Black phosphorus phthalocyanine hydrogel and preparation method and application thereof

Abstract

This invention belongs to the field of biomedical materials technology, and relates to a black phosphorus phthalocyanine hydrogel, its preparation method, and its application. The black phosphorus phthalocyanine hydrogel comprises black phosphorus phthalocyanine and a photocurable gel. The preparation method of black phosphorus phthalocyanine includes the following process: under acidic conditions, black phosphorus nanosheets are mixed with aminosilicon phthalocyanine, allowing the black phosphorus nanosheets and aminosilicon phthalocyanine to self-assemble to form black phosphorus phthalocyanine. This invention combines black phosphorus with aminosilicon phthalocyanine; under the action of near-infrared laser, black phosphorus and aminosilicon phthalocyanine can synergistically exert a bactericidal effect. Simultaneously, black phosphorus can degrade, and its product, phosphate, can promote bone tissue regeneration, thus more effectively treating peri-implantitis.

Description

Technical Field

[0001] This invention belongs to the field of biomedical materials technology, and relates to a black phosphorus phthalocyanine hydrogel, its preparation method and application. Background Technology

[0002] The information disclosed in this background section is intended only to enhance understanding of the overall background of the invention and is not necessarily to be construed as an admission or in any way implying that such information constitutes prior art known to those skilled in the art.

[0003] Peri-implantitis is a plaque-related pathological condition primarily caused by poor plaque control, mainly manifested as inflammation of the peri-implant mucosa and progressive bone loss. With the increasing number of dental implants and their extended service life, the incidence of peri-implantitis is also rising, ultimately leading to implant failure. Currently, there is still no effective treatment for peri-implantitis; for example, mechanical debridement can damage the implant surface structure, and antibiotic use can lead to increased drug resistance, among other problems.

[0004] The inventors' research found that photothermal therapy shows promising potential in the treatment of peri-implantitis, effectively reducing inflammation and removing plaque without damaging the implant surface structure. However, there is currently a lack of materials or drugs that can be used for photothermal therapy to treat peri-implantitis. Summary of the Invention

[0005] To address the shortcomings of existing technologies, the present invention aims to provide a black phosphorus phthalocyanine hydrogel, its preparation method, and its application. The present invention combines black phosphorus with aminosilicon phthalocyanine. Under the action of near-infrared laser, black phosphorus and aminosilicon phthalocyanine can synergistically exert a bactericidal effect. At the same time, black phosphorus can be degraded, and its product phosphate can promote bone tissue regeneration, thus more effectively treating peri-implantitis.

[0006] To achieve the above objectives, the technical solution of the present invention is as follows:

[0007] Firstly, a method for preparing black phosphorus phthalocyanine includes the following steps:

[0008] Under acidic conditions, black phosphorus nanosheets are mixed with aminosilicon phthalocyanine, allowing the black phosphorus nanosheets and aminosilicon phthalocyanine to self-assemble into black phosphorus phthalocyanine.

[0009] The black phosphorus nanosheets used in this invention have a large surface area and are negatively charged. Under acidic conditions, this facilitates the binding of protons to aminosilicon phthalocyanine, making the aminosilicon phthalocyanine positively charged. Thus, black phosphorus phthalocyanine is obtained by self-assembling the black phosphorus nanosheets and aminosilicon phthalocyanine through electrostatic forces.

[0010] On the other hand, a black phosphorus phthalocyanine is obtained by the above preparation method.

[0011] Thirdly, a black phosphorus phthalocyanine hydrogel comprising the aforementioned black phosphorus phthalocyanine and a matrix hydrogel.

[0012] Fourthly, a method for preparing the above-mentioned black phosphorus phthalocyanine hydrogel involves mixing the black phosphorus phthalocyanine with a matrix hydrogel until homogeneous.

[0013] Fifthly, the application of the aforementioned black phosphorus phthalocyanine or black phosphorus phthalocyanine hydrogel in the preparation of a drug for treating peri-implantitis.

[0014] The beneficial effects of this invention are as follows:

[0015] (1) The present invention can effectively utilize the electrons on the surface of black phosphorus by combining black phosphorus with aminosilyl phthalocyanine, thereby reducing the electron transfer between black phosphorus and oxygen molecules, thus effectively reducing the oxidation of black phosphorus and enhancing its stability.

[0016] (2) Black phosphorus has a negative charge on its surface. In this invention, amino-silicon phthalocyanine is introduced into the surface of black phosphorus, which can weaken the negative charge on the surface of black phosphorus and promote the interaction between black phosphorus and bacteria with a negative charge on its surface.

[0017] (3) In the black phosphorus phthalocyanine provided by the present invention, aminosilyl phthalocyanine has a good bactericidal effect under the action of laser, which can synergistically enhance the bactericidal performance of black phosphorus.

[0018] (4) In the black phosphorus phthalocyanine provided by the present invention, the dispersibility of black phosphorus can be increased by loading amino-silicon phthalocyanine onto the surface of black phosphorus, which can effectively improve its biocompatibility.

[0019] (5) In the black phosphorus phthalocyanine provided by the present invention, black phosphorus is a biodegradable two-dimensional nanosheet, and the degradation product is phosphate, which can promote the regeneration of bone tissue around the implant. The amino-silicon phthalocyanine and black phosphorus synergistically enhance osteogenic properties during the degradation process. Therefore, the present invention has the function of promoting bone tissue regeneration while being antibacterial and anti-inflammatory.

[0020] (6) The black phosphorus phthalocyanine provided by the present invention can effectively kill bacteria around the implant and reduce inflammatory response under the action of laser. At the same time, the application of black phosphorus and aminosilicon phthalocyanine can reduce the dependence on laser intensity and exert the bactericidal effect under the action of laser with lower energy intensity. Attached Figure Description

[0021] The accompanying drawings, which form part of this invention, are used to provide a further understanding of the invention. The illustrative embodiments of the invention and their descriptions are used to explain the invention and do not constitute an improper limitation of the invention.

[0022] Figure 1 This is a schematic diagram of the synthesis of BP-PC@GelMA in an embodiment of the present invention.

[0023] Figure 2 The images shown are characterization diagrams of the BP-PC prepared in the embodiments of the present invention; a is a scanning electron microscope image of BP and BP-PC, b is the result of the surface elemental distribution test of BP-PC, and c is the result of the zeta potential test.

[0024] Figure 3 The images show the surface morphology of BP-PC@GelMA in this embodiment of the invention; a is the surface morphology of the hydrogel, and b is the surface morphology of the hydrogel after one month of degradation.

[0025] Figure 4 The diagram shows the biosafety results of BP-PC@GelMA in this embodiment of the invention. a is the result of CCK8, and b is the result of cell live / dead fluorescence staining.

[0026] Figure 5 The diagram shows the bactericidal effect of BP-PC@GelMA in this embodiment of the invention. a represents the bactericidal rate of BP-PC@GelMA against Staphylococcus aureus, b indicates that the structure of Staphylococcus aureus was destroyed after treatment with BP-PC@GelMA, c indicates that Staphylococcus aureus died after treatment with BP-PC@GelMA, d represents the bactericidal rate of BP-PC@GelMA against Escherichia coli, e indicates that the structure of Escherichia coli was destroyed after treatment with BP-PC@GelMA, and f indicates that Escherichia coli died after treatment with BP-PC@GelMA.

[0027] Figure 6 The image shows the results of in vitro BP-PC@GelMA promoting bone tissue regeneration in an embodiment of the present invention. a) shows ALP and ARS staining, b) shows the bands in Western blotting, and c) shows the statistical results of Western blotting.

[0028] Figure 7 The image shows the results of BP-PC@GelMA promoting bone tissue regeneration in vivo in an embodiment of the present invention. A is the result of Micro-CT, B is the statistical result of Micro-CT, C is HE staining, and D is Masson staining. Detailed Implementation

[0029] It should be noted that the following detailed descriptions are exemplary and intended to provide further illustration of the invention. Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains.

[0030] It should be noted that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to limit the scope of exemplary embodiments according to the invention. As used herein, the singular form is intended to include the plural form as well, unless the context clearly indicates otherwise. Furthermore, it should be understood that when the terms "comprising" and / or "including" are used in this specification, they indicate the presence of features, steps, operations, devices, components, and / or combinations thereof.

[0031] This invention proposes a black phosphorus phthalocyanine hydrogel, its preparation method, and its application.

[0032] A typical embodiment of the present invention provides a method for preparing black phosphorus phthalocyanine, comprising the following steps:

[0033] Under acidic conditions, black phosphorus nanosheets are mixed with aminosilicon phthalocyanine, allowing the black phosphorus nanosheets and aminosilicon phthalocyanine to self-assemble into black phosphorus phthalocyanine.

[0034] In some embodiments, under anaerobic conditions, black phosphorus nanosheets and aminosilyl phthalocyanine are added to water, the pH is adjusted to acidic, and the mixture is thoroughly mixed. Specifically, thorough mixing is achieved by ultrasonication and stirring.

[0035] In one or more embodiments, after thorough mixing, the mixture is centrifuged and washed with anhydrous ethanol.

[0036] In some embodiments, the acidic conditions are: pH value of 3 to 5.

[0037] In some embodiments, the preparation method of black phosphorus nanosheets is as follows: under anaerobic conditions, black phosphorus powder is added to an organic solvent to prepare a black phosphorus crystal powder solution, followed by ice bath sonication, then two centrifugations, and finally washing and centrifugation. Specifically, the two centrifugation processes are as follows: first, centrifugation is performed at a speed of 3000-5000 rpm to collect the supernatant, and then centrifugation is performed at a speed of 9000-11000 rpm to obtain the precipitate. The washing and centrifugation process is as follows: washing is performed with anhydrous ethanol, followed by centrifugation.

[0038] In some embodiments, the preparation method of aminosilylphthalocyanine is as follows: dichlorosilylphthalocyanine is reacted with an amino long-chain compound in a solvent containing potassium carbonate and pyridine under an inert atmosphere. Specifically, the amino long-chain compound is... n is a natural number, for example, 2-(2-aminoethoxy)ethanol.

[0039] Another embodiment of the present invention provides a black phosphorus phthalocyanine obtained by the above preparation method.

[0040] A third embodiment of the present invention provides a black phosphorus phthalocyanine hydrogel comprising the above-mentioned black phosphorus phthalocyanine and a matrix hydrogel.

[0041] In some embodiments, the matrix hydrogel is methacrylamide gelatin (GelMA).

[0042] In some embodiments, the mass ratio of black phosphorus phthalocyanine to the matrix hydrogel is 1:500 to 1100.

[0043] A fourth embodiment of the present invention provides a method for preparing the above-mentioned black phosphorus phthalocyanine hydrogel, wherein the black phosphorus phthalocyanine is mixed evenly with the matrix hydrogel to obtain the product.

[0044] In some embodiments, the mixing temperature is 35–40°C.

[0045] In some embodiments, the mixture is thoroughly mixed and then cured. Specifically, when the matrix hydrogel is methacrylamide gelatin, the curing is either temporary cooling or permanent curing by blue-violet light irradiation. Temporary cooling is used for storage and facilitates subsequent use, typically involving cooling at 3–5°C for 5–15 minutes. For use, permanent curing is performed by blue-violet light irradiation for 2–5 minutes.

[0046] A fifth embodiment of the present invention provides the application of the above-mentioned black phosphorus phthalocyanine or black phosphorus phthalocyanine hydrogel in the preparation of a medicament for treating peri-implantitis.

[0047] To enable those skilled in the art to better understand the technical solution of the present invention, the technical solution of the present invention will be described in detail below with reference to specific embodiments.

[0048] Example

[0049] Synthesis of BP-PC@GelMA

[0050] Weigh 100 mL of N-methylpyrrolidone (NMP) in an anaerobic chamber. Weigh 100 mg of black phosphorus (BP) using a balance, place it in a 5 mL test tube, and then place the test tube in the anaerobic chamber. Grind the BP into powder in the anaerobic chamber, mix it with 100 mL of NMP, and transfer the mixture to a volumetric flask to prepare a 1 mg / mL black phosphorus crystalline powder solution. Seal the flask with sealing film to prevent oxidation. Place the volumetric flask in an ultrasonic cleaner and sonicate on ice for 10 hours. Aliquot the solution into 50 mL high-speed centrifuge tubes and centrifuge twice (4000 rpm for 15 min, collect the supernatant; 10000 rpm for 20 min, collect the precipitate). Wash the obtained powder with anhydrous ethanol and centrifuge again (10000 rpm) to obtain black phosphorus nanosheets, which are then stored anaerobically.

[0051] Dichlorosilyl phthalocyanine (SiPcCl2) (200.5 mg, 0.328 mmol), 2-(2-aminoethoxy)ethanol (1.33 mL, 13.08 mmol), K2CO3 (500 mg, 3.62 mmol), and pyridine (3 mL) were dissolved in 35 mL of anhydrous toluene. The mixture was then heated to 130 °C under a nitrogen atmosphere and stirred for 12 h. After heating, the reaction mixture was allowed to cool naturally to room temperature. The solvent in the reaction mixture was then removed by rotary evaporation. The mixture was dissolved in 80 mL of chloroform, washed 2-3 times with deionized water, filtered through filter paper, and distilled under reduced pressure using a rotary evaporator to obtain the crude product. Finally, the crude product was recrystallized from chloroform / n-hexane to obtain the bluish-green solid product, aminosilyl phthalocyanine (SiPC-NH2).

[0052] Weigh appropriate amounts of black phosphorus nanosheets (5 mg) and aminosilyl phthalocyanine (50 mg) in an anaerobic chamber, place them in a reagent bottle containing 10 mL of pure water, add an appropriate amount of hydrochloric acid to adjust the pH to an acidic environment (pH 3-5), mix well, seal, sonicate for 20 min, then stir on a magnetic stirrer for 5 h, centrifuge at 10000 rpm for 20 min, collect the precipitate, wash the powder with anhydrous ethanol, and centrifuge again (10000 rpm) to obtain black phosphorus phthalocyanine (BP-PC). Figure 1 As shown.

[0053] 0.1 g of methacrylamide gelatin (GelMA) was dissolved in 1 ml of solvent to prepare a photocurable gel, which was then placed in a 37°C incubator to prevent it from solidifying. 0.172 mg of black phosphorus phthalocyanine was mixed with the photocurable gel, heated to 37°C, and stirred until homogeneous to obtain a black phosphorus phthalocyanine hydrogel (BP-PC@GelMA). This hydrogel was then placed in a 4°C refrigerator and cooled for 10 minutes to temporarily solidify. For use, it was permanently cured by irradiation with blue-violet light for 4 minutes.

[0054] The prepared BP-PC was characterized by scanning electron microscopy and zeta potential analysis, and the results are as follows: Figure 2 As shown, the results demonstrate the successful synthesis of BP-PC.

[0055] Surface morphology of BP-PC@GelMA as follows Figure 3 As shown, the surface of BP-PC@GelMA has a porous structure.

[0056] Rat osteoblasts were co-cultured with BP-PC@GelMA, and the biosafety of BP-PC@GelMA was verified using CCK8 and cell liveness / death fluorescence staining. The results are as follows: Figure 4 As shown, BP-PC@GelMA does not exhibit significant cytotoxicity.

[0057] Through laser (2W / cm) 2BP-PC@GelMA was irradiated with 808 nm light to induce a temperature rise that killed bacteria. The bactericidal effect of BP-PC@GelMA on different bacteria was verified using colony counting, scanning electron microscopy, and fluorescent staining for bacterial viability and mortality. Results are as follows: Figure 5 As shown, BP-PC@GelMA has good bactericidal effects against both Staphylococcus aureus and Escherichia coli.

[0058] Rat osteoblasts were co-cultured with BP-PC@GelMA, and the osteogenic capacity of BP-PC@GelMA was verified using ALP staining, ARS staining, and Western blotting. Results are as follows: Figure 6 As shown, BP-PC@GelMA has a significant bone-promoting ability.

[0059] A rat model of skull defect was established to verify the osteogenic capacity of BP-PC@GelMA in vivo. Results are as follows: Figure 7 As shown, BP-PC@GelMA significantly promotes bone regeneration in rats.

[0060] The above description is merely a preferred embodiment of the present invention and is not intended to limit the invention. Various modifications and variations can be made to the present invention by those skilled in the art. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the scope of protection of the present invention.

Claims

1. A method for preparing black phosphorus phthalocyanine, characterized in that it includes... The process is as follows: Under acidic conditions, black phosphorus nanosheets are mixed with aminosilicon phthalocyanine, which allows the black phosphorus nanosheets and aminosilicon phthalocyanine to self-assemble into black phosphorus phthalocyanine. The acidic conditions have a pH value of 3 to 5.

2. The method for preparing black phosphorus phthalocyanine as described in claim 1, characterized in that, Under anaerobic conditions, black phosphorus nanosheets and aminosilyl phthalocyanine were added to water, the pH was adjusted to acidic, and the mixture was thoroughly mixed.

3. The method for preparing black phosphorus phthalocyanine as described in claim 2, characterized in that, After thorough mixing, centrifuge the mixture and wash it with anhydrous ethanol.

4. The method for preparing black phosphorus phthalocyanine as described in claim 1, characterized in that, The preparation method of black phosphorus nanosheets is as follows: under anaerobic conditions, black phosphorus powder is added to an organic solvent to form a black phosphorus crystal powder solution, which is then subjected to ice bath sonication, followed by two centrifugations, and finally washed and centrifuged.

5. A black phosphorus phthalocyanine, characterized in that, Obtained by the preparation method described in any one of claims 1 to 4.

6. A black phosphorus phthalocyanine hydrogel, characterized in that, It comprises the black phosphorus phthalocyanine and matrix hydrogel as described in claim 5.

7. The black phosphorus phthalocyanine hydrogel as described in claim 6, characterized in that, The matrix hydrogel is methacrylamide gelatin.

8. The black phosphorus phthalocyanine hydrogel as described in claim 6, characterized in that, The mass ratio of black phosphorus phthalocyanine to the matrix hydrogel is 1:500~1100.

9. A method for preparing the black phosphorus phthalocyanine hydrogel according to claim 6, characterized in that, The black phosphorus phthalocyanine is mixed evenly with the matrix hydrogel to obtain the final product.

10. The use of the black phosphorus phthalocyanine hydrogel of claim 5 or any one of claims 6-8 in the preparation of a medicament for treating peri-implantitis.

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