A supramolecular hybrid hydrogel continuously generating hydroxyl radicals and a preparation method thereof

Abstract

The application discloses a kind of supermolecular hybrid hydrogel continuously generating hydroxyl radical, wherein the supermolecular hybrid hydrogel is composed of sodium hyaluronate and a nanocomposite, the sodium hyaluronate is the substance forming the network structure of the supermolecular hybrid hydrogel, the nanocomposite is distributed in the network structure, and the nanocomposite is the complex of beta-cyclodextrin modified upconversion nanoparticle and (ferrocene methyl) trimethyl ammonium iodide;Wherein the mass ratio of beta-cyclodextrin modified upconversion nanoparticle and (ferrocene methyl) trimethyl ammonium iodide complex: sodium hyaluronate: deionized water is 52: (17.5-20): 1000.The supermolecular hybrid hydrogel of the application can catalyze the reverse Fenton reaction under the tumor microenvironment, in situ cyclic generation of hydroxyl radical, and has potential application in the treatment of malignant tumors.

Description

Technical Field

[0001] This invention relates to the field of polymer materials, specifically to a supramolecular hybrid hydrogel that continuously generates hydroxyl radicals and its preparation method. Background Technology

[0002] Reactive oxygen species (ROS) are a class of oxygen-containing compounds that are more chemically reactive than molecular oxygen, derived directly or indirectly from it. However, in recent years, with the deepening research on the biological functions of ROS, their role in killing tumor cells has gradually been recognized.

[0003] Hydroxyl radicals (·OH), as a type of reactive oxygen species, possess strong oxidizing properties and can damage biomolecules such as proteins, lipids, and DNA in tumor cells, leading to cancer cell death. By utilizing the Fenton reaction or a Fenton-like reaction, hydrogen peroxide in the tumor microenvironment (weakly acidic with excess H₂O₂) can be converted into highly toxic hydroxyl radicals (·OH), without requiring additional external intervention. This makes chemokinetic therapy (CDT) a novel strategy for effectively modulating the tumor microenvironment to achieve tumor treatment.

[0004] However, existing Fenton catalysis systems primarily deliver the metal catalyst (Fe2+) required for the Fenton reaction to the tumor site via intravenous injection, utilizing metal-based nanomaterials, organic framework materials, and carbon-based nanomaterials. 2+ The aforementioned delivery medium materials and the large amount of catalysts they bring are difficult to degrade in organisms and have serious toxic side effects and poor therapeutic effects, making them difficult to widely use in clinical practice.

[0005] Therefore, there is a need to develop a Fenton catalytic reaction promoter that can efficiently generate hydroxyl radicals in the microenvironment of malignant tumors and has low toxicity, in order to improve the clinical efficacy of chemokinetic therapy (CDT). Summary of the Invention

[0006] This invention provides a supramolecular hybrid hydrogel carrier material that continuously generates hydroxyl radicals. This hydrogel has high catalytic ability under the conditions of malignant tumor microenvironment, which can enhance the efficiency of Fenton reaction and achieve the purpose of continuous and efficient generation of ·OH, providing new ideas for biomaterial development and cancer treatment.

[0007] The first aspect of the present invention provides a supramolecular hybrid hydrogel that continuously generates hydroxyl radicals, the supramolecular hybrid hydrogel being composed of sodium hyaluronate and a nanocomposite, wherein the sodium hyaluronate is the substance forming the network structure of the supramolecular hybrid hydrogel, and the nanocomposite is distributed in the network structure. The nanocomposite is a complex of β-cyclodextrin-modified upconversion nanoparticles and (ferrocenemethyl)trimethylammonium iodide, and therefore, the nanocomposite is in a complex particle state; in the supramolecular hybrid hydrogel, the mass ratio of the β-cyclodextrin-modified upconversion nanoparticles and the (ferrocenemethyl)trimethylammonium iodide complex to sodium hyaluronate to deionized water is 52:(17.5-20):1000.

[0008] In the supramolecular hybrid hydrogel of the present invention, the raw material components include upconversion nanoparticles (NaYF4:Yb,Tm@NaYF4). After being assembled into the hydrogel, the upconversion nanoparticles still exhibit excellent ultraviolet emission properties, thus giving the hydrogel the characteristic of ultraviolet emission.

[0009] In the supramolecular hybrid hydrogel of the present invention, the upconversion nanoparticles are NaYF4:Yb,Tm@NaYF4 nanoparticles with a core-shell structure known in the prior art (NaYF4:Yb,Tm as the core component and NaYF4 as the shell component), wherein the molar ratio of Yb:Tm is 10:1, thereby ensuring that NaYF4:Yb,Tm is a particle with ultraviolet emission properties.

[0010] In the supramolecular hybrid hydrogel of this invention, the raw material component includes (ferrocenemethyl)trimethylammonium iodide (Fc). By incorporating Fc into the hydrogel material, the hydrogel can still function as a drug carrier, undergoing a phase transition in response to ROS, thereby releasing Fc and Fe. 2+ The released Fe 2+ It can trigger the Fenton reaction and generate hydroxyl radicals.

[0011] In the supramolecular hybrid hydrogel of this invention, the key to the formation of the supramolecular hybrid hydrogel lies not only in the mass ratio of sodium hyaluronate to water being (17.5-20):1000, but also in the mass ratio of the complex of β-cyclodextrin-modified upconversion nanoparticles and (ferrocenemethyl)trimethylammonium iodide, sodium hyaluronate, and deionized water being 52:(17.5-20):1000. If the ratio of sodium hyaluronate to deionized water in the raw materials does not reach (17.5-20):1000, the gel will not form; if it exceeds (17.5-20):1000, it cannot be injected and the expected effect cannot be achieved. More importantly, if the total amount of the complex of β-cyclodextrin-modified upconversion nanoparticles and (ferrocenemethyl)trimethylammonium iodide added is small, the positive charge provided by (ferrocenemethyl)trimethylammonium iodide will be insufficient, and it will not form a gel with the negatively charged sodium hyaluronate through electrostatic interaction.

[0012] In one embodiment of the supramolecular hybrid hydrogel of the present invention, in the nanocomposite, in the β-cyclodextrin-modified upconversion nanoparticles, the host-guest molar ratio of β-cyclodextrin to (ferrocenemethyl)trimethylammonium iodide is 1:1, that is, the mass ratio of β-cyclodextrin-modified upconversion nanoparticles to (ferrocenemethyl)trimethylammonium iodide is 50:2.

[0013] Different molecules assemble into nanocomposites in different ways. The different groups on (ferrocenemethyl)trimethylammonium iodide affect its assembly mechanism. Only when β-cyclodextrin and (ferrocenemethyl)trimethylammonium iodide are assembled in a 1:1 host-guest molar ratio can they fully assemble and form ROS-responsive bonds.

[0014] In this invention, in the nanocomposite, the β-cyclodextrin is the assembly host and the (ferrocenemethyl)trimethylammonium iodide is the assembly guest, and the nanocomposite is formed by the host-guest assembly of β-cyclodextrin and (ferrocenemethyl)trimethylammonium iodide.

[0015] In one embodiment of the supramolecular hybrid hydrogel of the present invention, the mass ratio of β-cyclodextrin-modified upconversion nanoparticles: (ferrocenemethyl)trimethylammonium iodide: sodium hyaluronate is 50:2:(17.5-20). By controlling the mass ratio of β-cyclodextrin-modified upconversion nanoparticles, (ferrocenemethyl)trimethylammonium iodide, and sodium hyaluronate within the above range, a supramolecular hybrid hydrogel with controllable mechanical properties can be obtained, that is, the injectability, adhesion, and other properties of the obtained supramolecular hybrid hydrogel can be controlled.

[0016] Another aspect of the present invention provides a method for preparing a supramolecular hybrid hydrogel, which can obtain the supramolecular hybrid hydrogel of the present invention through a very simple synthesis process. The method includes the following steps:

[0017] Step 1: Synthesis of β-cyclodextrin-modified upconversion nanoparticles;

[0018] Step 2: Assemble the β-cyclodextrin-modified upconversion nanoparticles with (ferrocenemethyl)trimethylammonium iodide to obtain a positively charged complex.

[0019] Step 3: Dissolve the nanocomposite in deionized water, add sodium hyaluronate, and form the supramolecular hybrid hydrogel through electrostatic interaction.

[0020] In a further embodiment of the method of the present invention, the following steps are included:

[0021] Step 1: Prepare ethylenediamine-modified β-cyclodextrin and prepare UV-emitting carboxylated NaYF4:Yb,Tm@NaYF4 with a Yb / Tm molar ratio of 10 / 1 by solvothermal method and ion exchange method. Then, use the ethylenediamine-modified β-cyclodextrin and the carboxylated NaYF4:Yb,Tm@NaYF4 to synthesize β-cyclodextrin-modified upconversion nanoparticles.

[0022] Step 2: The prepared β-cyclodextrin-modified upconversion nanoparticles were assembled with (ferrocenemethyl)trimethylammonium iodide at a host-guest molar ratio of 1:1 to obtain a nanocomposite. Only when assembled at a host-guest molar ratio of 1:1 can the β-cyclodextrin and (ferrocenemethyl)trimethylammonium iodide in the upconversion nanoparticles be fully assembled to form ROS-responsive bonds.

[0023] Step 3: Dissolve the nanocomposite in deionized water, add sodium hyaluronate, and form the supramolecular hybrid hydrogel through electrostatic interaction.

[0024] In a further embodiment of the method of the present invention, in step 3, the mass ratio of the nanocomposite to sodium hyaluronate to deionized water is 52:(17.5-20):1000. If the ratio of sodium hyaluronate to deionized water in the raw materials does not reach (17.5-20):1000, the gel will not form; if it exceeds (17.5-20):1000, it cannot be injected and the expected effect cannot be achieved. More importantly, if the total amount of the β-cyclodextrin-modified upconversion nanoparticles and the (ferrocenemethyl)trimethylammonium iodide nanocomposite is insufficient, the positive charge provided by (ferrocenemethyl)trimethylammonium iodide will be insufficient, and it will not form gel with the negatively charged sodium hyaluronate through electrostatic interaction.

[0025] In a further embodiment of the method of the present invention, the mass ratio of β-cyclodextrin-modified upconversion nanoparticles: (ferrocenemethyl)trimethylammonium iodide: sodium hyaluronate is 50:2:(17.5-20). By adjusting the ratio of the three components within the above range, a supramolecular hybrid hydrogel with tunable mechanical properties can be obtained.

[0026] In this invention, "β-CD" represents β-cyclodextrin; "UCs" represents "NaYF4:Yb,Tm@NaYF4"; "Fc" represents (ferrocenemethyl)trimethylammonium iodide; "HA-T" represents sodium hyaluronate; "UCs / CD" represents β-cyclodextrin-modified upconversion nanoparticles; "UCs / CD / Fc" represents a complex of β-cyclodextrin-modified upconversion nanoparticles (UCs / CD) and (ferrocenemethyl)trimethylammonium iodide (Fc); and "UCs / CD / Fc-HA" represents supramolecular hybrid hydrogel (abbreviated as UCFH).

[0027] The beneficial effects of this invention are as follows:

[0028] 1. The (ferrocenemethyl)trimethylammonium iodide describes the loading of Fe into the hydrogel material. 2+ In the malignant tumor microenvironment (pH 4.8-7.4 and H2O2 concentration 0.01mol / L-0.06mol / L), this Fe... 2+ It can be released from the hydrogel, thereby reacting with H2O2 to generate hydroxyl radicals through a Fenton reaction;

[0029] The (ferrocenemethyl)trimethylammonium iodide, as one of the raw materials, can be molecularly assembled with β-cyclodextrin-modified upconversion nanoparticles, with a host-guest molar ratio as high as 1:1; and

[0030] (Ferrocenemethyl)trimethylammonium iodide can make the surface of the molecular assembly complex of β-cyclodextrin-modified upconversion nanoparticles positively charged, and then form a hydrogel with negatively charged sodium hyaluronate through electrostatic interaction, making it possible to obtain the supramolecular hybrid hydrogel.

[0031] 2. NaYF4:Yb,Tm@NaYF4 in the supramolecular hybrid hydrogel of this invention can still convert near-infrared light into ultraviolet light (UCs / CD exhibits a blue-violet hue under 980nm near-infrared laser irradiation), thereby catalyzing the reverse Fenton reaction to form Fe. 2+ —Fe 3+ —Fe 2+ This cyclical system allows hydroxyl radicals (·OH) to be generated in situ within the malignant tumor microenvironment, thereby introducing less Fe into the organism. 2+This allows for the continuous and efficient generation of ·OH, maintaining or even enhancing the efficiency of the Fenton reaction.

[0032] 3. The supramolecular hybrid hydrogel of the present invention has properties such as injectability, adhesion and biocompatibility. It can be injected into malignant tumors at specific points through minimally invasive injection and adhere to the inner wall of the tumor. Therefore, it has stronger targeting and a longer duration of action in the malignant tumor microenvironment. Thus, it can efficiently generate hydroxyl radicals by introducing a low amount of UCs / CD / Fc into the body, reducing the toxic side effects on the body. It has broad application prospects in tumor treatment.

[0033] 4. The method for preparing supramolecular hybrid hydrogels in this invention forms supramolecular hybrid hydrogels through molecular assembly and intermolecular electrostatic forces. It involves few steps, readily available reaction conditions, and is very simple. Attached Figure Description

[0034] Figure 1 This is a schematic diagram illustrating the synthetic route of supramolecular hybrid hydrogels and their role in enhancing tumor CDT therapy.

[0035] Figure 2 (a) is a transmission electron microscope image of UCs; Figure 2 (b) is a transmission electron microscope image of UCs / CD.

[0036] Figure 3 The image shows the fluorescence emission spectrum of UCs / CD.

[0037] Figure 4 The UV-Vis absorption spectra of compounds Fc, CD, and CD / Fc inclusion complex are shown.

[0038] Figure 5 (a) The 1H NMR spectra of compounds Fc, CD and CD / Fc inclusion complex; Figure 5 (b) is the two-dimensional NOESY spectrum of the CD / Fc inclusion complex.

[0039] Figure 6 (a) Electron photograph of the formation process of supramolecular hybrid hydrogels; Figure 6 (b) Injectability testing of supramolecular hybrid hydrogels; Figure 6 (c) Viscosity test of supramolecular hybrid hydrogels.

[0040] Figure 7 (a) is a transmission electron microscope image of the supramolecular hybrid hydrogel; Figure 7 (b) is a scanning electron microscope image of the supramolecular hybrid hydrogel.

[0041] Figure 8 Rheological testing of supramolecular hybrid hydrogels. Figure 8 (a) Oscillation frequency scan curve; Figure 8(b) Graph showing the variation of shear stress and viscosity with shear rate.

[0042] Figure 9 Zeta potential diagrams for UCs, UCs / CD, UCs / CD / Fc, and UCFH supramolecular hybrid hydrogels.

[0043] Figure 10 Fourier transform infrared spectra of Fc, UCs / CD / Fc, HA, and UCFH supramolecular hybrid hydrogels.

[0044] Figure 11 The UV-Vis absorption spectra of supramolecular hybrid hydrogel (0.75 mg / mL) and 3,3',5,5'-tetramethylbenzidine (TMB) under different conditions are shown.

[0045] Figure 12 The UV-Vis absorption spectra of supramolecular hybrid hydrogel (0.75 mg / mL) and TMB at pH 4.8 are shown in H2O2 solutions of different concentrations (0.01-0.06 mol / L).

[0046] Figure 13 Images showing the color variations of supramolecular hybrid hydrogel (0.75 mg / mL) and TMB in H2O2 solutions of different concentrations (0.01-0.06 mol / L) at pH 4.8.

[0047] Figure 14 The UV-Vis absorption spectra of supramolecular hybrid hydrogels (0.25-1.25 mg / mL) and TMB in 0.04 mol / L H2O2 solution at pH 4.8 are shown.

[0048] Figure 15 (a) The UV-Vis absorption spectra of supramolecular hybrid hydrogel (0.75 mg / mL) and TMB at different reaction times in 0.04 mol / L H2O2 solution; Figure 15 (b) The curves showing the change in absorbance at 655 nm of the supramolecular hybrid hydrogel at different reaction times.

[0049] Figure 16 (a) The UV-Vis absorption spectra of supramolecular hybrid hydrogel (0.75 mg / mL) and TMB at different reaction times in 0.04 mol / L H2O2 solution under near-infrared light (980 nm) irradiation. Figure 16 (b) shows the change curves of the absorbance of the supramolecular hybrid hydrogel at 655 nm under near-infrared light (980 nm) irradiation and different reaction times. Detailed Implementation

[0050] The present invention will be further described below with reference to specific embodiments, but the present invention is not limited to the following embodiments. Those skilled in the art should understand that modifications or substitutions can be made to the details and form of the technical solutions of the present invention without departing from the spirit and scope of the present invention, but all such modifications and substitutions fall within the protection scope of the present invention.

[0051] Main instruments and equipment used in the experiment

[0052]

[0053] Main raw materials and reagents

[0054]

[0055]

[0056]

[0057] Example

[0058] 1. Synthesis of β-cyclodextrin-modified upconversion nanoparticles (UCs / CD)

[0059] (1) Synthesis of mono-(6-ethylenediamine-6-deoxy)-β-cyclodextrin

[0060] Add 1.5L of distilled water and 270g of β-CD (β-cyclodextrin) to a 5L beaker to obtain a white emulsion. Stir constantly. Dissolve 25.7g of NaOH in 77mL of distilled water, cool, and slowly add to the beaker containing the white emulsion. The solution becomes clear and pale yellow. Dissolve 39g of TSCI (p-toluenesulfonyl chloride) in 128mL of acetonitrile and slowly add to the clear, pale yellow reaction system at 1-2 drops per second. After the addition is complete, stir at room temperature for 2 hours, producing a small amount of white precipitate. Filter and discard the precipitate to obtain a pale yellow-green solution. Adjust the pH of the pale yellow-green solution to 7-8 with 2M HCl (25.7mL concentrated HCl added to 250mL of water), causing a white precipitate to form. Stir thoroughly and refrigerate overnight. Filter under reduced pressure to obtain the white precipitate. After recrystallizing the white precipitate twice, the mixture was filtered and dried to obtain a white solid, which is 6-β-CD-OTS (mono-(6-O-p-methylbenzenesulfonyl)-β-cyclodextrin).

[0061] 4 g of 6-β-CD-OTS was dissolved in 45 mL of ethylenediamine. The mixture was heated to 85 °C under N2 protection and refluxed for 48 h. The reaction product was then cooled to room temperature. 1000 mL of acetone was prepared, and the reaction product was added dropwise to the acetone, resulting in a precipitate. The precipitate was obtained by suction filtration. The obtained solid was dissolved in 10 mL of water, the pH was adjusted to 7, and the precipitate was added dropwise to acetone. The precipitate was then filtered, dried, and mono-(6-ethylenediamine-6-deoxy)-β-cyclodextrin was obtained.

[0062] (2) Synthesis of NaYF4:Yb,Tm@NaYF4(UCs)

[0063] Dissolve 1.60 mmol of YCl3·6H2O, 0.40 mmol of YbCl3·6H2O, and 0.04 mmol of TmCl3·6H2O in 2 mL of water. Add 15 mL of oleic acid and 30 mL of 1-octadecene to the solution and stir vigorously at room temperature for 1 h to ensure thorough mixing. Then heat the mixture to 120 °C under N2 protection and maintain the temperature for 15 min to remove water. Continue heating to 160 °C and stir for 1 h until the solution becomes a uniform, transparent yellow. Cool the solution to room temperature under N2 protection, then add 10 mL of a methanol mixture containing 296.3 mg of ammonium fluoride and 200 mg of sodium hydroxide and stir vigorously at room temperature for 2 h. Heat the solution to 65 °C to remove methanol, then slowly heat to 290 °C and react for 1.5 h. After cooling, add 20 mL of ethanol and centrifuge at 10000 rpm for 15 min to collect the crude product. The crude product was dispersed in 5 mL of cyclohexane, and precipitated by adding 15 mL of ethanol. The precipitate was washed by centrifugation at 10000 r / 15 min, and the washing process was repeated 4 times. The final centrifuged product was added to 20 mL of cyclohexane to obtain monodisperse NaYF4:Yb,Tm nanoparticles (Yb:Tm molar ratio of 10:1).

[0064] 800 μmol of YCl3·6H2O, 15 mL of oleic acid, and 30 mL of 1-octadecene were mixed and stirred vigorously for 1 h. The mixture was heated to 160 °C under N2 and maintained for 1 h until the solution turned yellow and transparent. Then, it was cooled to room temperature. Next, 5 mL of the NaYF4:Yb,Tm prepared above was added and stirring was continued for 30 min. Cyclohexane was removed by heating. After cooling, 10 mL of a methanol mixture containing 296.3 mg of ammonium fluoride and 200 mg of sodium hydroxide was added and stirred for 2 h. After removing the methanol by heating, the solution was slowly heated to 270 °C and reacted for 1.5 h before cooling. The washing process was the same as above. The resulting product was dispersed in 20 mL of cyclohexane to prepare UV-emitting core-shell upconversion nanoparticles NaYF4:Yb,Tm@NaYF4 (UCs).

[0065] (3) Synthesis of β-cyclodextrin modified upconversion nanoparticles (UCs / CD)

[0066] UV-emitting upconversion nanoparticles NaYF4:Yb,Tm@NaYF4 (UCs) prepared by a solvothermal method were carboxylated by ion exchange. 20 mL of dichloromethane / anhydrous ethanol (v / v = 1:1) solution was added to 100 mg of UCs, followed by the addition of 100 mg of glyphosate. The mixture was stirred vigorously overnight. 50 mL of n-hexane was added to the mixture, and after thorough stirring, the product was collected by centrifugation. The product was repeatedly washed with n-hexane, and the precipitate was dried under vacuum to obtain carboxylated UCs nanoparticles.

[0067] 60 mg of the carboxylated UCs nanoparticles prepared above were dissolved in 10 mL of dimethylformamide solution, and then 200 mg of mono-(6-ethylenediamine-6-deoxy)-β-cyclodextrin, 100.0 mg of N,N-diisopropylethylamine and 160.1 mg of benzotriazol-1-yl-oxytripyrrolidinephosphine hexafluorophosphate were added. The reaction was carried out under N2 protection and stirred at room temperature for 48 h. After the reaction was completed, the product was collected by centrifugation, dialyzed for 48 h to remove excess impurities, and then lyophilized to obtain UCs / CD.

[0068] Figure 2 (a) and Figure 2 (b) shows that upconversion nanoparticles UCs ​​with a particle size of approximately 50 nm were successfully synthesized. Further surface modification with CD yielded UCs / CD composite nanoparticles, which still possessed a hexagonal crystal structure.

[0069] Figure 3 This indicates that UCs / CD composite nanoparticles exhibit ultraviolet emission characteristics, with a significant absorption peak in the ultraviolet region.

[0070] 2. Synthesis of (ferrocenemethyl)trimethylammonium iodide (Fc) loaded composite nanoparticles (UCs / CD / Fc)

[0071] The prepared UCs / CD and Fc were further assembled at a host-guest molar ratio of 1:1. 50 mg of UCs / CD was dissolved in 10 mL of deionized water and ultrasonically mixed until homogeneous. 2 mg of Fc was added to the solution, and ultrasonication was continued for 30 min to obtain positively charged UCs / CD / Fc composite nanoparticles.

[0072] Figure 4 This indicates that compounds Fc and CD form a stable inclusion complex in a 1:1 molar ratio.

[0073] Figure 5One-dimensional 1H NMR spectrum (298K, 400MHz, D2O) and two-dimensional NOESY spectrum show that in aqueous solution, compound Fc enters the hydrophobic cavity of CD to form a stable CD / Fc assembly, further indicating that adding compound Fc to UCs / CD can form UCs / CD / Fc composite nanoparticles.

[0074] 3. Synthesis of supramolecular hybrid hydrogels (UCs / CD / Fc-HA, UCFH)

[0075] Dissolve 52 mg of UCs / CD / Fc in 1 mL of deionized water, add 18 mg of sodium hyaluronate (HA-T), and stir at room temperature for 12 h to form a UCFH supramolecular hybrid hydrogel under electrostatic interaction.

[0076] Figure 6 middle, Figure 6 (a) Figure 6 (b) and Figure 6 (c) This indicates that when sodium hyaluronate (HA) and deionized water are added to UCs / CD / Fc at the mass ratio of the present invention, a UCs / CD / Fc-HA (UCFH) supramolecular hydrogel is formed in situ under electrostatic interaction. UCFH is in an inverted, non-flowing gel state and has injectability and a certain degree of viscosity.

[0077] Figure 7 The transmission electron microscopy (TEM) images show spherical particles with a diameter of approximately 60 nm, which are UCs doped into the hydrogel. Scanning electron microscopy (SEM) reveals the three-dimensional cross-linked network structure of the supramolecular hybrid hydrogel, which possesses a large specific surface area and porosity, making it a potential drug carrier material.

[0078] Figure 8 The supramolecular hybrid hydrogel exhibits excellent mechanical properties and shear-thinning properties.

[0079] Figure 9 The changes in the zeta potential of the samples demonstrate that the supramolecular hybrid hydrogels are bound together through electrostatic interactions.

[0080] Figure 10 This indicates that the supramolecular hybrid hydrogel was successfully constructed.

[0081] Figure 11This demonstrates that the generation of ·OH (hydroxyl radicals) can be detected using the TMB method, specifically through an in vitro experiment simulating the tumor microenvironment (pH 4.8 + H₂O₂). The supramolecular hybrid hydrogel was added to ethanol solutions containing H₂O₂ at pH 7.4 and 4.8, and to an ethanol solution without H₂O₂ at pH 4.8, respectively, resulting in final sodium hyaluronate concentrations of 0.75 mg / mL and 0.04 mol / L (in this invention, "0.75 mg / mL" refers to the mass of sodium hyaluronate, and 1 mL refers to the volume of the aqueous solution). With the addition of H₂O₂ and acid, the ·OH generated by the supramolecular hybrid hydrogel can be oxidized to the oxidation state of TMB (signal peak at 652 nm). The figure shows that under the conditions of pH 4.8 + H₂O₂, the supramolecular hybrid hydrogel of this invention significantly generates ·OH (hydroxyl radicals).

[0082] Figure 12 The UV-Vis absorption spectra show the results of the determination of ·OH formation using the TMB colorimetric reaction. Ethanol solutions containing 0.8 mM TMB were mixed with supramolecular hybrid hydrogels and H2O2 solutions of different concentrations, resulting in a final sodium hyaluronate concentration of 0.75 mg / mL and an H2O2 concentration of 0.01–0.06 mol / L. The reactions were carried out at pH 4.8 for 8 min.

[0083] Figure 13 Images showing the color changes of solutions after mixing ethanol solutions containing TMB (0.8 mM) with supramolecular hybrid hydrogels and H2O2 solutions of different concentrations, resulting in a final sodium hyaluronate concentration of 0.75 mg / mL and an H2O2 concentration of 0.01-0.06 mol / L, under pH 4.8 conditions for 8 min.

[0084] Figure 14 The UV-Vis absorption spectra of ethanol solutions containing TMB (0.8 mM) mixed with supramolecular hybrid hydrogels containing H2O2 solutions of varying concentrations, resulting in a final H2O2 concentration of 0.04 mol / L and sodium hyaluronate concentrations of 0.25–1.25 mg / mL, were shown after 8 min of reaction at pH 4.8.

[0085] Figure 15 (a) shows the UV-Vis absorption spectra of an ethanol solution containing TMB (0.8 mM) mixed with H2O2 solution and supramolecular hybrid hydrogel, respectively, to achieve a final H2O2 concentration of 0.04 mol / L and a sodium hyaluronate concentration of 0.75 mg / mL, at different reaction times under pH 4.8 conditions (without near-infrared light (980 nm) irradiation). Figure 15(b) shows the variation curves of the absorbance at 655 nm of the supramolecular hybrid hydrogel at different reaction times.

[0086] Figure 16 (a) shows the UV-Vis absorption spectra of an ethanol solution containing TMB (0.8 mM) mixed with H2O2 solution and supramolecular hybrid hydrogel under near-infrared light (980 nm) irradiation, with the final H2O2 concentration being 0.04 mol / L and the sodium hyaluronate concentration being 0.75 mg / mL, at different reaction times under pH=4.8 conditions. Figure 16 (b) shows the variation curves of the absorbance of the supramolecular hybrid hydrogel at 655 nm under near-infrared light (980 nm) irradiation and different reaction times.

[0087] contrast Figure 15 b and Figure 16 As can be seen from the slope of b, the rate of ·OH generation of the supramolecular hybrid hydrogel of the present invention is accelerated under near-infrared light (980nm) irradiation compared to the case without near-infrared light (980nm) irradiation.

Claims

1. A supramolecular hybrid hydrogel that continuously generates hydroxyl radicals, characterized in that, The supramolecular hybrid hydrogel is composed of sodium hyaluronate and a nanocomposite. Sodium hyaluronate is the substance that forms the network structure of the supramolecular hybrid hydrogel. The nanocomposite is distributed in the network structure. The nanocomposite is a complex of β-cyclodextrin-modified upconversion nanoparticles and (ferrocenemethyl)trimethylammonium iodide. In the supramolecular hybrid hydrogel, the mass ratio of the complex of β-cyclodextrin-modified upconversion nanoparticles and (ferrocenemethyl)trimethylammonium iodide:sodium hyaluronate:deionized water is 52:(17.5-20):1000; The nanocomposite was prepared by the following steps: Step 1: Prepare ethylenediamine-modified β-cyclodextrin and prepare UV-emitting carboxylated NaYF4:Yb,Tm@NaYF4 with a Yb:Tm molar ratio of 10:1 by solvothermal method and ion exchange method. Then, use the ethylenediamine-modified β-cyclodextrin and the carboxylated NaYF4:Yb,Tm@NaYF4 to synthesize β-cyclodextrin-modified upconversion nanoparticles. Step 2: Assemble the prepared β-cyclodextrin-modified upconversion nanoparticles with (ferrocenemethyl)trimethylammonium iodide at a molar ratio of 1:1 to obtain a nanocomposite.

2. The supramolecular hybrid hydrogel according to claim 1, characterized in that, β-cyclodextrin modified upconversion nanoparticles: (ferrocenemethyl)trimethylammonium iodide:sodium hyaluronate in a mass ratio of 50:2:(17.5-20).

3. The supramolecular hybrid hydrogel according to claim 1 or 2, characterized in that, The supramolecular hybrid hydrogel is formed by first preparing β-cyclodextrin-modified upconversion nanoparticles, then assembling the prepared β-cyclodextrin-modified upconversion nanoparticles with (ferrocenemethyl)trimethylammonium iodide at a molar ratio of 1:1 to form the nanocomposite, and finally forming the supramolecular hybrid hydrogel by electrostatic interaction between the nanocomposite and sodium hyaluronate in deionized water at a mass ratio of nanocomposite:sodium hyaluronate:deionized water of 52:(17.5-20):1000.

4. The method for preparing the supramolecular hybrid hydrogel according to any one of claims 1-3, characterized in that, Includes the following steps: Step 1: Prepare ethylenediamine-modified β-cyclodextrin and prepare UV-emitting carboxylated NaYF4:Yb,Tm@NaYF4 with a Yb:Tm molar ratio of 10:1 by solvothermal method and ion exchange method. Then, use the ethylenediamine-modified β-cyclodextrin and the carboxylated NaYF4:Yb,Tm@NaYF4 to synthesize β-cyclodextrin-modified upconversion nanoparticles. Step 2: The prepared β-cyclodextrin-modified upconversion nanoparticles were assembled with (ferrocenemethyl)trimethylammonium iodide at a molar ratio of 1:1 to obtain a nanocomposite. Step 3: Dissolve the nanocomposite in deionized water, add sodium hyaluronate, and form the supramolecular hybrid hydrogel through electrostatic interaction. The mass ratio of the nanocomposite to sodium hyaluronate to deionized water is 52:(17.5-20):1000.

5. The method according to claim 4, characterized in that, The mass ratio of β-cyclodextrin-modified upconversion nanoparticles to sodium hyaluronate was 50:2:(17.5-20).

Images