Hydrogels and compositions and methods for forming hydrogels

Abstract

The invention relates to a hydrogel and a composition and method for forming the hydrogel, and belongs to the technical field of biomedical materials, the composition for forming the hydrogel comprises an inflammation targeting component and a dense cross-linking component, and the inflammation targeting component and the dense cross-linking component are kept separated from each other until being used, the inflammation targeting component has a targeting function and is microgel obtained after cross-linking reaction of polyvinyl alcohol, sodium alginate and calcium ions, the dense cross-linking component is a mixed solution containing cyclodextrin and / or a derivative thereof and a boron-containing compound, and the dense cross-linking component is used for cross-linking with the inflammation targeting component to form high-density hydrogel. By sequentially using the targeting function of the inflammation targeting component and the dense cross-linking function of the dense cross-linking component, the hydrogel with a targeted treatment effect and high compactness can be formed, so that the treatment effect of the hydrogel on inflammatory diseases is improved, and the hydrogel can be well applied to treatment of inflammatory bowel diseases, skin damage, operative wounds and other diseases.

Description

Technical Field

[0001] The present application relates to the technical field of biomedical materials, and in particular to a hydrogel and a composition and method for forming the hydrogel. Background Art

[0002] Inflammatory response can cause damage to the structure and function of the mucosa. The integrity of the mucosa is essential to health. The mucosa constitutes the first line of defense against the external environment, blocking the invasion of bacteria and other pathogens. A damaged mucosal barrier not only increases the risk of pathogen invasion, but may also lead to further aggravation of inflammation, forming a vicious cycle. Restoring the mucosal barrier damaged by inflammation is essential for maintaining the health of the body and immune balance.

[0003] Hydrogel is a class of high molecular weight polymers, and for mucosal injuries, hydrogel provides an innovative repair strategy. The three-dimensional network structure of hydrogel can mimic the natural extracellular matrix of the mucosa, providing an ideal scaffold for cell growth and regeneration of damaged tissues.

[0004] However, the therapeutic effect of existing hydrogels on inflammatory diseases still needs to be improved. Summary of the invention

[0005] The purpose of the present application is to provide a hydrogel and a composition and method for forming a hydrogel, so as to provide a hydrogel that can simultaneously achieve targeted therapeutic effects and high density, thereby improving the therapeutic effect of the hydrogel on inflammatory diseases.

[0006] The present application embodiment provides a composition for forming a hydrogel, which includes an inflammation targeting component and a dense cross-linking component, and the inflammation targeting component and the dense cross-linking component are kept separate from each other until use; wherein the inflammation targeting component has a targeting function and is a microgel obtained by cross-linking reaction of polyvinyl alcohol, sodium alginate and calcium ions; the dense cross-linking component is a mixed solution containing cyclodextrin and / or its derivatives and boron-containing compounds, and the dense cross-linking component is used to cross-link with the inflammation targeting component to form a highly dense hydrogel. By sequentially using the targeting function of the inflammation targeting component and the dense cross-linking function of the dense cross-linking component, a hydrogel with both targeted therapeutic effects and high density can be formed to improve the therapeutic effect of the hydrogel on inflammatory diseases (such as inflammatory bowel disease), and can be well applied in the treatment of inflammatory bowel disease, skin damage and surgical trauma.

[0007] Among them, the inflammation targeting component is specifically a microgel obtained by cross-linking reaction of polyvinyl alcohol solution, sodium alginate solution and calcium ion solution; wherein the mass fraction of polyvinyl alcohol in the polyvinyl alcohol solution is 5% to 20%, the mass fraction of sodium alginate in the sodium alginate solution is 1% to 10%, the mass fraction of calcium ions in the calcium ion solution is 5% to 30%, and the volume ratio of polyvinyl alcohol solution to sodium alginate solution is 2 / 1 to 8 / 1.

[0008] The molecular weight of polyvinyl alcohol is 47000-87000Da; the calcium ion is calcium chloride and / or its analogs, calcium chloride includes anhydrous calcium chloride and / or calcium chloride hydrate, and the analogs of calcium chloride are water-soluble calcium salts, including calcium bicarbonate and / or calcium dihydrogen phosphate.

[0009] Among them, the cyclodextrin includes at least one of α-cyclodextrin, β-cyclodextrin and γ-cyclodextrin; the derivatives of cyclodextrin include at least one of amino-modified cyclodextrin, carboxyl-modified cyclodextrin and thiol-modified cyclodextrin; the boron-containing compound includes at least one of boric acid, borax pentahydrate and anhydrous borax.

[0010] Wherein, in the dense cross-linking component, the mass ratio of cyclodextrin and / or its derivatives to the mass ratio of the boron-containing compound is 1 / 4 to 5 / 2.

[0011] An embodiment of the present application also provides a method for forming a hydrogel, which comprises: preparing an inflammation targeting component and a dense cross-linking component included in any of the above-mentioned compositions for forming a hydrogel; and cross-linking the inflammation targeting component and the dense cross-linking component to form a highly dense hydrogel.

[0012] Among them, the inflammation targeting component and the dense cross-linking component are cross-linked to form a highly dense hydrogel, including: sequentially using the targeting function of the inflammation targeting component and the dense cross-linking function of the dense cross-linking component to target the inflammation targeting component to the inflammatory site and then cross-link with the dense cross-linking component to form a hydrogel with both targeted therapeutic effect and high density.

[0013] Before targeting the inflammation targeting component to the inflammation site, the method further includes: loading the inflammation targeting component with a drug, wherein the drug includes at least one of a protein drug, an antibody drug, a nucleic acid drug, a chemical drug and a nano drug.

[0014] Among them, the steps of preparing the inflammation targeting component include: preparing polyvinyl alcohol solution, sodium alginate solution and calcium ion solution; after mixing the polyvinyl alcohol solution and the sodium alginate solution evenly, immersing them in a calcium ion solution for a cross-linking reaction, taking them out after the reaction is completed to obtain a gel; extruding the gel through a sieve to form a microgel with a particle size of micrometer level, and collecting the extruded microgel to obtain the inflammation targeting component.

[0015] The step of preparing the dense cross-linked component includes: adding anhydrous borax and β-cyclodextrin polymer into a solvent, then centrifuging, and taking the supernatant as the dense cross-linked component.

[0016] The embodiment of the present application also provides a hydrogel, which is formed by any of the above methods.

[0017] The present application provides a hydrogel and a composition and method for forming a hydrogel. The composition for forming a hydrogel includes an inflammation targeting component and a dense cross-linking component, and the inflammation targeting component and the dense cross-linking component are kept separate from each other until use, wherein the inflammation targeting component has a targeting function and is a microgel obtained by a cross-linking reaction of polyvinyl alcohol, sodium alginate and calcium ions, the dense cross-linking component is a mixed solution containing cyclodextrin and / or its derivatives and a boron-containing compound, and the dense cross-linking component is used to cross-link with the inflammation targeting component to form a highly dense hydrogel, so that by sequentially using the targeting function of the inflammation targeting component and the dense cross-linking function of the dense cross-linking component, a hydrogel with both targeted therapeutic effects and high density can be formed, so as to improve the therapeutic effect of the hydrogel on inflammatory diseases (such as inflammatory bowel disease), and can be well applied in the treatment of diseases such as inflammatory bowel disease, skin damage and surgical trauma. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] The technical solution and other beneficial effects of the present application will be made apparent by describing in detail the specific implementation methods of the present application in conjunction with the accompanying drawings.

[0019] Figure 1 is a schematic flow chart of a method for forming a hydrogel provided in an embodiment of the present application;

[0020] Figure 2 It is a schematic diagram of the changes that occur after a period of time when a dense cross-linked component is added to a suspension of an inflammation targeting component provided in an embodiment of the present application;

[0021] Figure 3 Graph showing the viscosity modulus of the inflammation targeting component and the high-density hydrogel provided in the examples of the present application.

[0022] Figure 4 These are photographs and scanning electron microscope images of the corresponding suspension of the inflammation targeting component before and after the addition of the dense cross-linking component provided in the examples of the present application;

[0023] Figure 5 is another schematic flow chart of the method for forming a hydrogel provided in an embodiment of the present application;

[0024] Figure 6 It is the in vitro inflammatory targeting fluorescence and quantification diagram of the inflammatory targeting component provided in the examples of the present application;

[0025] Figure 7 It is the in vitro inflammatory targeting fluorescence and quantification diagram of the inflammatory targeting component in human colon samples provided in the examples of the present application;

[0026] Figure 8 It is the in vivo inflammatory targeting fluorescence and quantification diagram of the inflammatory targeting component provided in the examples of the present application;

[0027] Fig. 9 is a schematic diagram of the structure of the Transwell device provided in the embodiment of the present application;

[0028] Fig.10 is a permeability diagram of the RhoB molecule provided in the examples of the present application;

[0029] Fig.11 This is a graph of the in vitro permeability measurement of the BSA protein provided in the examples of the present application;

[0030] Fig.12 This is a graph of the in vitro permeability measurement of Staphylococcus aureus provided in the examples of the present application;

[0031] Fig.13 This is a graph of the in vitro permeability measurement of LPS provided in the examples of the present application;

[0032] Fig.14 This is a diagram of the therapeutic effect based on hydrogel provided in the examples of the present application. DETAILED DESCRIPTION

[0033] The present application is further described in detail below in conjunction with the accompanying drawings and examples. It is particularly noted that the following examples are only used to illustrate the present application, but are not intended to limit the scope of the present application. Similarly, the following examples are only some embodiments of the present application rather than all embodiments, and all other embodiments obtained by ordinary technicians in the field without making creative work are within the scope of protection of this application.

[0034] In the following description of the present application, reference is made to “some embodiments”, which describe a subset of all possible embodiments, but it can be understood that “some embodiments” may be the same subset or different subsets of all possible embodiments and may be combined with each other without conflict.

[0035] In the following description of the present application, the terms "first\second\third" involved are merely used to distinguish similar objects and do not represent a specific ordering of the objects. It can be understood that "first\second\third" can be interchanged with a specific order or sequence where permitted, so that the embodiments of the present application described here can be implemented in an order other than that illustrated or described here.

[0036] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art to which this application belongs. The terms used herein are only for the purpose of describing the embodiments of this application and are not intended to limit this application.

[0037] Hydrogel is a class of high molecular weight polymers that provide an innovative repair strategy for mucosal injuries. The three-dimensional network structure of hydrogel can mimic the natural extracellular matrix of the mucosa, providing an ideal scaffold for cell growth and regeneration of damaged tissues. The targeting and high density of hydrogels provide significant advantages for the repair of mucosal injuries.

[0038] However, although targeted hydrogels and high-density hydrogels have been developed in the related art, no hydrogel that can achieve both properties has been developed. Therefore, a strategy is needed to form a hydrogel with both targeted and high-density properties to achieve the best effect in treating diseases.

[0039] In response to the above problems, the embodiments of the present application provide a hydrogel and a composition and method for forming a hydrogel. The composition for forming the hydrogel includes an inflammation targeting component and a dense cross-linking component, and the inflammation targeting component and the dense cross-linking component are kept separate from each other until use, wherein the inflammation targeting component has a targeting function and is a microgel obtained by a cross-linking reaction of polyvinyl alcohol, sodium alginate and calcium ions, the dense cross-linking component is a mixed solution containing cyclodextrin and / or its derivatives and a boron-containing compound, and the dense cross-linking component is used to cross-link with the inflammation targeting component to form a highly dense hydrogel, so that by sequentially using the targeting function of the inflammation targeting component and the dense cross-linking function of the dense cross-linking component, a hydrogel with both targeted therapeutic effects and high density can be formed, so as to improve the therapeutic effect of the hydrogel on inflammatory diseases, and can be well applied in the treatment of diseases such as inflammatory bowel disease, skin damage and surgical trauma.

[0040] The following is a detailed description with reference to specific embodiments. It should be noted that the serial numbers of the following embodiments are not intended to limit the preferred order of the embodiments.

[0041] The present application embodiment provides a composition for forming a hydrogel, which comprises an inflammation targeting component and a dense cross-linking component, and the inflammation targeting component and the dense cross-linking component are kept separate from each other until use. Among them, the inflammation targeting component has a targeting function and is a microgel obtained by cross-linking reaction of polyvinyl alcohol, sodium alginate and calcium ions. The dense cross-linking component is a mixed solution containing cyclodextrin and / or its derivatives and a boron-containing compound, and the dense cross-linking component is used to cross-link with the inflammation targeting component to form a highly dense hydrogel.

[0042] Furthermore, it should be noted that, in this embodiment, by using an inflammation targeting component with a targeting function and a dense cross-linking component that can be cross-linked with the inflammation targeting component to form a hydrogel, it is possible to first target the inflammation targeting component to the inflammation site, and then cross-link the dense cross-linking component with the inflammation targeting component targeted to the inflammation site to form a highly dense hydrogel, so that the hydrogel can simultaneously achieve targeted therapeutic effects and high density, thereby improving the therapeutic effect of the hydrogel on inflammatory diseases.

[0043] In this embodiment, the inflammation targeting component may be negatively charged and may have a positive charge targeting function. Furthermore, the positive charge targeting principle of the inflammation targeting component may be: electrostatic interaction between positive and negative charges.

[0044] Specifically, in the process of preparing the above-mentioned inflammation targeting component by cross-linking reaction using polyvinyl alcohol, sodium alginate and calcium ions, polyvinyl alcohol can be the main source of negative charge, sodium alginate can be used as a cross-linking monomer, and calcium ions can be used as a cross-linking agent. Moreover, compared with the preparation formula of common hydrogels at present, the inflammation targeting component in this embodiment has a higher positive charge targeting ability. Specifically, the negative charge of the inflammation targeting component in this embodiment can reach above -40mV.

[0045] In some embodiments, the microgel may be a gel ball with a particle size of micrometers, for example, a gel ball with a particle size of 150 micrometers.

[0046] In some embodiments, the above-mentioned inflammation targeting component can be specifically a microgel obtained by cross-linking a polyvinyl alcohol solution, a sodium alginate solution and a calcium ion solution. Among them, the mass fraction of polyvinyl alcohol in the polyvinyl alcohol solution can be 5% to 20%, for example, specifically 10% ± 5%. The mass fraction of sodium alginate in the sodium alginate solution can be 1% to 10%, for example, specifically 3% ± 5%. The mass fraction of calcium ions in the calcium ion solution can be 5% to 30%, for example, specifically 20% ± 10%. In addition, the volume ratio of the polyvinyl alcohol solution to the sodium alginate solution can be 2 / 1 to 8 / 1.

[0047] Specifically, the polyvinyl alcohol solution may be an aqueous solution of polyvinyl alcohol, the sodium alginate solution may be an aqueous solution of sodium alginate, and the calcium ion solution may be an aqueous solution of calcium ions.

[0048] In some embodiments, the molecular weight (e.g., weight average molecular weight) of the polyvinyl alcohol used to prepare the above-mentioned inflammation targeting component can be 47000-87000Da. The calcium ion used to prepare the above-mentioned inflammation targeting component can be calcium chloride and / or its analogs, wherein the calcium chloride can include anhydrous calcium chloride and / or calcium chloride hydrate, the analog of calcium chloride can include water-soluble calcium salts such as calcium bicarbonate and / or calcium dihydrogen phosphate, and the calcium chloride hydrate can include at least one of calcium chloride dihydrate, calcium chloride tetrahydrate and calcium chloride hexahydrate.

[0049] Moreover, in a specific implementation, the above-mentioned inflammation targeting component can be obtained by first uniformly mixing a polyvinyl alcohol solution (for example, a polyvinyl alcohol aqueous solution with a mass concentration of 10%±5%) and a sodium alginate solution (for example, a sodium alginate aqueous solution with a mass concentration of 3%±5%) in a volume ratio of 2 / 1 to 8 / 1, and then cross-linking with a calcium ion solution (for example, a calcium chloride aqueous solution with a mass concentration of 20%±10%).

[0050] In some embodiments, the dense cross-linking component may specifically be an aqueous solution containing cyclodextrin and / or its derivatives and a boron-containing compound.

[0051] In some embodiments, in the cyclodextrin and / or its derivatives used to prepare the dense cross-linked component, the cyclodextrin may include at least one of α-cyclodextrin, β-cyclodextrin and γ-cyclodextrin, and the derivative of cyclodextrin may include cyclodextrin modified with functional groups such as amino, carboxyl, and thiol. The boron-containing compound used to prepare the dense cross-linked component may include at least one of boric acid, borax pentahydrate, and anhydrous borax.

[0052] In some embodiments, in the dense cross-linked component, the mass ratio of cyclodextrin and / or its derivatives to the mass ratio of the boron-containing compound may be 1 / 4 to 5 / 2.

[0053] For example, in the dense cross-linked component, cyclodextrin and / or its derivatives may be specifically β-cyclodextrin polymer, and its mass may be 0.2±0.05 g, and the boron-containing compound may be specifically anhydrous borax, and its mass may be 0.18±0.04 g.

[0054] As can be seen from the above, the composition for forming a hydrogel provided in this embodiment includes an inflammation targeting component and a dense cross-linking component, and the inflammation targeting component and the dense cross-linking component are kept separate from each other until use, wherein the inflammation targeting component has a targeting function and is a microgel obtained by cross-linking reaction of polyvinyl alcohol, sodium alginate and calcium ions, the dense cross-linking component is a mixed solution containing cyclodextrin and / or its derivatives and boron-containing compounds, and the dense cross-linking component is used to cross-link with the inflammation targeting component to form a highly dense hydrogel, so that by sequentially using the targeting function of the inflammation targeting component and the dense cross-linking function of the dense cross-linking component, a hydrogel with both targeted therapeutic effects and high density can be formed, so as to improve the therapeutic effect of the hydrogel on inflammatory diseases, and can be well applied in the treatment of diseases such as inflammatory bowel disease, skin damage and surgical trauma.

[0055] According to the composition for forming a hydrogel described in the above embodiment, this embodiment will further describe the method for forming a hydrogel using the composition for forming a hydrogel described in the above embodiment.

[0056] See also Figure 1 , Figure 1 The schematic diagram of the process for forming a hydrogel provided in the embodiment of the present application is shown in FIG. Figure 1 As shown, the specific process of the method for forming a hydrogel can be as follows:

[0057] Step S101. Prepare inflammation targeting components and dense cross-linking components.

[0058] The inflammation targeting component may be the inflammation targeting component included in the composition for forming a hydrogel provided in any of the above embodiments, and the dense cross-linking component may be the dense cross-linking component included in the composition for forming a hydrogel provided in any of the above embodiments.

[0059] In some embodiments, the preparation process of the above-mentioned inflammation targeting component may include:

[0060] Step A1. Prepare polyvinyl alcohol solution, sodium alginate solution and calcium ion solution.

[0061] The polyvinyl alcohol solution may be an aqueous solution of polyvinyl alcohol, the sodium alginate solution may be an aqueous solution of sodium alginate, and the calcium ion solution may be an aqueous solution of calcium ions. The calcium ions may be calcium chloride and / or its analogs, wherein the calcium chloride may include anhydrous calcium chloride and / or calcium chloride hydrate, and the analogs of calcium chloride may include water-soluble calcium salts such as calcium bicarbonate and / or calcium dihydrogen phosphate.

[0062] Specifically, the mass fraction of polyvinyl alcohol in the polyvinyl alcohol solution may be 5% to 20%, the mass fraction of sodium alginate in the sodium alginate solution may be 1% to 10%, and the mass fraction of calcium ions in the calcium ion solution may be 5% to 30%.

[0063] Furthermore, in a specific implementation, a polyvinyl alcohol aqueous solution with a mass concentration of 10%±5%, a sodium alginate aqueous solution with a mass concentration of 3%±5%, and a calcium chloride aqueous solution with a mass concentration of 20%±10% can be prepared.

[0064] Step A2: After the polyvinyl alcohol solution and the sodium alginate solution are evenly mixed, they are immersed in a calcium ion solution for cross-linking reaction, and after the reaction is completed, they are taken out to obtain a gel.

[0065] The reaction time of the cross-linking reaction can be 0.5 to 1 hour. The volume ratio of the polyvinyl alcohol solution and the sodium alginate solution used in the cross-linking reaction can be 2 / 1 to 8 / 1.

[0066] Moreover, in a specific implementation, a polyvinyl alcohol aqueous solution with a mass concentration of 10%±5% and a sodium alginate aqueous solution with a mass concentration of 3%±5% can be uniformly mixed in a volume ratio of 2 / 1 to 8 / 1, and then immersed in a 20%±10% calcium chloride aqueous solution for cross-linking reaction. The reaction time is controlled at 0.5 to 1 hour. After the reaction is completed, the gel (i.e., a microgel) can be obtained.

[0067] Step A3: Extrude the gel through a sieve to form microgels with a particle size of micrometer level, and collect the extruded microgels to obtain inflammation targeting components.

[0068] The sieve may be a cell sieve, for example, a 100-mesh sterile cell sieve.

[0069] Moreover, in a specific implementation, a 100-mesh sterile cell sieve can be used to screen the gel obtained in the above step A2 to obtain a microgel suspension, wherein the microgel suspension includes microgels with a particle size of micrometer level. The microgels in the microgel suspension are then collected by centrifugation to obtain microgels with a particle size of micrometer level that are used as the above-mentioned inflammation targeting component.

[0070] It should be noted that, in the process of collecting the extruded microgel by centrifugation, in order to maintain the microspherical morphology and function of the microgel, the centrifugation time should not be too long. For example, the centrifugation time can be 1 min.

[0071] In some embodiments, the preparation process of the dense cross-linked component may include:

[0072] Anhydrous borax and beta-cyclodextrin polymer are added into a solvent, and then centrifuged, and the supernatant is a dense cross-linked component.

[0073] Wherein, the solvent may be water, for example, specifically ddH2O (Double Distilled Water). Cyclodextrin and / or its derivatives may include at least one of α-cyclodextrin, β-cyclodextrin and γ-cyclodextrin, for example, specifically β-cyclodextrin polymer (β-CDP). The boron-containing compound may include at least one of boric acid, borax pentahydrate and anhydrous borax, for example, specifically anhydrous borax. In the above-mentioned dense cross-linked component, the ratio of the mass of cyclodextrin and / or its derivatives to the mass of the boron-containing compound may be 1 / 4 to 5 / 2.

[0074] In addition, in a specific implementation, 0.2±0.05 g of β-cyclodextrin polymer can be added to 8±5 ml of dimethyl sulfoxide (DMSO), and then added with 0.18±0.04 g of anhydrous borax to 1.3±0.06 ml of ddH2O, and centrifuged at 10000 rpm for 1 min. The supernatant is the above-mentioned dense cross-linked component.

[0075] Step S102: cross-linking the inflammation targeting component and the dense cross-linking component to form a highly dense hydrogel.

[0076] Specifically, the dense cross-linked component can be added to the suspension of the inflammation targeting component at a volume ratio of 1:1, so that the dense cross-linked component and the inflammation targeting component are cross-linked to form a highly dense hydrogel.

[0077] For example, the changes that occur after a period of time when the dense cross-linked component is added to the suspension of the inflammation targeting component at a volume ratio of 1:1 can be seen in Figure 2 , specifically, in Figure 2 In the figure, from left to right, the changes that occur after 0 seconds, 5 seconds, and 13 seconds are shown, where: Figure 2 The white substance seen in the figure is a highly dense hydrogel formed by cross-linking of densely cross-linked components and inflammation-targeted components.

[0078] And, if Figure 3 As shown, the viscosity modulus of the highly dense hydrogel (ITTH hydrogel) formed by cross-linking the dense cross-linking component (CPC) and the inflammatory targeting component (PALMs) is always greater than the viscosity modulus of the inflammatory targeting component (PALMs), indicating that a new hydrogel is formed after the dense cross-linking component (CPC) is added to the suspension of the inflammatory targeting component (PALMs).

[0079] Furthermore, if Figure 4As shown, by comparing the scanning electron micrograph of the suspension (PALMs suspension) of the inflammatory targeting component before the addition of the dense cross-linking component (CPC) and the scanning electron micrograph of the suspension (PALMs suspension) of the inflammatory targeting component after the addition of the dense cross-linking component (CPC), it can be seen that the suspension (PALMs suspension) of the inflammatory targeting component has undergone significant changes before and after the addition of the dense cross-linking component (CPC), which also indicates that a new hydrogel is formed after the dense cross-linking component (CPC) is added to the suspension (PALMs suspension).

[0080] In some embodiments, the above step S102 can be specifically as follows: by sequentially using the targeting function of the inflammation targeting component and the dense cross-linking function of the dense cross-linking component, the inflammation targeting component is targeted to the inflammatory site and then cross-linked with the dense cross-linking component to form a hydrogel with both targeted therapeutic effect and high density, so as to improve the therapeutic effect of the hydrogel on inflammatory diseases (such as inflammatory bowel disease), and can be well applied in the treatment of diseases such as inflammatory bowel disease, skin damage and surgical trauma.

[0081] Specifically, Figure 5 As shown, the above step S102 may include:

[0082] Step S1021. Utilize the targeting function of the inflammation targeting component to target the inflammation targeting component to the inflammation site.

[0083] Moreover, in a specific implementation, before targeting the inflammation targeting component to the inflammation site, the above method for forming a hydrogel may also include: loading the inflammation targeting component with drugs, thereby targeting the drug-loaded inflammation targeting component to the inflammation site, achieving the purpose of transporting the drug to the inflammation site, so as to effectively treat the disease in combination with the drug.

[0084] Illustratively, the above-mentioned drugs may include at least one of protein drugs, antibody drugs, nucleic acid drugs, chemical drugs and nano drugs.

[0085] It should be noted that the inflammation targeting component in the examples of the present application has not only high ex vivo inflammatory targeting, but also high in vivo inflammatory targeting.

[0086] Specifically, in order to illustrate the ex vivo inflammatory targeting of the inflammation targeting component, the inventors of the present application conducted the following experiments:

[0087] A homemade intestinal fluid system was used to simulate the pH value and flow rate of the human intestine. Specifically, a whole section of the colon of a healthy mouse was cut and the lining was unfolded in a test tube along the longitudinal axis. Then, the test tube was connected to a peristaltic pump, and 1 to 3 ml of fluorescent dye-labeled inflammatory targeting components (PALMs) were injected. The peristaltic pump was run at a speed of 8 ml / min for 2 hours. Then, IVIS fluorescence imaging was performed on the colon section to observe and analyze the enrichment of microgels (i.e., inflammatory targeting components). The results can be seen in Figure 6 In addition, the same experiment was performed on the inflammatory bowel disease mouse samples (i.e., inflammatory bowel disease group or IBD) and the above healthy mouse samples (i.e., healthy group or Healthy), and the corresponding results can be found in Figure 6 Furthermore, the same experiment can be performed on healthy human samples (i.e., healthy group) and inflammatory bowel disease samples (i.e., inflammatory bowel disease group) and the above healthy mouse samples. The corresponding results can be found in Figure 7 The experimental results show that the enrichment of inflammatory targeting components (PALMs) on inflammatory tissues is significantly higher than that in the control group (ie, healthy group or Healthy).

[0088] Specifically, in order to illustrate the in vivo inflammatory targeting of the inflammation targeting component, the inventors of the present application conducted the following experiments:

[0089] C57 mice were randomly divided into two experimental groups, namely, a healthy group without experimental colitis model (i.e., control group) and an inflammatory disease group with experimental colitis model; then, 100 μl of PALMs suspension was infused into each mouse in the healthy group without experimental colitis model, and 100 μl of PALMs suspension was infused into each mouse in the experimental colitis model group; then, the fluorescence enrichment of the two experimental groups was observed by IVIS every 0.5 h, and the observation results can be seen in Figure 8 , and the results showed that the inflammation-targeting component has a high degree of positive charge targeting.

[0090] Step S1022. Cross-linking the dense cross-linked component with the inflammation targeting component targeted to the inflammation site to form a highly dense hydrogel.

[0091] Specifically, after the inflammation targeting component is targeted to the inflammation site by using the targeting function of the inflammation targeting component, that is, after the disease site is located by using the inflammation targeting component, the inflammation targeting component is cross-linked in situ by using the dense cross-linking component to form a highly dense hydrogel. In this way, the hydrogel can have both targeted therapeutic effects and high density, so as to effectively repair the inflammatory mucosal barrier and improve the therapeutic effect of inflammatory diseases, and has a wide range of application prospects, such as in the treatment of inflammatory bowel disease, chronic cervicitis, chronic sinusitis, skin damage and other diseases.

[0092] It should be noted that the highly dense hydrogel formed by cross-linking the dense cross-linked component and the inflammation targeting component in the embodiment of the present application can effectively resist the invasion of RhoB small molecules, BSA proteins, Staphylococcus aureus and LPS (endotoxin) because of its high density.

[0093] Specifically, in order to illustrate that the highly dense hydrogel formed by cross-linking the dense cross-linking component and the inflammation targeting component in the embodiment of the present application can resist the invasion of RhoB small molecules, BSA protein, Staphylococcus aureus and LPS, the inventors of the present application used Fig. 9 The following experiments were performed using the Transwell device shown:

[0094] (1) Experiments on the resistance of RhoB small molecule invasion by highly dense hydrogels formed by cross-linking dense cross-linked components with inflammatory targeting components

[0095] First, MC38 cells were placed on the lower layer of the semipermeable membrane of the Transwell device to simulate intestinal epithelial cells, and 1.2 mL of the inflammatory targeting component (PALMs) suspension was added to the semipermeable membrane of the Transwell device, and the mixture was naturally precipitated for 1 hour, and then 100 μL of the dense cross-linking component (CPC) was added and placed for 30 minutes to form a cross-linked membrane (i.e., a highly dense hydrogel) as the experimental group for detecting its permeability. In addition, three groups of control experiments were performed to obtain control group 1, control group 2, and control group 3, wherein: the difference between control group 1 and the experimental group was that only 1.2 mL of water was added to the semipermeable membrane of the Transwell device; the difference between control group 2 and the experimental group was that only 100 μL of the dense cross-linking component (CPC) was added to the semipermeable membrane of the Transwell device; the difference between control group 3 and the experimental group was that only 1.2 mL of the inflammatory targeting component (PALMs) suspension was added to the semipermeable membrane of the Transwell device.

[0096] Next, for each of the experimental group, control group 1, control group 2, and control group 3, 100 μL of RhoB solution was added to the upper layer of the semipermeable membrane of the Transwell device, and the colors of the upper and lower liquid layers of the semipermeable membrane were observed after 10 hours of release. In addition, the ratio of the fluorescence intensity of RhoB in the lower solution after 10 hours of release to the fluorescence intensity of RhoB in the upper solution at the initial moment was used as the standard value of RhoB permeability (i.e., standardized RhoB fluorescence intensity). The results can be seen in Fig.10 The results showed that the highly dense hydrogel (ITTH hydrogel) formed by cross-linking the dense cross-linking component (CPC) and the inflammatory targeting component (PALMs) can effectively prevent the leakage of RhoB molecules.

[0097] (2) Experiments on the resistance of BSA protein invasion by highly dense hydrogels formed by cross-linking dense cross-linked components with inflammation-targeting components

[0098] First, MC38 cells were placed on the lower layer of the semipermeable membrane of the Transwell device to simulate intestinal epithelial cells, and 1.2 mL of the inflammatory targeting component (PALMs) suspension was added to the semipermeable membrane of the Transwell device, and the mixture was naturally precipitated for 1 hour, and then 100 μL of the dense cross-linking component (CPC) was added and placed for 30 minutes to form a cross-linked membrane (i.e., a highly dense hydrogel) as the experimental group for detecting its permeability. In addition, three groups of control experiments were performed to obtain control group 1, control group 2, and control group 3, wherein: the difference between control group 1 and the experimental group was that only 1.2 mL of water was added to the semipermeable membrane of the Transwell device; the difference between control group 2 and the experimental group was that only 100 μL of the dense cross-linking component (CPC) was added to the semipermeable membrane of the Transwell device; the difference between control group 3 and the experimental group was that only 1.2 mL of the inflammatory targeting component (PALMs) suspension was added to the semipermeable membrane of the Transwell device.

[0099] Next, for each of the experimental group, control group 1, control group 2, and control group 3, 100 μL of 1 mg / mL BSA solution was added to the upper layer of the semipermeable membrane of the Transwell device, and the protein concentration of the upper and lower liquid layers of the semipermeable membrane was determined by the BSA quantitative method. In addition, the ratio of the protein concentration of the lower layer of the solution after 10 hours of release to the protein concentration of the upper layer at the initial time was used as the standard value of the BSA protein permeation amount (i.e., the standardized BSA amount). The results can be referred to Fig.11 The results showed that the highly dense hydrogel (ITTHhydrogel) formed by cross-linking the dense cross-linking component (CPC) and the inflammation targeting component (PALMs) could effectively prevent the leakage of BSA.

[0100] (3) Experiment on the resistance of Staphylococcus aureus invasion by highly dense hydrogels formed by cross-linking dense cross-linked components and inflammation-targeted components

[0101] First, MC38 cells were placed on the lower layer of the semipermeable membrane of the Transwell device to simulate intestinal epithelial cells, and 1.2 mL of the inflammatory targeting component (PALMs) suspension was added to the semipermeable membrane of the Transwell device, and the mixture was naturally precipitated for 1 hour, and then 100 μL of the dense cross-linking component (CPC) was added and placed for 30 minutes to form a cross-linked membrane (i.e., a highly dense hydrogel) as the experimental group for detecting its permeability. In addition, three groups of control experiments were performed to obtain control group 1, control group 2, and control group 3, wherein: the difference between control group 1 and the experimental group was that only 1.2 mL of water was added to the semipermeable membrane of the Transwell device; the difference between control group 2 and the experimental group was that only 100 μL of the dense cross-linking component (CPC) was added to the semipermeable membrane of the Transwell device; the difference between control group 3 and the experimental group was that only 1.2 mL of the inflammatory targeting component (PALMs) suspension was added to the semipermeable membrane of the Transwell device.

[0102] Next, for each of the experimental group, control group 1, control group 2, and control group 3, the E. coli or Staphylococcus aureus suspension was added to the upper layer of the semipermeable membrane of the Transwell device, and the absorbance of the E. coli or Staphylococcus aureus suspension added to each group at a wavelength of 600 nm was controlled to be the same. In addition, the number of bacteria in the solution at the lower layer of the semipermeable membrane was quantified by the absorbance at a wavelength of 600 nm (i.e., the absorbance of S. aureus at 600 nm). The results can be seen in Fig.12 The results showed that the highly dense hydrogel (ITTH hydrogel) formed by cross-linking the dense cross-linking component (CPC) and the inflammation targeting component (PALMs) can effectively prevent the leakage of Staphylococcus aureus.

[0103] (4) Experiment on the resistance of LPS invasion to the highly dense hydrogel formed by cross-linking the dense cross-linking component and the inflammation targeting component

[0104] MC38 cells used to simulate intestinal epithelium were placed on the lower layer of the semipermeable membrane of the Transwell device, and macrophage Raw264.7 cells were placed on the lower layer of the EP tube; 1.2 mL of the inflammatory targeting component (PALMs) suspension was added to the semipermeable membrane of the Transwell device, and it was naturally precipitated for 1 hour, and then 100 μL of the dense cross-linking component (CPC) was added and placed for 30 minutes to form a cross-linked membrane (i.e., a highly dense hydrogel) as the experimental group for detecting its permeability. In addition, three groups of control experiments were performed to obtain control group 1, control group 2 and control group 3, wherein: the difference between control group 1 and the experimental group was that only 1.2 mL of water was added to the semipermeable membrane of the Transwell device; the difference between control group 2 and the experimental group was that only 100 μL of the dense cross-linking component (CPC) was added to the semipermeable membrane of the Transwell device; the difference between control group 3 and the experimental group was that only 1.2 mL of the inflammatory targeting component (PALMs) suspension was added to the semipermeable membrane of the Transwell device.

[0105] Next, for each of the experimental group, control group 1, control group 2, and control group 3, LPS was added to the upper layer of the semipermeable membrane of the Transwell device. If LPS penetrates into the lower layer of the semipermeable membrane, it will react with the Raw264.7 cells to produce ROS (reactive oxygen species). In addition, the Raw264.7 cell nuclei at the bottom of the EP tube were stained with DAPI, and the generated ROS were stained with DHE. Then, the degree of activation of Raw264.7 was observed by fluorescence microscopy to reflect the amount of LPS infiltration. The results can be seen in Fig.13 , Fig.13 The red fluorescence and blue fluorescence in the image are the fluorescence of ROS and the fluorescence of Raw264.7 cell nuclei, respectively. The results show that the highly dense hydrogel (ITTHhydrogel) formed by cross-linking the dense cross-linked component (CPC) and the inflammatory targeting component (PALMs) can effectively prevent the leakage of LPS.

[0106] It should be noted that, compared with the hydrogel in the prior art, the highly dense hydrogel formed by cross-linking the dense cross-linked component and the inflammation targeting component in the embodiment of the present application has a better therapeutic effect on inflammatory diseases.

[0107] Specifically, in order to illustrate that the highly dense hydrogel formed by cross-linking the dense cross-linked component and the inflammation targeting component in the embodiment of the present application has a better therapeutic effect on inflammatory diseases, the inventors of the present application conducted the following experiments:

[0108] Potassium bismuth citrate (BPC) forms a microgel with a negative surface charge under acidic conditions and a microgel with a positive surface charge under alkaline conditions, which serves as a control for the study. The negative charge advantage of the highly dense hydrogel (ITTH hydrogel) formed by cross-linking the dense cross-linked component (CPC) and the inflammatory targeting component (PALMs) in this application under alkaline conditions was compared. 5-ASA, a first-line drug for the clinical treatment of inflammatory bowel disease (IBD), was used as a positive control group for treatment, and PALMs or CPC alone were used as negative control groups for treatment to compare the advantages of ITTH hydrogel in the treatment of IBD. In addition, the above five materials were delivered separately by enema.

[0109] like Fig.14 As shown ( Fig.14 (a. Changes in mouse body weight, b. Mouse spleen weight, c. Quantification of mouse colon length, d. Mouse colon length, e. Histological pathological staining evaluation of mice). ITTH hydrogel treatment significantly alleviated the trend of weight loss in mice, and the change in body weight at the end of treatment was significantly higher than that in the PALMs group, CPC group, and BPC group. The DSS group had severe splenomegaly, indicating a severe inflammatory response in the body, with a spleen weight of 0.134 g, while the ITTH hydrogel group and 5-ASA group both significantly inhibited the occurrence of splenomegaly, which were 0.094 g and 0.083 g, respectively (e.g. Fig.14 The average colon length in the DSS group was shortened to 5.49 cm, while that in the ITTH hydrogel group recovered to 7.508 cm, slightly longer than the 7.17 cm after 5-ASA treatment and significantly longer than that in the other treatment groups (e.g. Fig.14 Representative periodic acid-Schiff staining (PAS) results (as shown in c and d). Fig.14 As shown in e), ITTH hydrogel significantly restored more PAS-positive particles (stained purple) compared with other treatments, indicating that it promoted mucin secretion of goblet cells. These results indicate that ITTH hydrogel achieved the best effect and significantly reduced colonic inflammation.

[0110] As can be seen from the above, the method for forming a hydrogel provided in this embodiment prepares an inflammation targeting component and a dense cross-linking component, and cross-links the inflammation targeting component with the dense cross-linking component to form a highly dense hydrogel, thereby first targeting the inflammation targeting component to the inflammation site, and then cross-linking the dense cross-linking component with the inflammation targeting component targeted to the inflammation site to form a highly dense hydrogel, so that the hydrogel can simultaneously achieve targeted therapeutic effects and high density, thereby improving the therapeutic effect of the hydrogel on inflammatory diseases.

[0111] In order to better implement the method for forming a hydrogel provided in the embodiment of the present application, the embodiment of the present application also provides a hydrogel, which is formed by the method for forming a hydrogel provided in any of the above embodiments.

[0112] Specifically, the hydrogel in the embodiments of the present application can be used in the treatment of diseases such as inflammatory bowel disease, inflammatory bowel disease, chronic cervicitis, chronic sinusitis, skin damage, surgical trauma, etc.

[0113] Furthermore, the hydrogel in the embodiment of the present application is formed by the method for forming the hydrogel provided in any of the above embodiments, and therefore has all the same beneficial effects, which will not be described in detail in this embodiment.

[0114] In the above embodiments, the description of each embodiment has its own emphasis. For parts that are not described in detail in a certain embodiment, reference can be made to the relevant descriptions of other embodiments.

[0115] The above description is only a preferred embodiment of the present application and is not intended to limit the present application. Any modifications, equivalent substitutions and improvements made within the spirit and principles of the present application shall be included in the protection scope of the present application. The above description is only a preferred embodiment of the present application and is not intended to limit the present application. Any modifications, equivalent substitutions and improvements made within the spirit and principles of the present application shall be included in the protection scope of the present application.

Claims

1. A composition for forming a hydrogel, characterized in that comprising an inflammation targeting component and a dense cross-linking component, wherein the inflammation targeting component and the dense cross-linking component are kept separate from each other until use; Wherein, the inflammation targeting component has a targeting function and is a microgel obtained by cross-linking reaction of polyvinyl alcohol, sodium alginate and calcium ions; The dense cross-linking component is a mixed solution containing cyclodextrin and / or its derivatives and a boron-containing compound, and the dense cross-linking component is used to cross-link with the inflammation targeting component to form a high-density hydrogel.

2. The composition for forming a hydrogel according to claim 1, characterized in that The inflammation targeting component is specifically a microgel obtained by cross-linking a polyvinyl alcohol solution, a sodium alginate solution and a calcium ion solution; Among them, the mass fraction of the polyvinyl alcohol in the polyvinyl alcohol solution is 5% to 20%, the mass fraction of the sodium alginate in the sodium alginate solution is 1% to 10%, the mass fraction of the calcium ions in the calcium ion solution is 5% to 30%, and the volume ratio of the polyvinyl alcohol solution to the sodium alginate solution is 2 / 1 to 8 / 1.

3. The composition for forming a hydrogel according to claim 1, characterized in that The molecular weight of the polyvinyl alcohol is 47000-87000Da; The calcium ion is calcium chloride and / or its analogs, wherein the calcium chloride includes anhydrous calcium chloride and / or calcium chloride hydrate; the calcium chloride analogs are water-soluble calcium salts, including calcium bicarbonate and / or calcium dihydrogen phosphate.

4. The composition for forming a hydrogel according to claim 1, characterized in that The cyclodextrin includes at least one of α-cyclodextrin, β-cyclodextrin and γ-cyclodextrin; the derivative of the cyclodextrin includes at least one of amino-modified cyclodextrin, carboxyl-modified cyclodextrin and thiol-modified cyclodextrin; the boron-containing compound includes at least one of boric acid, borax pentahydrate and anhydrous borax.

5. The composition for forming a hydrogel according to claim 1, characterized in that In the dense cross-linking component, the mass ratio of the cyclodextrin and / or its derivative to the mass of the boron-containing compound is 1 / 4 to 5 / 2.

6. A method for forming a hydrogel, characterized in that include: The inflammation targeting component and the dense cross-linking component included in the composition for forming a hydrogel according to any one of claims 1 to 5 are prepared; The inflammation targeting component is cross-linked with the dense cross-linking component to form a highly dense hydrogel.

7. The method for forming a hydrogel according to claim 6, characterized in that The step of cross-linking the inflammation targeting component and the dense cross-linking component to form a highly dense hydrogel comprises: By sequentially using the targeting function of the inflammation targeting component and the dense cross-linking function of the dense cross-linking component, the inflammation targeting component is targeted to the inflammation site and then cross-linked with the dense cross-linking component to form a hydrogel with both targeted therapeutic effect and high density; Wherein, before targeting the inflammation targeting component to the inflammation site, the method further comprises: The inflammation targeting component is loaded with drugs, wherein the drugs include at least one of protein drugs, antibody drugs, nucleic acid drugs, chemical drugs and nano drugs.

8. The method for forming a hydrogel according to claim 6, characterized in that The steps of preparing the inflammation targeting component include: preparing polyvinyl alcohol solution, sodium alginate solution and calcium ion solution; After the polyvinyl alcohol solution and the sodium alginate solution are uniformly mixed, the mixture is immersed in the calcium ion solution for cross-linking reaction, and after the reaction is completed, the mixture is taken out to obtain a gel; The gel is extruded through a sieve to form a microgel with a particle size of micrometer level, and the extruded microgel is collected to obtain the inflammation targeting component.

9. The method for forming a composition for hydrogel according to claim 6, characterized in that: The steps of preparing the dense cross-linked component include: Anhydrous borax and beta-cyclodextrin polymer are added to a solvent, followed by centrifugation, and the supernatant is the dense cross-linked component.

10. A hydrogel, characterized in that: The method according to any one of claims 6 to 9 is used for the formation.

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