Preparation method of long afterglow nano probe platform capable of loading different modules

Abstract

A preparation method of a long afterglow nanoprobe platform capable of loading different modules. A chromium-doped zinc gallium indate long afterglow nanoprobe platform is synthesized by a hydrothermal method, a photosensitizer chlorin e6 module is loaded by using an amide condensation reaction, manganese dioxide modules with glutathione and pH / H2O2 dual response are prepared on the surface of the chromium-doped zinc gallium indate long afterglow nanoprobe platform by stirring in a potassium permanganate solution and adding into a sodium citrate solution, a polydopamine module is loaded in a tris-hydroxymethyl aminomethane hydrochloride buffer solution containing hydrochloric acid dopamine to improve biocompatibility, and a DOX module is loaded on the long afterglow nanoprobe platform in a π-π stacking manner, so that different functional modules of near-infrared imaging, fluorescence imaging, photodynamic effect, photothermal effect and drug loading can be loaded alone or simultaneously, multifunctional synergistic effect is realized, and the long afterglow nanoprobe platform is applied to the preparation of a tumor microenvironment prevention and monitoring preparation or a drug.

Description

Technical Field

[0001] This invention belongs to the field of preparation of long-persistence nanoprobe platforms capable of loading different modules, specifically involving the preparation method of chromium-doped zinc gallium indium oxide long-persistence nanoprobe platform, and the loading method of different modules, taking dihydroporphyrin E6, manganese dioxide, polydopamine and DOX as examples. Background Technology

[0002] Malignant tumors pose a significant threat to human health. They typically grow rapidly and are invasive, capable of spreading and metastasizing to distant sites. However, existing strategies for tumor microenvironment prevention and monitoring often cause considerable damage to normal cells, making the development of highly efficient and low-toxicity tumor microenvironment prevention and monitoring strategies urgently needed. Emerging modalities include photodynamic therapy, photothermal therapy, and chemokinetic therapy, which have attracted widespread attention due to their advantages such as low invasiveness, good tumor targeting, fewer side effects, and lower drug resistance. Despite significant progress, some challenges remain. For example, the thermal tolerance of the tumor microenvironment, due to the expression of heat shock proteins at high temperatures, limits the efficiency of the photothermal effect; and the hypoxic environment of solid tumors hinders the efficiency of photodynamic therapy. Therefore, multimodal synergistic detection of the tumor microenvironment is an effective strategy to overcome the limitations of single-modal approaches.

[0003] Dihydroporphyrin E6, used as a photosensitizer in photodynamic therapy, generates reactive oxygen species (ROS) under excitation by a specific wavelength of light, thereby inducing tumor cell apoptosis. This substance exhibits high quantum yield and low dark toxicity, with a maximum absorption wavelength of 640–660 nm, and good penetration into biological tissues under near-infrared light excitation. However, the tumor microenvironment (hypoxia, high concentrations of H2O2 and glutathione, low pH) limits the photodynamic effect of the photosensitizer.

[0004] Long-persistent light nanoparticles can store excitation light energy and then slowly release the captured charge carriers, allowing them to emit light continuously even without continuous excitation from a light source. Near-infrared long-persistent light nanoparticles offer advantages such as avoiding interference from background tissue fluorescence, avoiding in-situ excitation, and being re-exciteable, demonstrating significant advantages in bioimaging and tumor microenvironment monitoring. Summary of the Invention

[0005] The purpose of this invention is to address the limitation of the photodynamic effect of photosensitizers in the tumor microenvironment and to provide a standardized long-persistence nanoprobe platform that can individually or simultaneously load different functional modules such as near-infrared imaging, fluorescence imaging, photodynamic effect, photothermal effect, and drug loading, achieving multifunctional synergistic effects and fully utilizing its high biocompatibility for application in the preparation of tumor microenvironment prevention and monitoring agents or drugs.

[0006] Manganese dioxide materials exhibit pH / H2O2 responsiveness. Manganese dioxide reacts with H2O2 in tumor cells to generate oxygen, overcoming intracellular hypoxia. Furthermore, manganese dioxide materials can consume glutathione, effectively enhancing the photodynamic effect on tumor microenvironment prevention and monitoring. Polydopamine possesses excellent biocompatibility, biodegradability, and adhesiveness. It can effectively convert near-infrared light energy into heat energy to kill tumor cells, making it an excellent photothermal agent. It also possesses abundant surface groups, allowing chemotherapeutic drugs or dyes to be loaded onto its surface through π-π stacking. This not only enables photothermal effect prevention and monitoring but also improves drug loading rates.

[0007] The technical solution of the present invention:

[0008] This invention first provides a method for preparing a long-persistence nanoprobe platform capable of loading different modules. The platform is a chromium-doped zinc gallium indium oxide long-persistence nanoprobe platform, and the preparation method includes the following steps:

[0009] (1) Take an appropriate amount of dilute nitric acid solution into Ga2O3 powder, then heat under reflux at 110-130℃ for 10-13 hours. After cooling, add deionized water to prepare a 0.2 mol / L Ga2O3 solution. 3+ Solution. A mixed solution was obtained by dissolving Zn(NO3)2, In(NO3)3, and Cr(NO3)3 in deionized water. 2+ Solution, In 3+ Solution and Cr 3+ The molar ratio of the solution is 0.4:0.2:0.1. Then, different volumes of Zn are added sequentially according to a stoichiometric ratio of 1:1.99-x:x:0.01 (x = 0, 0.05, 0.1, 0.15, 0.2, 0.3, 0.4). 2+ Ga 3+ In 3+ and Cr 3+ Place the ionic solution in a container, then add an equal volume of ethylene glycol (V mixed solution:V ethylene glycol = 1:1). Stir the solution at room temperature for 20–40 min, then adjust the pH to 9–11 with concentrated ammonia and continue stirring for 1–3 h.

[0010] (2) Place the mixed solution obtained in step (1) in a reaction vessel and hydrothermally react at 160–180°C for 20–24 h. After the reaction is complete, allow it to cool naturally, wash with anhydrous ethanol, dry under vacuum at 60–80°C for 10–14 h, then grind it carefully in an agate mortar. Calcinate the ground sample at 600–900°C for 2–4 h (heating rate 4–8°C / min). After the reaction is complete, different In values ​​can be obtained. 3+ ZnGa content 1.99-x In x O4:Cr3+ (x = 0, 0.05, 0.1, 0.15, 0.2, 0.3, 0.4) Chromium-doped zinc gallium indium oxide long afterglow nanoprobe platform.

[0011] Under the experimental conditions described above, the pH value of the precursor mixture and the Zn were changed respectively. 2+ Ga 3+ In 3+ and Cr 3+ By adjusting the proportions, calcination temperature, and calcination time, the optimal preparation conditions can ultimately be determined.

[0012] Preferably, the chromium-doped zinc gallium indium oxide long afterglow nanoprobe platform ZnGa 1.99-x In x O4:Cr 3+ Among the given values, x = 0.05, 0.1, 0.15, 0.2, 0.3, 0.4, with x being the preferred value of 0.1.

[0013] Preferably, the hydrothermal reaction temperature is set to 170°C, and the hydrothermal treatment lasts for 24 hours.

[0014] (3) The present invention refines the chromium-doped zinc gallium indium carbonate long afterglow nanoprobe platform prepared above:

[0015] The chromium-doped zinc gallium indium carbonate long-afterglow nanoprobe platform obtained in step (2) is placed in an agate mortar and ground with a small amount of anhydrous ethanol for 20-40 min. Then it is transferred to a container, and 80-120 mL of 0.05 mol / L NaOH solution is added. After sonication for 4-8 min, it is stirred at room temperature for 20-24 h. The pH of the above solution is then adjusted to 6-8 with 0.1 mol / L HCl. After centrifugation at 2000-4000 rpm for 4-6 min, the supernatant is taken and centrifuged at 1000-2000 rpm for 7-9 min to obtain a refined chromium-doped zinc gallium indium carbonate long-afterglow nanoprobe platform.

[0016] This invention also provides a method for loading different modules onto a chromium-doped zinc gallium indium oxide long-afterglow nanoprobe platform. The loading method includes the following steps:

[0017] (1) Method for loading dihydroporphyrin e6 module onto chromium-doped zinc gallium indium oxide long afterglow nanoprobe platform

[0018] A refined chromium-doped zinc gallium indium carbonate (ZCI) long-afterglow nanoprobe platform was dispersed in N,N-dimethylformamide. Then, 3-aminopropyltriethoxysilane was added under stirring. The resulting reaction mixture was stirred, centrifuged after the reaction was complete, and washed with N,N-dimethylformamide to obtain an aminated chromium-doped ZCI long-afterglow nanoprobe platform. 1-Ethyl-(3-dimethylaminopropyl)carbodiimide, N-hydroxysuccinimide, and dihydroporphyrin e6 were dissolved in N,N-dimethylformamide to obtain a mixed solution. After vigorous stirring in the dark, the obtained aminated chromium-doped ZCI long-afterglow nanoprobe platform was added to the mixed solution and stirring continued. After stirring was complete, the mixture was centrifuged, and the precipitate was washed with N,N-dimethylformamide and vacuum dried to obtain a chromium-doped ZCI long-afterglow nanoprobe platform loaded with a dihydroporphyrin e6 module.

[0019] Furthermore, the refined chromium-doped zinc gallium indium long afterglow nanoprobe platform, 1-ethyl-(3-dimethylaminopropyl)carbodiimide, N-hydroxysuccinimide and dihydroporphyrin e6 have a mass ratio of 25:8:8:1.

[0020] (2) Method for loading manganese dioxide module onto chromium-doped zinc gallium indium oxide long afterglow nanoprobe platform

[0021] Potassium permanganate solution was added to sodium citrate solution under stirring. After stirring at room temperature, the mixture was transferred to 70-90℃ and reacted for 2-4 hours. After cooling, a refined chromium-doped zinc gallium indium nitrate long afterglow nanoprobe platform was added. After water bath centrifugation, the precipitate was washed with deionized water and vacuum dried to obtain a chromium-doped zinc gallium indium nitrate long afterglow nanoprobe platform loaded with manganese dioxide module.

[0022] Furthermore, the mass ratio of the potassium permanganate solution (8-12 mmol / L), sodium citrate solution (0.3 mg / mL), and the refined chromium-doped zinc gallium indium carbonate long afterglow nanoprobe platform is 1:4:5;

[0023] (3) Method for loading polydopamine modules onto a chromium-doped zinc gallium indium oxide long-afterglow nanoprobe platform

[0024] A refined chromium-doped zinc gallium indium long-afterglow nanoprobe platform was dispersed in a tris(hydroxymethyl)aminomethane hydrochloride buffer solution containing dopamine hydrochloride, stirred at room temperature in the dark, and the precipitate after centrifugation was washed with deionized water to obtain a chromium-doped zinc gallium indium long-afterglow nanoprobe platform loaded with a polydopamine module.

[0025] Furthermore, the mass ratio of the refined chromium-doped zinc gallium indium long afterglow nanoprobe platform to dopamine hydrochloride is 10:1.

[0026] (4) A method for simultaneously loading dihydroporphyrin E6, manganese dioxide, and polydopamine modules onto a chromium-doped zinc gallium indium carbonate long afterglow nanoprobe platform.

[0027] A refined chromium-doped zinc gallium indium carbonate (ZCI) long-afterglow nanoprobe platform was dispersed in N,N-dimethylformamide. Then, 3-aminopropyltriethoxysilane was added under stirring. The resulting reaction mixture was stirred, and after the reaction was completed, it was centrifuged and washed with N,N-dimethylformamide to obtain an aminated chromium-doped ZCI long-afterglow nanoprobe platform. 1-Ethyl-(3-dimethylaminopropyl)carbodiimide, N-hydroxysuccinimide, and dihydroporphyrin e6 were dissolved in N,N-dimethylformamide to obtain a mixed solution. After vigorous stirring in the dark, the obtained aminated chromium-doped ZCI long-afterglow nanoprobe platform was added to the mixed solution and stirring was continued. After stirring was completed, it was centrifuged, and the precipitate was washed with N,N-dimethylformamide and vacuum dried to obtain a chromium-doped ZCI long-afterglow nanoprobe platform loaded with a dihydroporphyrin e6 module.

[0028] Potassium permanganate solution was added to sodium citrate solution under stirring. After stirring at room temperature, the mixture was transferred to 70-90℃ and reacted for 2-4 hours. After cooling, a chromium-doped zinc gallium indium long-afterglow nanoprobe platform loaded with dihydroporphyrin e6 module was added. After water bath centrifugation, the precipitate was washed with deionized water and vacuum dried to obtain a chromium-doped zinc gallium indium long-afterglow nanoprobe platform loaded with dihydroporphyrin e6 and manganese dioxide modules.

[0029] A chromium-doped zinc gallium indium long-afterglow nanoprobe platform loaded with dihydroporphyrin e6 and manganese dioxide modules was dispersed in a tris(hydroxymethyl)aminomethane hydrochloride buffer solution containing dopamine hydrochloride. The mixture was stirred at room temperature in the dark, and the precipitate after centrifugation was washed with deionized water to obtain a chromium-doped zinc gallium indium long-afterglow nanoprobe platform simultaneously loaded with dihydroporphyrin e6, manganese dioxide and polydopamine modules.

[0030] Furthermore, the mass ratio of the refined chromium-doped zinc gallium indium long-afterglow nanoprobe platform, 1-ethyl-(3-dimethylaminopropyl)carbodiimide, N-hydroxysuccinimide, and dihydroporphyrin e6 is 25:8:8:1; the mass ratio of the potassium permanganate solution (8-12 mmol / L), sodium citrate solution (0.3 mg / mL), and the chromium-doped zinc gallium indium long-afterglow nanoprobe platform loaded with the dihydroporphyrin e6 module is 1:4:5; and the mass ratio of the chromium-doped zinc gallium indium long-afterglow nanoprobe platform loaded with the dihydroporphyrin e6 and manganese dioxide module and dopamine hydrochloride is 10:1.

[0031] (5) A method for simultaneously loading dihydroporphyrin E6, manganese dioxide, polydopamine, and DOX modules onto a chromium-doped zinc gallium indium carbonate long-afterglow nanoprobe platform.

[0032] The chromium-doped zinc gallium indium long-afterglow nanoprobe platform simultaneously loaded with dihydroporphyrin e6, manganese dioxide, and polydopamine modules obtained in step (4) and DOX were added to a container, followed by deionized water and phosphate buffer solution (pH = 7.4), and stirred in the dark for 24 h. To remove excess DOX, after the reaction was completed, the mixed solution was dialyzed in a dialysis bag for 9–11 h, and then freeze-dried to obtain the chromium-doped zinc gallium indium long-afterglow nanoprobe platform simultaneously loaded with dihydroporphyrin e6, manganese dioxide, polydopamine, and DOX modules.

[0033] Furthermore, the mass ratio of the chromium-doped zinc gallium indium long afterglow nanoprobe platform simultaneously loaded with dihydroporphyrin e6, manganese dioxide, and polydopamine modules to DOX is 15:2.7.

[0034] Advantages and beneficial effects of the present invention:

[0035] The preparation method of this invention exhibits good biocompatibility and water solubility. It allows for the application of a long-persistence nanoprobe platform with different modules. This platform continues to emit light after excitation ceases, with its emission wavelength falling within the transmission window of biological tissues, effectively avoiding interference from tissue autofluorescence. The reusable excitation capability and the elimination of the need for in-situ excitation further highlight the advantages of the long-persistence nanoprobe platform in fields such as bioimaging, sensing, and nanomedicine. The long-persistence nanoprobe platform, used in conjunction with biosensing and bioimaging, can improve the precision diagnosis and treatment of tumor cells, enhancing the fluorescence imaging capability, photodynamic effect, and photothermal effect of the modules. It can be applied to near-infrared imaging, drug delivery, and the preparation of tumor microenvironment prevention and monitoring agents or drugs. Attached Figure Description

[0036] Figure 1 Transmission electron microscopy (TEM) images of a chromium-doped zinc gallium indium carbonate long afterglow nanoprobe platform simultaneously loaded with dihydroporphyrin e6, manganese dioxide, polydopamine, and DOX modules.

[0037] Figure 2 (a) is a near-infrared afterglow image of a chromium-doped zinc gallium indium carbonate long afterglow nanoprobe platform simultaneously loaded with dihydroporphyrin e6, manganese dioxide, polydopamine, and DOX modules. (b) is the afterglow intensity map corresponding to each experimental group in (a).

[0038] Figure 3 Simultaneous loading of dihydroporphyrin E6, manganese dioxide, polydopamine, and DOX modules onto a chromium-doped zinc gallium indium oxide long-afterglow nanoprobe platform with different concentrations under an 808 nm near-infrared laser (2.0 W / cm²). 2 Temperature change curve over time under irradiation.

[0039] Figure 4The linear relationship between fluorescence intensity and DOX concentration was established for a chromium-doped zinc gallium indium oxide long afterglow nanoprobe platform simultaneously loaded with dihydroporphyrin E6, manganese dioxide, polydopamine, and DOX modules. Detailed Implementation

[0040] The present invention will be further described in detail below with reference to the accompanying drawings and embodiments. However, the present invention is not limited to these embodiments.

[0041] Example 1:

[0042] A method for preparing a long-persistence nanoprobe platform capable of loading different modules, wherein the platform is a chromium-doped zinc gallium indium oxide long-persistence nanoprobe platform, and its preparation includes the following steps:

[0043] Take an appropriate amount of dilute nitric acid solution and add it to 4.686 g of gallium oxide powder. Then heat and reflux at 120 °C for 12 h. After cooling, add deionized water to prepare a 0.2 mol / L Ga solution. 3+ Solution. A mixed solution was obtained by dissolving Zn(NO3)2, In(NO3)3, and Cr(NO3)3 in deionized water. 2+ Solution, In 3+ Solution and Cr 3+ The molar ratio of the solution is 0.4:0.2:0.1. Then, different volumes of Zn are added sequentially according to a stoichiometric ratio of 1:1.89:0.1:0.01 (x = 0.1). 2+ Ga 3+ In 3+ and Cr 3+ The ionic solution was placed in a flask, and 5 mL of ethylene glycol was added. The mixture was stirred at room temperature for 30 min, and the pH was adjusted to 10 with concentrated ammonia. Stirring was continued for 1 h. After stirring, the mixture was placed in a reaction vessel and hydrothermally reacted at 170 °C for 24 h. After cooling, the mixture was washed three times with anhydrous ethanol and dried under vacuum at 70 °C for 12 h. The product was carefully ground in an agate mortar, and the ground sample was calcined at 800 °C for 3 h (heating rate of 4–8 °C / min). After the reaction was completed, a chromium-doped zinc gallium indium carbonate long-afterglow nanoprobe platform was obtained.

[0044] Example 2:

[0045] A method for preparing a long-afterglow nanoprobe platform capable of loading different modules, wherein the platform is a chromium-doped zinc gallium indium nitride long-afterglow nanoprobe platform, which is basically the same as in Example 1, except that the pH of the mixed solution precursor is changed to 11.

[0046] Example 3:

[0047] A method for preparing a long-afterglow nanoprobe platform capable of loading different modules, wherein the platform is a chromium-doped zinc gallium indium oxide long-afterglow nanoprobe platform, which is basically the same as in Example 1, except that the calcination time is changed to 2 hours.

[0048] Example 4:

[0049] A method for preparing a long-afterglow nanoprobe platform capable of loading different modules, wherein the platform is a chromium-doped zinc gallium indium oxide long-afterglow nanoprobe platform, which is basically the same as in Example 1, except that the calcination temperature is changed to 700℃.

[0050] Example 5:

[0051] Refinement of the chromium-doped zinc gallium indium oxide long-afterglow nanoprobe platform:

[0052] 100 mg of the chromium-doped zinc gallium indium carbonate long-afterglow nanoprobe platform obtained in Example 4 was placed in an agate mortar and ground with a small amount of anhydrous ethanol for 30 min. Then, it was transferred to a flask, and 100 mL of 0.05 mol / L NaOH solution was added. After sonication for 5 min, the mixture was stirred at room temperature for 24 h. The pH of the solution was then adjusted to 7 with 0.1 mol / L HCl. After centrifugation at 3500 rpm for 5 min, the supernatant was centrifuged at 1100 rpm for 8 min to obtain a refined chromium-doped zinc gallium indium carbonate long-afterglow nanoprobe platform.

[0053] Example 6:

[0054] A method for loading different modules onto a chromium-doped zinc gallium indium oxide long-afterglow nanoprobe platform includes the following steps:

[0055] (1) Method for loading dihydroporphyrin e6 modules onto a chromium-doped zinc gallium indium oxide long-afterglow nanoprobe platform:

[0056] 50 mg of refined chromium-doped zinc gallium indium carbonate (ZCI) long-afterglow nanoprobe platform was dispersed in 20 mL of N,N-dimethylformamide and ultrasonically dispersed for 10 min. Then, 500 μL of 3-aminopropyltriethoxysilane was added dropwise with stirring, and the reaction mixture was vigorously stirred at 80 °C for 8 h. After the reaction was completed, the mixture was centrifuged at 1200 rpm for 8 min and washed with N,N-dimethylformamide to obtain the aminated chromium-doped ZCI long-afterglow nanoprobe platform. 16 mg of 1-ethyl-(3-dimethylaminopropyl)carbodiimide, 16 mg of N-hydroxysuccinimide, and 2 mg of dihydroporphyrin E6 were dissolved in 10 mL of N,N-dimethylformamide and vigorously stirred in the dark for 1 h. Then, 10 mg of the aminated chromium-doped ZCI long-afterglow nanoprobe platform was added to the above solution, and stirring was continued for 24 h. After stirring, centrifuge, wash the precipitate with N,N-dimethylformamide, and vacuum dry at 25°C to obtain a chromium-doped zinc gallium indium long-afterglow nanoprobe platform loaded with a dihydroporphyrin e6 module.

[0057] (2) Method for loading manganese dioxide modules onto a chromium-doped zinc gallium indium oxide long-afterglow nanoprobe platform:

[0058] 2 mL of potassium permanganate solution (10 mmol / L) was slowly added to 8 mL of sodium citrate solution (0.3 mg / mL) with stirring. The mixture was stirred at room temperature for 1 h, and then transferred to 80 °C for 3 h. After cooling, 10 mg of refined chromium-doped zinc gallium indium oxide long-afterglow nanoprobe platform was added to the above solution. After centrifugation in a water bath at 37 °C for 4 h, the precipitate was washed with deionized water and dried under vacuum at 37 °C to obtain the chromium-doped zinc gallium indium oxide long-afterglow nanoprobe platform loaded with manganese dioxide module.

[0059] (3) Method for loading polydopamine modules onto a chromium-doped zinc gallium indium oxide long-afterglow nanoprobe platform:

[0060] 10 mg of a finely refined chromium-doped zinc gallium indium carbonate long-afterglow nanoprobe platform was dispersed in 10 mL of tris(hydroxymethyl)aminomethane hydrochloride buffer solution (pH = 8.5, 10 × 10⁻⁶) containing 1 mg of dopamine hydrochloride. -3 In a solution of mol / L, after stirring for 1 h in the dark at room temperature, centrifuge and wash the precipitate three times with deionized water to obtain a chromium-doped zinc gallium indium carbonate long afterglow nanoprobe platform loaded with a polydopamine module.

[0061] (4) A method for simultaneously loading dihydroporphyrin E6, manganese dioxide, and polydopamine modules onto a chromium-doped zinc gallium indium carbonate long afterglow nanoprobe platform:

[0062] 50 mg of refined chromium-doped zinc gallium indium carbonate (ZCI) long-afterglow nanoprobe platform was dispersed in 20 mL of N,N-dimethylformamide and ultrasonically dispersed for 10 min. Then, 500 μL of 3-aminopropyltriethoxysilane was added dropwise with stirring, and the reaction mixture was vigorously stirred at 80 °C for 8 h. After the reaction was completed, the mixture was centrifuged at 1200 rpm for 8 min and washed with N,N-dimethylformamide to obtain the aminated chromium-doped ZCI long-afterglow nanoprobe platform. 16 mg of 1-ethyl-(3-dimethylaminopropyl)carbodiimide, 16 mg of N-hydroxysuccinimide, and 2 mg of dihydroporphyrin E6 were dissolved in 10 mL of N,N-dimethylformamide and vigorously stirred in the dark for 1 h. Then, 10 mg of the aminated chromium-doped ZCI long-afterglow nanoprobe platform was added to the above solution, and stirring was continued for 24 h. After stirring, the precipitate was centrifuged, washed with N,N-dimethylformamide, and vacuum dried at 25°C to obtain a chromium-doped zinc gallium indium long afterglow nanoprobe platform loaded with dihydroporphyrin e6 module.

[0063] 2 mL of potassium permanganate solution (10 mmol / L) was slowly added to 8 mL of sodium citrate solution (0.3 mg / mL) with stirring. The mixture was stirred at room temperature for 1 h, and then transferred to 80 °C for 3 h. After cooling, 10 mg of chromium-doped zinc gallium indium oxide long-afterglow nanoprobe platform loaded with dihydroporphyrin e6 module was added to the above solution. After water bath at 37 °C for 4 h, the mixture was centrifuged. The precipitate was washed with deionized water and dried under vacuum at 37 °C to obtain chromium-doped zinc gallium indium oxide long-afterglow nanoprobe platform loaded with dihydroporphyrin e6 and manganese dioxide module.

[0064] A chromium-doped zinc gallium indium carbonate long-afterglow nanoprobe platform loaded with 10 mg of dihydroporphyrin E6 and manganese dioxide modules was dispersed in 10 mL of tris(hydroxymethyl)aminomethane hydrochloride buffer solution (pH = 8.5, 10 × 10⁻⁶) containing 1 mg of dopamine hydrochloride. -3 In mol / L, after stirring for 1 h at room temperature in the dark, centrifuge, and washing the precipitate three times with deionized water, a chromium-doped zinc gallium indium carbonate long afterglow nanoprobe platform simultaneously loaded with dihydroporphyrin E6, manganese dioxide, and polydopamine modules can be obtained.

[0065] (5) A method for simultaneously loading dihydroporphyrin E6, manganese dioxide, polydopamine, and DOX modules onto a chromium-doped zinc gallium indium carbonate long-afterglow nanoprobe platform.

[0066] 15 mg of the chromium-doped zinc gallium indium carbonate long-afterglow nanoprobe platform simultaneously loaded with dihydroporphyrin e6, manganese dioxide, and polydopamine modules obtained in step (4) and 2.7 mg of DOX were added to a flask, followed by 8 mL of deionized water and 2 mL of phosphate buffer solution (pH = 7.4). The mixture was stirred in the dark for 24 h. To remove excess DOX, the mixture was dialyzed in a dialysis bag for 10 h after the reaction was completed. After freeze-drying, the chromium-doped zinc gallium indium carbonate long-afterglow nanoprobe platform simultaneously loaded with dihydroporphyrin e6, manganese dioxide, polydopamine, and DOX modules was obtained.

[0067] (6) Figure 1 The image shows a transmission electron microscope (TEM) image of a chromium-doped zinc gallium indium oxide (ZCI) long-afterglow nanoprobe platform simultaneously loaded with dihydroporphyrin e6, manganese dioxide, polydopamine, and DOX modules. The morphology is characterized by irregular spherical shapes with relatively uniform particle size. The nanoshell structure can be seen in the HR-TEM image in the upper right corner, indicating that the polydopamine and DOX modules were successfully modified onto the sample. The chromium-doped ZCI long-afterglow nanoprobe platform simultaneously loaded with dihydroporphyrin e6, manganese dioxide, polydopamine, and DOX modules was successfully prepared.

[0068] (7) In order to study the recovery of afterglow intensity of the chromium-doped zinc gallium indium carbonate long afterglow nanoprobe platform simultaneously loaded with dihydroporphyrin e6, manganese dioxide, polydopamine, and DOX modules under different conditions (different pH, presence of H2O2 and glutathione), Figure 2 (a) is a near-infrared afterglow image. The results show that the afterglow intensity at pH=5.3 is the strongest among all control groups, indicating that the afterglow recovery is the best when H2O2 and glutathione are present at the same time. It also shows that the chromium-doped zinc gallium indium oxide long afterglow nanoprobe platform loaded with dihydroporphyrin e6, manganese dioxide, polydopamine and DOX modules has dual response capability of pH / H2O2 and glutathione. Figure 2 (b) is Figure 2 (a) Afterglow intensity diagrams for each experimental group. It can be seen from the diagram that the afterglow intensity when pH=5.3 and H2O2 and glutathione are present is 3.38 times that when pH=7.4.

[0069] (8) In order to investigate the photothermal effect of a chromium-doped zinc gallium indium oxide long afterglow nanoprobe platform simultaneously loaded with dihydroporphyrin e6, manganese dioxide, polydopamine, and DOX modules. Figure 3 A chromium-doped zinc gallium indium nitride long-afterglow nanoprobe platform, simultaneously loaded with different concentrations of dihydroporphyrin E6, manganese dioxide, polydopamine, and DOX modules, was tested under 808 nm near-infrared laser light (2.0 W / cm²). 2Temperature change curve under irradiation over time. Phosphate buffer solutions with concentrations of 0, 62.5, 125, 250, 500, and 1000 g / mL and a pH of 7.4 were prepared for the long-afterglow nanoprobe platform, utilizing a power of 2.0 W / cm². 2 Irradiation of long-persistence nanoprobe platforms with different concentrations using 808nm laser light resulted in a gradual increase in temperature over time. Furthermore, the higher the concentration of the long-persistence nanoprobe platform, the more pronounced the temperature change, indicating that the presence of polydopamine enhances photothermal conversion performance.

[0070] (9) In order to study the DOX loading capacity of the chromium-doped zinc gallium indium oxide long afterglow nanoprobe platform simultaneously loading dihydroporphyrin e6, manganese dioxide and polydopamine modules. Figure 4 The linear regression equation loaded from DOX is y = -3.25 × 10⁻⁵ + 2.14 × 10⁻⁵x, with a correlation coefficient R₀. 2 =0.998, and based on the DOX concentration standard curve, the drug loading efficiency can be calculated to be 29.6%. This indicates that it has DOX loading capacity and is a carrier material that can be used to deliver drugs.

[0071] Although the present invention has been described through embodiments, these embodiments are not intended to limit the invention. Those skilled in the art can make various modifications and improvements within the spirit of the invention, such as adjusting the component ratios or time ranges. The effects of such adjustments are predictable and therefore also fall within the scope of protection of the present invention. Therefore, the scope of protection of the present invention should be determined by the scope defined by the same or equivalent technical features in the claims of this application.

Claims

1. A method for preparing a chromium-doped zinc gallium indium carbonate long-afterglow nanoprobe platform simultaneously loaded with dihydroporphyrin E6, manganese dioxide, and polydopamine modules, characterized in that, Includes the following steps: (1) Take an appropriate amount of dilute nitric acid solution into Ga2O3 powder, then heat it under reflux at 110~130℃ for 10~13h, cool it down and add deionized water to prepare a Ga2O3 solution with a concentration of 0.2mol / L. 3+ Solution; a mixed solution was obtained by dissolving Zn(NO3)2, In(NO3)3, and Cr(NO3)3 in deionized water, Zn 2+ Solution, In 3+ Solution and Cr 3+ The molar ratio of the solution was 0.4:0.2:0.1, and then different volumes of the Zn were added sequentially according to a stoichiometric ratio of 1:1.99-x:x:0.

01. 2+ Ga 3+ In 3+ and Cr 3+ An ionic solution is placed in a container, where x = 0.05, 0.1, 0.15, 0.2, 0.3, 0.4, and an equal volume of ethylene glycol is added, with V_solution:V_ethylene glycol = 1:

1. The solution is stirred at room temperature for 20-40 minutes, and then the pH is adjusted to 9-11 with concentrated ammonia. Stirring continues for 1-3 hours. (2) Place the mixed solution obtained in step (1) in a reaction vessel and carry out a hydrothermal reaction; after the reaction is completed, allow it to cool naturally, wash it with anhydrous ethanol, dry it under vacuum at 60~80℃ for 10~14h, then place it in an agate mortar and grind it carefully. Calcine the ground sample. After the reaction is completed, different In samples can be obtained. 3+ ZnGa content 1.99-x In x O4:Cr 3+ Chromium-doped zinc gallium indium oxide long afterglow nanoprobe platform; (3) Place the chromium-doped zinc gallium indium carbonate long-afterglow nanoprobe platform obtained in step (2) in an agate mortar and grind it with a small amount of anhydrous ethanol for 20-40 min. Then transfer it to a container, add 80-120 mL of 0.05 mol / L NaOH solution, sonicate for 4-8 min, stir at room temperature for 20-24 h, then adjust the pH of the above solution to 6-8 with 0.1 mol / L HCl, centrifuge at 2000-4000 rpm for 4-6 min, take the supernatant and centrifuge at 1000-2000 rpm for 7-9 min to obtain a refined chromium-doped zinc gallium indium carbonate long-afterglow nanoprobe platform: (4) The refined chromium-doped zinc gallium indium long-afterglow nanoprobe platform obtained in step (3) is dispersed in N,N-dimethylformamide, and then 3-aminopropyltriethoxysilane is added under stirring. The reaction mixture is stirred and centrifuged after the reaction is completed. After washing with N,N-dimethylformamide, an aminated chromium-doped zinc gallium indium long-afterglow nanoprobe platform is obtained. 1-ethyl-(3-dimethylaminopropyl)carbodiimide, N-hydroxysuccinimide and dihydroporphyrin e6 are dissolved in N,N-dimethylformamide to obtain a mixed solution. After vigorous stirring under dark conditions, the aminated chromium-doped zinc gallium indium long-afterglow nanoprobe platform is added to the mixed solution and stirring is continued. After stirring is completed, the mixture is centrifuged, the precipitate is washed with N,N-dimethylformamide, and vacuum dried to obtain a chromium-doped zinc gallium indium long-afterglow nanoprobe platform loaded with a dihydroporphyrin e6 module. Potassium permanganate solution was added to sodium citrate solution under stirring. After stirring at room temperature, the mixture was transferred to 70-90℃ and reacted for 2-4 hours. After cooling, a chromium-doped zinc gallium indium long-afterglow nanoprobe platform loaded with dihydroporphyrin e6 module was added. After water bath centrifugation, the precipitate was washed with deionized water and vacuum dried to obtain a chromium-doped zinc gallium indium long-afterglow nanoprobe platform loaded with dihydroporphyrin e6 and manganese dioxide modules. A chromium-doped zinc gallium indium long-afterglow nanoprobe platform loaded with dihydroporphyrin e6 and manganese dioxide modules was dispersed in a tris(hydroxymethyl)aminomethane hydrochloride buffer solution containing dopamine hydrochloride. The mixture was stirred at room temperature in the dark, and the precipitate after centrifugation was washed with deionized water to obtain a chromium-doped zinc gallium indium long-afterglow nanoprobe platform simultaneously loaded with dihydroporphyrin e6, manganese dioxide, and polydopamine modules.

2. The preparation method according to claim 1, characterized in that, The hydrothermal reaction in step (2) is carried out at a temperature of 160~180℃ for 20~24h. After grinding, the mixture is calcined at a high temperature of 600~900℃ for 2~4h, with a heating rate of 4~8℃ / min.

3. The preparation method according to claim 1, characterized in that, The refined chromium-doped zinc gallium indium long afterglow nanoprobe platform, 1-ethyl-(3-dimethylaminopropyl)carbodiimide, N-hydroxysuccinimide and dihydroporphyrin e6 in step (4) have a mass ratio of 25:8:8:

1.

4. The chromium-doped zinc gallium indium long afterglow nanoprobe platform simultaneously loaded with dihydroporphyrin E6, manganese dioxide, and polydopamine modules obtained by the preparation method according to any one of claims 1 to 3.

5. A method for preparing a chromium-doped zinc gallium indium carbonate long-afterglow nanoprobe platform simultaneously loaded with dihydroporphyrin E6, manganese dioxide, polydopamine, and DOX modules, characterized in that, The chromium-doped zinc gallium indium long-afterglow nanoprobe platform, which is simultaneously loaded with dihydroporphyrin e6, manganese dioxide, and polydopamine modules, obtained by the preparation method according to any one of claims 1 to 3, and DOX are added to a container, followed by the addition of deionized water and phosphate buffer solution. The mixture is stirred in the dark for 24 h. To remove excess DOX, the mixed solution is dialyzed in a dialysis bag for 9-11 h after the reaction is completed. After freeze-drying, the chromium-doped zinc gallium indium long-afterglow nanoprobe platform simultaneously loaded with dihydroporphyrin e6, manganese dioxide, polydopamine, and DOX modules is obtained.

6. The preparation method according to claim 5, characterized in that, The mass ratio of the chromium-doped zinc gallium indium oxide long afterglow nanoprobe platform simultaneously loaded with dihydroporphyrin e6, manganese dioxide, and polydopamine modules to DOX is 15:2.

7.

7. The chromium-doped zinc gallium indium long afterglow nanoprobe platform simultaneously loaded with dihydroporphyrin e6, manganese dioxide, polydopamine, and DOX modules obtained by the preparation method of claim 5 or 6.

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