiO2 enhanced its SDT performance after ultrasound irradiation compared with the pure phase single substance, causing U251 cells to undergo apoptosis or death.
Claims
1. A BiVO4@TiO2 nanocomposite sonosensitizer, characterized in that: The BiVO4@TiO2 nanocomposite sonosensitizer contains 40% TiO2 by mass percentage.
2. The method for preparing a BiVO4@TiO2 nanocomposite sonosensitizer according to claim 1, characterized in that: The method comprises the following steps: dissolving bismuth nitrate pentahydrate in 50 mL of HNO3 solution with a mass concentration of 10%, then adding citric acid thereto, stirring until colorless, to obtain solution A; dissolving ammonium metavanadate in 50 mL of deionized water, stirring at 80 DEG C until the solution is clear lemon yellow, then adding citric acid thereto, to obtain a brown solution, which is solution B; dropping solution A into solution B to form a dark blue mixture, then adding TiO2, adjusting the pH value to 6.5 with ammonia water, forming a gel under continuous stirring at 100 DEG C, drying, finally calcining, and grinding to obtain the BiVO4@TiO2 nanocomposite sonosensitizer.
3. The preparation method according to claim 2, characterized in that: The amount of bismuth nitrate pentahydrate added was 4.85 g, the amount of ammonium metavanadate added was 1.17 g, the amount of citric acid added twice was 4.2 g, and the amount of TiO2 added was 1.25 g.
4. The preparation method according to claim 2, characterized in that: The rate of adding solution A to solution B is 20-30 mL / min.
5. The preparation method according to claim 2, characterized in that: The drying condition is 100°C for 12 h; the calcination condition is 500°C for 2 h.
6. Use of the BiVO4@TiO2 nanocomposite sonosensitizer according to claim 1 in sonodynamic anti-tumor therapy.
7. The use according to claim 6, characterized in that: The anti-tumor effect is to inhibit human glioma cells.
8. The use according to claim 7, characterized in that: The method is as follows: BiVO4@TiO2 composite sonosensitizer diluted in liquid culture medium is added to human glioma cells in the logarithmic growth phase after 24 hours of culture, and then ultrasonically irradiated after 24 hours of incubation, and cultured for another 24 hours.
9. The use according to claim 8, characterized in that: The ultrasonic irradiation time is 1 min, and the ultrasonic frequency is 1.0 MHz.
10. The use according to claim 8, characterized in that: The concentration of the BiVO4@TiO2 composite sonosensitizer is 200 μg / mL.