CN115944732B - Bismuth-based porous nanomaterial and preparation method and application thereof - Google Patents
Bismuth-based porous nanomaterial and preparation method and application thereof Download PDFInfo
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Abstract
The invention provides a bismuth-based porous nanomaterial and a preparation method and application thereof, and particularly relates to the field of acoustic power treatment. The bismuth-based porous nanomaterial is a product obtained by taking bismuth nanospheres and selenate as raw materials to react, and can be used for preparing an acoustic sensitizer for acoustic power treatment. The sound sensitive agent provided by the invention has excellent biocompatibility and biosafety, low toxicity and low immunogenicity, and is 1W/cm 2 Active oxygen can be generated in a large amount under the condition of ultrasonic treatment for 1min, so that a large amount of tumor cells die, and the ultrasonic therapeutic effect is excellent. The sound-sensitive agent provided by the invention has the advantages of simple preparation process, mild reaction condition and low production cost, and has wide application prospect and clinical conversion value in the aspect of sound power treatment.
Description
Technical Field
The invention belongs to the field of acoustic power treatment, and particularly relates to a bismuth-based porous nanomaterial and a preparation method and application thereof.
Background
Cancer morbidity and mortality continue to rise in recent years, and have become a major public health problem that endangers human life and health. The means for treating cancer commonly used in clinic at present comprise surgical excision, chemotherapy, radiotherapy, immunotherapy and the like, but the problems of high postoperative recurrence and metastasis rate, obvious side effect and the like still exist. There is therefore a need for new treatments to overcome the challenges currently faced.
Sonodynamic therapy is a non-invasive tumor treatment that has evolved based on photodynamic therapyIn this way, apoptosis and necrosis can be induced by applying ultrasonic irradiation of a specific intensity, thereby killing tumor cells. The anti-tumor mechanism of sonodynamic therapy is complex, in which the generation of Reactive Oxygen Species (ROS) is the primary mechanism, and the acoustic energy caused by ultrasound causes a rapid increase in mechanical pressure as the ultrasound passes through a liquid medium, thereby generating microbubbles in tissue fluids and storing these energies in the microbubbles. The microbubble collapses and instantaneously releases a large amount of energy to generate a 'sonoluminescence phenomenon', and the sonosensitizer is activated after absorbing the energy and directly reacts with surrounding triplet oxygen to generate singlet oxygen @, which is a natural gas 1 O 2 ）， 1 O 2 Can destroy cell membrane and affect mitochondrial function, and induce apoptosis of tumor cells while leading to G2/M phase retardation of tumor cells, thereby achieving the purpose of treating cancer.
In view of the use of sonodynamic therapy and the inseparability of sonosensitizers, researchers have constructed a variety of sonosensitizers to enhance the efficacy of sonodynamic therapy. For example, hematoporphyrin, chlorophyll derivative, protoporphyrin IX and other organic molecules, but these organic sonosensitizers have problems of poor chemical stability, low bioavailability, rapid regression, poor tumor accumulation ability and the like, and severely limit the curative effect of sonodynamic therapy. Fortunately, inorganic sonosensitizers have advantages such as high acoustic stability, but the biosafety and ROS conversion efficiency of inorganic sonosensitizers need to be further improved. The Chinese patent application with publication number of CN113209290A discloses a bismuth/barium titanate heterojunction for enhancing acoustic power and resisting tumor, and the preparation method of the bismuth/barium titanate heterojunction comprises the following steps: firstly, synthesizing barium titanate piezoelectric nano particles through hydrothermal treatment, carrying out polarization treatment on the barium titanate nano particles through high-temperature high-pressure polarization, and then loading bismuth simple substances on the surfaces of the barium titanate piezoelectric nano particles by utilizing a one-step in-situ deposition method to form a bismuth/barium titanate heterojunction. The bismuth/barium titanate heterojunction promotes and regulates and controls the separation of carriers (electron hole pairs) through the coupling of a built-in electric field generated under the ultrasonic response of barium titanate and the bismuth heterojunction, and improves the active oxygen generation content so as to further enhance the curative effect of the acoustic power therapy. However, the preparation process of the bismuth/barium titanate heterojunction is complex, the reaction condition is harsh, the high-temperature high-pressure reaction and the high-temperature high-pressure polarization treatment process are involved, and the bismuth/barium titanate heterojunction is not suitable for industrial production.
The development of the sound-sensitive agent with simple preparation process, mild reaction conditions, high biological safety, high active oxygen yield and good sound power curative effect has important clinical application value.
Disclosure of Invention
The invention aims to provide a bismuth-based porous nanomaterial as well as a preparation method and application thereof.
The invention provides a bismuth-based porous nanomaterial, which is a product obtained by taking bismuth nanospheres and selenium hydrogenated salt as raw materials to react.
Further, the selenate is sodium selenate.
Further, the preparation method of the sodium selenate comprises the following steps: and (3) reacting sodium borohydride with selenium powder in water to obtain sodium selenate.
Further, the mass ratio of the sodium borohydride to the selenium powder is 1: (0.9-1.3).
Further, the mass ratio of the sodium borohydride to the selenium powder is 1:1.1.
further, the bismuth nanospheres are Bi 3+ And (3) reacting the salt or hydrate thereof, a reducing agent, a surfactant and a base to obtain the product.
Further, the Bi 3+ The salt is Bi (NO) 3 ) 3 The Bi is 3+ The hydrate of the salt is Bi (NO 3 ) 3 ·5H 2 O, wherein the reducing agent is glycol, the surfactant is polyvinylpyrrolidone, and the alkali is sodium hydroxide;
the Bi is 3+ The mass volume ratio of the salt, the reducing agent, the surfactant and the alkali is 1mg: (0.1-0.3) mL: (0.5-1.5) mg: (1.0-2.0) mg.
Further, the Bi 3+ The mass volume ratio of the salt, the reducing agent, the surfactant and the alkali is 1mg:0.2mL:1.1mg:1.5mg.
The invention provides a method for preparing the bismuth-based porous nanomaterial, which comprises the following steps:
(1) Reacting sodium borohydride with selenium powder in water to obtain sodium selenate solution;
（2）Bi 3+ reacting salt or hydrate thereof, a reducing agent, a surfactant and alkali to obtain bismuth nanospheres;
(3) And mixing the selenium hydride salt solution with bismuth nanospheres, and reacting to obtain the bismuth-based porous nanomaterial.
Further, the selenium powder and Bi 3+ The mass ratio of the salt or the hydrate thereof is (1.0-2.0): 1, a step of;
in the step (2), the temperature of the reaction is 130-170 ℃ and the time is 2-4 hours;
in the step (3), the reaction condition is stirring reaction for 20-40min at normal temperature.
Further, the selenium powder and Bi 3+ The mass ratio of the salt or the hydrate thereof is 1.4:1, a step of;
in the step (2), the temperature of the reaction is 150 ℃ and the time is 3 hours;
in the step (3), the reaction condition is stirring reaction for 30min at normal temperature.
The invention also provides application of the bismuth-based porous nanomaterial in preparation of an acoustic sensitizer for acoustic power treatment.
Compared with the acoustic sensitizer in the prior art, the acoustic sensitizer provided by the invention has the following beneficial effects:
1. the sound-sensitive agent BSNs provided by the invention has excellent biocompatibility and biosafety, low toxicity and low immunogenicity.
2. The sound-sensitive agent BSNs provided by the invention is 1W/cm 2 Active oxygen can be generated in a large amount under the condition of ultrasonic treatment for 1min, so that a large amount of tumor cells die, and the ultrasonic therapeutic effect is excellent.
3. The sound-sensitive agent BSNs provided by the invention has the advantages of simple preparation process, mild reaction conditions, low production cost, nontoxic and harmless raw materials and suitability for industrial production.
In conclusion, the sound-sensitive agent provided by the invention has wide application prospect and clinical conversion value in the aspect of sound power treatment.
It should be apparent that, in light of the foregoing, various modifications, substitutions and alterations can be made herein without departing from the spirit and scope of the invention as defined by the appended claims.
The above-described aspects of the present invention will be described in further detail below with reference to specific embodiments in the form of examples. It should not be understood that the scope of the above subject matter of the present invention is limited to the following examples only. All techniques implemented based on the above description of the invention are within the scope of the invention.
Drawings
FIG. 1 is a scanning electron microscope image of the BSNs acoustic sensitizer prepared in example 1.
FIG. 2 is an in vitro ROS assay result of BSNs sonosensitizer prepared in example 1.
Fig. 3 is a biosafety test result of BSNs prepared in example 1.
FIG. 4 is a graph showing the detection of intracellular ROS by BSNs sonosensitizer prepared in example 1.
Fig. 5 shows the apoptosis-inducing effect of the BSNs sonosensitizer prepared in example 1.
Detailed Description
The raw materials and equipment used in the invention are all known products and are obtained by purchasing commercial products.
The operations in the examples of the present invention were carried out at normal temperature (25.+ -. 5 ℃ C.) unless otherwise specified.
Example 1: preparation of sonosensitizers BSNs
Step 1, 242 mg Bi (NO) 3 ) 3 ·5H 2 O was added to 50 mL glycol and stirred until dissolved. 260 mg polyvinylpyrrolidone and 363mg NaOH were added under inert gas and reacted at a constant temperature of 150℃for 3 hours. After the reaction is finished, centrifuging at 10000 rpm, discarding supernatant, collecting precipitate, and washing for several times to obtain bismuth nanosphere black powder.
And step 2, adding 334 mg selenium powder into the three-neck flask, and protecting by vacuum argon. Weighing 304 mg sodium borohydride (NaBH) 4 ) Dissolving in ice water, adding into a three-neck flask after complete dissolution,stirring the ice bath until the solution is transparent, and obtaining NaHSe precursor solution.
And step 3, rapidly adding the NaHSe precursor solution into the bismuth nanosphere black powder in the step 1, vigorously stirring for 30min to stabilize the growth of the material, centrifuging at 10000 rpm, discarding supernatant, collecting precipitate, washing for 6 times by using PBS, dialyzing by using ultrapure water, and finally dispersing in PBS to obtain the PBS solution of BSNs.
The scanning electron microscope image of the acoustic sensitizer BSNs is shown in FIG. 1, and the BSNs can be seen to be porous nano-structures.
Example 2: preparation of sonosensitizers BSNs
Step 1, 200 mg Bi (NO) was stirred magnetically at 300 rpm 3 ) 3 ·5H 2 O was added to 20 mL glycol and stirred until dissolved. 100 mg polyvinylpyrrolidone and 300mg NaOH are added under the protection of inert gas, and the reaction is carried out for 3 hours at the constant temperature of 150 ℃. After the reaction is finished, centrifuging at 10000 rpm, discarding supernatant, collecting precipitate, and washing for several times to obtain bismuth nanosphere black powder.
And step 2, adding 200 mg selenium powder into the three-neck flask, and protecting by vacuum argon. 200 mg sodium borohydride (NaBH) was weighed 4 ) Dissolving in ice water, injecting into a three-neck flask after complete dissolution, stirring in an ice bath until the solution is transparent, and obtaining NaHSe precursor solution.
And step 3, rapidly adding the NaHSe precursor solution into the bismuth nanosphere black powder in the step 1, vigorously stirring for 20 min to stabilize the growth of the material, centrifuging at 10000 rpm, discarding supernatant, collecting precipitate, washing for 6 times by using PBS, dialyzing by using ultrapure water, and finally dispersing in PBS to obtain the PBS solution of BSNs.
Example 3: preparation of sonosensitizers BSNs
Step 1, 400mg Bi (NO) was stirred magnetically at 700 rpm 3 ) 3 ·5H 2 O was added to 50 mL glycol and stirred until dissolved. 300mg polyvinylpyrrolidone and 400mg NaOH were further added under inert gas protection, and the reaction was continued at a constant temperature of 150℃for 3 hours. After the reaction is finished, centrifuging at 10000 rpm, discarding supernatant, collecting precipitate, and washing for several times to obtain bismuth nanosphere black powder.
And step 2, adding 400mg selenium powder into the three-neck flask, and protecting by vacuum argon. 400mg sodium borohydride (NaBH) was weighed 4 ) Dissolving in ice water, injecting into a three-neck flask after complete dissolution, stirring in an ice bath until the solution is transparent, and obtaining NaHSe precursor solution.
And step 3, rapidly adding the NaHSe precursor solution into the bismuth nanosphere black powder in the step 1, vigorously stirring for 40min to stabilize the growth of the material, centrifuging at 10000 rpm, discarding supernatant, collecting precipitate, washing for 6 times by using PBS, dialyzing by using ultrapure water, and finally dispersing in PBS to obtain the PBS solution of BSNs.
The beneficial effects of the present invention are demonstrated by experimental examples below, and BSNs used in the following experiments were prepared as in example 1.
Experimental example 1: in vitro ROS detection
1. Experimental method
In vitro 1 O 2 The formation was detected by the 1, 3-diphenyl isobenzofuran (DPBF) reagent. 20. Mu.g/mL of BSNs was added to a DPBF (singlet oxygen indicating fluorescent probe, 60. Mu.M) solution, the DPBF+BSNs group was not subjected to any treatment, and the DPBF+BSNs+US group was 1W/cm 2 Different times (20 s-160 s) of ultrasound (i.e. US) irradiation, the absorbance at each group 420 nm was observed by uv spectrometer.
2. Experimental results
The experimental results are shown in FIG. 2. It can be seen that the absorbance at 420 nm after mixing of DPBF with BSNs has a significant decrease with increasing time of ultrasonic irradiation, indicating that BSNs produce significant amounts of singlet oxygen upon ultrasonic irradiation.
Experimental example 2: biosafety test
1. Experimental method
In 96-well plates, 100. Mu.L of liver cancer cell SMMC-7721 (1' -10) was added to each well 4 Per mL), after culturing cells 24 h, 100 μl of BSNs complete medium was added per well at different concentrations, each concentration gradient being set with 6 replicate wells. After incubation of the well plates with 24 h, incubation was continued for 2 h after addition of 10. Mu.L CCK-8 per well, without any treatment in the BSNs group, the BSNs+US group was used at 1W/cm 2 Is of (a) ultrasonic radiationThe incubation was continued for 30min after 1 min. Finally, the absorbance at 450 and nm was read using a microplate reader.
2. Experimental results
The experimental results are shown in FIG. 3. The BSNs provided by the invention are almost free from cytotoxicity and have high biological safety when being singly used; and the BSNs can effectively kill liver cancer cells under ultrasonic irradiation.
Experimental example 3: detection of intracellular ROS
1. Experimental method
In 24-well plates, 500. Mu.L of SMMC-7721 cells (1' -10) were added to each well 5 Per mL), after culturing cells 24 h, 500 μl of 75 μg/mL BSNs complete medium was added per well, and 3 multiplex wells were set. After incubation of the well plate with 24 h, BSNs that did not enter the cells were washed away, and 500. Mu.L of DCFH-DA probe (10. Mu.M) was added. The control group (i.e., CON group) and BSNs group were not subjected to any treatment, and the ultrasound group and BSNs+ ultrasound group were used at 1W/cm 2 After 1min, incubation was continued for 30min, and green fluorescence was observed using an inverted fluorescence microscope.
2. Experimental results
The experimental results are shown in FIG. 4. It can be seen that SMMC-7721 hepatoma cells incubated 24 h with BSNs, after 1min of ultrasound irradiation, significant green fluorescence could be observed, whereas the pure irradiation only had weak fluorescence, which indicates that BSNs produced a large amount of ROS under ultrasound irradiation and were captured by intracellular DCFH.
Experimental example 4: detection of apoptosis-causing effects
1. Experimental method
In 12-well plates, 1 mL SMMC-7721 cells (1' -10) were added to each well 5 Per mL), after culturing cells 24 h, 1 mL of 75 μg/mL BSNs complete medium was added to each well, and 3 multiplex wells were set. After incubation of the well plate with 24 h, the BSNs not entering the cells were washed away, and the control group and the BSNs group were not subjected to any treatment, and the ultrasound group and the BSNs+ ultrasound group were 1W/cm 2 Is irradiated by ultrasonic waves for 1 min. 5. Mu.L of Annexin-FITC and 5. Mu.L of PI were added according to the Annexin-FITC/PI apoptosis kit instructions, incubation was continued for 30min, and apoptosis was detected using a flow cytometer.
2. Experimental results
The experimental results are shown in FIG. 5. Comparison with the control group shows that the apoptosis rate (early apoptosis and late apoptosis) of the BSNs group is 15%, while the BSNs+ultrasound group uses 1W/cm 2 The apoptosis rate after 1min of ultrasonic irradiation reaches 45 percent. The sound-sensitive agent BSNs provided by the invention can cause a large number of tumor cells to die under the ultrasonic condition, and has excellent sound power curative effect.
Claims (8)
1. A bismuth-based porous nanomaterial characterized in that: the product is obtained by taking bismuth nanospheres and selenate as raw materials,
the bismuth nanospheres are products obtained by reacting Bi3+ salt or hydrate thereof, a reducing agent, a surfactant and alkali; the reaction temperature is 130-170 ℃ and the reaction time is 2-4 hours;
the Bi is 3+ The salt is Bi (NO) 3 ) 3 The Bi is 3+ The hydrate of the salt is Bi (NO 3 ) 3 ·5H 2 O, wherein the reducing agent is glycol;
the surfactant is polyvinylpyrrolidone, and the alkali is sodium hydroxide;
the Bi is 3+ The mass volume ratio of the salt, the reducing agent, the surfactant and the alkali is 1mg: (0.1-0.3) mL: (0.5-1.5) mg: (1.0-2.0) mg.
2. The bismuth-based porous nanomaterial of claim 1, wherein: the selenate is sodium selenate.
3. The bismuth-based porous nanomaterial of claim 2, characterized in that: the preparation method of the sodium selenate comprises the following steps: and (3) reacting sodium borohydride with selenium powder in water to obtain sodium selenate.
4. The bismuth-based porous nanomaterial of claim 3, wherein: the mass ratio of the sodium borohydride to the selenium powder is 1: (0.9-1.3).
5. A method of preparing the bismuth-based porous nanomaterial of any of claims 1-4, characterized by: the method comprises the following steps:
(1) Reacting sodium borohydride with selenium powder in water to obtain sodium selenate solution;
（2）Bi 3+ reacting salt or hydrate thereof, a reducing agent, a surfactant and alkali to obtain bismuth nanospheres;
(3) And mixing the selenium hydride salt solution with bismuth nanospheres, and reacting to obtain the bismuth-based porous nanomaterial.
6. The method according to claim 5, wherein: the selenium powder and Bi 3+ The mass ratio of the salt or the hydrate thereof is (1.0-2.0): 1, a step of;
in the step (2), the temperature of the reaction is 130-170 ℃ and the time is 2-4 hours;
in the step (3), the reaction condition is stirring reaction for 20-40min at normal temperature.
7. The method according to claim 6, wherein: the selenium powder and Bi 3+ The mass ratio of the salt or the hydrate thereof is 1.4:1, a step of;
in the step (2), the temperature of the reaction is 150 ℃ and the time is 3 hours;
in the step (3), the reaction condition is stirring reaction for 30min at normal temperature.
8. Use of the bismuth-based porous nanomaterial of any one of claims 1-4 for the preparation of an acoustic sensitizer for acoustic power therapy.
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| WO2011112992A1 (en) * | 2010-03-12 | 2011-09-15 | Energ2, Inc. | Mesoporous carbon materials comprising bifunctional catalysts |
| CN103754837A (en) * | 2013-12-17 | 2014-04-30 | 武汉工程大学 | Method for preparation of bismuth-containing nano-hollow ball by using porous bismuth oxide as template |
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