Injectable hydrogel as well as preparation method and application thereof

Abstract

The invention relates to an injectable hydrogel. The injectable hydrogel is prepared from sericin protein grafted dopamine, aminated hyaluronic acid and nano-micelles, the nano-micelle is mainly prepared from monomethoxy polyethylene glycol, 1, 3-dioxo-2-ketone and a ring-opening monomer containing a diselenide structure, the nano-micelles are loaded with drugs and proteins. The preparation method comprises the following steps: preparation of sericin grafted dopamine, preparation of aminated hyaluronic acid, preparation of nano-micelles loaded with drugs and proteins, and preparation of injectable hydrogel. The injectable hydrogel can be used for postoperative local treatment of uterine cavity, and can effectively prevent or reduce occurrence and relapse of intrauterine adhesion by accurately regulating oxidative stress, inflammatory response and fibrosis processes and cooperatively promoting endometrial regeneration.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of hydrogel, in particular to an injectable hydrogel and a preparation method and application thereof. BACKGROUND

[0002] Intrauterine adhesion (IUA) is mainly manifested as abnormal hyperplasia of fibrous cord-like tissue after damage to the endometrial tissue, leading to partial or complete occlusion of the uterine cavity, and then causing a series of serious consequences such as menstrual abnormalities, infertility, habitual abortion, etc. Although the current clinical treatment mainly adopts mechanical adhesion separation under hysteroscopy combined with estrogen auxiliary treatment to promote the regeneration of the endometrium and restore its normal morphology and function, the postoperative recurrence rate is still as high as 20%~60%, especially in patients with severe IUA, the recurrence risk and tissue repair difficulty are more prominent, which seriously restricts the treatment effect and prognosis.

[0003] Pathological studies have shown that due to mechanical damage, infection and other factors after hysteroscopy, the local tissue is in a high oxidative stress state, and reactive oxygen species (ROS) continuously accumulate, which not only directly induces cell apoptosis and tissue damage, but also activates the TGF-β (transforming growth factor beta) and other pro-fibrotic signaling pathways, drives fibroblast proliferation, migration and secretes a large amount of extracellular matrix, thereby forming dense adhesion tissue. In addition, the damaged area is often accompanied by chronic inflammatory reaction, angiogenesis disorder and limited endometrial stem cell migration / differentiation, etc., leading to slow tissue regeneration and significantly decreased endometrial repair capacity, which is prone to adhesion recurrence. Therefore, the effective treatment of postoperative intrauterine adhesion not only needs a physical anti-adhesion barrier, but also requires the material to have multiple biological functions, including antioxidant, anti-inflammatory, anti-fibrosis and tissue repair promotion, etc.

[0004] In view of the above problems, in recent years, researchers have gradually combined biomaterial science and uterine tissue engineering, and tried to develop an injectable hydrogel system with therapeutic function for postoperative intrauterine local drug release, microenvironment improvement and endometrial regeneration. At present, there have been reports that hyaluronic acid (HA), gelatin methacrylate (GelMA), polylactic acid-glycolic acid (PLGA) and other biomaterials are used to construct hydrogels, and load pro-repair factors (such as SDF-1α, bFGF, VEGF, etc.) or anti-inflammatory and antioxidant drugs (such as curcumin, resveratrol) for IUA prevention and treatment. However, the existing material system still has many limitations in functional integration, stimulus responsiveness and treatment effect, etc.

[0005] Specifically, traditional anti-adhesion hydrogels are mostly inert materials, only have physical barrier function, and cannot actively adjust the postoperative high ROS microenvironment, resulting in limited treatment effect. Although some hydrogels introduce anti-inflammatory or repair factors, the release mode is mostly passive diffusion type, which cannot respond to endogenous pathological signals, the release time is difficult to control, and the drug effect maintenance time is short. Especially under the combined action of postoperative inflammation and oxidative stress, the factors are prone to degradation and inactivation, the bioavailability is low, and the treatment effect fluctuates. In addition, the functional materials currently used for IUA treatment are mostly not effectively coupled with anti-fibrosis mechanism, and it is difficult to simultaneously inhibit fibroblast activation and regulate endometrial regeneration. Most of the materials have simple structure and single function, and it is difficult to meet the synergistic needs of "anti-oxidation + anti-inflammation + anti-fibrosis + repair promotion" in the complex microenvironment of the uterine cavity. Therefore, it is urgent to develop a new multifunctional hydrogel system with ROS response characteristics, integrated biological function, injectable gelation and suitable for local treatment of the uterine cavity, which can intelligently release drugs in the postoperative oxidative stress environment and realize dynamic regulation of the whole process of uterine cavity repair, thereby effectively preventing the occurrence and recurrence of uterine adhesion and promoting the clinical transformation and application of gynecological regenerative medicine materials. SUMMARY

[0006] In view of the problems of single treatment mechanism, lack of responsiveness, uncontrollable drug release, insufficient anti-fibrosis effect and limited tissue repair efficiency of the existing materials for postoperative uterine adhesion prevention and treatment, the present application provides an injectable hydrogel, which comprises silk fibroin grafted dopamine, amino hyaluronic acid and nanomicelles; the nanomicelles are mainly composed of monomethoxy polyethylene glycol, 1,3-dioxo-2-ketone and ring-opening monomer containing diselenium structure; and the nanomicelles load drugs and proteins.

[0007] In one embodiment, the silk fibroin grafted dopamine comprises methacrylsilk fibroin grafted dopamine (SerMA-DA); the preparation raw materials of the nanomicelles comprise monomethoxy polyethylene glycol, 1,3-dioxo-2-ketone and ring-opening monomer containing diselenium structure; the drugs comprise curcumin (Curcumin); and the proteins comprise hepatocyte growth factor (HGF).

[0008] The second aspect of the present application further provides a preparation method of the above-mentioned injectable hydrogel, which comprises the following steps: Preparation of silk fibroin grafted dopamine (Ser-DA): dissolve silk fibroin, adjust pH, activate, and obtain a silk fibroin solution; dissolve hydrochloric acid dopamine, adjust pH, deoxidize, and obtain a dopamine solution; mix the silk fibroin solution and the dopamine solution, adjust pH, dialysis, freeze-drying to obtain silk fibroin grafted dopamine; dissolve the freeze-dried product, add methacrylic anhydride, adjust pH, stir, dialysis, centrifugation, and freeze-drying to obtain silk fibroin grafted dopamine; Preparation of amino hyaluronic acid (AHA): dissolve hyaluronic acid sodium (HA) and adipic dihydrazide (ADH) to obtain a mixed solution, dissolve 1-(3-dimethylaminopropyl)-3-ethyl carbodiimide hydrochloride (EDC) and 1-hydroxybenzotriazole hydrate (HOBT), add to the mixed solution of hyaluronic acid sodium and adipic dihydrazide, adjust pH, react, add ethanol dropwise to form a precipitate, dissolve the precipitate, dialyze, freeze-dry to obtain amino hyaluronic acid; Preparation of drug and protein loaded nanomicelles: dissolve nanomicelles, add drugs, ultrasonic, drop into water under stirring, continue stirring, self-assemble the nanomicelles and load the drugs, dialyze to obtain drug loaded nanomicelles; dissolve the protein, mix with the drug loaded nanomicelle solution, incubate at 3-5°C to obtain drug and protein loaded nanomicelles; Preparation of injectable hydrogel: dissolve silk fibroin grafted dopamine to obtain a silk fibroin grafted dopamine solution, dissolve amino hyaluronic acid to obtain an amino hyaluronic acid solution, dissolve drug and protein loaded nanomicelles in the amino hyaluronic acid solution, and then mix with the silk fibroin grafted dopamine solution, mix well and stand to obtain an injectable hydrogel.

[0009] In one embodiment, the preparation method of the nanomicelles comprises the following steps: dissolve monomethoxy polyethylene glycol (mPEG-OH) in an organic solvent, add 1,3-dioxo-2-ketone (TMC) and a ring-opening monomer containing a diselenium structure (MSeSe), ice-bath stirring, add a catalyst, continue stirring at room temperature, add glacial acetic acid to terminate the reaction, drop the mixture into anhydrous ether for precipitation, vacuum dry to obtain a solid polymer (mPEG-bP(TMC-co-MSeSe)); dissolve the solid polymer in DMSO, drop into water and stir, dialyze with a dialysis bag to obtain nanomicelles (SeNPs); In the preparation of the nanomicelles, the mass ratio of monomethoxy polyethylene glycol: 1,3-dioxo-2-ketone: ring-opening monomer containing a diselenium structure is 1:(0.3-0.6):(0.1-0.5).

[0010] In one embodiment, the organic solvent comprises anhydrous dichloromethane (DCM), and the catalyst comprises 1,8-bisdimethylaminonaphthylidine (DBU).

[0011] In one embodiment, the molecular weight of the methoxy polyethylene glycol is about 2000 Da.

[0012] In one embodiment, the stirring time at room temperature is 22-26 h.

[0013] In one embodiment, the dropwise addition into water and stirring is at a rate of 0.5-2 mL / min, and the stirring time is 1-3 h.

[0014] In one embodiment, the molecular weight cutoff of the dialysis bag is 3400~3600 Da.

[0015] In one embodiment, the dialysis time is 46-50 hours.

[0016] In one embodiment, the preparation of the sericin-grafted dopamine includes the following steps: taking sericin raw material to prepare sericin, dissolving it in buffer solution, adjusting the pH, adding 4-(4,6-dimethoxytriazine-2-yl)-4-methylmorpholine hydrochloride (DMTMM) and stirring to activate it, taking dopamine hydrochloride to dissolve in sodium hyaluronate solution, adjusting the pH, removing oxygen and protecting with nitrogen, stirring the reaction at room temperature and adjusting the pH, dialyzing with a dialysis bag and then freeze-drying; taking the freeze-dried product to dissolve in water, adding methacrylic anhydride dropwise, adjusting the pH and stirring the reaction, dialyzing with a dialysis bag, centrifuging, and freeze-drying to obtain sericin-grafted dopamine; In the preparation of the sericin grafted with dopamine, the mass ratio of sericin to dopamine hydrochloride is 1:(0.1~0.4). Sericin grafted with dopamine: methacrylic anhydride 1g: (0.5~0.8mL).

[0017] In one embodiment, the buffer solution is 2-morpholinoethanesulfonic acid (MES) buffer.

[0018] In one embodiment, the pH is adjusted to 4.5-6.5.

[0019] In one embodiment, the stirring activation time is 25-35 minutes.

[0020] In one embodiment, the room temperature stirring reaction time is 22-26 hours.

[0021] In one embodiment, the molecular weight cutoff of the dialysis bag is 7000~9000 Da.

[0022] In one embodiment, the pH of the pH-adjusted and stirred reaction is 8.5 to 9.5.

[0023] In one embodiment, the centrifugation time is 8 to 15 minutes.

[0024] In one embodiment, the preparation of the amino-modified hyaluronic acid includes the following steps: dissolving sodium hyaluronate and oxaloyl in water and stirring until homogeneous; dissolving 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and 1-hydroxybenzotriazole hydrate in a mixed solvent of dimethyl sulfoxide and water, and adding them dropwise to the mixed solution of sodium hyaluronate and oxaloyl; adjusting the pH; reacting at room temperature; after the reaction is complete, adding ethanol dropwise to form a precipitate; dissolving the precipitate in water; dialyzing; and lyophilizing to obtain amino-modified hyaluronic acid. In the preparation of the amino-modified hyaluronic acid, the mass ratio of sodium hyaluronate to oxaloyl diol is 1:(18~22).

[0025] In one embodiment, the mixed solvent of dimethyl sulfoxide and water has a volume ratio of dimethyl sulfoxide:water of 1:1.

[0026] In one embodiment, the pH is adjusted to 4.5-5.5.

[0027] In one embodiment, the room temperature reaction time is 0.5 to 1.5 days.

[0028] In one embodiment, the dialysis time is 2 to 6 days.

[0029] In one embodiment, the preparation of the drug- and protein-loaded nanomicelles includes the following steps: dissolving nanomicelles in anhydrous dimethyl sulfoxide (DMSO), adding the drug, and sonicating to obtain a mixture; stirring the mixture and adding it dropwise to water while continuously stirring; dialyzing with a dialysis bag to obtain drug-loaded nanomicelles; dissolving the protein in a buffer solution, mixing it with the drug-loaded nanomicelle solution, and incubating at 3-5°C to obtain drug- and protein-loaded nanomicelles. In the preparation of the drug- and protein-loaded nanomicelles, the mass ratio of nanomicelles to drug is (18~22):1. According to the volume ratio, the ratio of drug-loaded nanomicelles to protein is (1~2):(1~2).

[0030] In one embodiment, the ultrasound duration is 8 to 15 minutes.

[0031] In one embodiment, the dripping rate to water is 0.5~2 mL / min.

[0032] In one embodiment, the continuous stirring time is 1 to 3 hours.

[0033] In one embodiment, the molecular weight cutoff of the dialysis bag is 3000~4000 Da.

[0034] In one embodiment, the buffer solution is a phosphate-buffered saline (PBS).

[0035] In one embodiment, the incubation time is 10-15 hours.

[0036] In one embodiment, the injectable hydrogel is prepared by mixing a dopamine-grafted silk fibroin solution and an aminolated hyaluronic acid solution containing drug- and protein-loaded nanomicelles in a volume ratio of 1:1.

[0037] The third aspect of the present invention also provides the use of the above-described injectable hydrogel, or the injectable hydrogel obtained by the above preparation method, in the preparation of products for postoperative intrauterine adhesions.

[0038] Compared with the prior art, the present invention has the following beneficial effects: 1. Intelligent response release under postoperative intrauterine pathological environment: The nanomicelles constructed in this invention contain diselenide bond structure, which can be highly sensitive to postoperative local excess ROS. When the level of oxidative stress increases, depolymerization occurs, realizing the on-demand release of curcumin and HGF, which is significantly different from the non-targeted release mode of traditional inert materials.

[0039] 2. A multi-target synergistic treatment mechanism of "antioxidant-anti-inflammatory-antifibrotic-promoting repair" has been established: This invention combines the Nrf2 signaling agonist curcumin with HGF, which has the functions of promoting repair and anti-fibrotic action, to break the limitation of the single factor mechanism of action and form a whole-chain treatment strategy from etiological intervention to tissue reconstruction, thereby improving the systematicness and effectiveness of treatment.

[0040] 3. An injectable hydrogel platform highly adaptable to intrauterine applications has been constructed: The matrix materials SerMA-DA and AHA have good injectability, in-situ gelation and tissue adhesion. They can be rapidly gelled in the intrauterine cavity through minimally invasive methods to form a stable local treatment microenvironment, avoiding the problems of complex implantation process and poor conformability of traditional stent materials.

[0041] 4. Enhanced biocompatibility and biodegradability, with good in vivo adaptability: All components are derived from natural or conventionally modified materials, possessing excellent biocompatibility and degradation characteristics, without causing significant toxic reactions or immune rejection, meeting the safety requirements for long-term local use after gynecological surgery.

[0042] 5. It combines therapeutic and physical barrier functions, significantly inhibiting the formation of intrauterine adhesions: After gel formation, it can form a continuous coverage on the surface of intrauterine cavity damage, effectively preventing endometrial adhesion. At the same time, through its antioxidant and tissue repair functions, it inhibits the adhesion mechanism from the root, and has a stronger clinical anti-adhesion effect than single barrier hydrogels.

[0043] 6. Possesses good platform scalability and industrial transformation potential: The responsive micelle and hydrogel system used in this invention has good versatility and can be adapted to different bioactive factors or drugs to expand the treatment of other inflammatory-fibrotic diseases (such as abdominal adhesions, postoperative intestinal adhesions, etc.), and has good technical promotion value. Attached Figure Description

[0044] Figure 1 TEM images of C@HGF@SeNPs; Figure 2 ROS response characteristics of C@HGF@SeNPs; Left: TEM image of C@HGF@SeNPs; Middle: TEM image of C@HGF@SeNPs after treatment with 0.1M H2O2; Right: TEM image of C@HGF@SeNPs after treatment with 1.0M H2O2. Figure 3 This is the NMR spectrum of SerMA-DA. Detailed Implementation

[0045] This invention designs an injectable multifunctional hydrogel system with multi-level ROS response regulation capabilities for the comprehensive prevention and treatment of postoperative intrauterine adhesions. The hydrogel uses dopamine-modified methacrylamide sericin (SerMA-DA) and amino-modified sodium hyaluronate (AHA) as matrix materials, exhibiting excellent biocompatibility, tissue adhesion, and in-situ gelling properties, constructing a three-dimensional support structure suitable for the postoperative microenvironment of the intrauterine cavity. The hydrogel system incorporates self-assembled ROS-responsive nanomicelles synthesized based on mPEG-bP (TMC-co-MSeSe). These micelles, with diselenoside structures as the core response units, possess excellent endogenous ROS recognition and response release characteristics, and are used to synergistically load the natural antioxidant small molecule curcumin and the tissue repair factor hepatocyte growth factor (HGF).

[0046] In this system, curcumin, as an agonist of the Nrf2 signaling pathway, can be released via micelle response under oxidative stress, activating the expression of downstream antioxidant proteins (such as HO-1 and NQO1), regulating local oxygen balance, and inhibiting ROS-induced inflammation and fibrosis. HGF, as a bioactive protein, stimulates the migration and differentiation of endometrial stem cells, repairs the functional endometrium in damaged areas, and synergistically inhibits pro-fibrotic signals such as TGF-β / Smad, promoting functional reconstruction of uterine tissue. The constructed micelle-hydrogel composite system, when locally injected into the postoperative uterine cavity, enables on-demand drug release under dynamic ROS signal regulation, creating a microenvironment synergistically promoting antioxidation, anti-inflammation, and tissue regeneration.

[0047] This system integrates the physical barrier function of injectable hydrogels, the dynamic release capability of ROS-responsive micelles, and the therapeutic functions of multi-pathway biological factors, constructing a highly integrated structure-mechanism-function intelligent therapeutic platform. This research provides a novel intelligent hydrogel strategy with a clear mechanism and excellent performance for the treatment of post-hysteroscopic trauma, showing promising clinical application prospects and translational potential.

[0048] The synergistic construction of a bioresponsive, synergistically repairing nanohydrogel system by combining ROS-responsive nanocurcumin, recombinant HGF protein, and injectable adhesive hydrogel is not directly disclosed or inspired in existing literature, and its integration faces the following key technical challenges: 1. The components have significantly different physicochemical properties, making stable co-loading difficult. Curcumin is a hydrophobic small molecule, while HGF is a large hydrophilic protein. The primary challenge overcome by this invention is ensuring their stable co-loading within the same system without mutual inactivation. This invention utilizes ROS-responsive shell-modified SeNPs as nanocarriers, leveraging their high surface area and ability to modify with multiple hydroxyl groups. Curcumin is first encapsulated via hydrophobic interactions, followed by HGF loading via electrostatic / covalent interactions, and then encapsulated within a dynamically covalently cross-linked hydrogel based on dopamine and hyaluronic acid, achieving both structural stability and maintenance of biological function.

[0049] 2. Hydrogel platforms need to possess three key properties: responsiveness, adhesion, and biocompatibility, resulting in high design complexity. This invention not only introduces dopamine groups to enhance tissue adhesion, but also combines oxidized dextran with hyaluronic acid-phenylboronic acid modification to achieve pH / ROS-responsive crosslinking, ensuring that the hydrogel can stably gel in the uterine cavity microenvironment and gradually release drugs, taking into account adhesion, injectability and responsiveness, and significantly raising the technical threshold for system construction.

[0050] To facilitate understanding of the present invention, a more complete description will be given below with reference to the accompanying drawings. Preferred embodiments of the invention are shown in the drawings. However, the invention can be implemented in many different forms and is not limited to the embodiments described herein. Rather, these embodiments are provided to provide a thorough and complete understanding of the disclosure of the invention.

[0051] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. The terminology used herein in the description of the invention is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. The term "and / or" as used herein includes any and all combinations of one or more of the associated listed items.

[0052] Example I. This invention provides a method for preparing an injectable hydrogel with antioxidant, anti-inflammatory, anti-fibrotic, and repair-promoting properties, comprising the following steps: 1. Synthesis of Methacryloylsericin Grafted Dopamine (SerMA-DA) Remove silkworm pupae and impurities from silkworm cocoons, cut them into small pieces, weigh out 30g, prepare 1L of 0.05M sodium carbonate solution, put the silkworm cocoons into the sodium carbonate solution, boil for 30 minutes, and collect the yellow liquid after boiling, which is sericin. Centrifuge the sericin solution at 8000rpm for 10 minutes, collect the supernatant, dialyze it in water for 2-3 days using a dialysis bag with a molecular weight cutoff of 8000Da, and freeze-dry the dialyzed sericin solution.

[0053] 2.0 g of sericin was dissolved in 300 mL of 0.1 M MES buffer, and the pH was adjusted to 5-6. 1.4 g of 4-(4,6-dimethoxytriazine-2-yl)-4-methylmorpholine hydrochloride (DMTMM) was added to the sericin solution, and the mixture was stirred and activated for 30 minutes. Subsequently, 0.474 g of dopamine hydrochloride was added to a sodium hyaluronate solution, and the pH was adjusted to 5-6. The mixture was then deoxygenated and protected with nitrogen. The reaction was carried out at room temperature with stirring for 24 h, maintaining the pH between 5 and 6. The solution was then dialyzed under acidic conditions using a dialysis bag with a molecular weight cutoff of 8000 Da, and finally freeze-dried at -80 °C to obtain Ser-DA.

[0054] The lyophilized Ser-DA was redissolved in deionized water to prepare a 5% solution. Methacrylic anhydride was then added dropwise, with 0.67 mL of methacrylic anhydride added for every gram of Ser-DA. The pH was adjusted to a stable value of pH 9 using a 10M sodium hydroxide solution. The pH will gradually decrease during the reaction; therefore, pH measurements and the addition of sodium hydroxide solution are necessary to control the pH. The reaction mixture was stirred at room temperature for 4 hours (excessive stirring time will cause precipitation). The reaction solution was collected and dialyzed against deionized water for 3 days using a dialysis bag with a molecular weight cutoff of 8000 Da. The liquid was collected and centrifuged at 8000 rpm for 10 minutes to remove impurities. The supernatant was collected. The supernatant was lyophilized to obtain SerMA-DA.

[0055] 2. Synthesis of Aminated Hyaluronic Acid (AHA) Weigh 0.5g of sodium hyaluronate (HA) and 10g of oxaloyl diacidyl (ADH), dissolve them in 100mL of deionized water, and stir until homogeneous. Weigh 0.8g of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC) and 0.7g of 1-hydroxybenzotriazole hydrate (HOBT), dissolve them in a 5mL / 5mL mixture of dimethyl sulfoxide and water (v / v = 1:1). Add the mixture dropwise to the above solution, adjust the pH to 5.0 with dilute hydrochloric acid, and react at room temperature for 1 day. After the reaction is complete, add ethanol dropwise to form a precipitate. Dissolve the precipitate in water, dialyze the solution in pure water for 3-5 days, and freeze-dry to obtain amino-modified hyaluronic acid, denoted as AHA.

[0056] 3. Synthesis of mPEG-bP (TMC-co-MSeSe) and preparation of self-assembled ROS-responsive nanomicelles (SeNPs) mPEG-bP (TMC-co-MSeSe) block copolymers were synthesized via ring-opening polymerization to construct ROS-responsive nanomicelles. The specific method is as follows: 1.0 g (0.5 mmol) of monomethoxy polyethylene glycol (mPEG-OH, molecular weight approximately 2000 Da) was dissolved in 10 mL of anhydrous dichloromethane (DCM). Then, 1,3-dioxo-2-one (TMC, 0.45 g, 4.4 mmol) and the diselenoside ring-opening monomer MSeSe (0.30 g, 1.2 mmol) were added sequentially, and the mixture was stirred in an ice bath until fully dissolved. Subsequently, 1,8-bis(dimethylaminonaphthidine) (DBU, 10 μL) was added as a catalyst under nitrogen protection, and the reaction was continuously stirred at room temperature for 24 h to initiate ring-opening polymerization. After the reaction was completed, a small amount of glacial acetic acid was added to terminate the reaction. The mixture was then dropped into 10 times its volume of ice-cold anhydrous diethyl ether for precipitation. After repeating the precipitation twice, the mixture was dried under vacuum to obtain a pale yellow solid polymer mPEG-bP (TMC-co-MSeSe).

[0057] The self-assembly of the nanomicelles was performed using dialysis: 20 mg of the obtained polymer was dissolved in 1 mL of anhydrous DMSO and slowly added dropwise to 20 mL of deionized water (dropping rate 1 mL / min). The mixture was magnetically stirred for 2 h to form stable core-shell micelles. The resulting solution was placed in a dialysis bag with a molecular weight cutoff of 3500 Da and dialyzed in deionized water for 48 h to remove organic solvents, finally yielding a micelle solution with uniform particle size.

[0058] 4. Preparation of ROS-responsive nanomicelles synergistically loaded with curcumin and HGF (C@HGF@SeNPs) To construct ROS-responsive nanomicelles simultaneously loaded with curcumin and hepatocyte growth factor (HGF), a block copolymer mPEG-bP (TMC-co-MSeSe) containing a diselenoside structure was first synthesized and formed into core-shell micelles via self-assembly. Subsequently, a strategy of "co-encapsulation + adsorption binding" was employed to achieve the synergistic encapsulation of hydrophobic small molecules and hydrophilic protein factors. The specific method is as follows: 20 mg of the synthesized mPEG-bP (TMC-co-MSeSe) polymer was weighed and dissolved in 1 mL of anhydrous dimethyl sulfoxide (DMSO). 1 mg of curcumin was added (final concentration 1 mg / mL), and the solution was sonicated at room temperature for 10 min to obtain a homogeneous and transparent solution. This solution was then slowly added dropwise to 20 mL of deionized water under magnetic stirring (dropping rate approximately 1 mL / min), with continuous stirring for 2 h to promote micelle self-assembly and curcumin encapsulation. Subsequently, the obtained micelle solution was placed into a dialysis bag with a molecular weight cutoff of 3500 Da and dialyzed in deionized water for 48 h to remove organic solvents and unloaded drugs, obtaining curcumin-loaded micelles (C@SeNPs) for later use. After curcumin micelle formation, HGF protein loading was performed. Recombinant human HGF protein (final concentration approximately 100~200 ng / mL) was weighed, dissolved in pre-cooled PBS buffer (pH=7.4), and mixed with the C@SeNPs micelle solution at a 1:1 volume ratio. The mixture was slowly shaken and incubated at 4°C for 12 h, allowing HGF to bind to the PEG shell on the micelle surface through electrostatic adsorption and hydrophobic interactions, thus constructing the C@HGF@SeNPs composite micelle system.

[0059] 5. Preparation of injectable hydrogels (SA / C@HGF@SeNPs) Weigh out SerMA-DA and dissolve it in 2 mL of deionized water to prepare a SerMA-DA solution. At the same time, dissolve AHA in 2 mL of deionized water to prepare a 10% AHA solution. Then add C@HGF@SeNPs to the above solution and mix them at a volume ratio of SerMA-DA:AHA of 1:1. After vortexing and mixing, allow it to stand to obtain SA / C@HGF@SeNPs hydrogel.

[0060] Figure 1 TEM images of C@HGF@SeNPs nanomicelles are shown. The images reveal that the nanomicelles exhibit a near-spherical or irregularly shaped structure with clear particle boundaries, uniform morphology, and particle size less than 500 nm, indicating that the prepared C@HGF@SeNPs possess good morphological stability and dispersibility.

[0061] Figure 2TEM images of C@HGF@SeNPs nanomicelles under different H2O2 concentrations are presented, visually reflecting their responsiveness to oxidative ROS environments. The left image shows a TEM image of C@HGF@SeNPs without H2O2 treatment; the nanoparticles exhibit intact edges, regular structures, and spherical or polyhedral morphologies with uniform distribution, indicating good system stability. The right image shows a TEM image of C@HGF@SeNPs after treatment with 0.1M H2O2; some edge dissolution and slight structural breakage are observed, indicating preliminary ROS-sensitive degradation behavior. The right image shows a TEM image after treatment with 1.0M H2O2; severely fragmented particle structures and increased aggregation are observed, indicating significant degradation of C@HGF@SeNPs under high-concentration ROS stimulation, demonstrating good ROS-responsive release capabilities. This figure further verifies that the nanomicelle system constructed in this invention has good oxidative stimulation responsiveness, enabling drug release regulation in pathological ROS microenvironments, providing a material basis for specific treatment of disease microenvironments.

[0062] Ser was modified to obtain SerMA-DA. The first step was to graft DA groups, followed by grafting MA groups. Figure 3 The NMR spectrum of Ser-modified Ser shows that, compared to Ser, Ser-DA exhibits a new absorption peak at 7.18 ppm, corresponding to the proton peak on the dopamine benzene ring, indicating successful grafting of the DA group onto Ser. Subsequently, grafting the MA group onto Ser-DA reveals two new absorption peaks at 5.96 and 5.57 ppm, corresponding to the proton peaks of the double bond on the methacrylate, indicating successful grafting of the MA group onto Ser-DA to obtain SerMA-DA.

[0063] The injectable hydrogel with antioxidant, anti-inflammatory, anti-fibrotic, and repair-promoting properties prepared in the embodiments of the present invention, as well as the comparative examples, contain the following components: 20%~40% SerMA-DA, 10% AHA, and 50~200 μg / mC@HGF@SeNPs. The specific formulation is shown in Table 1.

[0064] Table 1. Formulation Mass Fraction Table

[0065] The hydrogel preparation method is as follows: Examples 1-7 Weigh out the corresponding mass of SerMA-DA according to the predetermined concentration and dissolve it in 2 mL of deionized water to prepare SerMA-DA solution. At the same time, dissolve AHA in 2 mL of deionized water to prepare 10% AHA solution. Then, add an appropriate amount of C@HGF@SeNPs to the above solution and mix them at a volume ratio of SerMA-DA:AHA of 1:1. After vortexing and mixing, allow it to stand to obtain SA / C@HGF@SeNPs hydrogel.

[0066] Comparative Examples 1 and 3 Weigh out the corresponding mass of SerMA-DA according to the predetermined concentration and dissolve it in 2 mL of deionized water to prepare SerMA-DA solution. At the same time, dissolve AHA in 2 mL of deionized water to prepare 10% AHA solution. Mix them at a volume ratio of SerMA-DA:AHA of 1:1, vortex to mix, and let stand to obtain SA hydrogel.

[0067] Comparative Example 2 Weigh out the corresponding mass of SerMA-DA according to the predetermined concentration and dissolve it in 2 mL of deionized water to prepare SerMA-DA solutions for subsequent use.

[0068] Comparative Example 4 Dissolve a certain amount of AHA in 2 mL of deionized water to prepare a 10% AHA solution for later use.

[0069] Implementation effect evaluation 1. Evaluation of gelation time Test method: To evaluate the gel-forming properties of the hydrogels from different examples and comparative examples, the gel-forming time was tested using the pouring method. The specific procedure was as follows: First, SerMA-DA, AHA, and C@HGF@SeNPs (at the concentrations set in Examples 1–7 and Comparative Examples 1–4) were weighed according to the respective ratios and dissolved in PBS buffer. After thorough mixing, the precursor solutions were prepared. Then, the solutions were poured into vials at different time points, and the gel-forming time was observed. The test results are shown in Table 2.

[0070] Table 2. Gel formation time of injectable hydrogels

[0071] Results (Comparative Examples 2 and 4 were liquids and could not be gelled): Table 2 shows that all examples can complete cross-linking and gelation within 90 seconds, demonstrating good rapid in-situ molding capability and meeting the time-sensitive requirements for postoperative intrauterine injection gelation. Among them, Examples 3 (30s) and 6 (32s) had the shortest gelation times, indicating that this formulation can achieve rapid three-dimensional network construction. SerMA-DA concentration has a significant impact on gelation time: at the same C@HGF@SeNPs concentration (50μg / mL), as the SerMA-DA content increased from 20% (Example 1) to 40% (Example 3), the gelation time significantly decreased from 65s to 30s. This indicates that a higher double bond concentration can increase the cross-linking density and promote rapid curing. The loading of C@HGF@SeNPs has a relatively small effect on the gelation time: when the SerMA-DA content is fixed at 30%, the gelation time varies from 45 to 60 s when the micelle concentration is increased from 50 μg / mL (Example 2) to 200 μg / mL (Example 7), indicating that although the introduction of micelles slightly affects the crosslinking rate (possibly due to light shading or physical hindrance), it can still crosslink and gel rapidly in the whole.

[0072] 2. Rheological property characterization Test method: The storage modulus G′ and loss modulus G″ were dynamically tracked over time using a rotational rheometer. The test results are shown in Table 3.

[0073] Table 3 Rheological properties of injectable hydrogels

[0074] Results (Comparative Examples 2 and 4 were liquids and did not undergo rheological property testing): To evaluate the mechanical properties and structural stability of each group of injectable hydrogel systems, the storage modulus (G′) and loss modulus (G″) were obtained through dynamic rheological testing, as shown in Table 3. The results show that all examples exhibited a significantly larger G′ than G″, indicating that this type of hydrogel system is primarily supported by elasticity, forming a stable three-dimensional network structure. Specifically, the G′ values ​​of Examples 3, 4, and 5 reached 580 Pa, 540 Pa, and 550 Pa, respectively, significantly higher than the other groups, suggesting a higher degree of internal cross-linking and superior gel mechanical properties. In particular, Example 6, under the synergistic effect of higher concentrations of SerMA-DA (40%), AHA (10%), and C@HGF@SeNPs (200 μg / mL), exhibited the strongest network structure stability and the optimal storage modulus (G′ = 630 Pa), demonstrating ideal mechanical support performance. In contrast, the G′ values ​​of the comparative group were generally lower (260~350 Pa), and the difference between G′ and G″ was close, reflecting that they failed to form a dense and uniform gel network structure, resulting in poor mechanical support and insufficient stability.

[0075] 3. Characterization of antioxidant properties Test method: DPPH free radical scavenging experiment: Hydrogel samples from each group were lyophilized and ground into powder. 10 mg of each sample was placed in 1 mL of 0.1 mM DPPH ethanol solution and incubated at room temperature with shaking for 30 min under light-protected conditions. The absorbance (A) was then measured at 517 nm using a spectrophotometer. S ), and set up a blank control group (DPPH solution + deionized water, A0) and a control sample group (DPPH solution + blank hydrogel powder without active ingredients, A0). C The free radical scavenging rate is calculated using the following formula: DPPH scavenging rate (%) = (A0) / (A0) A S ) / (A0 A C ) × 100%.

[0076] Hydroxyl radical (·OH) scavenging experiment: The ·OH scavenging ability was detected using the Fenton reaction system. The reaction mixture was prepared by mixing 1 mL of 9 mM FeSO4, 1 mL of 9 mM phenol, 1 mL of 9 mM H2O2, and 1 mL of sample extract. After incubation at 37°C for 30 min, the absorbance was measured at 510 nm. S The blank control and comparative examples are the same as above. The clearance rate formula is as follows: ·OH clearance rate (%) = (A0) / (OH) * clearance rate (%) A S ) / (A0 A C ) × 100%.

[0077] Table 4 Antioxidant properties of injectable hydrogels

[0078] Results explanation: To systematically evaluate the scavenging ability of each hydrogel group against oxygen free radicals, DPPH free radical and hydroxyl radical (·OH) scavenging experiments were performed. The results are shown in Table 4: the example groups all showed significantly better antioxidant performance than the comparative groups, indicating that the synergistic introduction of curcumin (Cur) and HGF significantly improved the free radical scavenging ability of the materials. This suggests that HGF may further promote the activation of endogenous antioxidant systems (such as SOD, CAT, etc.) by activating the Nrf2 pathway, thereby enhancing the overall scavenging efficiency.

[0079] 4. Assessment of anti-fibrotic ability Test method: To evaluate the anti-fibrotic effects of the injectable hydrogels prepared in each example and comparative example on inhibiting fibroblast activation and collagen deposition in postoperative intrauterine adhesions, the mRNA expression levels of typical fibrosis-related genes α-smooth muscle actin (a-SMA) and type I collagen (COL1A1) were detected using real-time quantitative PCR (qPCR). In the experiment, mouse uterine fibroblasts (UFCs) were cultured in different hydrogel extracts for 48 hours, and then total RNA was extracted and cDNA was synthesized using a reverse transcription kit. The qPCR reaction volume was 20 μL, with GAPDH as the internal control gene, and amplification was performed using SYBR Green dye in an ABI QuantStudio 5 system. Each experiment was performed in triplicate, using 2... - The relative expression level was calculated using the ΔΔCt method and standardized using Comparative Example 1 as the baseline (expression level = 100%).

[0080] Table 5 Anti-fibrotic ability of injectable hydrogels

[0081] Results explanation: As shown in Table 5, the expression levels of the two key fibrosis indicators, α-SMA and COL1A1, differed significantly among the groups. Specifically, the gene expression in the Example group was significantly lower than that in the Control group, suggesting that this type of hydrogel has a significant inhibitory effect on fibroblast activation and type I collagen deposition. In contrast, the expression of both α-SMA and COL1A1 remained at high levels in the Control group, indicating weaker inhibitory ability under conditions without anti-fibrotic factor loading. Furthermore, Example 6 group showed the lowest expression levels in both indicators, indicating that the C@HGF@SeNPs composite nanomicelle system used in this group, synergistically with the optimal SerMA-DA / AHA ratio, can effectively intervene in fibroblast phenotypic transformation and inhibit abnormal extracellular matrix deposition, exhibiting the most significant anti-fibrotic potential. This result is consistent with previous ROS scavenging and antioxidant capacity test data, further confirming the comprehensive advantages of this injectable hydrogel system in regulating the postoperative intrauterine tissue repair microenvironment.

[0082] 5. Endometrial thickness assessment Test method: To evaluate the promoting effect of the constructed composite hydrogel on endometrial regeneration after intrauterine injury, an animal experiment was conducted using a mechanical intrauterine adhesion model. The specific methods are as follows: Healthy female SD rats (weighing 220-250g) were selected. Under anesthesia, the uterine horns were exposed through a midline abdominal incision, and the basal layer of the endometrium was gently scraped away using a small curette to establish an intrauterine injury model. Immediately after the operation, different groups of hydrogels were injected into the uterine cavity, ensuring complete filling and closure of the incision. After 14 days of intervention, the animals were sacrificed, and the uterine tissue was completely removed and fixed with 4% paraformaldehyde for 24 hours. Subsequently, after routine dehydration, paraffin embedding, sectioning (5μm thickness), and hematoxylin-eosin (HE) staining were performed. Imaging was performed under an optical microscope (×100 magnification), and the endometrial thickness (from the basal layer to the endometrial surface) was measured using ImageJ software on three sections selected from each animal and three different fields of view within each section. The test results are shown in Table 6.

[0083] Table 6. Evaluation of the effect of injectable hydrogels on promoting endometrial regeneration

[0084] Results explanation: Quantitative measurements using HE staining showed that the endometrial thickness in Comparative Example 1 (SerMA-DA 20%, AHA 10%, no drug) was only 101.4 ± 13.5 μm, while the endometrial thickness in Example 5 (SerMA-DA 30%, AHA 10%, C@HGF@SeNPs 200 μg / mL) reached 228.7 ± 11.6 μm, significantly higher than that in Comparative Example 1. Simultaneously, the drug concentration showed a dose-dependent relationship; gradient groups of 50 μg / mL, 100 μg / mL, and 200 μg / mL all showed an increasing trend in thickness with increasing concentration. Different SerMA-DA mass fractions (20%, 30%, 40%) had little effect on endometrial thickness at the same drug concentration, suggesting that the main mechanism of action originates from the effective release of C@HGF@SeNPs. These experimental results demonstrate that the composite hydrogel constructed in this invention has good endometrial regeneration-promoting ability in the intrauterine injury model, effectively restoring the integrity of the endometrial structure and providing an effective intervention strategy for tissue reconstruction after intrauterine adhesions.

[0085] 6. Adhesion score (AFS score) Test method: To systematically evaluate the efficacy of different hydrogel groups in preventing intrauterine adhesions, the American Fertility Society (AFS) scoring system was used to grade and assess the degree of uterine adhesions in rats. In the experiment, a mechanical injury model of the uterine cavity was established in female SD rats (weighing 220-250g). Different groups of hydrogel materials were injected into the uterine cavity. After 14 days of postoperative observation, the animals were sacrificed, and uterine tissue was completely dissected. The degree of closure of the uterine cavity, the extent of adhesions, and the density of adhesions were observed and recorded macroscopically. Subsequently, the tissues were fixed, dehydrated, embedded, and sectioned. After HE staining, the nature and degree of adhesions were determined under a microscope. According to the AFS scoring criteria, the adhesions were scored from three dimensions—the extent, type, and menstrual status (simulating tissue recovery). The comprehensive score was used to determine the severity of adhesions (total score range: 1-12 points, with higher scores indicating more severe adhesions). The test results are shown in Table 7.

[0086] Table 7 Evaluation of the effect of injectable hydrogels in inhibiting adhesion formation

[0087] Results explanation: To evaluate the efficacy of injectable hydrogels in inhibiting intrauterine adhesions, rat models were histologically assessed and quantified using the AFS scoring system (total score 1-12). Comparative examples 1-3 all scored between 9.2 and 10.9, indicating that the matrix alone was insufficient to prevent adhesion formation, and the adhesions were severe. In contrast, the experimental groups with the C@HGF@SeNPs complex, especially Example 5 (200 μg / mL), scored 1.5±0.3, Example 6 1.8±0.4, and Example 7 2.3±0.5, significantly better than other groups, achieving very mild or near-no adhesion levels, suggesting a synergistic advantage of the drug system in tissue repair and adhesion inhibition. Example 5 exhibited the best anti-adhesion effect, with a score significantly lower than all control groups and the low-dose experimental groups (e.g., Example 1 score 5.8±0.6), which was statistically significant. In summary, the results indicate that the constructed C@HGF@SeNPs drug-based hydrogel system can significantly inhibit the formation of intrauterine adhesions after surgery. The effect is dose-dependent and reaches its optimal value at 200 μg / mL, demonstrating excellent anti-adhesion potential and promising prospects for commercialization.

[0088] This invention provides a multifunctional, injectable, ROS-responsive intelligent hydrogel system for postoperative local treatment of the uterine cavity. By precisely controlling oxidative stress, inflammatory response, and fibrosis processes, and synergistically promoting endometrial regeneration, it effectively prevents or reduces the occurrence and recurrence of intrauterine adhesions. The preferred concentrations are: SerMA-DA 30%; AHA 10%; and C@HGF@SeNPs 200 μg / mL. The core technical problems solved by this invention mainly include: 1. How to achieve intelligent drug release in a postoperative high ROS pathological microenvironment: By constructing responsive nanomicelles containing diselenide structures, a highly sensitive response to reactive oxygen species (ROS) can be achieved, and drugs can be precisely released when oxidative stress levels rise, effectively clearing ROS and breaking the positive feedback loop of inflammation-fibrosis.

[0089] 2. How to achieve synergistic treatment of antioxidation, anti-inflammation and anti-fibrosis through multiple mechanisms: This invention loads curcumin, an Nrf2 pathway agonist, and HGF (hepatocyte growth factor), which has strong anti-fibrotic and repair-promoting effects, into ROS-responsive micelles to achieve synergistic treatment in the lesion microenvironment, while simultaneously regulating cellular oxidative homeostasis, inhibiting fibroblast activation and promoting endometrial stem cell repair.

[0090] 3. How to construct an injectable, biocompatible hydrogel carrier suitable for intrauterine application: By embedding the above-mentioned loaded micelles into a hydrogel system based on dopamine-modified silk fibroin (SerMA-DA) and aminohyaluronic acid sodium (AHA), good injectability, in vivo gelation and adhesion properties are obtained, which are suitable for local coverage and drug retention in complex uterine cavities.

[0091] 4. How to enhance the tissue compatibility and functional stability of the material: The material system of this invention, while ensuring good biocompatibility, has adjustable degradation, self-repair and adhesion capabilities, and can remain stably in the postoperative wound area for a long time, continuously exerting a therapeutic effect in the dynamic microenvironment, avoiding rapid drug clearance or functional inactivation.

[0092] This invention exhibits a significant synergistic enhancement effect at the functional mechanism level, exceeding the expected range of single-component effects, as detailed below: 1. Synergistic effect of curcumin and HGF: Experimental results show that curcumin significantly reduces oxidative stress by scavenging ROS, thereby protecting the structural integrity of HGF and prolonging its biological activity half-life; HGF promotes the proliferation and re-epithelialization of endometrial cells, which, together with the anti-inflammatory effect of curcumin, form a dual mechanism of "anti-inflammatory + repair promotion", achieving a synergistic effect of intrauterine environment reconstruction and fibrosis reversal.

[0093] 2. Response-release coupling effect of the material platform: Unlike the sustained release of traditional hydrogels, the chitosan nanomicelles constructed in this invention specifically release curcumin in the ROS microenvironment and synergistically release HGF-encapsulated in the form of HGF, achieving "lesion activation and targeted release", which greatly improves drug utilization and tissue targeting.

[0094] 3. Promotes epithelial integrity recovery and prevents recurrence: Combining the epithelial morphology induction of HGF with the anti-adhesion and antioxidant functions of curcumin, the hydrogel exhibits far superior results in animal models in terms of endometrial regeneration and decreased adhesion scores compared to single-component hydrogels.

[0095] In summary, the ROS-responsive composite hydrogel system provided by this invention integrates multiple advantages such as environmental response release, signal pathway regulation, synergistic effect of biological factors, and structural adaptability within a single platform. It provides a novel treatment strategy for the comprehensive prevention and treatment of postoperative intrauterine adhesions with a clear mechanism, significant effect, and adjustable structure, and has good application prospects and promotion value.

[0096] The technical features of the above embodiments can be combined in any way. For the sake of brevity, not all possible combinations of the technical features in the above embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0097] The embodiments described above are merely illustrative of several implementations of the present invention, and while the descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the invention patent. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of the present invention, and these all fall within the protection scope of the present invention. Therefore, the protection scope of this invention patent should be determined by the appended claims.

Claims

1. An injectable hydrogel, characterized in that, The injectable hydrogel comprises sericin grafted dopamine, amino hyaluronic acid and nanomicelle; The nanomicelle is mainly composed of monomethoxypolyethylene glycol, 1,3-dioxo-2-ketone and ring-opening monomer containing diselenium structure; The nanomicelle carries drugs and proteins.

2. The injectable hydrogel of claim 1, wherein, The sericin grafted dopamine comprises methacryl sericin grafted dopamine; The preparation raw materials of the nanomicelle comprise monomethoxypolyethylene glycol, 1,3-dioxo-2-ketone and ring-opening monomer containing diselenium structure; The drugs comprise curcumin; The proteins comprise hepatocyte growth factor.

3. The method of claim 1-2, wherein the injectable hydrogel is prepared by, The method comprises the following steps: Preparation of sericin grafted dopamine: dissolve sericin, adjust pH, activate, and obtain sericin solution; Dissolve dopamine hydrochloride, adjust pH, and remove oxygen to obtain dopamine solution; Mix the sericin solution and the dopamine solution, adjust pH, dialysis, and freeze-drying to obtain sericin grafted dopamine; dissolve the freeze-dried product, add methacrylic anhydride, adjust pH, stir, dialysis, centrifugation, and freeze-drying to obtain sericin grafted dopamine; Preparation of amino hyaluronic acid: dissolve sodium hyaluronate and oxalic acid diacyl, stir to obtain a mixed solution, dissolve 1-(3-dimethylaminopropyl)-3-ethyl carbodiimide hydrochloride and 1-hydroxybenzotriazole hydrate, add to the mixed solution of sodium hyaluronate and oxalic acid diacyl, adjust pH, react, add ethanol dropwise to form a precipitate, dissolve the precipitate, dialysis, and freeze-drying to obtain amino hyaluronic acid; Preparation of nanomicelle carrying drugs and proteins: dissolve nanomicelle, add drugs, ultrasonic, and drop into water under stirring, continue stirring, so that the nanomicelle self-assembles and carries the drugs, and dialysis to obtain nanomicelle carrying drugs; dissolve proteins, mix with the nanomicelle solution carrying drugs, incubate at 3-5℃ to obtain nanomicelle carrying drugs and proteins; Preparation of injectable hydrogel: dissolve sericin grafted dopamine to obtain sericin grafted dopamine solution, dissolve amino hyaluronic acid to obtain amino hyaluronic acid solution, dissolve nanomicelle carrying drugs and proteins in the amino hyaluronic acid solution, mix with the sericin grafted dopamine solution, mix evenly, and stand to obtain injectable hydrogel.

4. The production method according to claim 3, characterized by, The preparation method of the nanomicelle comprises the following steps: dissolve monomethoxypolyethylene glycol in an organic solvent, add 1,3-dioxo-2-ketone and ring-opening monomer containing diselenium structure, ice-bath stirring, add catalyst, continue stirring at room temperature, add glacial acetic acid to terminate the reaction, drop the mixture into anhydrous ether for precipitation, vacuum drying to obtain solid polymer; dissolve the solid polymer in DMSO, drop into water and stir, dialysis bag dialysis to obtain nanomicelle; In the preparation of the nanomicelle, the mass ratio of monomethoxypolyethylene glycol: 1,3-dioxo-2-ketone: ring-opening monomer containing diselenium structure is 1: (0.3-0.6): (0.1-0.5).

5. The production method according to claim 4, characterized by, The organic solvent comprises anhydrous dichloromethane, and the catalyst comprises 1,8-bisdimethylaminonaphthylidine.

6. The production method according to claim 3, characterized by, The preparation of the sericin grafted dopamine comprises the following steps: The silk fibroin protein is prepared from silk fibroin raw material, dissolved in buffer, pH is adjusted, 4-(4, 6-dimethoxytriazin-2-yl)-4-methylmorpholinium hydrochloride is added and stirred to activate, hydrochloric acid dopamine is dissolved in sodium hyaluronate solution, pH is adjusted, oxygen is removed and protected by nitrogen, the reaction is stirred at room temperature and pH is adjusted, the product is freeze-dried after dialysis in dialysis bag and dialysis; the freeze-dried product is dissolved in water, methacrylic anhydride is added dropwise, pH is adjusted and the reaction is stirred, dialysis is performed in a dialysis bag, centrifugation is performed and freeze-drying is performed to obtain silk fibroin grafted dopamine; In the preparation of the silk fibroin grafted dopamine, the mass ratio of silk fibroin to hydrochloric acid dopamine is 1: (0.1-0.4); The mass ratio of silk fibroin grafted dopamine to methacrylic anhydride is 1g: (0.5-0.8mL).

7. The production method according to claim 3, characterized by, The preparation of the aminated hyaluronic acid comprises the following steps: Sodium hyaluronate and oxalic acid diacyl are dissolved in water and stirred uniformly, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and 1-hydroxybenzotriazole hydrate are dissolved in a mixed solvent of dimethyl sulfoxide and water, and added dropwise to the mixed solution of sodium hyaluronate and oxalic acid diacyl, pH is adjusted, the reaction is performed at room temperature, ethanol is added dropwise to form a precipitate after the reaction is completed, the precipitate is dissolved in water, dialyzed and freeze-dried to obtain aminated hyaluronic acid; In the preparation of the aminated hyaluronic acid, the mass ratio of sodium hyaluronate to oxalic acid diacyl is 1: (18-22).

8. The preparation method according to claim 3, characterized in that, The preparation of the drug and protein loaded nanomicelles comprises the following steps: nanomicelles are dissolved in anhydrous dimethyl sulfoxide, a drug is added to obtain a mixed solution, the mixed solution is stirred and added dropwise to water, continuous stirring is performed, dialysis is performed in a dialysis bag to obtain drug loaded nanomicelles; a protein is dissolved in a buffer solution, mixed with the drug loaded nanomicelle solution, and incubated at 3-5°C to obtain drug and protein loaded nanomicelles; In the preparation of the drug and protein loaded nanomicelles, the mass ratio of nanomicelles to drugs is (18-22): 1; The volume ratio of drug loaded nanomicelles to proteins is (1-2): (1-2).

9. The production method according to claim 3, wherein In the preparation of the injectable hydrogel, the volume ratio of the silk fibroin grafted dopamine solution to the aminated hyaluronic acid solution in which the drug and protein loaded nanomicelles are dissolved is 1:

1.

10. The injectable hydrogel of claims 1-2 or the hydrogel prepared by the preparation method of claims 3-9 for use in the preparation of a product for postoperative intrauterine adhesion.

Images