A hydrogel loaded with cyclodextrin aggregates and its preparation method and application

Abstract

The application belongs to the technical field of medicine, and particularly relates to a hydrogel loaded with cyclodextrin aggregates and a preparation method and application thereof. The hydrogel loaded with cyclodextrin aggregates comprises a hydrogel matrix and cyclodextrin aggregates loaded on the hydrogel matrix; the cyclodextrin aggregates are hydrophobic group modified sulfobutyl beta-cyclodextrin; and the hydrophobic group is one of n-dodecyl and 2-methylundecyl. The hydrogel has abundant negative charges, can effectively enhance the combination capacity of the hydrogel with positively charged polyamines, the cyclodextrin cavity is matched with the size of polyamine molecules, the efficiency of combination with polyamines is improved, the capture and removal of excessive polyamines in the local inflammatory area of periodontal are realized in the form of the hydrogel. Meanwhile, the hydrogel loaded with cyclodextrin aggregates can significantly improve the inflammatory microenvironment, effectively reduce the level of cell oxidative stress, maintain immune homeostasis, and provide a new idea for the treatment of periodontal inflammatory areas.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology, specifically relating to a hydrogel loaded with cyclodextrin aggregates, its preparation method, and its application. Background Technology

[0002] Periodontitis is a chronic inflammatory disease primarily caused by the irritation of periodontal tissues by bacteria in dental plaque. During the pathogenesis of periodontitis, when pathogenic bacteria invade the periodontal tissues, they trigger a series of immune responses, including immune cell infiltration and upregulation of the expression of inflammatory factors, leading to an imbalance in immune homeostasis and causing chronic damage to the periodontal tissues. Many inflammatory stimuli are involved in this process. Clearing these inflammatory stimuli helps maintain immune homeostasis and achieves the goal of treating periodontitis.

[0003] Polyamines, as metabolic products of bacteria and cells, are also inflammatory stimuli that can promote the occurrence and development of local inflammatory responses. Polyamines mainly include spermine, spermidine, and putrescine, and are a class of biological metabolites with two or more amino groups, widely present in eukaryotic and prokaryotic cells. They carry a positive charge and participate in life processes such as cell growth, tissue regeneration, and inflammation repair. Regulating the homeostasis of polyamine concentration in the microenvironment is closely related to maintaining bodily health. Changes in polyamine concentration also occur during the development of periodontitis. Studies have found that the polyamine concentration in the saliva of normal healthy individuals is only 10-20 μM, while the polyamine concentration in the gingival crevicular fluid of patients with periodontitis is significantly elevated. After systematic periodontal treatment, the concentration decreases significantly, but remains higher than normal. Therefore, regulating polyamine concentration can control the occurrence and development of periodontitis. In addition to mechanical treatment, adjuvant medications can be used to further remove excess polyamines from periodontal pockets and maintain the balance of periodontal immune homeostasis.

[0004] Previous studies have primarily focused on inhibiting the activity of the rate-limiting enzymes that synthesize polyamines, thereby suppressing polyamine synthesis. However, this approach suffers from drawbacks such as significant side effects and limited applicability. Therefore, finding a suitable, inexpensive, and biosafety-friendly small molecule material, and modifying it to effectively capture locally excess polyamines to achieve polyamine removal and treat periodontitis, has become an urgent problem to be solved. Summary of the Invention

[0005] The purpose of this invention is to provide a hydrogel loaded with cyclodextrin aggregates, its preparation method and application, thereby overcoming the shortcomings of the prior art. The modified cyclodextrin has more negative charge, which effectively enhances its binding ability with positively charged polyamines. In the form of a hydrogel, it can capture and remove excess polyamines in the local inflammatory area of ​​periodontium, thereby achieving the purpose of removing polyamines and treating periodontitis.

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

[0007] In a first aspect, the present invention provides a hydrogel loaded with cyclodextrin aggregates, comprising a hydrogel matrix and cyclodextrin aggregates loaded on the hydrogel matrix;

[0008] The cyclodextrin aggregates are sulfobutyl β-cyclodextrins modified with hydrophobic groups;

[0009] The hydrophobic group is -(CH2). 11 One of CH3 and -(CH2)9CHCH3CH3.

[0010] In some other embodiments, the molar ratio of sulfobutyl β-cyclodextrin to hydrophobic groups in the cyclodextrin aggregate is 1:(1-6).

[0011] In some other embodiments, the hydrogel matrix is ​​one or more of the following: methacrylamide dextran hydrogel, methacrylamide carboxymethyl chitosan hydrogel, methacrylamide gelatin hydrogel, and methacrylamide sodium alginate hydrogel.

[0012] In a second aspect, the present invention provides a method for preparing a hydrogel containing the cyclodextrin aggregates described in the first aspect, comprising the following steps:

[0013] (1) Alkyl alcohol, p-toluenesulfonyl chloride and catalyst are added to the first solvent to react and prepare p-toluenesulfonic acid alkyl ester;

[0014] (2) After mixing sulfobutyl-β-cyclodextrin with a second solvent, potassium iodide and sodium hydride are added to carry out the first step reaction, and p-toluenesulfonic acid alkyl ester is added to carry out the second step reaction to obtain alkyl-sulfobutyl-β-cyclodextrin.

[0015] (3) A mixed solution was prepared by adding a photoinitiator to a phosphate buffer solution of alkyl-sulfobutyl-β-cyclodextrin, and the hydrogel matrix was added to the mixed solution to carry out a photocrosslinking reaction to obtain a hydrogel loaded with cyclodextrin aggregates.

[0016] In some other embodiments, in step (1), the catalyst is triethylamine and 4-dimethylaminopyridine;

[0017] The molar ratio of the alkanol, p-toluenesulfonyl chloride, triethylamine, and 4-dimethylaminopyridine is (1.2-12):(1-10):(3-30):(0.1-1);

[0018] The alkanol is one of n-dodecanool and 2-methylundecanool;

[0019] The alkyl group is -(CH2). 11One of CH3 and -(CH2)9CHCH3CH3;

[0020] The ratio of p-toluenesulfonyl chloride to the first solvent is (1-10) g: (10-50) mL;

[0021] The first solvent is one of dichloromethane, N,N-dimethylformamide, and dimethyl sulfoxide;

[0022] The reaction is carried out at a temperature of 25-30℃ for 15-30 hours.

[0023] In some other embodiments, step (1) further includes purification after the reaction is completed, wherein the purification process consists of extraction, drying, filtration and rotary evaporation in sequence;

[0024] The extraction process involves adding a saturated Na2S2O3 solution to the organic phase, extracting the organic phase, separating the organic phase, extracting the aqueous phase with dichloromethane, and then combining the organic phases.

[0025] The drying process involves adding MgSO4.

[0026] The filtration process involves adding n-hexane before filtration.

[0027] In some other embodiments, in step (2), the molar ratio of the sulfobutyl-β-cyclodextrin, potassium iodide, sodium hydride and alkyl p-toluenesulfonate is 1:(14-20):(120-150):(20-30);

[0028] The mixing ratio of the sulfobutyl-β-cyclodextrin to the second solvent is (0.1-1) g : (20-50) mL;

[0029] The second solvent is one or two of N,N-dimethylformamide, dimethyl sulfoxide, and acetonitrile;

[0030] The first step reaction is to react in an ice bath for 0.5-1.5 hours, and then continue the reaction for 6-7 hours after removing the ice bath;

[0031] The second step reaction is carried out at a temperature of 25-30℃ for 30-35 hours.

[0032] In some other embodiments, step (2) further includes a purification process after the second reaction is completed, wherein the purification process involves removing unreacted NaH and solvent, followed by concentration and lyophilization.

[0033] The method for removing NaH is to add methanol dropwise to the product until no more bubbles are generated;

[0034] The method for removing the solvent is dialysis.

[0035] In some other embodiments, in step (3), the concentration of alkyl-sulfobutyl-β-cyclodextrin in the mixed solution is 0-500 μM, and the concentration of photoinitiator is 0.2-0.3 wt%.

[0036] The photoinitiator is lithium phenyl-2,4,6-trimethylbenzoyl phosphate;

[0037] The ratio of the hydrogel matrix to the mixed solution is (45-55) mg: 1 mL;

[0038] The hydrogel matrix is ​​one or more of the following: methacrylamide dextran hydrogel, methacrylamide carboxymethyl chitosan hydrogel, methacrylamide gelatin hydrogel, and methacrylamide sodium alginate hydrogel.

[0039] The photocrosslinking reaction is cured at room temperature under ultraviolet light for 5-10 seconds.

[0040] The ultraviolet light has a wavelength of 405 nm and an irradiance of 25 mW / cm². 2 .

[0041] Thirdly, the present invention provides the use of the hydrogel containing the cyclodextrin aggregates described in the first aspect in the preparation of a medicament for treating periodontitis.

[0042] The beneficial effects of this invention are:

[0043] 1. The hydrogel containing cyclodextrin aggregates prepared in this invention has abundant negative charge, which effectively enhances its binding ability with positively charged polyamines. At the same time, the size of the cyclodextrin cavity matches that of the polyamine molecule, improving the efficiency of binding with polyamines. In the form of hydrogel, the capture and removal of excess polyamines in the local inflammatory area of ​​periodontium are realized.

[0044] 2. The hydrogel containing cyclodextrin aggregates prepared in this invention can significantly improve the inflammatory microenvironment, effectively reduce cellular oxidative stress levels by removing excess polyamines, maintain immune homeostasis, and achieve the purpose of treating periodontitis. Attached Figure Description

[0045] 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.

[0046] Figure 1 Preparation of SCDC in Example 1 of the present invention 12 The reaction pathway;

[0047] Figure 2 SCDC prepared in Example 1 of this invention 12 ESI-MS results;

[0048] Figure 3 SCDC prepared in Example 1 of this invention 12 One-dimensional proton NMR spectrum results;

[0049] Figure 4 β-CD and SCDC in Embodiment 1 of the present invention 12 Zeta potential diagram;

[0050] Figure 5 The DexMA hydrogel and DexMA@SCDC in Example 2 of this invention 12 Scanning electron microscopy structure of hydrogels;

[0051] Figure 6 The DexMA hydrogel and DexMA@SCDC in Example 2 of this invention 12 Porosity of hydrogels;

[0052] Figure 7 The DexMA hydrogel and DexMA@SCDC in Example 2 of this invention 12 The swelling ratio of the hydrogel;

[0053] Figure 8 The DexMA hydrogel and DexMA@SCDC in Example 2 of this invention 12 Rheological properties of hydrogels;

[0054] Figure 9 The DexMA hydrogel and DexMA@SCDC in Example 2 of this invention 12 The degradation properties of hydrogels;

[0055] Figure 10 SCDC in Embodiment 1 of the present invention 12 A graph showing the effect of polyamine scavenging capacity in artificial saliva;

[0056] Figure 11 SCDC in Embodiment 1 of the present invention 12 The effect of clearing intracellular ROS (Image showing the effect of the drug);

[0057] Figure 12 The DexMA hydrogel and DexMA@SCDC in Example 2 of this invention 12 Statistical graph of the vertical distance between the cementoenamel junction (CEJ) and alveolar ridge (ABC) after treatment with hydrogel in rats with periodontitis. Detailed Implementation

[0058] Unless otherwise specified, the experimental methods described in the following examples are conventional methods; unless otherwise specified, the reagents and materials are commercially available.

[0059] The inventive concept adopted in this invention is as follows:

[0060] β-Cyclodextrin, a naturally occurring glucose derivative, possesses a rigid cyclic structure, size matching with polyamines, and exhibits good biocompatibility and stability, making it widely used in drug development for various diseases. Sulfobutyl-β-cyclodextrin (without specific requirements for the degree of substitution, and can be a mixture of sulfobutyl-β-cyclodextrins with different degrees of substitution), as a derivative of β-cyclodextrin, exhibits good water solubility and biocompatibility, and is widely used in pharmaceutical excipients, vaccine carriers, and other fields. Its internal cavity structure matches the size of polyamine molecules, allowing for binding. To better capture polyamines, hydrophobic groups are modified into sulfobutyl-β-cyclodextrin to obtain cyclodextrin aggregates. These cyclodextrin aggregates are then photocrosslinked and cured with a hydrogel matrix, possessing abundant negative charges, effectively enhancing its binding ability with positively charged polyamines. Simultaneously, the cyclodextrin cavity size matches the polyamine molecule size, improving the efficiency of polyamine binding. This hydrogel form enables the capture and removal of excess polyamines in locally inflammatory areas of the periodontium.

[0061] Example 1

[0062] A method for preparing a hydrogel loaded with cyclodextrin aggregates, such as Figure 1 As shown, the steps are as follows:

[0063] (1) Using 30 mL of dichloromethane (CH2Cl2) as solvent, 12.74 g (90 mM) of triethylamine (Et3N) and 0.363 g (3 mM) of 4-dimethylaminopyridine (DMAP) as catalysts, 5.58 g (36 mM) of n-dodecyl alcohol (C) was added as raw material. 12 H 24 OH) and 6.86 g (30 mM) p-toluenesulfonyl chloride (TsCl) were reacted at room temperature for 16 hours.

[0064] A saturated Na₂S₂O₃ solution was added to the organic phase, and after phase separation extraction, the organic phase was separated. The aqueous phase was extracted with dichloromethane, and the organic phases were combined. A small amount of MgSO₄ was added for drying, and the mixture was filtered and rotary evaporated to obtain a pale yellow oily substance with a precipitate. Hexane was added, and a precipitate formed. After filtration, the filtrate was obtained, and rotary evaporation yielded a pale yellow liquid, which was the product, dodecyl p-toluenesulfonate (TsC). 12 ).

[0065] (2) Dissolve 0.3 g (0.1 mM) sulfobutyl-β-cyclodextrin (SEB6-β-CD) in 30 mL of dimethyl sulfoxide (DMSO) and 3 mL of N,N-dimethylformamide (DMF), then add 0.24 g (1.45 mM) potassium iodide (KI) and 0.3 g (12.5 mM) sodium hydride (NaH) sequentially. React in an ice bath for 1 hour, then remove the ice bath and continue the reaction for 6-7 hours. Add 0.8 g (2.35 mM) TsC 12 The reaction was continued for 32 hours. If any solid precipitated during this period, DMSO and DMF solvents were added until the precipitated solid was redissolved. The final product, dodecyl-sulfobutyl-β-cyclodextrin (SCDC), was obtained. 12 )

[0066] (3) Post-treatment: Add methanol dropwise to the final product until no more bubbles are generated to remove excess NaH. Remove methanol by rotary evaporation. Dialyze to remove inorganic small molecules and organic solvents for 48 hours. Collect the dialyzed liquid, remove the solvent water by rotary evaporation again, and concentrate. Collect the concentrated product and freeze-dry it.

[0067] Example 2

[0068] A method for preparing a hydrogel loaded with cyclodextrin aggregates is as described in Example 1, except that SCDC is obtained according to step (3) of Example 1. 12 Then, it was added to a pre-gel solution of DexMA (model EFL-DexMA-500K) containing 0.25% photoinitiator LAP, and then subjected to ultraviolet light (405nm, 25mW / cm²). 2 After 5 seconds of irradiation, a stable DexMA@SCDC structure is formed. 12 Hydrogel.

[0069] Example 3

[0070] Unlike Example 2, methacrylamide carboxymethyl chitosan hydrogel (CMCSMA) was used.

[0071] The methacryloyl dextran hydrogel (DexMA) was replaced with EFL-CMCSMA-200K, and the other preparation methods were the same as in Example 2.

[0072] Example 4

[0073] Unlike Example 2, methacrylamide gelatin hydrogel (GelMA, model number 1) was used.

[0074] The EFL-GM-90K hydrogel was replaced with methacrylamide dextran hydrogel (DexMA), and the other preparation methods were the same as in Example 2.

[0075] Example 5

[0076] Unlike Example 2, methacrylamide sodium alginate hydrogel (AlgMA, model EFL-AlgMA-300K) was used instead of methacrylamide dextran hydrogel (DexMA), while the other preparation methods were the same as in Example 2.

[0077] Example 6

[0078] Unlike Example 2, 2-methylundecyl alcohol was used instead of n-dodecyl alcohol in Example 1, while the other preparation methods were the same as in Example 2.

[0079] Comparative Example 1

[0080] Unlike Example 2, no modification treatment was performed on the sulfobutyl-β-cyclodextrin; that is, the sulfobutyl-β-cyclodextrin was directly modified.

[0081] Cyclodextrin was reacted with methacrylamide dextran hydrogel, and other preparation methods were the same as in Example 2.

[0082] Comparative Example 2

[0083] Unlike Example 2, β-cyclodextrin was used instead of sulfobutyl-β-cyclodextrin, while the other preparation methods were the same as in Example 2.

[0084] Comparative Example 3

[0085] Unlike Example 2, butanol was used instead of dodecanol; the other preparation methods were the same as in Example 2.

[0086] Consistent.

[0087] Performance testing

[0088] (1) Performance testing of cyclodextrin aggregates

[0089] Figure 2 For SCDC 12 ESI-MS results, Figure 3 For SCDC 12 One-dimensional proton NMR spectrum results. From Figure 2 and Figure 3 It can be seen that dodecyl-sulfobutyl-β-cyclodextrin (SCDC) was successfully prepared. 12 ).

[0090] Figure 4 β-CD and SCDC in Embodiment 1 of the present invention 12 Zeta potential plot; where β-CD is β-cyclodextrin, SCDC 12 It is a cyclodextrin aggregate. From Figure 4 It can be seen that after β-cyclodextrin is modified with sulfonyl, SCDC 12The presence of a stronger negative charge demonstrates that cyclodextrin aggregates are more likely to bind to positively charged polyamines.

[0091] (2) DexMA hydrogel and DexMA@SCDC 12 Structural performance testing of hydrogels

[0092] Figure 5 The DexMA hydrogel and DexMA@SCDC in Example 2 of this invention 12 Scanning electron microscopy structure of the hydrogel. From Figure 5 It can be seen that both have a loose and porous structure, and are loaded with SCDC. 12 It has no significant effect on the internal structure of the hydrogel.

[0093] Figure 6 The DexMA hydrogel and DexMA@SCDC in Example 2 of this invention 12 Porosity of hydrogels. From Figure 6 It can be seen that the porosity of both is approximately 45%, and the loading of SCDC is... 12 It has no significant effect on the porosity of the hydrogel.

[0094] Figure 7 The DexMA hydrogel and DexMA@SCDC in Example 2 of this invention 12 Swelling rate of hydrogels. The swelling rate is tested by calculating the change in mass of the hydrogel. The dry weight of the lyophilized hydrogel is W0. At 37℃, the lyophilized hydrogel sample is immersed in PBS solution, and weighed at different time points, recorded as W1. Swelling rate (%) = (W1 - W0) / W0 × 100%. Figure 7 It can be seen that both reached a high swelling rate within 4 hours, and the swelling rates at different time points were similar, indicating that the loading of SCDC... 12 It has no significant effect on the swelling properties of the hydrogel.

[0095] Figure 8 The DexMA hydrogel and DexMA@SCDC in Example 2 of this invention 12 Rheological properties of hydrogels. The method for testing rheological properties was as follows: a disc-shaped hydrogel sample with a diameter of approximately 10 mm and a thickness of approximately 2 mm was prepared. At room temperature, the storage modulus (G') and loss modulus (G”) of the hydrogel under different stress and strain conditions were measured using a rheometer at an angular velocity of 5 rad / s. From... Figure 8 It can be seen that the trends of G' and G” are similar under different stress and strain conditions, indicating that the load SCDC 12 It has no significant effect on the rheological properties of the hydrogel.

[0096] Figure 9The DexMA hydrogel and DexMA@SCDC in Example 2 of this invention 12 The degradation performance of the hydrogel was tested as follows: The prepared hydrogel sample was weighed, and the initial mass was recorded. The sample was then immersed in artificial saliva, shaken at 70 rpm at 37°C, with the solution changed every 8 hours. The sample was removed and weighed again at different time points, and the remaining mass of the hydrogel was recorded. The in vitro degradation rate was calculated by determining the ratio of the remaining mass to the initial mass of the hydrogel. Figure 9 It can be seen that both exhibit good degradation performance, with a degradation time of approximately 15 days in artificial saliva, indicating that the loading of SCDC... 12 It has no significant effect on the degradation performance of the hydrogel.

[0097] (3) Effect of cyclodextrin aggregates on the polyamine scavenging ability of artificial saliva

[0098] At room temperature, using artificial saliva (pH 6.8, Leagene, CZ0190) as a solvent, 20 μM spermine, spermidine, and putrescine were reacted with 20, 40, 100, and 200 μM SCDC, respectively. 12 Mix for 1 hour, then determine the residual polyamine content using a polyamine assay kit (Abcam, ab239728). Residual ratio (%) = final polyamine concentration / initial polyamine concentration × 100%.

[0099] Figure 10 SCDC in Embodiment 1 of the present invention 12 The effect of polyamine clearance in artificial saliva; where Spermine represents the spermine group, Spermidine the spermidine group, and Putrescine the putrescine group; Residual ratio (%) represents the residual ratio; NC is the blank control group, and 1:1, 1:2, 1:5, and 1:10 represent the ratios of polyamines to SCDC. 12 Concentration ratio. From Figure 10 It can be seen that in artificial saliva, SCDC 12 The ability to scavenge polyamines is related to the type and concentration ratio of the polyamines; spermine and putrescine are more readily scavenged by SCDC. 12 Combined with removal, and with SCDC 12 The efficiency of polyamine binding is improved with the increase of relative concentration.

[0100] In Example 2, SCDC 12 Loading with DexMA hydrogel will facilitate intrapocket injection during periodontal treatment for periodontitis. Because DexMA hydrogel possesses stable physicochemical properties and good biocompatibility, and can be slowly degraded by enzymes in the human body, loading with it can achieve SCDC (slightly concentrated cavitation) treatment. 12 The sustained-release process within the periodontal pocket. Simultaneously, the effects of loading on SCDC...12 The effect on polyamine scavenging ability is not significant.

[0101] In Examples 3, 4, and 5, similar to Example 2, SCDC is used. 12 Loaded in CMCSMA, GelMA, or AlgMA hydrogels, all of which possess stable physicochemical properties and good biocompatibility, are biodegradable in vivo, and are resistant to SCDC. 12 The polyamine scavenging ability was not significantly affected.

[0102] In Example 6, 2-methylundecyl alcohol was used instead of n-dodecyl alcohol in Example 1, and the properties of the prepared product were similar.

[0103] In Comparative Example 1, since sulfobutyl-β-cyclodextrin does not have a hydrophobic chain, its self-assembly performance in solution is not strong. Compared with the product in Example 1, it has a poorer specific surface area, reactivity, and dispersibility in solution, which is not conducive to its binding with polyamines.

[0104] In Comparative Example 2, since β-cyclodextrin does not have a negatively charged sulfonyl group, its electrostatic attraction to positively charged polyamines is weak, resulting in a weaker ability to bind to polyamines and a poorer polyamine scavenging ability compared to the product in Example 1.

[0105] In Comparative Example 3, the use of butanol instead of dodecyl alcohol resulted in a product with poor self-assembly properties. The length of the hydrophobic chain affects the self-assembly performance of the product; the longer the chain, the stronger the hydrophobic interaction between molecules, resulting in higher aggregation ability and greater stability of the self-assembled structure. However, a carbon chain length greater than 12 reduces solubility and limits self-assembly. Therefore, a suitable hydrophobic chain length can balance hydrophobicity and hydrophilicity, achieving efficient self-assembly.

[0106] (4) The role of cyclodextrin aggregates in scavenging intracellular ROS

[0107] ROS production in human gingival fibroblasts (HGFs) stimulated with spermine was assessed using a reactive oxygen species (ROS) detection kit (S0033S, Beyotime). HGFs cells were cultured at 2 × 10⁶ cells / year. 4 Cells were seeded at a density of 10 cells / well in 6-well plates and cultured for 24 hours. After stimulating the cells with 10 μM spermine for 24 hours, they were then seeded with SCDC at concentrations of 0, 10, 20, and 40 μM, respectively. 12 After 24 hours of culture, the production of intracellular ROS was assessed using a reactive oxygen species (ROS) detection kit, and the fluorescence intensity of intracellular ROS in each group was observed using a fluorescence microscope (Leica, Germany).

[0108] Figure 11 SCDC in Embodiment 1 of the present invention 12The effect of scavenging intracellular ROS is illustrated in the image; where Control represents the blank control group, 0 μM represents the spermine-stimulated inflammation-simulated group, and 10 μM, 20 μM, and 40 μM represent the experimental groups after spermine stimulation followed by the addition of different concentrations of cyclodextrin aggregates; Merged represents the fusion state, DCFH represents the DCFH fluorescent probe signal channel, and Bright Field represents the bright field image. Figure 11 It can be seen that, compared with the control group, the 0 μM group showed increased fluorescence intensity, enhanced intracellular oxidative stress response of HGFs, and significantly increased intracellular ROS levels upon stimulation with high concentrations of polyamines. In contrast, the 10 μM, 20 μM, and 40 μM groups, after stimulation with different concentrations of SCDC... 12 The fluorescence intensity and ROS level decreased significantly after treatment, indicating that the cyclodextrin aggregates enhanced the ROS scavenging ability by capturing and scavenging polyamines.

[0109] (5) Hydrogels loaded with cyclodextrin aggregates were used to statistically analyze the vertical distance between the cementum-enamel junction (CEJ) and alveolar ridge (ABC) in rats with periodontitis after treatment.

[0110] The rats were infected by ligating their second molars and injecting Porphyromonas gingivalis into the gingival sulcus (injection of bacterial solution every 3 days for 2 weeks, with a bacterial concentration of approximately 1*10). 9 A rat periodontitis model was established using CFU / mL, and then drugs were injected into the gingival sulcus or periodontal pocket (PBS was injected into the healthy group and the periodontitis group, DexMA gel was injected into the gel-only group, and DexMA@SCDC was injected into the material group). 12 Rats were treated with a gel for 28 days (twice a week, 10 μL each time). Bone loss in the second molar alveolar bone (vertical distance between CEJ and ABC) was measured using Micro-CT in each group of rats, and the DexMA@SCDC gel was analyzed. 12 Improvement in periodontal inflammation.

[0111] Figure 12 The DexMA hydrogel and DexMA@SCDC in Example 2 of this invention 12 Statistical graph of the vertical distance between the cementum-enamel junction (CEJ) and alveolar ridge crest (ABC) after treatment with hydrogels in rats with periodontitis; where Normal represents the normal group, Periodontitis represents the periodontitis group, DexMA represents the methacrylamide dextran hydrogel group, and DexMA@SCDC represents the DexMA@SCDC group. 12 The hydrogel experimental group is loaded with cyclodextrin aggregates; Mesial represents the mesial surface, and Distal represents the distal surface; CEJ-ABC (μm) is the vertical distance between the cementum-enamel junction and the alveolar ridge crest. From Figure 12It can be seen that, compared with the Normal group, the vertical distance between the cementum-enamel junction (CEJ) and the alveolar ridge crest (ABC) was significantly increased in the Periodontitis group and the DexMA group, while the DexMA@SCDC group showed a significantly increased vertical distance. 12 The vertical distance between the cementoenamel junction (CEJ) and alveolar ridge crest (ABC) was significantly reduced in the group compared to the Periodontitis group, indicating that the hydrogel loaded with cyclodextrin aggregates can effectively alleviate bone loss caused by periodontal inflammation, reduce bone resorption height, and alleviate inflammatory bone loss.

[0112] 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 hydrogel loaded with cyclodextrin aggregates, characterized in that, Includes the hydrogel matrix and cyclodextrin aggregates supported on the hydrogel matrix; The cyclodextrin aggregates are sulfobutyl β-cyclodextrins modified with hydrophobic groups; The hydrophobic group is -(CH2). 11 One of CH3 and -(CH2)9CHCH3CH3; In the cyclodextrin aggregate, the molar ratio of sulfobutyl β-cyclodextrin to hydrophobic groups is 1:(1-6).

2. The hydrogel supporting cyclodextrin aggregates according to claim 1, characterized in that, The hydrogel matrix is ​​one or more of the following: methacrylamide dextran hydrogel, methacrylamide carboxymethyl chitosan hydrogel, methacrylamide gelatin hydrogel, and methacrylamide sodium alginate hydrogel.

3. A method for preparing a hydrogel containing supported cyclodextrin aggregates according to any one of claims 1-2, characterized in that, Includes the following steps: (1) Alkyl alcohol, p-toluenesulfonyl chloride and catalyst are added to the first solvent to react and prepare p-toluenesulfonic acid alkyl ester; (2) After mixing sulfobutyl-β-cyclodextrin with a second solvent, potassium iodide and sodium hydride are added to carry out the first step reaction, and p-toluenesulfonic acid alkyl ester is added to carry out the second step reaction to obtain alkyl-sulfobutyl-β-cyclodextrin; (3) A mixed solution was prepared by adding the photoinitiator to a phosphate buffer solution of alkyl-sulfobutyl-β-cyclodextrin, and the hydrogel matrix was added to the mixed solution to carry out a photocrosslinking reaction to obtain a hydrogel loaded with cyclodextrin aggregates.

4. The method for preparing a hydrogel loaded with cyclodextrin aggregates according to claim 3, characterized in that, In step (1), the catalyst is triethylamine and 4-dimethylaminopyridine; The molar ratio of the alkanol, p-toluenesulfonyl chloride, triethylamine, and 4-dimethylaminopyridine is (1.2-12):(1-10):(3-30):(0.1-1). The alkanol is one of n-dodecanool and 2-methylundecanool; The alkyl group is -(CH2). 11 One of CH3 and -(CH2)9CHCH3CH3; The ratio of p-toluenesulfonyl chloride to the first solvent is (1-10) g: (10-50) mL; The first solvent is one of dichloromethane, N,N-dimethylformamide, and dimethyl sulfoxide; The reaction is carried out at a temperature of 25-30℃ for 15-30 hours.

5. The method for preparing a hydrogel loaded with cyclodextrin aggregates according to claim 3, characterized in that, In step (1), after the reaction is completed, a purification process is also included, which consists of extraction, drying, filtration and rotary evaporation in sequence. The extraction process involves adding a saturated Na2S2O3 solution to the organic phase, extracting the organic phase, separating the organic phase, extracting the aqueous phase with dichloromethane, and then combining the organic phases. The drying process involves adding MgSO4. The filtration process involves adding n-hexane before filtration.

6. The method for preparing a hydrogel loaded with cyclodextrin aggregates according to claim 3, characterized in that, In step (2), the molar ratio of the sulfobutyl-β-cyclodextrin, potassium iodide, sodium hydride and p-toluenesulfonate alkyl ester is 1:(14-20):(120-150):(20-30). The mixing ratio of the sulfobutyl-β-cyclodextrin to the second solvent is (0.1-1) g : (20-50) mL; The second solvent is one or two of N,N-dimethylformamide, dimethyl sulfoxide, and acetonitrile; The first step reaction is to react in an ice bath for 0.5-1.5 hours, and then continue the reaction for 6-7 hours after removing the ice bath; The second step reaction is carried out at a temperature of 25-30 ℃ for 30-35 h.

7. The method for preparing a hydrogel loaded with cyclodextrin aggregates according to claim 3, characterized in that, In step (2), after the second reaction is completed, a purification process is also included, which involves removing unreacted NaH and solvent, followed by concentration and lyophilization. The method for removing NaH is to add methanol dropwise to the product until no more bubbles are generated; The method for removing the solvent is dialysis.

8. The method for preparing a hydrogel loaded with cyclodextrin aggregates according to claim 3, characterized in that, In step (3), the concentration of alkyl-sulfobutyl-β-cyclodextrin in the mixed solution is 0-500 μM, and the concentration of the photoinitiator is 0.2-0.3 wt%. The photoinitiator is lithium phenyl-2,4,6-trimethylbenzoyl phosphate; The ratio of the hydrogel matrix to the mixed solution is (45-55) mg: 1 mL; The hydrogel matrix is ​​one or more of the following: methacrylamide dextran hydrogel, methacrylamide carboxymethyl chitosan hydrogel, methacrylamide gelatin hydrogel, and methacrylamide sodium alginate hydrogel. The photocrosslinking reaction is cured at room temperature under ultraviolet light for 5-10 seconds; The ultraviolet light has a wavelength of 405 nm and an irradiance of 25 mW / cm². 2 .

9. The use of a hydrogel containing cyclodextrin aggregates as described in any one of claims 1-2 in the preparation of a medicament for treating periodontitis.

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