Zeolite imidazole skeleton-8 composite hydrogel loaded with sulforaphane and preparation method of zeolite imidazole skeleton-8 composite hydrogel

Abstract

The invention discloses a preparation method of SFN-loaded ZIF-8 composite hydrogel. The preparation method specifically comprises the following steps: step 1, preparing ZIF-8 nanoparticles; step 2, preparing ZIF-8 nano particles loaded with the SFN; and step 3, preparing the ZIF-8 composite hydrogel loaded with the SFN. SFN is adopted as a medicine, ZIF-8 nano-particles with osteogenesis promoting and antibacterial effects are adopted as a carrier, SFN-loaded ZIF-8 nano-drug-loaded particles are synthesized, the SFN-loaded ZIF-8 nano-particles are combined with the hydrogel, the local concentration of SFN at the periodontitis diseased region can be effectively improved, the bioavailability of SFN can be improved, the hydrogel has injectability, and the hydrogel can be used for preparing the medicine for treating periodontitis. The drug-loaded nanoparticles are sent to the deep part of a periodontal pocket, and only a minimally invasive wound is generated. The invention also discloses the SFN-loaded ZIF-8 composite hydrogel and an application of the SFN-loaded ZIF-8 composite hydrogel in preparation of a medicine for treating periodontitis.

Description

Technical Field

[0001] The present invention belongs to the field of oral science technology and relates to a sulforaphane-loaded zeolite imidazole skeleton-8 composite hydrogel. The present invention also relates to a preparation method and application of the zeolite imidazole skeleton-8 composite hydrogel. Background Art

[0002] Periodontitis is a chronic inflammatory disease caused by dental plaque, which is mainly manifested by inflammation and destruction of periodontal tissues, leading to alveolar bone resorption and tooth mobility. Currently, the main strategy for the treatment of periodontitis is to remove the causative factors through mechanical debridement, such as periodontal cleaning and root planing. However, mechanical treatment methods cannot completely remove dental plaque in complex anatomical structures (periodontal pockets and root bifurcations). Local delivery of antibiotics, such as minocycline ointment and doxycycline gel, has become an auxiliary treatment method for reducing infection and inflammation in clinical practice, but excessive use of antibiotics may lead to drug resistance.

[0003] Sulforaphane (SFN) is an isothiocyanate, a sulfur-containing compound, extracted from cruciferous vegetables. Current research on SFN has revealed that it is a natural antioxidant and anti-inflammatory agent, exhibiting significant anti-inflammatory and antioxidant effects. However, SFN's poor water solubility and low bioavailability limit its application in the treatment of periodontitis.

[0004] The high porosity and large surface area of ​​zeolitic imidazolate framework-8 (ZIF-8) nanoparticles make them suitable for loading drugs or biopolymers, preventing rapid degradation of the encapsulated materials and thus improving the bioavailability of the bioactive ingredients. Furthermore, ZIF-8 is pH-responsive, releasing zinc ions under acidic conditions, enhancing osteoinductive activity, promoting angiogenesis, and exerting antibacterial effects. However, SFN-loaded ZIF-8 drug delivery systems cannot be noninvasively delivered into deep periodontal pockets. Therefore, a new drug carrier capable of effective localized drug release is currently lacking in the treatment of periodontitis. Summary of the Invention

[0005] The purpose of the present invention is to provide a SFN-loaded ZIF-8 composite hydrogel, which solves the problem in the prior art of lacking a new drug carrier capable of effectively locally releasing drugs in the treatment of periodontitis.

[0006] The second object of the present invention is to provide a method for preparing the above-mentioned SFN-loaded ZIF-8 composite hydrogel.

[0007] The third object of the present invention is to provide the use of the above-mentioned SFN-loaded ZIF-8 composite hydrogel in the preparation of drugs for treating periodontitis.

[0008] The technical solution adopted by the present invention is a method for preparing a ZIF-8 composite hydrogel loaded with SFN, which specifically comprises the following steps: Step 1, preparing ZIF-8 nanoparticles; Step 2, preparing SFN-loaded ZIF-8 nanoparticles; Step 3: Prepare SFN-loaded ZIF-8 composite hydrogel.

[0009] The present invention is also characterized in that: Step 1 specifically includes the following steps: Step 1.1, preparing anhydrous zinc acetate methanol solution; Step 1.2, prepare 2-methylimidazole methanol solution; Step 1.3, add 2-methylimidazole methanol solution to anhydrous zinc acetate methanol solution, stir for 30-60 minutes to obtain a milky white suspension, let it stand for 12-24 hours, and then centrifuge at a centrifuge speed of 8000-10000 rpm for 7-10 minutes to obtain a white precipitate, wash the white precipitate with anhydrous methanol 3-5 times, and then dry the precipitate at 60°C to obtain ZIF-8 nanoparticles.

[0010] In step 1.1, 50-70 mg of anhydrous zinc acetate was weighed and dissolved in 10-20 ml of anhydrous methanol to obtain an anhydrous zinc acetate methanol solution; In step 1.2, 500-700 mg of 2-methylimidazole was weighed and dissolved in 10-20 ml of anhydrous methanol to obtain a 2-methylimidazole methanol solution.

[0011] Step 2 specifically includes the following steps: Step 2.1, preparing SFN methanol solution; Step 2.2, add ZIF-8 to the SFN methanol solution, stir and shake in the dark, so that the SFN molecules are adsorbed in the ZIF-8 pores, and centrifuge the obtained solution at a speed of 8000~10000rpm for 7~10min to obtain a white precipitate. Wash the white precipitate with anhydrous methanol 3~5 times, and then dry the precipitate at 60°C to obtain drug-loaded ZIF-8 nanoparticles.

[0012] In step 2.1, at 25-28°C, add 6-10 mg of SFN to 3-5 ml of anhydrous methanol and mix well to obtain a SFN methanol solution. In step 2.2, weigh 50-70 mg of ZIF-8.

[0013] Step 3 specifically includes the following steps: Step 3.1, preparing an oxidized hyaluronic acid solution; Step 3.2, preparing carboxymethyl chitosan solution; In step 3.3, the SFN-loaded ZIF-8 nanoparticles are added to the carboxymethyl chitosan solution, and then mixed evenly with an equal volume of oxidized hyaluronic acid solution to obtain a composite hydrogel of the SFN-loaded ZIF-8 nanoparticles.

[0014] In step 3.1, the mass concentration of the oxidized hyaluronic acid solution is 4% to 5%; in step 3.2, the mass concentration of the carboxymethyl chitosan solution is 2% to 3%; in step 3.3, the concentration of the drug-loaded nanoparticles in the carboxymethyl chitosan solution is 0.5 to 2 mg / mL.

[0015] The second technical solution adopted by the present invention is that the SFN-loaded ZIF-8 composite hydrogel is obtained by adopting the above-mentioned preparation method.

[0016] The third technical solution adopted by the present invention is the use of SFN-loaded ZIF-8 composite hydrogel and the use of SFN-loaded ZIF-8 composite hydrogel in the preparation of drugs for treating periodontitis.

[0017] The beneficial effects of the present invention are: (1) The present invention uses SFN as a drug and ZIF-8 nanoparticles that promote osteogenesis and have antibacterial effects as a carrier to synthesize SFN-loaded ZIF-8 nanoparticles. By combining the SFN-loaded ZIF-8 nanoparticles with hydrogels, the local concentration of SFN in the periodontitis lesion site can be effectively increased, thereby improving the bioavailability of SFN.

[0018] (2) In the present invention, the amino group of carboxymethyl chitosan and the aldehyde group of oxidized hyaluronic acid react with the Schiff base to form an imine bond. At the same time, ZIF-8 has pH response. The combined use of the two makes the composite hydrogel have good sustained-release properties, achieving controllable and precise release of SFN and zinc ions, which is suitable for the acidic periodontitis environment, achieving controlled release of drugs, and can prolong the action time of SFN, exerting multiple functions of anti-inflammatory, antioxidant, antibacterial and periodontal bone repair.

[0019] (3) The hydrogel in the present invention is injectable and can deliver drug-loaded nanoparticles deep into the periodontal pocket to fill the bone defects caused by periodontitis, resulting in only minimal wounds and improving patient compliance. DETAILED DESCRIPTION

[0020] The following describes it in detail with reference to specific implementation methods.

[0021] The preparation method of SFN-loaded ZIF-8 composite hydrogel specifically comprises the following steps: Step 1, preparing ZIF-8 nanoparticles, the specific steps are as follows: Step 1.1, prepare anhydrous zinc acetate methanol solution, Weigh 50-70 mg of anhydrous zinc acetate and add it to 10-20 ml of anhydrous methanol to dissolve it to obtain an anhydrous zinc acetate methanol solution.

[0022] Step 1.2, prepare 2-methylimidazole methanol solution, Weigh 500-700 mg of 2-methylimidazole and dissolve it in 10-20 ml of anhydrous methanol to obtain a 2-methylimidazole methanol solution.

[0023] Step 1.3, add 2-methylimidazole methanol solution to anhydrous zinc acetate methanol solution, stir for 30-60 minutes to obtain a milky white suspension, let it stand for 12-24 hours, and then centrifuge at a centrifuge speed of 8000-10000 rpm for 7-10 minutes to obtain a white precipitate, wash the white precipitate with anhydrous methanol 3-5 times, and then dry the precipitate at 60°C to obtain ZIF-8 nanoparticles.

[0024] Step 2, preparing SFN-loaded ZIF-8 nanoparticles, specifically comprising the following steps: Step 2.1, prepare SFN methanol solution, At 25-28°C, add 6-10 mg of SFN to 3-5 ml of anhydrous methanol and mix well to obtain a SFN methanol solution.

[0025] Step 2.2, weigh 50-70 mg of ZIF-8, add ZIF-8 to the SFN methanol solution, stir and shake in the dark, so that the SFN molecules are adsorbed in the ZIF-8 pores, and centrifuge the obtained solution at a speed of 8000-10000 rpm for 7-10 min to obtain a white precipitate. Wash the white precipitate with anhydrous methanol 3-5 times, and then dry the precipitate at 60°C to obtain drug-loaded ZIF-8 nanoparticles.

[0026] Step 3, preparing the SFN-loaded ZIF-8 composite hydrogel, specifically comprising the following steps: Step 3.1, prepare oxidized hyaluronic acid solution, The oxidized hyaluronic acid was dissolved in deionized water to obtain an oxidized hyaluronic acid solution with a mass concentration of 4% to 5%.

[0027] Step 3.2, prepare carboxymethyl chitosan solution, Carboxymethyl chitosan was dissolved in deionized water to obtain a carboxymethyl chitosan solution with a mass concentration of 2% to 3%.

[0028] In step 3.3, SFN-loaded ZIF-8 nanoparticles are added to a carboxymethyl chitosan solution at a concentration of 0.5 to 2 mg / mL. The solution is then mixed with an equal volume of oxidized hyaluronic acid solution to produce a composite hydrogel of SFN-loaded ZIF-8 nanoparticles.

[0029] The SFN-loaded ZIF-8 composite hydrogel was obtained by the above-mentioned preparation method.

[0030] Application of SFN-loaded ZIF-8 composite hydrogel, and application of SFN-loaded ZIF-8 composite hydrogel in the preparation of drugs for treating periodontitis.

[0031] Example 1 The preparation method of SFN-loaded ZIF-8 composite hydrogel specifically comprises the following steps: Step 1, preparing ZIF-8 nanoparticles; Step 2, preparing SFN-loaded ZIF-8 nanoparticles; Step 3: Prepare SFN-loaded ZIF-8 composite hydrogel.

[0032] Example 2 The preparation method of SFN-loaded ZIF-8 composite hydrogel specifically comprises the following steps: Step 1, preparing ZIF-8 nanoparticles; Step 2, preparing SFN-loaded ZIF-8 nanoparticles; Step 3: Prepare SFN-loaded ZIF-8 composite hydrogel.

[0033] Step 1 specifically includes the following steps: Step 1.1, preparing anhydrous zinc acetate methanol solution; Step 1.2, prepare 2-methylimidazole methanol solution; Step 1.3, add 2-methylimidazole methanol solution to anhydrous zinc acetate methanol solution, stir for 30-60 minutes to obtain a milky white suspension, let it stand for 12-24 hours, and then centrifuge at a centrifuge speed of 8000-10000 rpm for 7-10 minutes to obtain a white precipitate, wash the white precipitate with anhydrous methanol 3-5 times, and then dry the precipitate at 60°C to obtain ZIF-8 nanoparticles.

[0034] Example 3 The preparation method of SFN-loaded ZIF-8 composite hydrogel specifically comprises the following steps: Step 1, preparing ZIF-8 nanoparticles; Step 2, preparing SFN-loaded ZIF-8 nanoparticles; Step 3: Prepare SFN-loaded ZIF-8 composite hydrogel.

[0035] Step 1 specifically includes the following steps: Step 1.1, preparing anhydrous zinc acetate methanol solution; Step 1.2, prepare 2-methylimidazole methanol solution; Step 1.3, add 2-methylimidazole methanol solution to anhydrous zinc acetate methanol solution, stir for 30-60 minutes to obtain a milky white suspension, let it stand for 12-24 hours, and then centrifuge at a centrifuge speed of 8000-10000 rpm for 7-10 minutes to obtain a white precipitate, wash the white precipitate with anhydrous methanol 3-5 times, and then dry the precipitate at 60°C to obtain ZIF-8 nanoparticles.

[0036] In step 1.1, 50-70 mg of anhydrous zinc acetate was weighed and dissolved in 10-20 ml of anhydrous methanol to obtain an anhydrous zinc acetate methanol solution; In step 1.2, 500-700 mg of 2-methylimidazole was weighed and dissolved in 10-20 ml of anhydrous methanol to obtain a 2-methylimidazole methanol solution.

[0037] Example 4 The preparation method of SFN-loaded ZIF-8 composite hydrogel specifically comprises the following steps: Step 1, preparing ZIF-8 nanoparticles; Step 2, preparing SFN-loaded ZIF-8 nanoparticles; Step 3: Prepare SFN-loaded ZIF-8 composite hydrogel.

[0038] Step 2 specifically includes the following steps: Step 2.1, preparing SFN methanol solution; Step 2.2, add ZIF-8 to the SFN methanol solution, stir and shake in the dark, so that the SFN molecules are adsorbed in the ZIF-8 pores, and centrifuge the obtained solution at a speed of 8000~10000rpm for 7~10min to obtain a white precipitate. Wash the white precipitate with anhydrous methanol 3~5 times, and then dry the precipitate at 60°C to obtain drug-loaded ZIF-8 nanoparticles.

[0039] Example 5 The preparation method of SFN-loaded ZIF-8 composite hydrogel specifically comprises the following steps: Step 1, preparing ZIF-8 nanoparticles; Step 2, preparing SFN-loaded ZIF-8 nanoparticles; Step 3: Prepare SFN-loaded ZIF-8 composite hydrogel.

[0040] Step 2 specifically includes the following steps: Step 2.1, preparing SFN methanol solution; Step 2.2, add ZIF-8 to the SFN methanol solution, stir and shake in the dark, so that the SFN molecules are adsorbed in the ZIF-8 pores, and centrifuge the obtained solution at a speed of 8000~10000rpm for 7~10min to obtain a white precipitate. Wash the white precipitate with anhydrous methanol 3~5 times, and then dry the precipitate at 60°C to obtain drug-loaded ZIF-8 nanoparticles.

[0041] In step 2.1, at 25-28°C, add 6-10 mg of SFN to 3-5 ml of anhydrous methanol and mix well to obtain a SFN methanol solution. In step 2.2, weigh 50-70 mg of ZIF-8.

[0042] Example 6 The preparation method of SFN-loaded ZIF-8 composite hydrogel specifically comprises the following steps: Step 1, preparing ZIF-8 nanoparticles; Step 2, preparing SFN-loaded ZIF-8 nanoparticles; Step 3: Prepare SFN-loaded ZIF-8 composite hydrogel.

[0043] Step 3 specifically includes the following steps: Step 3.1, preparing an oxidized hyaluronic acid solution; Step 3.2, preparing carboxymethyl chitosan solution; In step 3.3, the SFN-loaded ZIF-8 nanoparticles are added to the carboxymethyl chitosan solution, and then mixed evenly with an equal volume of oxidized hyaluronic acid solution to obtain a composite hydrogel of the SFN-loaded ZIF-8 nanoparticles.

[0044] Example 7 The preparation method of SFN-loaded ZIF-8 composite hydrogel specifically comprises the following steps: Step 1, preparing ZIF-8 nanoparticles; Step 2, preparing SFN-loaded ZIF-8 nanoparticles; Step 3: Prepare SFN-loaded ZIF-8 composite hydrogel.

[0045] Step 3 specifically includes the following steps: Step 3.1, preparing an oxidized hyaluronic acid solution; Step 3.2, preparing carboxymethyl chitosan solution; In step 3.3, the SFN-loaded ZIF-8 nanoparticles are added to the carboxymethyl chitosan solution, and then mixed evenly with an equal volume of oxidized hyaluronic acid solution to obtain a composite hydrogel of the SFN-loaded ZIF-8 nanoparticles.

[0046] In step 3.1, the mass concentration of the oxidized hyaluronic acid solution is 4% to 5%; in step 3.2, the mass concentration of the carboxymethyl chitosan solution is 2% to 3%; in step 3.3, the concentration of the drug-loaded nanoparticles in the carboxymethyl chitosan solution is 0.5 to 2 mg / mL.

[0047] Example 8 The SFN-loaded ZIF-8 composite hydrogel is obtained by using any of the above preparation methods.

[0048] Example 9 Application of SFN-loaded ZIF-8 composite hydrogel, and application of SFN-loaded ZIF-8 composite hydrogel in the preparation of drugs for treating periodontitis.

Claims

1. A method for preparing SFN-loaded ZIF-8 composite hydrogel, characterized in that: The specific steps include: Step 1, preparing ZIF-8 nanoparticles; Step 2, preparing SFN-loaded ZIF-8 nanoparticles; Step 3: Prepare SFN-loaded ZIF-8 composite hydrogel.

2. The preparation method of the SFN-loaded ZIF-8 composite hydrogel according to claim 1, wherein The step 1 specifically includes the following steps: Step 1.1, preparing anhydrous zinc acetate methanol solution; Step 1.2, prepare 2-methylimidazole methanol solution; Step 1.3, add 2-methylimidazole methanol solution to anhydrous zinc acetate methanol solution, stir for 30-60 minutes to obtain a milky white suspension, let it stand for 12-24 hours, and then centrifuge at a centrifuge speed of 8000-10000 rpm for 7-10 minutes to obtain a white precipitate, wash the white precipitate with anhydrous methanol 3-5 times, and then dry the precipitate at 60°C to obtain ZIF-8 nanoparticles.

3. The preparation method of the SFN-loaded ZIF-8 composite hydrogel according to claim 2, wherein In step 1.1, 50-70 mg of anhydrous zinc acetate is weighed and added to 10-20 ml of anhydrous methanol to dissolve to obtain an anhydrous zinc acetate methanol solution; In the step 1.2, 500-700 mg of 2-methylimidazole is weighed and added to 10-20 ml of anhydrous methanol to dissolve to obtain a 2-methylimidazole methanol solution.

4. The method for preparing the SFN-loaded ZIF-8 composite hydrogel according to claim 2, wherein The step 2 specifically includes the following steps: Step 2.1, preparing SFN methanol solution; Step 2.2, add ZIF-8 to the SFN methanol solution, stir and shake in the dark, so that the SFN molecules are adsorbed in the ZIF-8 pores, and centrifuge the obtained solution at a speed of 8000~10000rpm for 7~10min to obtain a white precipitate. Wash the white precipitate with anhydrous methanol 3~5 times, and then dry the precipitate at 60°C to obtain drug-loaded ZIF-8 nanoparticles.

5. The method for preparing the SFN-loaded ZIF-8 composite hydrogel according to claim 4, wherein In the step 2.1, 6-10 mg of SFN is added to 3-5 ml of anhydrous methanol at 25-28° C. and mixed to obtain a SFN methanol solution. In the step 2.2, 50-70 mg of ZIF-8 is weighed.

6. The method for preparing the SFN-loaded ZIF-8 composite hydrogel according to claim 4, wherein The step 3 specifically includes the following steps: Step 3.1, preparing an oxidized hyaluronic acid solution; Step 3.2, preparing carboxymethyl chitosan solution; In step 3.3, the SFN-loaded ZIF-8 nanoparticles are added to the carboxymethyl chitosan solution, and then mixed evenly with an equal volume of oxidized hyaluronic acid solution to obtain a composite hydrogel of the SFN-loaded ZIF-8 nanoparticles.

7. The method for preparing the SFN-loaded ZIF-8 composite hydrogel according to claim 6, wherein: In step 3.1, the mass concentration of the oxidized hyaluronic acid solution is 4% to 5%; in step 3.2, the mass concentration of the carboxymethyl chitosan solution is 2% to 3%; in step 3.3, the concentration of the drug-loaded nanoparticles in the carboxymethyl chitosan solution is 0.5 to 2 mg / mL.

8. SFN-loaded ZIF-8 composite hydrogel, characterized in that: The method is obtained by the preparation method according to any one of claims 1 to 7.

9. Application of SFN-loaded ZIF-8 composite hydrogel, characterized in that: Application of SFN-loaded ZIF-8 composite hydrogel in the preparation of drugs for the treatment of periodontitis.