Near-infrared light response nano heterojunction with antibacterial and anti-inflammatory functions and preparation method thereof
By preparing near-infrared light-responsive Prussian blue@polymanganese phthalocyanine nanoheterojunctions and combining the photothermal effect with reactive oxygen species scavenging function, the problems of antibiotic resistance and inflammation caused by photodynamic therapy were solved, achieving a temporal antibacterial and anti-inflammatory effect.
Patent Information
- Application Number
- CN202510923361.1
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-07-04
- Publication Date
- 2025-10-17
AI Technical Summary
Existing antibiotics have drug resistance problems in treating bacterial infections, and the reactive oxygen species produced during the sterilization process of photodynamic therapy make the inflammatory response difficult to control.
A near-infrared light-responsive Prussian blue@polymanganese phthalocyanine nanoheterojunction was prepared, and the physical and chemical properties of the two were combined to use near-infrared light to control the generation and removal of reactive oxygen species, thereby achieving antibacterial and anti-inflammatory functions.
It effectively kills bacteria under near-infrared light irradiation, removes reactive oxygen species after the light stops, reduces inflammatory responses, and restores redox homeostasis. It has a wide source of raw materials, a simple preparation process, and good biocompatibility.
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Figure CN120789246A_ABST
Abstract
Description
TECHNICAL FIELD
[0001] The application belongs to the technical field of nanomaterials, and relates to a near-infrared light-responsive nano-heterojunction with antibacterial and anti-inflammatory functions and a preparation method thereof.
[0002] BACKGROUND
[0003] Bacterial infection can pose a significant threat to human life and health. Since the middle of the last century, antibiotics have been widely used to treat bacterial infections due to their excellent bactericidal effect. However, the abuse of antibiotics has led to the development of bacterial drug resistance, greatly reducing the therapeutic effect and posing a more serious threat to human health. Therefore, there is an urgent need to develop new non-antibiotic treatment strategies. Phototherapy, represented by photodynamic therapy and photothermal therapy, has shown great potential in the field of antibacterial therapy due to its non-invasiveness, spatiotemporal controllability, and lack of drug resistance. However, it is worth noting that reactive oxygen species generated during photodynamic therapy not only cause bacterial death but also cause oxidative stress to adjacent tissues and cells. The presence of excessive and continuous reactive oxygen species, as well as bacterial proliferation, can induce severe inflammation and disrupt the redox balance. Therefore, it is necessary to achieve controlled reactive oxygen species regulation during treatment, first generating sufficient reactive oxygen species to kill bacteria, and then removing excess reactive oxygen species to inhibit inflammation and restore the redox balance.
[0004] Constructing a heterojunction is an effective strategy that can achieve multi-modal therapy using one material through band engineering and interface engineering design. A heterojunction is composed of two semiconductors with different band structures. The unique interface structure can promote charge transport and effective separation of electrons and holes, providing new possibilities for the design and functionalization of new materials. Therefore, it is of great significance to develop a heterojunction material with both antibacterial and anti-inflammatory functions. SUMMARY
[0005] In view of the deficiencies in the prior art, the application provides a near-infrared light-responsive nano-heterojunction with antibacterial and anti-inflammatory functions and a preparation method thereof. The near-infrared light-responsive nano-heterojunction with antibacterial and anti-inflammatory functions of the application has excellent reactive oxygen species generation capacity and photothermal effect under near-infrared light irradiation, can effectively kill bacteria, and can efficiently remove reactive oxygen species after stopping near-infrared light irradiation, thereby reducing inflammation and restoring the redox balance.
[0006] The application achieves the following technical solutions:
[0007] A near-infrared light-responsive nano-heterojunction with antibacterial and anti-inflammatory functions, and a preparation method thereof, includes the following steps:
[0008] 1) Potassium ferricyanide, polyvinylpyrrolidone are added to dilute hydrochloric acid, stirred to fully dissolve, aged, after the end centrifugal collection of precipitate, washed with deionized water, drying, to get Prussian blue PB nanoparticles.
[0009] 2) The PB nanoparticles obtained in step 1) are dispersed in ethylene glycol, 1,2,4,5-tetracyano benzene, manganese chloride and catalyst 1,8-diazabicyclo[5.4.0]undec-7-ene are added, polymerized under high temperature conditions, after the end centrifugal collection of precipitate, washed with ethanol, hydrochloric acid, ethanol, water in turn, drying, to get Prussian blue@polyphthalocyanine manganese nano-heterojunction.
[0010] In the above technical solution, further, the concentration ratio of potassium ferricyanide, polyvinylpyrrolidone and dilute hydrochloric acid in step 1) is 10:1.3:10.
[0011] Further, the aging temperature in step 1) is 75-85℃, and the aging time is 18-22h.
[0012] Further, the concentration ratio of PB nanoparticles, 1,2,4,5-tetracyano benzene and manganese chloride in step 2) is 0.5mg / mL:12.5-25mM:6-12.5mM, and the concentration of hydrochloric acid is 0.75M.
[0013] Further, the polymerization reaction temperature in step 2) is 175-185℃, and the reaction time is 20-40min.
[0014] The present application provides a near-infrared light responsive nano-heterojunction with antibacterial and anti-inflammatory functions based on the above method.
[0015] The principle of the present application is:
[0016] The present invention polymerizes polyphthalocyanine manganese (MnPPc) on the surface of Prussian blue nanoparticles by in situ polymerization to obtain Prussian blue @ polyphthalocyanine manganese (PB@MnPPc) nanoheterojunction. PB nanoparticles are a type of coordination polymer nanoparticle coordinated by iron and cyanide, have rich redox potentials, and are also an excellent photothermal agent. MnPPc is a nanoenzyme with excellent superoxide dismutase-like activity and catalase-like activity, and has a highly conjugated structure, which is conducive to charge transfer. By utilizing the physicochemical properties of the two, the two are combined to form a nanoheterojunction, which not only retains the photothermal effect and active oxygen scavenging function of the two, but also gives the new property of generating singlet oxygen under near-infrared light irradiation, thereby achieving the dual functions of antibacterial and anti-inflammatory through the control of near-infrared light. Experiments show that the nanoheterojunction obtained by the present invention has excellent singlet oxygen production ability and photothermal effect under 808nm near-infrared light irradiation, thereby being able to effectively kill bacteria; when the near-infrared light irradiation is stopped, the nanoheterojunction has excellent active oxygen scavenging ability, and can effectively scavenge hydrogen peroxide, superoxide anion radicals, and hydroxyl radicals, showing good anti-inflammatory effects.
[0017] The beneficial effects of the present invention are:
[0018] The raw materials are widely available, the preparation process is simple and controllable, and can be scaled up; it has excellent photodynamic and photothermal properties; it has outstanding active oxygen scavenging effects; it can achieve time-sequential antibacterial and anti-inflammatory dual functions through the control of near-infrared light; it has good stability under physiological conditions, good biocompatibility at the cellular level, and has practical application prospects. BRIEF DESCRIPTION OF THE DRAWINGS
[0019] Figure 1 is a transmission electron microscope image of the PB@MnPPc nanoheterojunction in the embodiment.
[0020] Figure 2 It is a high-angle annular dark-field imaging image of the PB@MnPPc nanoheterojunction in the embodiment and the corresponding distribution image of C, N, Fe, and Mn elements.
[0021] Figure 3 This is the hydrodynamic size of the PB@MnPPc nanoheterojunction in the embodiment after being treated in water and cell culture medium for 0 to 14 days.
[0022] Figure 4 In the embodiment, near infrared light (808nm, 0.5W / cm 2 ) Changes in the absorbance of the singlet oxygen detection probe (ABDA) at 399 nm after irradiation for 0 to 15 min.
[0023] Figure 5is the temperature-rising and temperature-falling curve of PB@MnPPc under near-infrared light (808nm, 0.5W / cm 2 ) irradiation, and the linear data of -lnθ and time obtained during the temperature-falling period.
[0024] Figure 6 is the scavenging ability of PB@MnPPc nano-heterojunctions of different concentrations on active oxygen, a is the inhibition ability on superoxide anion free radicals (·O2 - ), b is the scavenging ability on hydroxyl radicals (·OH), c is the scavenging ability on hydrogen peroxide (H2O2), and d is the case of decomposing H2O2 to produce O2.
[0025] Figure 7 is the comparison chart of the in-vitro antibacterial effect of PB@MnPPc nano-heterojunctions under near-infrared light (808nm, 0.5W / cm 2 ) irradiation for 10min.
[0026] Figure 8 is the active oxygen scavenging effect chart of PB@MnPPc nano-heterojunctions in mouse fibroblasts.
[0027] Figure 9 is the in-vitro cytotoxicity result chart of PB@MnPPc nano-heterojunctions after being co-incubated with mouse fibroblasts for 24h. DETAILED DESCRIPTION
[0028] In order to make the objects, technical solutions and advantages of the embodiments of the present application clearer, the technical solutions in the embodiments of the present application will be described clearly and completely below with reference to the drawings in the embodiments of the present application. Obviously, the described embodiments are some but not all of the embodiments of the present application. It should be understood that these embodiments are only used to explain the present application and not to limit the scope of the present application. Any changes, modifications, substitutions, combinations and simplifications made without departing from the spirit and principle of the present application shall be equivalent replacement modes and shall be included in the protection scope of the present application. In addition, it should be understood that after reading the content of the present application, those skilled in the art can make various changes or modifications to the present application, and these equivalent forms also fall within the scope defined by the claims of the present application and are within the protection scope of the present application.
[0029] The term “embodiment” mentioned in this document means that the specific features, structures or properties described in combination with the embodiment can be included in at least one embodiment of the present application. The term “embodiment” appearing at various places in the specification does not necessarily refer to the same embodiment, nor does it particularly limit its independence or association with other embodiments. In principle, in this application, as long as there is no technical contradiction or conflict, each technical feature mentioned in each embodiment can be combined in any way to form a corresponding implementable technical solution.
[0030] Unless otherwise defined, the meanings of the technical terms used herein are the same as those commonly understood by those skilled in the art to which the present application belongs; the use of related terms herein is only for the purpose of describing specific embodiments, and is not intended to limit the present application.
[0031] Unless otherwise specified, the reagents, instruments, equipment, etc. used in the following embodiments are reagents, instruments and equipment commonly used by those skilled in the art.
[0032] Embodiment
[0033] The present embodiment provides a near-infrared light responsive nano-heterojunction with antibacterial and anti-inflammatory functions, comprising the following steps:
[0034] 1) Add 526 mg of potassium ferricyanide and 12 g of polyvinylpyrrolidone to 160 mL of 10 mM dilute hydrochloric acid, stir to dissolve thoroughly, and age at 80℃ for 20 h. After completion, centrifuge at 12000 rpm for 10 min to collect the precipitate, wash with deionized water for 3 times, and dry to obtain PB nanoparticles.
[0035] 2) Disperse 10 mg of PB nanoparticles obtained in step 1) in 20 mL of ethylene glycol, add 19 mM of 1,2,4,5-tetracyanobenzene, 9.5 mM of manganese chloride and 67 μL of catalyst 1,8-diazabicyclo[5.4.0]undec-7-ene, and react at 180℃ for 30 min. After completion, centrifuge at 10000 rpm for 10 min to collect the precipitate, and wash with ethanol, 0.75 M hydrochloric acid, ethanol and water in turn, and dry to obtain PB@MnPPc nano-heterojunction.
[0036] The transmission electron microscope image of the near-infrared light responsive nano-heterojunction with antibacterial and anti-inflammatory functions prepared by the method of the present application is shown in Figure 1 As can be seen, the core-shell structure has a whole size of about 200 nm and a shell thickness of about 30 nm. Figure 2 The HAADF image and the C, N, Fe, Mn element distribution of the PB@MnPPc nano-heterojunction obtained under high-resolution transmission electron microscope further verify the structure of PB as the core and MnPPc as the shell. As shown in Figure 3As shown in Figure 3, the hydrodynamic size of PB@MnPPc in water and cell culture medium remained stable from 0 to 14 days, indicating that the PB@MnPPc nanoheterojunction has good stability under physiological conditions.
[0037] The singlet oxygen generation capability of PB@MnPPc nanoheterojunction under near-infrared light irradiation is shown in Fig. Figure 4 The particle concentration was 2.7 μg / mL, the H2O2 concentration was 60 μM, and the test solution was irradiated with an 808 nm laser at a power of 0.5 W / cm 2 The total irradiation time is 15 minutes. ABDA is a singlet oxygen detection probe. After binding to singlet oxygen, its absorbance will decrease. Figure 4 It can be seen that the absorbance of ABDA in the three groups of H2O+NIR, PB+NIR, and PB@MnPPc-NIR basically does not change with time. The absorbance of the MnPPc+NIR group decreased by about 23% after 15 minutes of irradiation, and the absorbance of the PB@MnPPc+NIR group decreased by about 83% after 15 minutes of irradiation, indicating that under near-infrared light irradiation, compared with PB and MnPPc alone, the singlet oxygen production capacity of the PB@MnPPc nanoheterojunction is greatly improved. The absorbance of the PB@MnPPc+H2O2+NIR group decreased by about 95% after 15 minutes of irradiation. This is because the catalase-like activity of PB@MnPPc itself will decompose H2O2 and produce O2, indicating that PB@MnPPc has a further enhanced photodynamic effect in the presence of H2O2. Figure 4 It can be seen that the PB@MnPPc nanoheterojunction prepared by the method of the present invention has excellent singlet oxygen generation ability under 808nm near-infrared light irradiation.
[0038] The photothermal performance of PB@MnPPc nanoheterojunction is shown in Figure 5 Using near infrared light (808nm, 0.5W / cm 2 ) irradiated 100 μg / mL PB@MnPPc dispersion for 10 min, then allowed to cool naturally, and monitored the temperature change of the liquid and the photothermal conversion. Figure 5 As shown, the calculated photothermal conversion efficiency of the PB@MnPPc nanoheterojunction is 36.7%, indicating that the PB@MnPPc nanoheterojunction prepared by the method of the present invention has excellent photothermal conversion capability.
[0039] The active oxygen scavenging ability of PB@MnPPc nanoheterojunction is shown in Figure 6 . Figure 6 a is the activity of PB@MnPPc to·O2 at different concentrations measured using the total superoxide dismutase activity detection kit (WST-8 method) -The inhibition rate of PB@MnPPc on ·O2 is more than 80%, indicating that the PB@MnPPc nano heterojunction has excellent superoxide anion radical scavenging capacity. - The inhibition rate of PB@MnPPc on ·O2 is more than 80%, indicating that the PB@MnPPc nano heterojunction has excellent superoxide anion radical scavenging capacity. Figure 6 In the b, the removal rate of PB@MnPPc on ·OH is detected by using terephthalic acid at different concentrations, and when the concentration reaches 10 μg / mL, the removal rate of PB@MnPPc on ·OH is more than 70%, indicating that the PB@MnPPc nano heterojunction has good hydroxyl radical scavenging capacity. Figure 6 In the c, the removal rate of PB@MnPPc on H2O2 is detected by using potassium iodide at different concentrations, and when the concentration reaches 10 μg / mL, the removal rate of PB@MnPPc on H2O2 is more than 80%, indicating that the PB@MnPPc nano heterojunction has excellent hydrogen peroxide scavenging capacity. Figure 6 In the d, the condition that PB@MnPPc at different concentrations decomposes H2O2 to produce O2 is measured by using a dissolved oxygen meter, and in a 10 mM H2O2 solution, after 5 min of adding 10 μg / mL PB@MnPPc, the content of dissolved oxygen in the solution increases by more than 10 mg / mL, combined with the result of Figure 6 In the c, it is proved that PB@MnPPc has excellent peroxidase-like activity. Figure 6 It can be seen that the PB@MnPPc nano heterojunction prepared by the method has excellent active oxygen scavenging function.
[0040] Gram-positive S. aureus and gram-negative E. coli are selected as model bacteria for in vitro antibacterial experiments, and the antibacterial effect of PB@MnPPc under near-infrared light irradiation is evaluated by agar plate counting method. 0.1M pH=5.5 acetic acid-acetate buffer solution and 100 μM H2O2 are used to simulate the microenvironment of the bacterial infection site, 10 6 CFU / mL bacteria are added, 100 μg / mL PB@MnPPc is added to the S. aureus system (80 μg / mL PB@MnPPc is added to the E. coli system), incubated for 1 h, irradiated under near-infrared light (808 nm, 0.5 W / cm 2 ) for 10 min, and then incubated for 1 h, diluted to an appropriate concentration, spread on an agar plate, and incubated at 37°C for 16 h before counting. As shown in Figure 7 PB@MnPPc+NIR and H2O2+PB@MnPPc+NIR both show significant bactericidal effect on S. aureus and E. coli, indicating that the PB@MnPPc nano heterojunction indeed has excellent bactericidal function under near-infrared light irradiation.
[0041] The effect of PB@MnPPc nano-heterojunction on the clearance of intracellular reactive oxygen species in vitro cell experiments is shown in Figure 8 . After PB@MnPPc was co-cultured with mouse fibroblasts for 4 h, 100 μΜ H2O2 was added for co-incubation for 1 h, and then the intracellular reactive oxygen level was detected using a reactive oxygen fluorescence probe (DCFH-DA), and the results are shown in Figure 8 . The positive control group has a higher reactive oxygen level, and the reactive oxygen level of the PB@MnPPc treated group is significantly lower than that of the positive control group, indicating that PB@MnPPc nano-heterojunction can effectively clear the intracellular reactive oxygen and relieve inflammation.
[0042] The cytotoxicity of PB@MnPPc nano-heterojunction is shown in Figure 9 . After different concentrations of PB@MnPPc were co-cultured with mouse fibroblasts for 24 h, the cell viability was detected using a cytotoxicity detection kit (MTT method), and it was found that when the concentration of PB@MnPPc reached 100 μg / mL, the relative cell viability could still be maintained above 95%, indicating that PB@MnPPc has good biocompatibility.
[0043] The above-described embodiments are only some of the preferred schemes of the present application, and are not intended to limit the present application. Those of ordinary skill in the relevant art can make various changes and modifications without departing from the spirit and scope of the present application. Therefore, any technical scheme obtained by equivalent replacement or equivalent transformation falls within the protection scope of the present application.
Claims
1. A method for preparing a near-infrared light-responsive nanoheterojunction with antibacterial and anti-inflammatory functions, characterized in that: The following steps are involved: 1) dissolving potassium ferrocyanide and polyvinyl pyrrolidone in dilute hydrochloric acid for aging treatment. After aging, collecting the precipitate by centrifugation, washing with deionized water, and then drying to obtain Prussian blue PB nanoparticles; 2) The PB nanoparticles obtained in step 1) were dispersed in ethylene glycol, and 1,2,4,5-tetracyanobenzene, manganese chloride, and a catalyst, 1,8-diazabicyclo[5.4.0]undec-7-ene, were added to carry out a polymerization reaction. After the reaction, the precipitate was collected by centrifugation, washed sequentially with ethanol, hydrochloric acid, ethanol, and water, and then dried to obtain a Prussian blue@manganese polyphthalocyanine (PB@MnPPc) nanoheterojunction.
2. The method for preparing a near-infrared light-responsive nanoheterojunction with antibacterial and anti-inflammatory functions according to claim 1, characterized in that: The concentration ratio of potassium ferrocyanide, polyvinyl pyrrolidone and dilute hydrochloric acid in step 1) is 10:1.3:
10.
3. The method for preparing a near-infrared light-responsive nanoheterojunction with antibacterial and anti-inflammatory functions according to claim 1, characterized in that: The temperature of the aging treatment in step 1) is 75-85° C., and the aging time is 18-22 hours.
4. The method for preparing a near-infrared light-responsive nanoheterojunction with antibacterial and anti-inflammatory functions according to claim 1, characterized in that: The concentration ratio of the PB nanoparticles, 1,2,4,5-tetracyanobenzene, and manganese chloride in step 2) is 0.5 mg / mL: 12.5-25 mM: 6-12.5 mM, and the concentration of the hydrochloric acid is 0.75 M.
5. The method for preparing a near-infrared light-responsive nanoheterojunction with antibacterial and anti-inflammatory functions according to claim 1, characterized in that: The polymerization reaction temperature in step 2) is 175-185° C., and the reaction time is 20-40 min.
6. A near-infrared light-responsive nanoheterojunction with antibacterial and anti-inflammatory functions, characterized in that: The method is prepared by any one of claims 1 to 5.