Barium titanate composite nanomaterial for ultrasonic piezoelectric catalysis and treatment of drug-resistant recurrent glioma

Abstract

The invention discloses a barium titanate composite nanomaterial for ultrasonic piezoelectric catalysis and treatment of drug-resistant recurrent glioma, and belongs to the field of glioma treatment. The preparation method comprises the following steps: firstly, preparing BaTiO3 nano-particles, then sequentially wrapping the surfaces of the BaTiO3 nano-particles with polydopamine (PDA), and loading peroxymonosulfate to obtain the composite piezoelectric material. The obtained composite piezoelectric material can regulate and control the internal and external electronic structures of the piezoelectric material at the same time to enhance activation of peroxymonosulfate and generate. SO4, so that synergistic ultrasonic treatment of glioblastoma is realized; besides, in a slightly acidic tumor microenvironment, imine (-NH-) protonation in PDA enhances adsorption of peroxymonosulfate through hydrogen-bond interaction, further promotes generation of. SO4 <->, finally causes mitochondrial dysfunction, effectively and synergistically enhances the chemotherapy effect of temozolomide on glioblastoma, and overcomes the drug resistance of glioblastoma.

Description

Technical Field

[0001] This invention relates to the field of glioma treatment, specifically to a barium titanate composite nanomaterial for ultrasonic piezoelectric catalysis and treatment of drug-resistant and recurrent gliomas. Background Technology

[0002] Glioblastoma is a primary brain tumor of the central nervous system with a high mortality rate; the median survival after initial diagnosis is only 15 months. The core of glioblastoma treatment is surgery, followed by radiotherapy and adjuvant chemotherapy. Temozolomide, as a lipophilic drug, can easily cross the blood-brain barrier to reach brain tissue and is the first-line chemotherapy drug for treating glioblastoma. The main problem with using temozolomide to treat glioblastoma is the development of temozolomide resistance. Temozolomide and other conventional chemotherapy drugs exert their killing effect on tumor cells by damaging DNA and inducing programmed cell death. In contrast, DNA repair pathways in temozolomide-resistant cells are often impaired; therefore, drug-resistant glioblastoma cells recover from the damage induced by temozolomide through alternative repair pathways, leading to decreased sensitivity of glioblastoma cells to temozolomide and ultimately promoting glioblastoma recurrence.

[0003] The generation of reactive oxygen species (ROS) is a key mechanism leading to cell death. Therefore, "nanocatalytic medicine," a technology utilizing nanomaterials to catalyze the generation of ROS within tumors, has become an important field in cancer treatment. However, the efficacy of currently developed nanomaterials is limited. Mitochondria reduce the effectiveness of ROS-based methods through dihydroorotate dehydrogenase. Furthermore, the hypoxic tumor microenvironment in glioblastoma restricts oxygen (common substrates of ROS, such as hydroxyl radicals (·OH) and singlet oxygen (·OH). 1 The supply of O2 further limited cell death induction. Summary of the Invention

[0004] [Technical Issues] This invention provides a novel piezoelectric material that, upon ultrasonic activation, promotes the death of recurrent glioblastoma cells, overcoming the drug resistance of glioblastoma and improving a novel treatment approach for recurrent gliomas after temozolomide treatment. This paves the way for the development of next-generation nanomedicines for cancer treatment.

[0005] [Technical Solution] This invention provides a method for preparing barium titanate composite nanomaterials, comprising the following steps: (1) Preparation of BaTiO3 nanoparticles (BT): Barium hydroxide and titanium dioxide were dispersed in water and heated to react for a period of time to obtain BT precursor; then the BT precursor was washed with acetic acid, washed with deionized water, centrifuged, dried and ground to obtain BT nanoparticles; (2) Synthesis of BT@PDA nanoparticles: Alkali solution was added to Tris-HCl aqueous solution and the pH was controlled to 8-9. Then, the BT nanoparticles obtained in step (1) were added, ultrasonically treated, and then dopamine hydrochloride was added. The mixture was reacted. After the reaction was completed, the solid and liquid were separated, the solid was collected, washed and dried to obtain BT nanoparticles with polydopamine on the surface, which were denoted as BT@PDA. (3) Synthesis of BT@PDA@PMS nanoparticles: The BT@PDA obtained in step (2) is mixed with persulfate to obtain BT@PDA nanoparticles loaded with persulfate.

[0006] In one embodiment of the present invention, in step (1), the molar ratio of barium hydroxide to titanium dioxide is 1.1 to 1.5:1.

[0007] In one embodiment of the present invention, in step (1), the dispersion concentration of titanium dioxide relative to water is 0.2-0.5 mol / L; specifically, 0.25 mol / L may be selected.

[0008] In one embodiment of the present invention, in step (1), the heating reaction is carried out at 140-170°C for 4-7 hours.

[0009] In one embodiment of the present invention, in step (2), the alkaline solution is an aqueous solution of any one or more of the following alkaline reagents: sodium hydroxide, potassium hydroxide, sodium carbonate, potassium carbonate, and ammonia.

[0010] In one embodiment of the present invention, in step (2), the mass ratio of BT nanoparticles to dopamine hydrochloride is (3-8):1; specifically, 5:1 may be selected.

[0011] In one embodiment of the present invention, in step (2), ultrasonic treatment is performed for 10-20 min.

[0012] In one embodiment of the present invention, in step (2), the mixing reaction is carried out for 15-30 h. Specifically, 24 h may be selected.

[0013] In one embodiment of the present invention, in step (3), persulfate is dissolved in water, then BT@PDA is added, the mixture is stirred and reacted for a period of time, and after the reaction is completed, the solid and liquid are separated, the solid is collected, washed, and dried to obtain the composite piezoelectric material; or, Persulfate and BT@PDA were mixed and ground in an agate mortar for a period of time to ensure that BT@PDA and persulfate were fully and uniformly mixed to obtain a composite piezoelectric material.

[0014] In one embodiment of the present invention, in step (3), the persulfate is potassium persulfate and / or sodium persulfate.

[0015] In one embodiment of the present invention, in step (3), the mass ratio of BT@PDA to persulfate is (1-3):1. Specifically, 1.2:1 may be selected.

[0016] In one embodiment of the present invention, in step (3), the concentration of persulfate dissolved in water is 15-30 mg / mL. Specifically, 20 mg / mL may be selected.

[0017] In one embodiment of the present invention, in step (3), grinding is performed for 1-2 hours.

[0018] This invention provides a barium titanate composite nanomaterial prepared based on the above method.

[0019] The present invention also provides the application of the above-mentioned barium titanate composite nanomaterials in piezoelectric catalysis.

[0020] This invention also provides the application of the above-mentioned barium titanate composite nanomaterials in the catalytic degradation of rhodamine B.

[0021] The present invention also provides the use of the above-mentioned barium titanate composite nanomaterial in the preparation of a medicament for ultrasound treatment of recurrent gliomas resistant to temozolomide.

[0022] Beneficial effects: This invention provides a barium titanate composite nanomaterial and introduces a dual-modulation piezoelectric catalysis strategy, which can simultaneously regulate the internal and external electronic structure of the piezoelectric material to enhance the activation of PMS and the generation of SO₂. 4⁻ This strategy enables synergistic ultrasound therapy for glioblastoma. It utilizes barium titanate nanoparticles coated with polydopamine (PDA) and loaded with PMS (BT@PDA@PMS nanoparticles) to enhance chemosensitivity. Furthermore, in the slightly acidic tumor microenvironment beyond the BT nanoparticles, the protonation of imine (-NH-) in PDA enhances PMS adsorption via hydrogen bonding, further promoting the adsorption of ·SO₄²⁻. 4- The generation of these substances ultimately leads to mitochondrial dysfunction. This synergistic mechanism effectively enhances the chemotherapeutic effect of temozolomide against glioblastoma. This innovative piezoelectric catalytic strategy overcomes drug resistance in glioblastoma and paves the way for the development of next-generation nanomedicines for cancer treatment. Attached Figure Description

[0023] Figure 1 This is a schematic diagram of the synthesis process of BT@PDA@PMS piezoelectric materials.

[0024] Figure 2The image shows the X-ray diffraction (XRD) pattern of the barium titanate precursor in Example 1.

[0025] Figure 3 The image shows the XRD pattern of the BT nanoparticles (Ba:Ti=1.2:1) finally obtained in Example 1.

[0026] Figure 4 The image shows a SEM image of barium titanate with a barium-to-titanium ratio of 1.1:1 to 1.5:1 in Example 1.

[0027] Figure 5 The image shows the PFM diagram of the BT nanoparticles (Ba:Ti=1.2:1) finally obtained in Example 1.

[0028] Figure 6 The concentration curves of BT, BT@PDA, and BT@PDA@PMS for RhB degradation in Example 2 are shown.

[0029] Figure 7 The cell viability after treatment with TMZ and BT@PDA@PMS in Example 3 is shown.

[0030] Figure 8 The results show the mitochondrial morphology of cells after treatment with TMZ and BT@PDA@PMS in Example 4.

[0031] Figure 9 This is a comparison of tumor size before and after BT@PDA@PMS ultrasound treatment in Example 5.

[0032] Figure 10 The results of HE staining of brain tissue after BT@PDA@PMS ultrasound treatment in Example 5 are shown.

[0033] Figure 11 The survival rates of mice in different groups during the 28-day tumorigenesis experiment in Example 5 are shown. The PBS group refers to mice without the BT@PDA@PMS material of this invention, while the PMS group refers to mice with the BT@PDA@PMS material of this invention. Detailed Implementation

[0034] Example 1: Preparation and characterization of BT@PDA@PMS 1. Preparation method A schematic diagram of the synthesis process of BT@PDA@PMS piezoelectric materials is shown below. Figure 1 As shown. Specifically, it includes the following steps: (1) Synthesis of BaTiO3 nanocrystals (BT) by a modified adsorption coprecipitation method: Weigh 0.05 mol of titanium dioxide and weigh barium hydroxide according to a barium-titanium molar ratio of 1.1:1~1.5:1 (1.1:1, 1.2:1, 1.3:1, 1.4:1, 1.5:1). Disperse the barium hydroxide in 200 ml of water in a 250 ml round-bottom flask and heat to 170 °C for 7 h to obtain a barium titanate precursor. Then add 50 ml of acetic acid and react at 50 °C for 5 h to remove barium carbonate impurities. Finally, wash with deionized water and centrifuge ten times, and dry at 50 °C for 12 h to obtain barium titanate (BT). Then grind the BT into BT nanoparticles using an agate mortar.

[0035] (2) Synthesis of BT@PDA@PMS nanoparticles: First, 0.1 g of Tris-HCl was dissolved in 50 mL of water, and then sodium hydroxide solution was added dropwise until the pH value reached 8.5. 0.5 g of BT (Ba:Ti=1.2:1) powder prepared in step (1) was added to the solution. After sonication for 10 minutes, 0.1 g of dopamine hydrochloride was added and the mixture was stirred for 24 h. The solid was separated from the liquid, centrifuged to collect the solid, washed with water 5 times, and then dried in an oven to obtain BT nanoparticles coated with polydopamine, denoted as BT@PDA.

[0036] Subsequently, 0.6 g of BT@PDA nanoparticles and 0.5 g of potassium peroxymonosulfate (PMS) were ground in an agate mortar for 1 h to ensure that the BT@PDA nanoparticles and PMS were thoroughly and uniformly mixed, thus obtaining BT@PDA nanoparticles loaded with PMS, denoted as BT@PDA@PMS.

[0037] Preparation of BT@PMS (Comparative Example): 0.6 g of BT nanoparticles and 0.5 g of potassium peroxymonosulfate (PMS) were ground in an agate mortar for 1 h to ensure that the BT nanoparticles and PMS were thoroughly and uniformly mixed, thus obtaining BT nanoparticles loaded with PMS, denoted as BT@PMS.

[0038] 2. Characterization Methods: X-ray diffraction (XRD) was used to analyze the phase structure of the material, field emission scanning electron microscopy (SEM) was used to observe the microstructure of the material, and piezoelectric microscopy (PFM) was used to evaluate the piezoelectric properties of the material.

[0039] Results: The crystal structure of BT was confirmed by XRD results. Figure 2 The XRD of the barium titanate precursor with a barium-to-titanium ratio of 1.1:1 to 1.5:1 in step (1) shows the presence of barium carbonate impurity. Figure 3The image shows the XRD pattern of barium titanate (Ba:Ti = 1.2:1) after washing with acetic acid. Impurities have been removed, and the image shows a single perovskite phase. Figure 4 The images show SEM images of barium titanate with a barium-to-titanium ratio of 1.1:1 to 1.5:1. The barium titanate with a barium-to-titanium ratio of 1.2:1 has the most regular morphology and the best dispersibility. Figure 5 This is a piezoelectric force microscopy (PFM) image of barium titanate (Ba:Ti=1.2:1), demonstrating that barium titanate has good piezoelectric properties.

[0040] Example 2: Study on the piezoelectric catalytic performance of BT@PDA@PMS 1. Method: The piezoelectric catalytic performance of the material was tested using a Rhodamine B (RhB) degradation solution as a model reaction. 110 mg of BT, 110 mg of BT@PDA, 110 mg of BT@PMS, and 110 mg of BT@PDA@PMS powder were dispersed in 50 mL of RhB aqueous solution (initial concentration 10 ppm). The suspension was then stirred in the dark for 1 hour to reach adsorption-desorption equilibrium between the dye and the material. Ultrasonic testing (power 2.4 W, sound intensity 0.8 W / cm²) was used to assess the piezoelectric catalytic performance. 2 The suspension was treated with a duty cycle of 30%, and 5 ml of the suspension was taken every 2 minutes, filtered, and the UV-Vis absorption spectrum was measured at 554 nm.

[0041] 2. Results: Figure 6 These are the concentration curves of RhB degradation by BT, BT@PDA, and BT@PDA@PMS. BT@PDA@PMS has the highest efficiency in degrading RhB.

[0042] Degradation rate within 10 minutes (min) -1 = (1-C / C0) / 10.

[0043] The specific degradation rate results are shown in Table 1.

[0044] Table 1

[0045] As shown in Table 1, the sum of the degradation rate constants of BT@PDA and BT@PMS is 0.09355 min. -1 The rate constant of BT@PDA@PMS is 3.3 times that of PMS. This demonstrates a synergistic effect among the three components.

[0046] Example 3: Therapeutic effect of BT@PDA@PMS on temozolomide-resistant glioma cells 1. Method: GL261 cells were purchased from Mirror Cell Technology (Shanghai) Co., Ltd., catalog number iCell-0059a.

[0047] After seeding standard GL261 cells in 24-well plates, they were treated with 400 μM temozolomide for 72 hours. The culture medium was then discarded, and the cells were washed twice with PBS, followed by replacement with standard culture medium. This process was repeated after the cells reached confluence. After 10 temozolomide stimulations, the IC50 of both cell types was determined using CCK-8 assays, and the resistance coefficients were calculated. Cells with a resistance coefficient exceeding 10 were defined as temozolomide-resistant GL261 / TMZ cells. Temozolomide-resistant GL261 / TMZ cells were then seeded in 96-well plates. After cell attachment, the cells were treated with 400 μM temozolomide (TMZ), 150 μg / mL BT@PDA, and 150 μg / mL BT@PDA@PMS for 24 hours, respectively, followed by sonication at 2.4°C / w for 2 minutes. Subsequently, 10 μL of CCK-8 solution was added to the culture medium, and the cells were incubated for 1 hour. Optical density (OD) was measured at 450 nm using a Multiskan FC microplate reader.

[0048] 2. Results Figure 7 The results show the cell survival rates after treatment with TMZ, TMZ+BT@PDA, TMZ+BT@PMS, and TMZ+BT@PDA@PMS, suggesting that sonication treatment with BT@PDA@PMS promotes cell death in GL261 / TMZ cells.

[0049] The specific cell survival rate results are shown in Table 2.

[0050] Table 2

[0051] As shown in Table 2, TMZ alone had no significant killing effect on temozolomide-resistant GL261 / TMZ cells. The synergistic use of BT@PDA enhanced the killing effect of temozolomide, and further, the use of BT@PDA@PMS significantly improved the killing effect of temozolomide. Moreover, the inhibition rate of TMZ+BT@PDA@PMS in this invention was slightly better than the sum of the effects of TMZ+BT@PDA and TMZ+BT@PMS, indicating a synergistic effect among the components of the composite material of this invention.

[0052] Example 4: Effects of BT@PDA@PMS on mitochondrial structure 1. Method: Temozolomide-resistant GL261 / TMZ cells were seeded in 6-well plates. After cell attachment, the cells were treated with 400 μM temozolomide (TMZ), 150 μg / mL BT@PDA, and 150 μg / mL BT@PDA@PMS for 24 hours, respectively, followed by sonication at 2.4°C / w for 2 minutes. After 12 hours, GL261 / TMZ cells were collected in 1 mL culture dishes, fixed with 2% glyceraldehyde solution, and then fixed in 1% osmium tetroxide solution. Subsequently, the samples were dehydrated in ethanol containing 3% ethylene glycol acetate, embedded in epoxy resin and propylene peroxide, and cured overnight. They were then cut into 70 nm thick sections, stained with lead chloride, and finally observed under an HT7700 transmission electron microscope.

[0053] 2. Results: Figure 8 The results show the morphology of mitochondria in cells after treatment with TMZ, BT@PDA, and BT@PDA@PMS, suggesting that sonication of BT@PDA@PMS disrupts the normal structure of mitochondria and promotes lipid deposition.

[0054] Example 5: Therapeutic effect of BT@PDA@PMS on temozolomide-resistant gliomas in situ 1. Method: 150 μg BT@PDA@PMS was dispersed in 1 mL PBS to obtain BT@PDA@PMS injection solution.

[0055] A recurrent glioma orthotopic model was established in C57 / BL6 mice using stereotactic injection of GL261 / TMZ cells. On day 7 post-surgery, MRI confirmed tumor growth. BT@PDA@PMS injection solution was then injected into the center of the glioblastoma using a stereotactic injection device. The tumor site was treated with ultrasound for 2 minutes daily for 3 consecutive days, 24 hours later. One week later, a follow-up MRI confirmed tumor size. Mice were then euthanized under anesthesia, and brain tissue was harvested for HE staining.

[0056] 2. Results: Figure 9 This is a comparison of tumor size before and after BT@PDA@PMS ultrasound treatment. Figure 10 This is the HE staining result of brain tissue after BT@PDA@PMS ultrasound treatment.

[0057] Figure 11 The survival rate of mice in different groups during the 28-day tumorigenesis experiment is shown in Table 3. Specific results are shown in Table 3.

[0058] Table 3

[0059] As shown in Table 3, the piezoelectric material BT@PDA@PMS of this invention significantly reduced the size of gliomas and significantly increased the survival rate of mice after ultrasound treatment. This indicates that BT@PDA@PMS of this invention has a significant therapeutic effect on temozolomide-resistant gliomas in situ.

[0060] The embodiments provided above are not intended to limit the scope of the invention, nor are the described steps intended to limit the order of execution. Any obvious modifications made to the invention by those skilled in the art based on existing common knowledge also fall within the scope of protection defined by the claims.

Claims

1. A method for preparing barium titanate composite nanomaterials, characterized in that, Includes the following steps: (1) Preparation of BaTiO3 nanoparticles: Barium hydroxide and titanium dioxide were dispersed in water and heated to react for a period of time to obtain BT precursor; The BT precursor was then washed with acetic acid, followed by washing with deionized water, centrifugation, drying, and grinding to obtain BT nanoparticles. (2) Synthesis of BT@PDA nanoparticles: Alkali solution was added to Tris-HCl aqueous solution and the pH was controlled to 8-9. Then, the BT nanoparticles obtained in step (1) were added, ultrasonically treated, and then dopamine hydrochloride was added. The mixture was reacted. After the reaction was completed, the solid and liquid were separated, the solid was collected, washed and dried to obtain BT nanoparticles with polydopamine on the surface, which were denoted as BT@PDA. (3) Synthesis of BT@PDA@PMS nanoparticles: The BT@PDA obtained in step (2) is mixed with persulfate to obtain BT@PDA nanoparticles loaded with persulfate.

2. The method according to claim 1, characterized in that, In step (1), the barium-titanium molar ratio in barium hydroxide and titanium dioxide is 1.1~1.5:1; the dispersion concentration of titanium dioxide relative to water is 0.2-0.5 mol / L.

3. The method according to claim 1, characterized in that, In step (1), the heating reaction is carried out at 140-170℃ for 4-7 hours.

4. The method according to claim 1, characterized in that, In step (2), the mass ratio of BT nanoparticles to dopamine hydrochloride is (3-8):

1.

5. The method according to claim 1, characterized in that, In step (3), the persulfate is potassium persulfate and / or sodium persulfate; the mass ratio of BT@PDA to persulfate is (1-3):

1.

6. The method according to any one of claims 1-5, characterized in that, In step (3), persulfate is dissolved in water, then BT@PDA is added, and the mixture is stirred and reacted for a period of time. After the reaction is complete, the solid and liquid are separated, the solid is collected, washed, and dried to obtain the composite piezoelectric material; or, Persulfate and BT@PDA were mixed and ground in an agate mortar for a period of time to ensure that BT@PDA and persulfate were fully and uniformly mixed to obtain a composite piezoelectric material.

7. A barium titanate composite nanomaterial prepared by any one of claims 1-6.

8. The application of the barium titanate composite nanomaterial according to claim 7 in piezoelectric catalysis.

9. The application of the barium titanate composite nanomaterial according to claim 7 in the catalytic degradation of Rhodamine B.

10. Use of the barium titanate composite nanomaterial of claim 7 in the preparation of a medicament for ultrasound treatment of recurrent gliomas resistant to temozolomide.

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