Self-adaptive repair hydrogel as well as preparation method and application thereof

Abstract

The invention relates to the technical field of medical instruments, in particular to self-adaptive repair hydrogel as well as a preparation method and application thereof. The invention provides self-adaptive repair hydrogel. The self-adaptive repair hydrogel comprises an adhesive hydrogel functional layer and a non-adhesive hydrogel functional layer which are sequentially stacked, the adhesive hydrogel functional layer and the non-adhesive hydrogel functional layer are combined through an amido bond; the non-adhesive hydrogel functional layer comprises chitin hydrogel with a network structure and a polyphenol compound loaded in the chitin hydrogel structure; the adhesive hydrogel comprises viscous gel with a network structure and an antioxidant loaded in the network structure of the viscous gel. The self-adaptive repairing hydrogel has good reconstruction and recovery efficiency and effect of the epithelial barrier.

Description

Technical Field

[0001] The present invention relates to the technical field of medical devices, and in particular to an adaptive repair hydrogel and a preparation method and application thereof. Background Art

[0002] After intubation, incision and other surgical operations, the trachea may be damaged, leading to organ stenosis, and in severe cases, it may even cause suffocation and endanger the patient's life. At present, the main methods for treating tracheal stenosis include drug therapy and bronchoscopic interventional therapy. Drug therapy mainly controls the proliferation of fibrous tissue through the anti-inflammatory effect of glucocorticoids and the anti-infective effect of antibiotics. Bronchoscopy interventional therapy removes hyperplasia to clear the airway by methods such as thermal ablation, cryotherapy and stent implantation. The above methods are widely used in clinical practice, but they still face many problems such as 1) the complex tracheal environment and the risk of repeated infection; 2) the injured site is directly exposed to the organ environment, aggravating the inflammatory response; 3) the plasticity of tracheal epithelial cells is low and the barrier reconstruction is difficult. Therefore, it will lead to repeated proliferation of granulation tissue after treatment, which will lead to frequent recurrence of tracheal stenosis. Therefore, comprehensive reconstruction of the trachea and restoration of tracheal function by bioengineering means is gradually becoming a potential treatment strategy.

[0003] However, it is still a challenging task to make the organ reconstructed through bioengineering match the natural trachea in composition, structure and function. The trachea is composed of three key components: cartilage, blood vessels and epithelium, which play a vital role in maintaining tracheal patency, delivering oxygen and nutrients, and preventing bacterial adhesion. In order to achieve comprehensive repair of the trachea, it is necessary to promote the regeneration of cartilage, blood vessels and epithelium at the same time. The current bioengineering trachea technology mainly focuses on the construction of cartilage, but often ignores the reconstruction of epithelial function, which leads to epithelial loss and dysfunction after transplantation, increases the risk of complications such as restenosis, and may lead to the failure of tracheal reconstruction. Therefore, restoring the function of the epithelium is crucial to reduce fatal complications and reduce respiratory interference in common clinical situations. However, the complex epithelial environment, severe inflammatory response at the site of injury, and lack of epithelial cell stemness make it difficult to have good epithelial reconstruction efficiency and effect. Summary of the invention

[0004] In view of this, the object of the present invention is to provide an adaptive repair hydrogel and a preparation method and application thereof, wherein the adaptive repair hydrogel has good efficiency and effect in rebuilding and restoring the epithelial barrier.

[0005] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

[0006] The present invention provides an adaptive repair hydrogel, comprising an adhesive hydrogel functional layer and a non-adhesive hydrogel functional layer stacked in sequence;

[0007] The adhesive hydrogel functional layer and the non-adhesive hydrogel functional layer are bonded via an amide bond;

[0008] The non-adhesive hydrogel functional layer includes a chitosan hydrogel having a network structure and a polyphenol compound loaded in the chitosan hydrogel structure;

[0009] The adhesive hydrogel comprises an adhesive gel having a network structure and an antioxidant loaded in the adhesive gel network structure.

[0010] Preferably, the viscous gel having a network structure comprises carboxyl groups and / or catechol groups.

[0011] Preferably, the viscous gel having a network structure includes one or more of polyaspartic acid hydrogel, polylipoic acid hydrogel, acrylic acid / vinyl pyrrolidone copolymer succinimide ester hydrogel, lipoic acid / polyethylene glycol copolymer succinimide ester hydrogel, polyaspartic acid hydrogel, polysebacic acid glyceryl hydrogel and linear paste polysebacic acid glyceryl gel;

[0012] The antioxidant comprises one or more of L-arginine, quercetin, curcumin, Prussian blue, tartaric acid, sodium urate and natural polyphenols.

[0013] Preferably, the viscous gel having a network structure comprises polyaspartic acid;

[0014] The antioxidant includes L-arginine.

[0015] Preferably, the mass ratio of the antioxidant to the viscous gel having a network structure is (1-10):100.

[0016] Preferably, the polyphenolic compound comprises tannic acid and / or gallic acid.

[0017] Preferably, the mass ratio of chitosan to polyphenol compound is (85-95):(5-15).

[0018] Preferably, the polyphenolic compound comprises tannic acid.

[0019] The present invention also provides a method for preparing the adaptive repair hydrogel described in the above technical solution, comprising the following steps:

[0020] The chitosan solution and the polyphenol compound solution are mixed and self-assembled to obtain a non-adhesive hydrogel functional layer;

[0021] Mixing a precursor for preparing a viscous gel, an antioxidant, and a photoinitiator to obtain a mixed system;

[0022] After the mixed system is dripped onto the surface of the non-adhesive hydrogel functional layer, ultraviolet light is irradiated to obtain the adaptive repair hydrogel.

[0023] The present invention also provides the use of the adaptive repair hydrogel described in the above technical solution or the adaptive repair hydrogel prepared by the preparation method described in the above technical solution in repair and regeneration medical devices.

[0024] The present invention provides an adaptive repair hydrogel, comprising an adhesive hydrogel functional layer and a non-adhesive hydrogel functional layer stacked in sequence; the adhesive hydrogel functional layer and the non-adhesive hydrogel functional layer are bonded by amide bonds; the non-adhesive hydrogel functional layer comprises a chitosan hydrogel having a network structure and a polyphenol compound loaded in the chitosan hydrogel structure; the adhesive hydrogel comprises a viscous gel having a network structure and an antioxidant loaded in the viscous gel network structure. The adaptive repair hydrogel of the present invention has two distinct functional surfaces, which achieve the repair goal together; wherein the adhesive hydrogel functional layer can enable the adaptive repair hydrogel to quickly adhere to a damaged area filled with mucus (such as a damaged area in the airway), providing direct contact and repair for the epithelium; because chitosan has good anti-pollution performance, it can resist the complex environment of the trachea and protect the damaged area, resulting in the surface of the non-adhesive hydrogel functional layer having excellent anti-pollution performance, forming a stable and biodegradable protective barrier in the air, effectively resisting various pathogenic factors, such as respiratory pollutants, physical and chemical factors, etc. The adaptive repair hydrogel significantly reduces the activation and infiltration of inflammatory cells, effectively controls the transitional immune response, prevents secondary damage to the epithelium when it is exposed to the airway environment, and promotes the recovery of epithelial stem cell characteristics, ultimately achieving the reconstruction and recovery of the epithelial barrier. In addition, the adaptive repair hydrogel of the present invention can be used not only for the inner wall of the trachea, but also for a variety of epithelial tissues including the skin. From the general environment of the skin surface to the special environment inside the trachea, it can repair the damaged epithelium in a targeted manner and achieve the comprehensive recovery of the epithelial barrier. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 This is a schematic diagram of the preparation process of the adaptive repair hydrogel described in Example 1;

[0026] Figure 2 This is a cross-sectional SEM image of the adaptive repair hydrogel described in Example 1;

[0027] Figure 3 The adhesion properties of the adaptive repair hydrogel on two different surfaces in Example 1 (a) and the adhesion of the LA@PA layer in the adaptive repair hydrogel on the tracheal surface of pigs (b) and rabbits (c);

[0028] Figure 4 The curves of the survival rate of the rabbits in the rabbit tracheal epithelial scraping injury model of the adaptive repair hydrogel described in Example 1, the PA described in Comparative Example 1, the PA-C described in Comparative Example 2, and the LA@PA-C described in Comparative Example 3 versus time (a) and the bronchoscopic examination results of the PA described in Comparative Example 1 and the adaptive repair hydrogel described in Example 1 on the 28th day (b);

[0029] Figure 5 It is the construction of comprehensive tracheal transplantation, the curve of survival rate changing over time, and the curve of respiratory rate changing over time. DETAILED DESCRIPTION

[0030] The present invention provides an adaptive repair hydrogel, comprising an adhesive hydrogel functional layer and a non-adhesive hydrogel functional layer stacked in sequence;

[0031] The adhesive hydrogel functional layer and the non-adhesive hydrogel functional layer are bonded via an amide bond;

[0032] The non-adhesive hydrogel functional layer includes a chitosan hydrogel having a network structure and a polyphenol compound loaded in the chitosan hydrogel structure;

[0033] The adhesive hydrogel comprises an adhesive gel having a network structure and an antioxidant loaded in the adhesive gel network structure.

[0034] In the present invention, the thickness of the adhesive hydrogel functional layer is preferably 0.1-5 mm, more preferably 0.5 mm. In an embodiment of the present invention, the thickness of the adhesive hydrogel functional layer may be 0.5 mm.

[0035] In the present invention, the adhesive hydrogel functional layer preferably comprises an adhesive gel having a network structure and an antioxidant loaded in the adhesive gel network structure. In the present invention, the mass ratio of the antioxidant to the adhesive gel having a network structure is preferably (1-10):100, more preferably (3-7):100. In an embodiment of the present invention, the mass ratio of the antioxidant to the adhesive gel having a network structure may be 5:100.

[0036] In the present invention, the viscous gel with a network structure preferably includes a carboxyl group and / or a catechol group. In the present invention, the viscous gel with a network structure preferably includes one or more of polyaspartic acid (PA) hydrogel, polylipoic acid hydrogel, acrylic acid / vinyl pyrrolidone copolymer succinimide ester hydrogel, lipoic acid / polyethylene glycol copolymer succinimide ester hydrogel, polyaspartic acid hydrogel, polysebacic acid glyceride hydrogel and linear paste polysebacic acid glyceride gel, more preferably including polyaspartic acid hydrogel; when the viscous gel with a network structure is two or more of the above-mentioned specific selections, the present invention does not have any special restrictions on the ratio of the above-mentioned specific substances, and can be mixed in any ratio. In an embodiment of the present invention, the viscous gel with a network structure can be a polyaspartic acid hydrogel.

[0037] In the present invention, the antioxidant preferably includes one or more of L-arginine (LA), quercetin, curcumin, Prussian blue, tartaric acid, sodium urate and natural polyphenols, and more preferably includes L-arginine; when the antioxidant is two or more of the above specific selections, the present invention has no special restrictions on the ratio of the above specific substances, and can be mixed in any ratio. In an embodiment of the present invention, the antioxidant can be L-arginine.

[0038] In the present invention, the thickness of the non-adhesive hydrogel functional layer is preferably 0.1 to 5 mm, more preferably 0.5 to 1 mm. In an embodiment of the present invention, the thickness of the non-adhesive hydrogel functional layer may be 1 mm.

[0039] In the present invention, the non-adhesive hydrogel functional layer comprises a chitosan hydrogel having a network structure and a polyphenol compound loaded in the chitosan hydrogel structure. In the present invention, the mass ratio of the chitosan hydrogel to the polyphenol compound is preferably (85-95): (5-15), more preferably (88-92): (8-12). In an embodiment of the present invention, the mass ratio of the chitosan hydrogel to the polyphenol compound can be 90:10.

[0040] In the present invention, the polyphenol compound preferably includes tannic acid (TA) and / or gallic acid, more preferably includes tannic acid; when the polyphenol compound is tannic acid and gallic acid, the present invention has no special limitation on the ratio of tannic acid and gallic acid, and they can be mixed in any ratio. In an embodiment of the present invention, the polyphenol compound can be tannic acid.

[0041] In the present invention, the above-mentioned viscous gel represented by the polyaspartic acid has good adhesion to epithelial tissues such as organs and skin, and constitutes the matrix of the adhesion layer; the chitin is a low-viscosity gel, which constitutes the matrix of the non-adhesion layer, and has good anti-pollution performance, can resist the complex environment of the trachea, and protect the damaged part. At the same time, the combination of the two can still show good antibacterial properties. At the same time, the polyphenolic compounds represented by tannic acid can remove ROS (reactive oxygen); the polyphenolic compounds represented by tannic acid and the antioxidants represented by LA are respectively loaded in the non-adhesive hydrogel functional layer and the adhesive hydrogel functional layer to form an adaptive repair hydrogel so that the non-adhesive hydrogel functional layer has multiple non-cervical cancer, hydrophilic and hydrophobic effects, and the adhesive functional layer plays a sustained release role, thereby realizing a differential release mode: the antioxidant represented by LA can be quickly released at the damaged part, reduce oxidative stress and regulate immune response in the early stage, and the polyphenolic compounds represented by tannic acid can be released long-term and lastingly, promote the stemness of basal stem cells and differentiate and repair the multifunctional epithelial cell layer. The adaptive repair hydrogel reprogrammed the oxidative stress-innate immune cascade axis and promoted the regeneration of epithelial cells with multiple functions (such as ciliated cells, mucus cells, etc.), thereby protecting the integrity and barrier function of epithelial cells.

[0042] The present invention also provides a method for preparing the adaptive repair hydrogel described in the above technical solution, comprising the following steps:

[0043] The chitosan solution and the polyphenol compound solution are mixed and self-assembled to obtain a non-adhesive hydrogel functional layer;

[0044] Mixing a precursor for preparing a viscous gel, an antioxidant, and a photoinitiator to obtain a mixed system;

[0045] After the mixed system is dripped onto the surface of the non-adhesive hydrogel functional layer, ultraviolet light is irradiated to obtain the adaptive repair hydrogel.

[0046] In the present invention, unless otherwise specified, all preparation raw materials are commercially available products well known to those skilled in the art.

[0047] The present invention mixes a chitosan solution and a polyphenol compound solution, performs self-assembly, and obtains a non-adhesive hydrogel functional layer.

[0048] In the present invention, the mass concentration of the chitosan solution is preferably 3% to 5%, more preferably 3.5% to 4.5%. In an embodiment of the present invention, the mass concentration of the chitosan solution may be 4.31%.

[0049] In the present invention, the method for preparing the chitosan solution preferably comprises the following steps:

[0050] Chitosan and alkali solution are mixed, and the obtained suspension is subjected to a first freeze-thaw cycle. Water is then added to the obtained suspension and the suspension is subjected to a second freeze-thaw cycle to obtain the chitosan solution.

[0051] In the present invention, the mass concentration of the alkali solution is preferably 20% to 40%, more preferably 28% to 38%. In an embodiment of the present invention, the mass concentration of the alkali solution may be 36.7%. In the present invention, the alkali solution is preferably a sodium hydroxide solution.

[0052] In the present invention, the mass ratio of chitosan to alkali solution is preferably (1-5):20, more preferably 3:20. In an embodiment of the present invention, the mass ratio of chitosan to alkali solution may be 3:20. In the present invention, the alkali solution is used to promote dissolution.

[0053] The present invention does not have any special limitation on the mixing process, and the mixing process may be carried out using a process well known to those skilled in the art.

[0054] After the mixing, the present invention also preferably includes leaving the mixture at room temperature overnight. In the present invention, the role of leaving the mixture at room temperature overnight is to further promote dissolution.

[0055] In the present invention, the freezing temperature in the first freeze-thaw cycle is preferably -160 to -20°C, more preferably -60 to -30°C; the melting temperature is preferably 4 to 40°C, more preferably 20 to 30°C. In the present invention, the number of cycles of the first freeze-thaw cycle is preferably 3 to 10 times, more preferably 3 to 5 times. In an embodiment of the present invention, the freezing temperature in the first freeze-thaw cycle may be -40°C, the melting temperature may be 20°C, and the number of cycles of the first freeze-thaw cycle may be 5 times. In the present invention, the process of the first freeze-thaw cycle can promote the dissolution of chitin.

[0056] In the present invention, the mass concentration of the polyphenol compound in the polyphenol compound solution is preferably 10%. In the present invention, the polyphenol compound solution is preferably prepared by mixing the polyphenol compound and water. In the present invention, the water is preferably deionized water.

[0057] In the present invention, the mass ratio of chitosan in the chitosan solution to the polyphenolic compound in the polyphenolic compound solution is preferably (85-95):(5-15), more preferably (88-92):(8-12). In an embodiment of the present invention, the mass ratio of chitosan in the chitosan solution to the polyphenolic compound in the polyphenolic compound solution can be 90:10.

[0058] In the present invention, the self-assembly temperature is preferably 60-80°C, more preferably 65-75°C; the self-assembly time is preferably 6-12h, more preferably 6-8h. In an embodiment of the present invention, the self-assembly temperature may be 65°C, and the time may be 8h.

[0059] In the present invention, the polyphenol compound can induce the chitosan to self-assemble to form a hydrogel.

[0060] After the self-assembly is completed, the present invention also preferably includes acid neutralization and cleaning in sequence. The present invention has no special limitation on the acid neutralization and cleaning process, and the process well known to those skilled in the art can be used.

[0061] After obtaining the non-adhesive hydrogel functional layer, the present invention mixes the precursor for preparing the viscous gel, the antioxidant and the photoinitiator to obtain a mixed system.

[0062] The present invention does not have any special limitation on the precursor of the viscous gel, and a precursor that can prepare a corresponding type of viscous gel known to those skilled in the art can be used. In an embodiment of the present invention, the viscous gel can be polyaspartic acid; the preparation of the precursor of the polyaspartic acid can be: after mixing 0.45g of aspartic acid and 0.55g of morpholineethanesulfonic acid buffer, 50mg of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide and 50mg of N-hydroxysuccinimide are added to carry out a substitution reaction to obtain N-acryloyl aspartic acid.

[0063] In the present invention, the antioxidant preferably includes one or more of L-arginine (LA), quercetin, curcumin, Prussian blue, tartaric acid, sodium urate and natural polyphenols, and more preferably includes L-arginine; when the antioxidant is two or more of the above specific selections, the present invention has no special restrictions on the ratio of the above specific substances, and can be mixed in any ratio. In an embodiment of the present invention, the antioxidant can be L-arginine.

[0064] In the present invention, the photoinitiator preferably includes one or more of IRGACURE 2959, 1173 (2-hydroxy-2-methyl-1-phenylpropanone), 184 (1-hydroxycyclohexyl phenyl ketone), 907 (2-methyl-2-(4-morpholinyl)-1-[4-(methylthio)phenyl]-1-propanone), TPO (2,4,6-trimethylbenzoyl-diphenylphosphine oxide), TPO-L (ethyl 2,4,6-trimethylbenzoylphenylphosphonate), IHT-PI 910 (2-dimethylamino-2-benzyl-1-[4-(4-morpholinyl)phenyl]-1-butanone), 659 (2-hydroxy-2-methyl-1-[4-(2-hydroxyethoxy)phenyl]-1-propanone) and MBF (methyl benzoylformate), and more preferably IRGACURE 2959; When the photoinitiator is two or more of the above-mentioned specific selections, the present invention does not have any special restrictions on the ratio of the above-mentioned specific substances, and they can be mixed in any ratio.

[0065] In the present invention, the mass ratio of the precursor, antioxidant and photoinitiator for preparing the viscous gel is preferably 100:(1-10):(0.1-0.5), more preferably 100:(4-8):(0.1-0.3). In an embodiment of the present invention, the mass ratio of the precursor, antioxidant and photoinitiator for preparing the viscous gel can be 100:5:0.1.

[0066] In the present invention, the mixing temperature is preferably 10-50°C, more preferably 20-30°C; the mixing time is preferably 1-100 min, more preferably 10-30 min. In an embodiment of the present invention, the temperature of reaction A can be 20°C, and the time can be 30 min.

[0067] After obtaining the mixed system, the present invention drips the mixed system onto the surface of the non-adhesive hydrogel functional layer and then irradiates the surface with ultraviolet light to obtain the adaptive repair hydrogel.

[0068] The present invention has no special limitation on the dropping process, and the dropping process can be carried out by a process well known to those skilled in the art.

[0069] In the present invention, the wavelength of the ultraviolet light irradiation is preferably 365 nm; the duration of the ultraviolet light irradiation is preferably 1 to 30 min, more preferably 10 to 20 min. In an embodiment of the present invention, the duration of the ultraviolet light irradiation may be 20 min.

[0070] The present invention also provides the use of the adaptive repair hydrogel described in the above technical solution or the adaptive repair hydrogel prepared by the preparation method described in the above technical solution in repair and regeneration medical devices. In the present invention, the repair and regeneration medical device is preferably a tracheal repair and regeneration medical device, a wound repair and regeneration medical device, a bone repair and regeneration medical device, or a fistula repair and regeneration medical device. The present invention does not have any special restrictions on the method of application, and the method well known to those skilled in the art can be used.

[0071] The technical solutions in the present invention will be described clearly and completely below in conjunction with the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0072] Example 1

[0073] like Figure 1 As shown, 4.5 g of chitosan powder was dissolved in 30 g of a sodium hydroxide solution with a mass concentration of 36.7%, and the suspension was subjected to repeated freeze-thaw cycles for 3 times (wherein the freezing temperature was -40°C and the melting temperature was 20°C), and then 70 g of deionized water was added, and the suspension was subjected to repeated freeze-thaw cycles for 5 times (wherein the freezing temperature was -40°C and the melting temperature was 20°C), to obtain a transparent chitosan solution with a mass concentration of 4.31%;

[0074] Dissolve 10 g of tannic acid in 90 g of deionized water to obtain a tannic acid solution with a mass concentration of 10%;

[0075] The chitosan solution and the tannic acid solution were mixed in a mass ratio of 9:1, and self-assembled overnight (8 h) at 65° C., neutralized with acid, and then thoroughly washed with water to obtain a non-adhesive hydrogel functional layer (denoted as TA@C, with a thickness of 1 mm);

[0076] 0.45 g of aspartic acid was dissolved in 0.55 g of morpholineethanesulfonic acid (MES) buffer solution, and 50 mg of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDC) and 50 mg of N-hydroxysuccinimide (NHS) were added, and the mixture was incubated overnight (temperature was 4° C., time was 12 h) to obtain N-acryloyl aspartic acid;

[0077] 1 mg of a photoinitiator (IRGACURE 2959), 50 mg of L-arginine and 1 g of the N-acryloyl aspartic acid were mixed and reacted at room temperature of 20 degrees for 30 minutes to obtain a mixed system;

[0078] After dropping the mixed system on the surface of the non-adhesive hydrogel functional layer, the mixture was irradiated with ultraviolet light at a wavelength of 365 nm for 20 minutes to obtain the adaptive repair hydrogel (denoted as LA@PA-TA@C hydrogel, and the thickness of the adhesive hydrogel functional layer (denoted as LA@PA) was 0.5 mm).

[0079] Figure 2 is a cross-sectional SEM image of the adaptive repair hydrogel. Figure 2 It can be seen that the two-sided structure can be clearly identified from the SEM image. In the structure, the LA@PA and TA@C constitute the anti-adhesion surface and the tissue adhesion surface respectively. Through the characteristics of their respective surface structures and composition differences, the two-sided nature of the hydrogel is significantly demonstrated.

[0080] Figure 3 The adhesion properties of the adaptive repair hydrogel on two different surfaces (a) and the adhesion of the LA@PA layer in the adaptive repair hydrogel on the tracheal surface of pigs (b) and rabbits (c), respectively. Figure 3 As shown in a, the two different surfaces of the adaptive repair hydrogel exhibited completely different adhesion properties. The LA@PA layer (marked by white arrows) had excellent adhesion to the skin and could firmly adhere to the skin surface, while the TA@C layer (marked by black arrows) had weak adhesion and could hardly adhere to the skin. This difference fully demonstrated the dual nature of the hydrogel adhesion. Figure 3 As shown in Figures b and c, even when facing the mucus-rich surface in the tracheal cavity, the LA@PA layer in the adaptive hydrogel can fit firmly and remain stable when undergoing changes such as stretching, compression, and rotation, which is consistent with the need for stability in tracheal physiological activities. At the same time, the adaptive hydrogel exhibits excellent adhesion properties on both stratified squamous epithelial surfaces (skin) and pseudostratified ciliated columnar epithelial surfaces (trachea), so it is widely applicable to various types of epithelial tissues.

[0081] Comparative Example 1

[0082] 0.45 g of aspartic acid was dissolved in 0.55 g of morpholineethanesulfonic acid (MES) buffer solution, and 50 mg of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDC) and 50 mg of N-hydroxysuccinimide (NHS) were added, and the mixture was incubated overnight (temperature was 4° C., time was 12 h) to obtain N-acryloyl aspartic acid;

[0083] 1 mg of a photoinitiator (IRGACURE 2959) and 1 g of the N-acryloyl aspartic acid were mixed and reacted at room temperature of 20 degrees for 30 minutes to obtain a mixed system;

[0084] The mixed system was irradiated with ultraviolet light of 365 nm wavelength for 20 min to obtain the hydrogel (denoted as PA, with a thickness of 0.5 mm).

[0085] Comparative Example 2

[0086] 4.5 g of chitosan powder was dissolved in 30 g of sodium hydroxide solution with a mass concentration of 36.7%, and the suspension was subjected to repeated freeze-thaw cycles for 3 times (wherein the freezing temperature was -40°C and the melting temperature was 20°C). Then, 70 g of deionized water was added and repeated freeze-thaw cycles were performed for 5 times (wherein the freezing temperature was -40°C and the melting temperature was 20°C) to obtain a transparent chitosan solution with a mass concentration of 4.31%.

[0087] The chitosan solution was self-assembled at 65° C. overnight, neutralized with acid, and then thoroughly washed with water to obtain a chitosan hydrogel layer (denoted as C, with a thickness of 1 mm);

[0088] 0.45 g of aspartic acid was dissolved in 0.55 g of morpholineethanesulfonic acid (MES) buffer solution, and 50 mg of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDC) and N-hydroxysuccinimide (NHS) were added, and the mixture was incubated overnight (temperature was 4° C., time was 12 h) to obtain N-acryloyl aspartic acid;

[0089] 1 mg of a photoinitiator (IRGACURE 2959) and 1 g of the N-acryloyl aspartic acid were mixed and reacted at room temperature of 20 degrees for 30 minutes to obtain a mixed system;

[0090] After the mixed system was dropped onto the surface of the chitosan hydrogel layer, the layer was irradiated with ultraviolet light at a wavelength of 365 nm for 20 minutes to obtain a hydrogel (denoted as PA-C, with a thickness of the PA layer of 0.5 mm).

[0091] Comparative Example 3

[0092] Referring to Example 1, the difference is that TA is not added to the non-adhesive hydrogel functional layer, and the obtained hydrogel is recorded as LA@PA-C.

[0093] Figure 4The following are the curves of the change in survival rate over time of the adaptive repair hydrogel described in Example 1, the PA described in Comparative Example 1, the PA-C described in Comparative Example 2, and the LA@PA-C described in Comparative Example 3 in the rabbit tracheal epithelial scraping injury model (the epithelium and most of the submucosal layer at the injured site were removed on the 0th day, and on the 7th day, extensive epithelial dysfunction and stenosis appeared in the entire trachea, and its pathological changes were very similar to human epithelial dysfunction, and then the corresponding treatment was started) and the bronchoscopic examination results of the PA described in Comparative Example 1 and the adaptive repair hydrogel described in Example 1 on the 28th day (b); Figure 4 It can be seen that in the rabbit tracheal epithelial scratch injury model, Figure 4 From a in the figure, it can be seen that, except for the adaptive repair hydrogel described in Example 1, the other hydrogels died due to severe shortness of breath symptoms; Figure 4 As shown in b, on the 28th day, bronchoscopic examination revealed that the PA treatment described in Comparative Example 1 resulted in severe stenosis and sputum adhesion, while the adaptive repair hydrogel treatment described in Example 1 achieved good tracheal patency, restored natural-like smooth breathing, and no sputum retention, successfully achieving complete tracheal repair including structure and function, and effectively restoring respiratory function;

[0094] To demonstrate the ability of the adaptive repair hydrogel described in the present invention to be used in combination with standardized vascular and cartilage repair methods to address the long-standing problem of complete tracheal repair (especially epithelial dysfunction): In this standardized process, the constructed engineered cartilage (TEC) was implanted into the anterior neck muscle to promote cartilage maturation and conventional prevascularization for 28 days, after which a comprehensive tracheal transplant (denoted as TEC / LA@PA-TA@C, as shown in Figure 1) was formed by adhering the adaptive repair hydrogel described in Example 1 to the vascularized TEC. Figure 5 28 days after tracheal transplantation, the survival rate of the comprehensive transplantation exceeded 83%, while the survival rate of the control group (specifically, the adaptive repair hydrogel was not applied) was less than 18% (as shown in A). Figure 5 At the same time, the rabbit's respiratory state remained stable, and its respiratory function gradually recovered to a state close to its natural state (as shown in B in Figure 1). Figure 5 As shown in C in the figure); As can be seen from the above, TEC / LA@PA-TA@C showed tracheal patency without stenosis and formed a new structure similar to natural cartilage at the transplantation site, while the control group showed stenosis and lack of cartilage regeneration. The adaptive repair hydrogel provides an innovative method for rescuing tracheal epithelial dysfunction. By applying standard vascular and cartilage regeneration techniques and re-epithelialization techniques, complete structural and functional repair of the trachea was achieved.

[0095] The above description is only a preferred embodiment of the present invention and does not limit the present invention in any form. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be regarded as the protection scope of the present invention.

Claims

1. An adaptive repair hydrogel, characterized in that: It includes an adhesive hydrogel functional layer and a non-adhesive hydrogel functional layer stacked in sequence; The adhesive hydrogel functional layer and the non-adhesive hydrogel functional layer are bonded via an amide bond; The non-adhesive hydrogel functional layer includes a chitosan hydrogel having a network structure and a polyphenol compound loaded in the chitosan hydrogel structure; The adhesive hydrogel comprises an adhesive gel having a network structure and an antioxidant loaded in the adhesive gel network structure.

2. The adaptive repair hydrogel according to claim 1, characterized in that The viscous gel having a network structure includes carboxyl groups and / or catechol groups.

3. The adaptive repair hydrogel according to claim 2, characterized in that: The viscous gel with a network structure includes one or more of polyaspartic acid hydrogel, polylipoic acid hydrogel, acrylic acid / vinyl pyrrolidone copolymer succinimide ester hydrogel, lipoic acid / polyethylene glycol copolymer succinimide ester hydrogel, polyaspartic acid hydrogel, polysebacic acid glyceryl hydrogel and linear paste polysebacic acid glyceryl gel; The antioxidant comprises one or more of L-arginine, quercetin, curcumin, Prussian blue, tartaric acid, sodium urate and natural polyphenols.

4. The adaptive repair hydrogel according to claim 2 or 3, characterized in that: The viscous gel having a network structure comprises polyaspartic acid; The antioxidant includes L-arginine.

5. The adaptive repair hydrogel according to claim 4, characterized in that: The mass ratio of the antioxidant to the viscous gel with a network structure is (1-10):

100.

6. The adaptive repair hydrogel according to claim 1, characterized in that: The polyphenolic compounds include tannic acid and / or gallic acid.

7. The adaptive repair hydrogel according to claim 1 or 6, characterized in that: The mass ratio of the chitosan hydrogel to the polyphenol compound is (85-95):(5-15).

8. The self-adaptive repair hydrogel according to claim 7, characterized in that: The polyphenolic compounds include tannic acid.

9. The method for preparing the adaptive repair hydrogel according to any one of claims 1 to 8, characterized in that: The following steps are involved: The chitosan solution and the polyphenol compound solution are mixed and self-assembled to obtain a non-adhesive hydrogel functional layer; Mixing a precursor for preparing a viscous gel, an antioxidant, and a photoinitiator to obtain a mixed system; After the mixed system is dripped onto the surface of the non-adhesive hydrogel functional layer, ultraviolet light is irradiated to obtain the adaptive repair hydrogel.

10. Use of the adaptive repair hydrogel according to any one of claims 1 to 8 or the adaptive repair hydrogel prepared by the preparation method according to claim 9 in repair and regeneration medical devices.

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