Acoustic power driven Janus hydrogel as well as preparation method and application thereof

Abstract

The invention discloses sound power driven Janus hydrogel as well as a preparation method and application thereof. The hydrogel comprises silver peroxide and a hydrogel matrix, the hydrogel matrix is prepared from methacrylated silk fibroin, gamma-polyglutamic acid-N-hydroxysuccinimide ester and a photoinitiator. Through component synergy and structural innovation, the prepared hydrogel can release ROS according to ultrasonic drive, destroys bacterial biofilms, and cooperates with Ag < + > slowly released by Ag2O2 to achieve an efficient sterilization effect; after the hydrogel is cross-linked by ultraviolet light, a Janus structure of which one side is adhered and the other side is anti-adhesion can be formed. Pharmacodynamic experiment results show that the sonodynamic-driven Janus hydrogel prepared by the invention has an excellent antibacterial effect, can be used for promoting wound healing, repairing alveolar bone defects and the like, and also provides a new choice for repairing infections such as periodontitis, oral mucosa inflammation and the like or skin wounds.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of gel biomaterials, and particularly relates to a sonodynamic driving Janus hydrogel and a preparation method and application thereof. BACKGROUND

[0002] Periodontitis is a common chronic inflammatory disease. Bacterial infection is the main pathogenic factor of periodontitis. Plaque microorganisms and their toxic metabolites can act on periodontal tissues for a long time, and neutrophils, lymphocytes and the like will infiltrate into the infection site, causing these tissues to be progressively destroyed, resulting in gum swelling, attachment loss, alveolar bone resorption and periodontal pocket formation, and if not timely intervention, it will lead to tooth loosening or even shedding. The current traditional antibacterial treatment of periodontitis includes subgingival scaling and the use of antibacterial drugs. However, these methods have significant limitations. First, subgingival scaling requires high patient cooperation, but due to the complexity of the process and the possibility of causing discomfort, the patient's compliance is poor, resulting in unstable treatment effect and easy re-colonization of bacterial biofilm. In addition, local antibacterial drugs such as minocycline hydrochloride, metronidazole and chlorhexidine used in clinical practice are effective for free bacteria, but have difficulty in playing a role for bacteria wrapped in biofilm. In addition, long-term use of antibiotics can also lead to increased bacterial resistance. Therefore, the key to the antibacterial treatment of periodontitis is how to remove bacterial biofilm, enhance host immunity and inhibit subsequent bacterial adhesion, so as to effectively inhibit the progression of the disease.

[0003] In view of the characteristics that periodontal bacterial biofilm is difficult to remove, antibacterial sonodynamic therapy (aSDT) is a new antibacterial technology, whose principle is to use ultrasound to activate a photosensitizer to produce reactive oxygen species to kill bacteria. Compared with traditional antibiotic therapy, aSDT has excellent tissue penetration, high biocompatibility, and no drug resistance, etc. This therapy concentrates ultrasonic energy on the infection site to activate the photosensitizer to produce highly cytotoxic ROS, thereby inducing bacterial death. However, when used alone, it has the defects of limited ROS action range and easy damage to normal tissues.

[0004] The human oral environment is a complex microbial ecosystem that is both moist and open. This environment not only contains a large amount of saliva, but also allows the presence of bacteria and other microorganisms. The flowability of saliva and the openness of the oral cavity pose a risk of drug washout for local drug administration, making it difficult to maintain the concentration of the drug in the target treatment area. In addition, after the bacterial biofilm is effectively removed in time, free bacteria will also quickly colonize on the surface of the periodontal tissue.

[0005] Janus hydrogels are a class of hydrogels with two faces of different functions, and their asymmetric chemical or physical properties endow them with unique properties and broad application prospects. In the fields of material science and biomedical engineering, Janus hydrogels are concerned because they can realize different adhesion properties by adjusting the functional groups on the two faces. However, the existing Janus hydrogels are mostly focused on wound healing or organ repair, and there is no literature report on the combination of Janus hydrogels and sonodynamic technology to prevent bacterial adhesion in periodontitis. SUMMARY

[0006] To solve the above problems, the purpose of the present application is to provide a kind of sonodynamic driven Janus hydrogel and its preparation method and application, the sonodynamic driven Janus hydrogel prepared by the present application has excellent antibacterial effect, can significantly repair alveolar bone defect, and can also improve the efficiency of skin wound healing, which provides a new choice for periodontitis, oral mucosa inflammation and other infections or skin wound repair.

[0007] The present application is realized by the following technical solutions: A kind of sonodynamic driven Janus hydrogel, the hydrogel includes silver peroxide (Ag2O2) and hydrogel matrix;The hydrogel matrix is composed of methacrylated silk fibroin (SFMA) and gamma-polyglutamic acid-N-hydroxysuccinimide ester (γ-PGA-NHS) and photoinitiator.

[0008] Gamma-polyglutamic acid (γ-PGA), also known as natto bacteria glue, is a water-soluble, biodegradable and non-toxic natural anionic polyamino acid, which is usually produced on a large scale by fermentation engineering. In order to increase its adhesion, NHS modification is carried out, and the prepared γ-PGA-NHS has excellent cell compatibility and high reactivity to primary amines.

[0009] In the present application, methacrylated silk fibroin (SFMA) and gamma-polyglutamic acid-N-hydroxysuccinimide ester (γ-PGA-NHS) are selected as the hydrogel matrix, wherein SFMA has good biocompatibility, biodegradability and tissue repair ability, and γ-PGA-NHS has good tissue adhesion ability. Double crosslinking system is a common and effective strategy to realize enhanced mechanical strength and strong adhesion. After ultraviolet irradiation, a double crosslinking network is formed, in which the NHS ester group of part of γ-PGA-NHS combines with the amino group on the surface of SFMA to crosslink, in addition, SFMA will crosslink under the action of photoinitiator after light irradiation. By rapidly solidifying into a double crosslinking structure, sufficient mechanical strength is provided while strong adhesion is formed, in addition, a physical barrier is formed, and finally the effect of inhibiting subsequent bacterial adhesion is realized.

[0010] As a preferred embodiment of the present application, the hydrogel also comprises titanium carbide nanosheets, and silver peroxide is loaded on the titanium carbide nanosheets.

[0011] The titanium carbide nanosheet is a two-dimensional layered material, which has excellent ultrasonic response performance and high specific surface area. In the present application, the titanium carbide nanosheet is used as a carrier of silver peroxide, which can not only improve the stability and dispersibility of Ag2O2, but also has certain sonodynamic performance itself, and cooperates with Ag2O2 to synergistically enhance the sonodynamic effect.

[0012] As a preferred embodiment of the present application, the titanium carbide nanosheet is also loaded with caffeic acid phenethyl ester (CAPE), which can not only inhibit the release of inflammatory factors, but also remove residual ROS to avoid tissue damage.

[0013] As a preferred embodiment of the present application, in the hydrogel, the mass ratio of the photoinitiator, the titanium carbide nanosheet loaded with caffeic acid phenethyl ester and silver peroxide, the methylacrylated silk fibroin, and the gamma-polyglutamic acid-N-hydroxysuccinimide ester is 1:40:40-120:40.

[0014] As a preferred embodiment of the present application, the photoinitiator is at least one of lithium phenyl-2,4,6-trimethylbenzoylphosphinate (LAP), 2,4,6-trimethylbenzoyl-diphenylphosphine oxide, or 2-hydroxy-1-[4-(2-hydroxyethoxy)phenyl]-2-methyl-1-propanone.

[0015] The present application also provides a preparation method of the above-mentioned sonodynamic Janus hydrogel, which comprises the following steps: (1) preparing a precursor solution A: dissolving silver peroxide, a photoinitiator, and methylacrylated silk fibroin (SFMA) in deionized water, mixing uniformly to obtain the precursor solution A; (2) preparing a precursor solution B: dissolving gamma-polyglutamic acid-N-hydroxysuccinimide ester (gamma-PGA-NHS) in a NaHCO3 solution to obtain the precursor solution B; (3) ultraviolet light curing: mixing the precursor solution A and the precursor solution B, and then performing ultrasonic treatment and ultraviolet light curing to obtain the sonodynamic Janus hydrogel.

[0016] As a preferred embodiment of the present application, in step (1), the titanium carbide nanosheet is modified by amino silane to obtain Ti3C2-NH2; then, the Ti3C2-NH2 is dissolved in an ethanol solution, and silver peroxide and / or caffeic acid phenethyl ester are added in sequence, and after stirring, the titanium carbide nanosheet loaded with silver peroxide and / or caffeic acid phenethyl ester is obtained by centrifugation and washing precipitation.

[0017] As a preferred embodiment of the present application, the temperature of the stirring is 25-37 DEG C, and the stirring time is 4-24 hours.

[0018] As a preferred embodiment of the present application, the mass ratio of the Ti3C2-NH2 to the caffeic acid phenethyl ester is 1:15-30, and the mass ratio of the Ti3C2-NH2 to the silver peroxide is 1:1-3.

[0019] As a preferred embodiment of the present application, the power of the ultrasonic is 1~1.2 W / cm 2 , the duty cycle is 50%, the frequency is 1~1.5 MHz, and the time is 2~3 min.

[0020] As a preferred embodiment of the present application, the wavelength of the ultraviolet light is 365~405 nm, and the curing time is 6~10 s.

[0021] The present application also provides the use of the above-mentioned sonodynamic Janus hydrogel in the preparation of drugs for preventing or treating periodontitis or oral mucosa inflammation.

[0022] The sonodynamic Janus hydrogel of the present application releases ROS under ultrasonic driving, destroys the bacterial biofilm, and then is cured by ultraviolet light, so that an adhesion surface (covalently combined with the periodontal soft tissue) is formed on the side of the hydrogel close to the tissue, and an anti-adhesion surface is formed on the side facing the oral cavity, forming an anti-adhesion barrier to inhibit the subsequent adhesion of free bacteria and prevent the generation of bacterial biofilm.

[0023] The present application also provides the use of the above-mentioned sonodynamic Janus hydrogel in the preparation of skin wound repair materials, which is particularly suitable for the repair of bacterial infection skin wounds.

[0024] Compared with the prior art, the present application has the following advantages: The sonodynamic Janus hydrogel provided by the present application can release ROS under ultrasonic driving, destroy the bacterial biofilm, and synergistically achieve high bactericidal effect with the slow-release Ag + of Ag2O2.

[0025] The results of pharmacodynamic experiments show that after the sonodynamic Janus hydrogel prepared by the present application acts on periodontitis rats, it can significantly inhibit Porphyromonas gingivalis and Fusobacterium nucleatum, has excellent antibacterial effect, and can significantly repair alveolar bone defects, providing a new choice for periodontitis or oral mucosa inflammation and the like infections.

[0026] In addition, the sonodynamic Janus hydrogel of the present application can also improve the skin wound healing efficiency, reduce bacterial infection and inflammatory reaction, and is suitable for skin wound repair materials. BRIEF DESCRIPTION OF DRAWINGS

[0027] Figure 1 SEM and EDS of Ti3C2, Ag2O2, TAg in the embodiments of the present application 、 SEM and EDS of TCAg in the embodiments of the present application Figure 2 SEM and EDS of Ti3C2, Ti3C2-NH2, Ag2O2, TAg in the embodiments of the present application 、 Potential diagram of TCAg in the embodiments of the present application Figure 3 Sonodynamic diagram of Ti3C2, TAg, TCAg in the embodiments of the present application Figure 4 NMR hydrogen spectrum of SFMA, γ-PGA-NHS in the embodiments of the present application Figure 5 SEM of TCAg@SMP in the embodiments of the present application Figure 6 Asymmetric adhesion of TCAg@SMP in the embodiments of the present application Figure 7 Evaluation of TCAg@SMP in the embodiments of the present application for inhibiting bacterial adhesion Figure 8 Bacterial spread plate of Ti3C2, Ag2O2, TCAg in the embodiments of the present application Figure 9 SEM of Ti3C2, Ag2O2, TCAg in the embodiments of the present application Figure 10 Changes in periodontal bacterial levels of periodontitis rats after the effect of the gel in the embodiments of the present application Figure 11 Repair of alveolar bone of periodontitis rats after the effect of the gel in the embodiments of the present application Figure 12 Changes in wound healing of rats from day 0 to day 14 after the effect of the gel in the embodiments of the present application DETAILED DESCRIPTION

[0028] The technical solutions of the present application will be described clearly and completely below in combination with the embodiments of the present application. The described embodiments are only some of the embodiments of the present application, not all. Based on the embodiments in the present application, all other embodiments obtained by those of ordinary skill in the art without creative labor fall within the scope of protection of the present application.

[0029] (1) Preparation of carbon nanosheets (Ti3C2): LiF powder (1.0 g) was added into 20 mL of 9 M HC1 solution and mixed for 30 min. Then, 1.0 g of Ti3AlC2 powder was added slowly and the mixture was reacted at 45 °C for 24 h. The product was washed with water by centrifugation until neutral. The multilayer Ti3C2 was obtained by vacuum drying at 60 °C for 12 h. Then, the multilayer Ti3C2 was dispersed in deionized water and ultrasonicated for 4 h at 400 W. The supernatant was collected by centrifugation at 3500 rpm for 60 min and freeze-dried to obtain the single-layer Ti3C2.

[0030] (2) Preparation of Ag2O2: PVP (0.5 g) was dissolved in an aqueous solution containing AgN03(5 mL, 20 mM). Then, NaOH (5 mL, 20 mM) and H202(100 μL, 30%) were added successively. After stirring for 30 min, Ag2O2 nanoparticles were collected by centrifugation and washed with ultrapure water and ethanol for several times. Finally, the product was freeze-dried by a freeze-dryer at low temperature to obtain Ag2O2.

[0031] (3) Preparation of methacrylated silk fibroin (SFMA): 10 g of silk fibroin (SF) was dissolved in 50 mL of 9.3 M LiBr solution and stirred until completely dissolved. Then, 5 mL of glycidyl methacrylate was added slowly and the mixture was reacted at 60 °C for 3 h. The reaction product was collected and dialyzed in ultrapure water for 3 days. Finally, the reaction solution was freeze-dried to obtain SFMA, which was stored at -20 °C.

[0032] (4) Preparation of γ-PGA-NHS: 30 wt% γ-PGA-Na + The aqueous solution was first dialyzed against 0.01 M HC1 for 24 h, then against ultrapure water for 24 h, and finally freeze-dried to obtain the acidic form of γ-PGA. The freeze-dried γ-PGA (1 g) was weighed into a conical flask containing anhydrous DMSO, which was immediately sealed after being filled with nitrogen. The mixture was stirred at room temperature until dissolved, then 1.44 g of EDC was added and stirred until dissolved. Then, 4.46 g of NHS was added and stirred until dissolved. After 24 h of reaction, anhydrous ethanol solution was added to precipitate γ-PGA-NHS, which was centrifuged at 3000 rpm for 5 min to remove the supernatant, then washed with anhydrous ethanol for 3 times and n-hexane solution for 1 time. Finally, the product was vacuum dried to obtain γ-PGA-NHS. Example 1:

[0033] Preparation of precursor solution A: silver peroxide (Ag2O2), photoinitiator LAP and methacrylated silk fibroin (SFMA) were dissolved in deionized water, mixed uniformly to obtain precursor solution A; Preparation of precursor solution B: γ-polyglutamic acid-N-hydroxysuccinimide ester (γ-PGA-NHS) was dissolved in 7% NaHCO3 solution with pH 8.3 to obtain precursor solution B; Dual syringe loading: precursor solution A and precursor solution B were loaded into the two lumens of the dual syringe respectively; the mass ratio of LAP: Ag2O2: SFMA: γ-PGA-NHS was 1:40:40:40. Example 2:

[0034] Preparation of titanium carbide nanosheet loaded with silver peroxide (TAg): 3-aminopropyltriethoxysilane (APTES) (100 μL) was added to Ti3C2 (1 mg / mL, 20 mL) and stirred at 50°C for 24 h to generate Ti3C2-NH2; Ti3C2-NH2 and Ag2O2 were dissolved in 10 mL deionized water at a mass ratio of 1:0.5 and stirred, and the precipitate was washed with deionized water to obtain titanium carbide nanosheet loaded with silver peroxide (TAg); Preparation of precursor solution A: titanium carbide nanosheet loaded with silver peroxide (TAg), photoinitiator LAP and methacrylated silk fibroin (SFMA) were dissolved in deionized water, mixed uniformly to obtain precursor solution A; Preparation of precursor solution B: γ-polyglutamic acid-N-hydroxysuccinimide ester (γ-PGA-NHS) was dissolved in 7% NaHCO3 solution with pH 8.3 to obtain precursor solution B; Dual syringe loading: precursor solution A and precursor solution B were loaded into the two lumens of the dual syringe respectively to obtain TAg@ SMP; the mass ratio of LAP: TAg: SFMA: γ-PGA-NHS was 1:40:40:40. Example 3:

[0035] Preparation of titanium carbide nanosheet (TCAg) loaded with caffeic acid phenethyl ester and silver peroxide: 3-aminopropyltriethoxysilane (APTES) (100 μL) was added to Ti3C2 (1 mg / mL, 20 mL) and stirred at 50℃ for 24 h to form Ti3C2-NH2; Ti3C2-NH2 was dissolved in 10 mL of deionized water with CAPE at a mass ratio of 1:15, stirred overnight, and then Ag2O2 (Ti3C2-NH2 and Ag2O2 mass ratio 1:0.5) was added. The precipitate was washed with deionized water to obtain TCAg nanosheets; Preparation of precursor solution A: titanium carbide nanosheet (TCAg) loaded with caffeic acid phenethyl ester (CAPE) and silver peroxide (Ag2O2), photoinitiator LAP, and methacrylated silk fibroin (SFMA) were dissolved in deionized water, mixed uniformly, and obtained as precursor solution A; Preparation of precursor solution B: γ-polyglutamic acid-N-hydroxysuccinimide ester (γ-PGA-NHS) was dissolved in a 7% NaHCO3 solution at pH 8.3 to obtain precursor solution B; Double syringe loading: precursor solution A and precursor solution B were loaded into the two lumens of a double syringe to obtain TCAg@SMP-10; the mass ratio of LAP: TCAg: SFMA: γ-PGA-NHS was 1:40:40:40. Example 4:

[0036] The mass ratio of Ti3C2-NH2 to CAPE was 1:20, and the mass ratio of Ti3C2-NH2 to Ag2O2 was 1:1; the mass ratio of LAP: TCAg: SFMA: γ-PGA-NHS was 1:40:80:40 to obtain TCAg@SMP-20, and the rest was the same as in Example 3. Example 5:

[0037] The mass ratio of Ti3C2-NH2 to CAPE was 1:25, and the mass ratio of Ti3C2-NH2 to Ag2O2 was 1:2; the mass ratio of LAP: TCAg: SFMA: γ-PGA-NHS was 1:40:120:40 to obtain TCAg@SMP-30, and the rest was the same as in Example 3.

[0038] Example 6: Process optimization The present application investigated the ratio of Ti3C2-NH2 to CAPE and the ratio of Ti3C2-NH2 to Ag2O2 by single factor investigation method, and the results are shown in Tables 1-2: Table 1 Effect of different ratios of Ti3C2-NH2: CAPE on the particle size and drug loading of nanosheets (n = 3, mean ± SD) Ti3C2-NH2: CAPE Size (nm) Drug loading (%) 1：15 518.24±0.66 90.45±0.63 1：30 522.68±0.70 95.25±0.46 1：45 530.76±0.62 89.60±0.77 As can be seen from Table 1, as the ratio of Ti3C2-NH2: CAPE increases, the drug loading of TCAg nanosheets gradually increases, but when it increases to 1:45, the drug loading decreases instead, and when the ratio of Ti3C2-NH2: Ag2O2 is 1:30, the drug loading is the highest, so the mass ratio of Ti3C2-NH2: CAPE is preferably 1:15-30, and more preferably 1:30.

[0039] Table 2 Effect of different ratios of Ti3C2-NH2: Ag2O2 on the particle size and drug loading of nanosheets (n = 3, mean ± SD) [Ti3C2-NH2: Ag2O2] Size (nm) Drug loading (%) 1：0.5 562.12±0.52 84.8±1.54 1：1.5 545.25±0.67 91.30±0.74 1：3.0 578.64±0.77 88.55±1.12 As can be seen from Table 2, as the ratio of Ti3C2-NH2: Ag2O2 increases, the drug loading of TCAg nanosheets gradually increases, but when it increases to 1:3, the drug loading decreases instead, and when the ratio of Ti3C2-NH2: Ag2O2 is 1:1.5, the drug loading is the highest, so the mass ratio of Ti3C2-NH2: CAPE is preferably 1:3, and preferably 1:1.5.

[0040] Example 7: Performance characterization (1) SEM and EDS characterization of Ti3C2, Ag2O2, TAg, and TCAg The morphology of Ti3C2, Ag2O2, TAg, and TCAg was observed by scanning electron microscopy (SEM) and transmission electron microscopy (TEM). The SEM results are shown in Figure 1 A, Ti3C2 is a flaky structure with a sharp edge like a knife blade, Ag2O2 is a spherical particle Figure 1 B), the transmission electron microscopy results show that TAg nanosheets are a two-dimensional flaky structure with spherical particles distributed on the surface Figure 1 C), and TCAg is a thickened flaky structure with spherical particles scattered on the surface, and a transparent paste is seen on the periphery Figure 1 D). The EDS characterization results show that Ti, Ag, C, and O elements are uniformly distributed on TCAg Figure 1 E).

[0041] (2) Potential characterization of Ti3C2, Ti3C2-NH2, Ag2O2, TAg, and TCAg The potential changes of Ti3C2, Ti3C2-NH2, Ag2O2, TAg, and TCAg were measured, and the results are shown in Figure 2The results show that Ti3C2 possesses a negative charge (-38.7 ± 1.1 mV) due to the presence of surface hydroxyl groups. The silanol groups of APTES react with the hydroxyl groups of Ti3C2, covalently anchoring the amino groups to the Ti3C2 surface, thereby causing the potential of Ti3C2-NH2 to become positive. This positive surface charge facilitates the subsequent loading of negatively charged Ag2O2 nanoparticles (-14.6 ± 0.3 mV). The potential of TAg is 32.6 ± 1.9 mV, and the potential of TCAg is 21.8 ± 2.4 mV.

[0042] (3) Acoustodynamic properties of Ti3C2, TAG, and TCAg Detection of ROS using 1,3-diphenylisobenzofuran (DPBF) as a molecular probe ( 1 The acoustic-dynamic properties of Ti3C2, TAg, and TCAg were evaluated using US stimulation (1 W / cm²). The acoustic-dynamic properties of Ti3C2, TAg, and TCAg were measured under US stimulation (1 W / cm²). 2 The ability to generate ROS at 50% duty cycle and 1 MHz, such as Figure 3 As shown, the decrease in the characteristic absorption peak of DPBF of Ti3C2 under ultrasonic treatment over time is relatively limited. Figure 3 (A); However, after loading Ag2O2 to form TAG, the characteristic absorbance value of DPBF showed a significant time-dependent decrease, indicating that TAG has the ability to generate reactive oxygen species in a time-dependent manner. Figure 3 Similarly, TCAg also exhibits excellent ROS generation performance, indicating that it has good acoustic dynamic effects. Figure 3 (C)

[0043] (4) Characterization of SFMA and γ-PGA-NHS proton NMR spectra use 1 SFMA was characterized by H-NMR ( Figure 4 Compared to SF, SFMA exhibits two new signal peaks at 6.06 ppm and 5.58 ppm, which are signals generated by two hydrogen nuclei in -CH=CH2, indicating successful synthesis of SFMA. 1 γ-PGA-NHS was characterized by H-NMR ( Figure 4 In the γ-PGA-NHS spectrum (Cb), compared with the γ-PGA spectrum, the peak with a chemical shift of 12.6 ppm (carboxyl proton peak) disappeared, while a new characteristic peak with a chemical shift of 2.7 ppm (NHS ester proton peak) was added, confirming that the carboxyl group of γ-PGA was successfully activated into NHS ester.

[0044] (5) Microstructure characterization of TCAg@SMP prepared in Examples 3-5 The TCAg@SMP mixture prepared in the examples was sputter-coated with gold and then observed using a scanning electron microscope. The results showed that the gel network gradually became denser with increasing SFMA concentration. Figure 5 Furthermore, the presence of nanoparticles can be observed within the gel mesh. Figure 5 ).

[0045] (6) Characterization of the asymmetric adhesion of TCAg@SMP prepared in Examples 3-5 Adhesion-Crosslinking: The adhesion strength of TCAg@SMP was tested using a standardized shear test. Pigskin was cleaned three times before the test and cut into 50 mm × 50 mm squares. An appropriate amount of TCAg@SMP was applied to a piece of pigskin, and then another piece of pigskin was used to cover it, recording the contact area. After irradiating the hydrogel precursor with ultraviolet light (405 nm) for 10 s, the adhesion strength was measured using an electronic universal testing machine (material testing system: MTS standard43, 50N load cell, 10 mm / min). -1 The adhesion strength is tested at the rate of adhesion.

[0046] Crosslinking followed by adhesion: The adhesion strength of TCAg@SMP was tested using a standardized shear test. Before the test, pigskin was cleaned three times and cut into 50 mm × 50 mm squares. An appropriate amount of TCAg@SMP precursor was coated onto a piece of pigskin. The hydrogel precursor was irradiated with ultraviolet light (405 nm) for 10 seconds, and then another piece of pigskin was used to cover it, and the contact area was recorded. An electronic universal testing machine (material testing system: MTS standard43, 50N load cell, 10 mm / min) was used. -1 The adhesion strength is tested at the rate of adhesion.

[0047] Experimental results show that the TCAg@SMP hydrogel exhibits asymmetric adhesion properties. TCAg@SMP-10 represents Example 1, TCAg@SMP-20 represents Example 2, and TCAg@SMP-30 represents Example 3. Figure 6 As shown in Figure A, the pre-adhesive-post-crosslinked TCAg@SMP exhibits strong adhesion to porcine skin tissue. This is attributed to the residual NHS ester groups in the hydrogel, which can form covalent bonds with the amine groups in biological tissue, thereby enhancing interfacial adhesion. The hydrogel prepared in Example 3 showed an adhesion strength of 26.72 ± 0.86 kPa, and the adhesion strength gradually increased with increasing SFMA concentration. Conversely, when TCAg@SMP was photocrosslinked before application to the tissue surface, the resulting hydrogel exhibited extremely low adhesion, with the hydrogel prepared in Example 3 showing an adhesion strength of 5.15 ± 0.23 kPa. Figure 6 (B) This is because the cross-linked network formed after photocuring restricts the movement of NHS ester groups on the hydrogel surface.

[0048] Example 8: Evaluation of the inhibitory effect of TCAg@SMP prepared in Example 3 on bacterial adhesion. Add 1 mL of Porphyromonas gingivalis ( Porphyromonas gingivalis, P.g ) and Fusobacterium nucleatum ( Fusobacterium nucleatum, F.n ) suspension (total 10) 8 CFU / mL was placed in a confocal microplate and incubated for 48 h. The bacterial biofilm was then stained with a mixture of SYTO9 (2.5 μM) and propyl iodide (2.5 μM). Images of the stained bacteria (live and dead) were captured using a laser confocal microscope. The bacteria were then sonicated with TCAg@SMP and cured under UV light. The bacterial biofilm was stained again with a mixture of SYTO9 (2.5 μM) and propyl iodide (2.5 μM). Observation was performed using a laser confocal microscope. Then, 100 μL of Pg and Fn suspension (total 10) was added. 8 After culturing for 24 h, the bacterial biofilm was stained with a mixture of SYTO 9 (2.5 μM) and propyl iodide (2.5 μM). Observation was performed using a laser confocal microscope. The control group used confocal dishes without TCAg@SMP, with the remaining procedures the same.

[0049] Experimental results showed that, based on live / dead cell staining, a large number of dead bacteria were found on the glass slides treated with TCAg@SMP hydrogel after sonication. Figure 7 (A-②), while the untreated control showed bacterial biofilm formation ( Figure 7 (B-②). Subsequently, photocrosslinking was performed using TCAg@SMP hydrogel, followed by application of the bacterial suspension to evaluate its anti-adhesion properties. The results showed that a large amount of bacteria proliferated and aggregated on the glass slide (B-②). Figure 7 (B-③). In contrast, in the photocrosslinked TCAg@SMP hydrogel, the NHS ester groups are immobilized in the hydrogel network, resulting in reduced bacterial adhesion ( Figure 7 (A-③). This indicates that TCAg@SMP hydrogel has a potential role in preventing subsequent bacterial adhesion in periodontitis.

[0050] Example 9: Antibacterial performance test of Ti3C2, Ag2O2, and TCAg CFU flat plate coating: In order to make CFU flat plate coating: P.g A single biofilm was formed, and the microbial concentration was adjusted to 10 in a 6-well plate. 8CFU / mL, with round glass climbing sheets at the bottom of each well. All biofilms were cultured in an anaerobic incubator at 37°C for 96 h. Five groups were established: Control group, CAPE group, Ti3C2 group, Ag2O2 group, TCAg group. Different nanomaterials and ultrasound (frequency 1 MHZ, duty cycle 50%, power 1 W / cm 2 ) were used to treat the biofilm for 3 days. Cell scraper was used to remove the attached biofilm. Then it was diluted by 10 times continuously. 200 μL of diluted bacterial solution was taken to Columbia blood agar.

[0051] Figure 8 The colony forming unit (CFU) count of P.g P. gingivalis in all test groups showed that TCAg exhibited the highest bacterial removal efficiency.

[0052] SEM bacterial morphology observation: In order to form a single biofilm of P. gingivalis, the microbial concentration was adjusted to 10 8 CFU / mL in a 6-well plate, with round glass climbing sheets at the bottom of each well. All biofilms were cultured in an anaerobic incubator at 37°C for 96 h. Five groups were established: Control group, CAPE group, Ti3C2 group, Ag2O2 group, TCAg group. Different nanomaterials and ultrasound (frequency 1 MHZ, duty cycle 50%, power 1 W / cm 2 ) were used to treat the biofilm for 3 days. Then the glass climbing sheets were taken out and placed in glutaraldehyde fixing solution for fixation, then dehydrated and dried with different concentrations of ethanol, and then observed by scanning electron microscope after gold spraying.

[0053] Figure 9 The CAPE group, Ti3C2 group, Ag2O2 group, and TCAg group all showed different degrees of bacterial biofilm destruction ability, which could cause the atrophy or even rupture of the bacterial biofilm. Among them, the TCAg group had the best destruction effect.

[0054] Example 10: Pharmacodynamic evaluation of TAg@SMP and TCAg@SMP prepared in Examples 2 and 3 for treating periodontitis rats The periodontitis rat model was constructed by ligation with silk thread combined with injection of P. gingivalis and Fusobacterium nucleatum. The specific operation is as follows: after one week of adaptive feeding of healthy SPF rats, except for the normal group, the rest of the groups were anesthetized with 3% sodium pentobarbital (30 mg / kg) by intraperitoneal injection. The rats were fixed in supine position, and the head and limbs were fixed. The left upper molar was fully exposed by pulling the upper and lower jaws, and then the left upper molar of the rat was ligated with 4-0 silk suture and injected with P. gingivalis and Fusobacterium nucleatum (the concentration was 10 7CFU / mL, 100 μL). Check if the silk thread has fallen off during the period, if it has slipped, re-ligate in time, and observe if it forms a periodontal pocket. The successful modeling rats are randomly divided into groups: normal group, model group, positive drug group (PERIO), TAg@SMP group, TCAg@SMP group. Every 3 days, 100 μL of the corresponding preparation is injected submucosally on the buccal and palatal side of the central alveolar ridge of the maxillary first molar of the rats. The normal group and the model group are injected with the same volume of normal saline. After administration of TAg@SMP group and TCAg@SMP group, ultrasonic treatment (frequency 1 MHZ, duty cycle 50%, power 1 W / cm 2 ) is performed for 2 minutes, followed by ultraviolet light curing. The continuous administration is performed for 4 weeks. After 4 weeks, the rats are sacrificed by intraperitoneal injection of an overdose of sodium pentobarbital, and the left maxillary bone is separated. The excess soft tissue is removed with a precision cutter, and the specimen is fixed in 4% paraformaldehyde for 48 h. The tooth specimen is taken out and placed in a 10% EDT solution for decalcification, gradient ethanol dehydration, paraffin embedding, sectioning, and Fish staining.

[0055] Fish fluorescence in situ hybridization sections are prepared, Figure 10 It is shown that the model group shows a large number of Porphyromonas gingivalis (red) and Fusobacterium nucleatum (green) invading the gingival tissue, while the TCAg@SMP group exhibits the lowest amount of Porphyromonas gingivalis (red) and Fusobacterium nucleatum (green) invading the gingival tissue. It is shown that TCAg@SMP has good antibacterial effect.

[0056] In order to study the alveolar bone repair, the maxillary bone is scanned using Micro-CT, and the rat maxillary first molar alveolar bone is observed. The results are shown in Figure 11 The model group shows significant alveolar bone defect, root exposure, and furcation, indicating that the rat periodontitis model is successfully constructed. The TAg@SMP group has a certain degree of alveolar bone repair, and the distance from the enamel cementum junction to the alveolar crest is significantly shortened. The TCAg@SMP group has significant alveolar bone recovery, close to the normal group.

[0057] Example 11: Wound healing effect of TAg@SMP and TCAg@SMP gels prepared in Examples 2 and 3 on MRSA infected mouse wound model A mouse infected wound model is established, and a wound area with a diameter of about 8 mm is created on the back area of C57BL / 6J mice (male, 8-10 weeks), followed by inoculation of 20 μL of MRSA bacteria. The mice are randomly divided into groups, and are administered with PBS (200 μL), positive drug group Betadine gel (200 μL), sterile normal saline (200 μL), TAg@SMP (200 μL), and TCAg@SMP (200 μL), respectively. After administration, ultrasonic treatment (frequency 1 MHZ, duty cycle 50%, power 1 W / cm 2) 2 minutes later, and wound changes were closely monitored throughout the experiment.

[0058] Results are shown in Figure 12 By monitoring the whole wound healing process, it was found that TAg@SMP gel improved the efficiency of wound healing, and the wound began to close at an accelerated rate on the 7th day, in addition, the wound healing rate of TCAg@SMP was the fastest, significantly better than other groups.

Claims

1. An acoustically powered Janus hydrogel, characterized in that, The hydrogel comprises silver peroxide and a hydrogel matrix; the hydrogel matrix is composed of methacrylated silk fibroin, gamma-polyglutamic acid-N-hydroxysuccinimide ester and a photoinitiator.

2. The sonodynamically driven Janus hydrogel according to claim 1, wherein, The hydrogel further comprises titanium carbide nanosheets, and the silver peroxide is loaded on the titanium carbide nanosheets.

3. The sonodynamically driven Janus hydrogel according to claim 2, wherein, The titanium carbide nanosheets are further loaded with caffeic acid phenethyl ester.

4. The sonodynamically driven Janus hydrogel according to claim 3, wherein, In the hydrogel, the mass ratio of the photoinitiator, the titanium carbide nanosheets loaded with caffeic acid phenethyl ester and silver peroxide, methacrylated silk fibroin and gamma-polyglutamic acid-N-hydroxysuccinimide ester is 1:40:40-120:

40.

5. The sonodynamically driven Janus hydrogel according to claim 1, wherein, The photoinitiator is at least one of lithium phenyl-2,4,6-trimethylbenzoylphosphinate, 2,4,6-trimethylbenzoyl-diphenylphosphine oxide or 2-hydroxy-1-[4-(2-hydroxyethoxy)phenyl]-2-methyl-1-propanone.

6. The method of preparing an acoustically powered Janus hydrogel according to any one of claims 1-5, wherein: The method comprises the following steps: (1) preparing precursor solution A: dissolving silver peroxide, a photoinitiator and methacrylated silk fibroin in deionized water, mixing uniformly to obtain precursor solution A; (2) preparing precursor solution B: dissolving gamma-polyglutamic acid-N-hydroxysuccinimide ester in a NaHCO3 solution to obtain precursor solution B; (3) ultraviolet light curing: mixing the precursor solution A and the precursor solution B, ultraviolet light curing after ultrasonic treatment to obtain the sonodynamic Janus hydrogel.

7. The method of claim 6, wherein: In step (1), the titanium carbide nanosheets are modified by amino silane to obtain Ti3C2-NH2; then the Ti3C2-NH2 is dissolved in an ethanol solution, and silver peroxide and / or caffeic acid phenethyl ester are added in sequence, and after stirring, the titanium carbide nanosheets loaded with silver peroxide and / or caffeic acid phenethyl ester are obtained by centrifugal washing and precipitation.

8. The method of claim 7, wherein: The stirring temperature is 25-37°C, and the stirring time is 4-24 hours; the mass ratio of the Ti3C2-NH2 to the caffeic acid phenethyl ester is 1:15-30; and the mass ratio of the Ti3C2-NH2 to the silver peroxide is 1:1-3.

9. Use of the sonodynamic Janus hydrogel according to any one of claims 1-5 in the preparation of a drug for preventing or treating periodontitis or oral mucositis.

10. Use of the sonodynamic Janus hydrogel according to any one of claims 1-5 in the preparation of a skin wound repair material.

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