Mesoscopic phase enhanced polyphenol-based coacervate adhesive and method of making

Abstract

The application discloses a mesoscopic phase reinforced polyphenol-based condensate adhesive and a preparation method thereof, and relates to the technical field of medical tissue adhesives. The preparation method of the adhesive comprises the following steps: step one, configuring a polyphenol compound into liquid A; step two, configuring an epoxy-based polymer material into liquid B; step three, diluting a silanization reagent with acid deionized water, and naming the dilution liquid C; step four, mixing liquid B and liquid C, and then adding the mixture into liquid A, and obtaining the mesoscopic phase reinforced polyphenol-based condensate adhesive after oscillation and centrifugation. After the silanization reagent is hydrolyzed, the silanization reagent is assembled into a mesoscopic phase structure of an organic-inorganic hybrid composite phase body around the epoxy-based polymer material phase, so that the hydrogen bond and the hydrophobic interaction between the polyphenol compound are strengthened. The prepared adhesive has the degradable purpose, has strong adhesion on the surface of a tissue, and can be used as a good medical tissue adhesive.

Description

Technical Field

[0001] This invention relates to the field of medical tissue adhesives, and more particularly to a mesoscopic phase-reinforced polyphenolic aggregate adhesive and its preparation method. Background Technology

[0002] As is well known, surgical patients often suffer from pain during sutures and wound healing. The advent and development of medical adhesives promise to help alleviate postoperative pain. Currently, tissue adhesives on the market are mainly divided into chemically synthesized materials and natural repair materials. Chemically synthesized materials include cyanoacrylate, polyethylene glycol polymer hydrogels, dendritic macromolecules, and urethane-based adhesives, while natural repair materials include fibrin adhesives, collagen adhesives, gelatin-based adhesives, and polysaccharide adhesives. However, both have their drawbacks. Chemically synthesized materials experience exothermic reactions or polymerization processes that can cause thermal damage, degradation that produces toxicity and may trigger inflammatory reactions, and the light curing or curing time is too long. Natural repair materials, on the other hand, often lack wet adhesion, have potential allergic reactions, and tend to have lower mechanical strength and adhesion.

[0003] In nature, mussels use their adhesive secretion, mussel adhesive protein (MAP), to adhere to underwater rocks and boats, remaining stable even in turbulent currents. MAP is rich in 3,4-dihydroxyphenylalanine (DOPA), considered a key factor in their strong underwater adhesion. Inspired by mussels, polyphenolic compounds can enhance adhesion to human tissues through various covalent and non-covalent interactions formed by their high-density phenolic hydroxyl groups. However, polyphenolic adhesives have relatively weak bulk mechanical properties, and their interfacial adhesion is easily affected by the physiological environment, leading to decreased adhesion performance or even failure. For example, the adhesive prepared in the published patent CN120285274A, a polyphenolic tissue adhesive for promoting wound healing and its preparation method, exhibits low wet adhesion strength, failing to meet the requirements for hemostasis and adhesion in organs such as the liver. Doping organic polymer hydrogels with inorganic materials such as silicon nanoparticles to enhance the mechanical properties of the hydrogel is common, providing a potential approach to indirectly improve adhesion performance by strengthening the cohesive force of the adhesive. However, at higher silicon content, the compatibility between particles and polymer chains deteriorates, leading to a decrease in performance. Furthermore, the formation of nanoparticles of suitable size and morphology in situ requires control of reaction conditions such as thermal hydrolysis, pH adjustment, and the addition of co-surfactants.

[0004] Therefore, this invention innovatively proposes a convenient and effective silanization reinforcement strategy based on organic-inorganic hybridization. By adding pre-hydrolyzed siloxanes to the surface of epoxy polymers, an in-situ Si-O-Si network is formed, which connects and anchors polymer chains within a certain range to form ordered silanized units. The subsequently added polyphenol material separates the silanized units into individual micro-regions through hydrogen bonding and hydrophobic interactions. We call this micro-ordered structure assembled from the silanized polymer network and polyphenols a "mesoscopic phase" reinforcer. The presence of the mesoscopic phase can share energy dissipation, thereby improving the strength of the adhesive matrix during the adhesion and fracture process, enhancing the adhesion performance of tissue adhesives, and meeting the needs of wound and tissue adhesion. Summary of the Invention

[0005] To overcome the technical deficiencies of existing tissue adhesives, this invention provides a mesophase-reinforced polyphenol-based aggregate adhesive and its preparation method. The mesophase-reinforced polyphenol-based aggregate adhesive is based on a polyphenol-epoxy organic polymer-silanizing agent. Related studies have shown that polyphenol materials and epoxy polymer materials can easily form aggregates with certain adhesive properties through hydrogen bonding and hydrophobic interactions. However, adhesives formed solely by polyphenol-epoxy polymer materials exhibit poor stability and require improvement in adhesive performance. The primary objective of this invention is to propose a novel, convenient, and effective silanization-enhanced strategy based on organic-inorganic hybridization, thereby providing a method for preparing a tissue adhesive with good biocompatibility, good tissue adhesion, and stable bonding. Through blending of epoxy polymer materials and silanizing agents, the silanizing agent can use the epoxy polymer material as a template and growth site, and after hydrolysis, assemble in situ to form silanized structural units. Building upon this foundation, the addition of polyphenolic materials introduces a high density of phenolic hydroxyl groups, enabling them to form hydrogen bonds and hydrophobic interactions with structural units. This further stabilizes and forms a stable micro-phase structure. This micro-ordered structure, assembled from a silicified polymer network and polyphenols, is called a "mesoscopic phase" reinforcement. The presence of this reinforcement plays a crucial role in enhancing the adhesive's adhesion properties. After freeze-drying and grinding into powder, the powdered material retains high viscosity even after self-gelling upon absorbing water. This adhesive not only exhibits excellent antibacterial and antioxidant properties but also demonstrates superior performance in liver tissue adhesion and hemostasis models.

[0006] Specifically, the following technical solutions are included: In a first aspect, a method for preparing a mesoscopically reinforced polyphenolic aggregate adhesive is provided, comprising the following steps: Step 1: Prepare a solution of the polyphenol compound, stir it evenly, and set it aside for later use. This solution is called Solution A. Step 2: Prepare a solution of the epoxy-based polymer material for later use; this solution is called Solution B. Step 3: Dilute the silanizing reagent with acidic deionized water and set aside; this solution is called solution C. Step 4: Mix the B solution prepared in Step 2 and the C solution prepared in Step 3 and react for 1 hour. Then add the mixture to the A solution prepared in Step 1 to obtain a mixture. After shaking and centrifugation, discard the supernatant to obtain a mesoscopic phase-enhanced polyphenolic aggregate adhesive.

[0007] Preferably, in step one, the polyphenol compound is added to deionized water to prepare a solution; in step two, the epoxy-based polymer material is added to deionized water to prepare a solution.

[0008] Furthermore, in step one, the polyphenolic compound is selected from at least one of catechol, dopamine, 1-(3,4-dihydroxyphenyl)-2-aminoethanol, flavonol, ellagic acid, salicumin, hesperidin, kaempferol, morin, catechin, luteolin, quercetin, pyrogallol, gallic acid, baicalin, 1,2,4-phenylpyrogallol, tannic acid, epigallocatechin gallate, epicatechin gallate, and epigallocatechin.

[0009] Furthermore, in step one, the concentration of the polyphenol compound in the solution is 100-600 mg / mL.

[0010] Furthermore, in step two, the epoxy polymer material is selected from at least one of polyethylene glycol (PEG), polyethylene oxide, polypropylene oxide, Pluronic F127 (PF127), and Pluronic F68 (PF68).

[0011] Furthermore, in step two, the concentration of the epoxy polymer material in the solution is 100-400 mg / mL.

[0012] Furthermore, in step three, the silanizing agent is selected from at least one of methyltrimethoxysilane, methyldiethoxysilane, ethyltriethoxysilane, dimethyldiethoxysilane, methyltriethoxysilane, tetraethyl orthosilicate (TEOS), tetramethyl orthosilicate (TMOS), bis(trimethylsilyl)acetamide (BSA), 3-aminopropyltriethoxysilane (APTES), 3-mercaptopropyltrimethoxysilane (MPTMS), trimethylchlorosilane (TMCS), diethylenetriaminepropyltrimethoxysilane, chlorotriethoxysilane (TECS), N-aminoethyl-γ-aminopropyltrimethoxysilane, octyltrimethoxysilane, cyclohexylmethyldimethoxysilane, trimethoxysilane, triethoxysilane, propyltriethoxysilane, benzyltriethoxysilane, vinyltrimethoxysilane, and isobutyltriethoxysilane.

[0013] More preferably, the silanizing agent A is selected from at least one of triethoxysilane, dimethyldiethoxysilane, tetraethyl orthosilicate (TEOS), tetramethyl orthosilicate (TMOS), 3-aminopropyltriethoxysilane (APTES), diethylenetriaminopropyltrimethoxysilane, vinyltrimethoxysilane, N-aminoethyl-γ-aminopropyltrimethoxysilane, isobutyltriethoxysilane, and propyltriethoxysilane.

[0014] Furthermore, in step three, the pH of the acidic deionized water is 3-4.

[0015] Furthermore, in step four, the volume ratio of liquid B to liquid A is 1:(0.1-5).

[0016] Furthermore, in step four, the concentration of the silanizing agent in the mixture is 0.1-10 mM.

[0017] In a second aspect, a mesophase-reinforced polyphenolic aggregate adhesive is provided, which is prepared by the method for preparing the mesophase-reinforced polyphenolic aggregate adhesive described in the first aspect.

[0018] Compared with the prior art, the present invention provides a mesoscopic phase-reinforced polyphenol-based aggregate adhesive and its preparation method, which has the following beneficial effects: This invention utilizes polyphenolic compounds and epoxy-based polymers as the aggregate matrix, and pre-introduces a silanizing agent into the epoxy polymer. After hydrolysis, the silanizing agent assembles around the epoxy polymer to form an organic-inorganic hybrid composite phase. The introduction of this dense composite phase further enhances the hydrogen bonding and hydrophobic interactions with the polyphenolic compounds. Polyphenolic compounds are natural organic compounds, and various epoxy-based polymers are widely used in medical materials due to their biodegradability and high biocompatibility. In simulated tissue adhesion on pig skin, the introduction of the silicon-based material significantly improved the adhesion strength, reaching a maximum of 88.5 kPa. Furthermore, this adhesive meets the requirements for organ hemostasis, exhibits strong adhesion to organ tissue surfaces, and also possesses antibacterial and antioxidant properties, making it suitable as a good medical tissue adhesive. Attached Figure Description

[0019] To more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings used in the following description of the embodiments will be briefly introduced. Obviously, the drawings described below are some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0020] Figure 1 These are SEM images of different silicide concentrations from Examples 1-3 and Comparative Examples 1-2 of the present invention; Figure 2 These are organ hemostasis and occlusion test diagrams of Examples 1-3, Comparative Examples 1-5, and the control group of the present invention; Figure 3 The graphs show the liver blood loss test results for Examples 1-3, Comparative Examples 1-5, and the control group of the present invention. Figure 4 The SAXS spectra of Embodiment 1 and Comparative Example 2 of the present invention are shown below. Figure 5 These are the Fourier transform infrared spectra of Embodiment 1 and Comparative Example 2 of the present invention; Figure 6 This is a graph showing the cell viability assay of Example 1 of the present invention; Figure 7 This is a graph showing the hemolysis rate determination in Example 1 of the present invention; Figure 8 This is a graph showing the antibacterial performance test results of Example 1 of the present invention; Figure 9 This is a graph showing the antioxidant performance measurement of Example 1 of the present invention; Figure 10 The burst pressure measurement diagrams are for Embodiment 1 and Comparative Example 2 of the present invention. Detailed Implementation

[0021] The technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only some, not all, of the embodiments of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0022] It should be understood that, when used in this specification and the appended claims, the terms "comprising" and "including" indicate the presence of the described features, integrals, steps, operations, elements and / or components, but do not exclude the presence or addition of one or more other features, integrals, steps, operations, elements, components and / or collections thereof.

[0023] It should also be understood that the terminology used in this specification is for the purpose of describing particular embodiments only and is not intended to limit the invention. As used in this specification and the appended claims, the singular forms “a,” “an,” and “the” are intended to include the plural forms unless the context clearly indicates otherwise.

[0024] It should also be further understood that the term "and / or" as used in this specification and the appended claims refers to any combination of one or more of the associated listed items and all possible combinations, and includes such combinations.

[0025] To better understand the technical content of the present invention, the technical solution of the present invention will be further introduced and explained below with reference to specific embodiments.

[0026] Example 1 A mesoscopically reinforced polyphenolic aggregate adhesive comprises a polyphenol compound, an epoxy polymer, and a silanizing agent. The polyphenol compound is tannic acid, the epoxy polymer is PF127, and the silanizing agent is tetraethyl orthosilicate (TEOS).

[0027] A method for preparing a mesoscopically reinforced polyphenolic aggregate adhesive includes the following steps: Step 1: Add the polyphenol compound to deionized water to prepare a solution with a concentration of 500 mg / mL. Stir well and set aside. This solution is called Solution A. Step 2: Add the epoxy-based polymer material to deionized water to prepare a solution with a concentration of 200 mg / mL, and set it aside for later use. This solution is called Solution B. Step 3: Dilute the silanizing reagent 10 times with acidic deionized water at pH 3 and set aside; this solution is called solution C. Step 4: Mix the B solution prepared in Step 2 and the C solution prepared in Step 3 at a volume ratio of 9.1:1, react for 1 hour, and then add it to the A solution prepared in Step 1. The volume ratio of A solution to B solution is 4:1 to obtain a mixed solution. The final concentration of the silanizing reagent is 10 mM. After shaking and centrifugation, discard the supernatant to obtain the mesoscopic phase-enhanced polyphenolic aggregate adhesive of Example 1.

[0028] Example 2 A mesoscopically reinforced polyphenolic aggregate adhesive comprises a polyphenol compound, an epoxy polymer, and a silanizing agent. The polyphenol compound is tannic acid, the epoxy polymer is PF127, and the silanizing agent is tetraethyl orthosilicate (TEOS).

[0029] A method for preparing a mesoscopically reinforced polyphenolic aggregate adhesive includes the following steps: Step 1: Add the polyphenol compound to deionized water to prepare a solution with a concentration of 500 mg / mL. Stir well and set aside. This solution is called Solution A. Step 2: Add the epoxy-based polymer material to deionized water to prepare a solution with a concentration of 200 mg / mL, and set it aside for later use. This solution is called Solution B. Step 3: Dilute the silanizing reagent 100 times with acidic deionized water at pH 3 and set aside; this solution is called solution C. Step 4: Mix the B solution prepared in Step 2 and the C solution prepared in Step 3 at a volume ratio of 9.1:1, react for 1 hour, and then add it to the A solution prepared in Step 1. The volume ratio of A solution to B solution is 4:1 to obtain a mixed solution. The final concentration of the silanizing reagent is 1 mM. After shaking and centrifugation, discard the supernatant to obtain the mesoscopic phase-enhanced polyphenolic aggregate adhesive of Example 2.

[0030] Example 3 A mesoscopically reinforced polyphenolic aggregate adhesive comprises a polyphenol compound, an epoxy polymer, and a silanizing agent. The polyphenol compound is tannic acid, the epoxy polymer is PF127, and the silanizing agent is tetraethyl orthosilicate (TEOS).

[0031] A method for preparing a mesoscopically reinforced polyphenolic aggregate adhesive includes the following steps: Step 1: Add the polyphenol compound to deionized water to prepare a solution with a concentration of 500 mg / mL. Stir well and set aside. This solution is called Solution A. Step 2: Add the epoxy-based polymer material to deionized water to prepare a solution with a concentration of 200 mg / mL, and set it aside for later use. This solution is called Solution B. Step 3: Dilute the silanizing reagent 10 times with acidic deionized water at pH 3 and set aside; this solution is called solution C. Step 4: Mix the B solution prepared in Step 2 and the C solution prepared in Step 3 at a volume ratio of 18.2:1, react for 1 hour, and then add it to the A solution prepared in Step 1. The volume ratio of A solution to B solution is 4:1 to obtain a mixed solution. The final concentration of the silanizing agent is 5mM. After shaking and centrifugation, discard the supernatant to obtain the mesoscopic phase-enhanced polyphenolic aggregate adhesive of Example 3.

[0032] Comparative Example 1 A mesoscopically reinforced polyphenolic aggregate adhesive comprises a polyphenol compound, an epoxy polymer, and a silanizing agent. The polyphenol compound is tannic acid, the epoxy polymer is PF127, and the silanizing agent is tetraethyl orthosilicate (TEOS).

[0033] A method for preparing a mesoscopically reinforced polyphenolic aggregate adhesive includes the following steps: Step 1: Add the polyphenol compound to deionized water to prepare a solution with a concentration of 500 mg / mL. Stir well and set aside. This solution is called Solution A. Step 2: Add the epoxy-based polymer material to deionized water to prepare a solution with a concentration of 200 mg / mL, and set it aside for later use. This solution is called Solution B. Step 3: Dilute the silanizing reagent 5 times with acidic deionized water at pH 3, and set aside as solution C; Step 4: Mix the B solution prepared in Step 2 and the C solution prepared in Step 3 at a volume ratio of 9.1:1, react for 1 hour, and then add it to the A solution prepared in Step 1. The volume ratio of A solution to B solution is 4:1 to obtain a mixed solution. The final concentration of the silanizing reagent is 20mM. After shaking and centrifugation, discard the supernatant to obtain the mesoscopic phase-enhanced polyphenolic aggregate adhesive of Comparative Example 1.

[0034] Comparative Example 2 A polyphenol-based aggregate adhesive without mesophase reinforcement, which differs from Example 1 in that it consists only of polyphenol compounds and epoxy polymer materials, while all other conditions are the same.

[0035] Comparative Example 3 Comparative Example 3 provides a method for preparing a mesoscopic phase-enhanced polyphenolic aggregate adhesive. The difference from Example 1 is that in step four, the volume ratio of liquid B to liquid A is 1:0.05, while all other conditions are the same.

[0036] Comparative Example 4 Comparative Example 4 provides a method for preparing a mesoscopic phase-enhanced polyphenolic aggregate adhesive. The difference from Example 1 is that in step four, the volume ratio of liquid B to liquid A is 1:6, while all other conditions are the same.

[0037] Comparative Example 5 Comparative Example 5 refers to patent CN120285274A, a polyphenolic tissue adhesive for promoting wound healing and its preparation method, and provides a method for preparing a polyphenolic aggregate adhesive without mesophase reinforcement. Compared with Example 1, the difference is that in step three, the silanizing agent is replaced with recombinant mussel adhesive protein, which is diluted with deionized water to a concentration of 10 mg / mL and set aside as solution C. All other conditions are the same.

[0038] Performance Testing—Adhesion Performance Testing: To investigate the adhesion performance of the cohesive adhesive, pigskin was used to simulate human skin tissue, and its adhesion properties were evaluated through an lap shear test. After cleaning and degreasing, the pigskin was cut into strips of 1cm × 4cm, and 100 μL of adhesive was directly applied to the surface of the pigskin. Another skin sample was superimposed on one side of the material coating to establish a 10 × 10 mm contact area. The assembly was compressed under 200g for 30 min, followed by an lap shear test at a tensile rate of 10cm / min. The shear adhesion strength was calculated by dividing the maximum stress by the contact area. All tests were conducted under ambient conditions, with three samples per experimental group.

[0039] Adhesion performance tests were conducted on Examples 1-3 and Comparative Examples 1-5, and the test results are shown in Table 1 below: Table 1. Adhesion performance test results of Examples 1-3 and Comparative Examples 1-5

[0040] As shown in Table 1, the adhesive prepared by the method for preparing the mesoscopic phase-reinforced polyphenol-based aggregate adhesive of this invention exhibits strong adhesion. Furthermore, the addition of epoxy-based organic polymer materials allows the hydrogel to autodegrade after a period of time, and the introduction of natural polyphenol materials also imparts better biocompatibility to the system. Due to the excessive silicon introduced in Comparative Example 1, over-silicification resulted in the formation of nanoparticles (such as…). Figure 1 As shown, Figure 1 The images show SEM images of different silanization concentrations in Examples 1-3 and Comparative Examples 1-2 of this invention. These images indicate decreased compatibility and uniformity between silicon and the polymer chains, leading to reduced adhesion performance. Comparative Example 2, lacking a silanizing agent in its preparation method, failed to assemble a mesoscopic organic-inorganic hybrid composite phase around the epoxy polymer phase, resulting in poor adhesive adhesion performance. Comparative Examples 3 and 4, not using liquids A and B within the volume ratio range specified in this invention, failed to achieve optimal system performance. Even when using an adhesive component instead of the silanizing agent in Comparative Example 5, the significant improvement in adhesion strength achieved by introducing the silanizing agent could not be realized.

[0041] To verify that the agglomerated adhesive formed by this system still possesses strong adhesive properties even when in powder form after absorbing water, the adhesive materials obtained from each group were placed in a -80°C freezer overnight, then freeze-dried overnight in a vacuum freeze dryer, and finally thoroughly ground in a mortar to form a powdered adhesive hemostatic material. Adhesion strength tests were performed on the powdered materials of each group: 20 mg of powder material was added to a moist pigskin surface to establish a 10 × 10 mm contact area. The components were compressed at 200 g weight for 30 min, followed by an overlap shear test at a tensile rate of 10 cm / min. Shear adhesion strength was calculated by dividing the maximum stress by the contact area. All tests were conducted under environmental conditions, with three samples from each experimental group.

[0042] Table 2 Adhesion strength of self-gelling powder materials in each group

[0043] Performance Testing—Organ Hemostasis and Occlusion Test: To investigate the hemostatic and occlusive effects of cohesive adhesives on organs, a mouse liver was used as a blood loss model. The abdominal cavity of anesthetized mice (BalB / C, weighing 28-33g, aged 8-10 weeks) was opened, and a pre-weighed (W0) medical gauze was placed under the liver. An incision (1cm in length, 0.5cm in depth) was made in the liver using surgical scissors. After 3 seconds of bleeding, powdered materials from different experimental groups were added to the wound surface. The control group received no material treatment. A small amount of physiological saline was dripped onto the gauze, which adhered to the wound surface for hemostasis and occlusion. One minute later, the gauze that had absorbed blood was weighed (Wt), and the blood loss was calculated using the formula: W = Wt - W0. Figure 2 These are organ hemostasis and occlusion test diagrams for Examples 1-3, Comparative Examples 1-5, and the control group of the present invention. Figure 3 The graphs show the liver blood loss test results for Examples 1-3, Comparative Examples 1-5, and the control group of the present invention.

[0044] Depend on Figures 2-3 The test results show that the adhesive prepared by the method of preparing the mesoscopic phase-reinforced polyphenolic aggregate adhesive in this embodiment of the invention has strong wet adhesion ability and can meet the requirements of liver hemostasis. The control group mice died of shock due to the lack of hemostasis treatment. Example 1, due to the strong adhesion of the mesoscopic phase-reinforced polyphenolic aggregate adhesive, can seal in time, and the added silicon network helps promote coagulation, thus exhibiting the best hemostatic performance and the least blood loss. Comparative Example 1 did not use the optimal concentration of silanizing reagent, Comparative Example 2 did not add silanizing reagent, and Comparative Examples 3 and 4 did not use solutions A and B within the volume ratio range specified in this invention; therefore, the test results of their adhesives for liver hemostasis and sealing were poor. Even when using an adhesive component to replace the silanizing reagent in this invention, Comparative Example 5 could not achieve the technical effect of significantly improving adhesion strength and enhancing hemostasis by introducing silanizing reagent, as described in this invention.

[0045] Structural characterization—scanning electron microscopy, small-angle X-ray diffraction, and Fourier transform infrared spectroscopy: To further demonstrate the existence of the "mesoscopic phase" reinforcing body, the tissue adhesive was subjected to scanning electron microscopy and small-angle X-ray diffraction. Figure 1As shown, with increasing silicide concentration, the pores of the aggregates gradually become more uniform and smooth (1-10 mM). The silicide network around the polymer chains gradually plays a role in making the system more uniform. However, when over-silicided (20 mM), the hydrolyzed TEOS forms silica particles that fill the pores, and the pore diameter also increases significantly. This indicates that at higher concentrations, silicon tends to form nanoparticles rather than a uniform mesoscopic silicide network with the polymer chains. This may be a factor that reduces the uniformity and cohesion of the material under high concentration conditions, leading to a decrease in adhesion performance. Electron microscopy images of adhesives prepared at low concentrations (1-10 mM) show that no obvious silicon particles were formed in any group. This corresponds to the trend in Table 1 where the adhesion performance first increases and then decreases as the silanization concentration increases from low to high (1 mM-20 mM). This invention, by introducing an appropriate dose of pre-hydrolyzed silanizing reagent to react with epoxy polymers, allows for further hydrolysis of orthosilicic acid to form Si-O-Si bonds. Amorphous silica surrounds the polymer chains, forming a stable and controllable silanized network that does not tend to generate silica particles. After TA assembly, an ordered structure of a "mesoscopic phase" reinforced body is formed. Furthermore, it can be seen that the porosity formed in Example 1 (10 mM) is the smallest, indicating that the network crosslinking density formed under this condition is the largest, further illustrating the reinforcing mechanism of the mesoscopic phase reinforced body.

[0046] To further illustrate the size of the ordered structure formed by the "mesoscopic phase" reinforcer, the aggregate adhesives prepared in Example 1 (10 mM) and Comparative Example 2 (0 mM) were tested by small-angle X-ray diffraction. The one-dimensional data obtained were analyzed and calculated using Origin 2021 and SASfit version 0.94.12 software. Figure 4 These are the SAXS spectra of Embodiment 1 and Comparative Example 2 of the present invention. Figure 4 It can be seen that, compared to Comparative Example 2, at 0.1 nm -1 The SAXS spectrum of nearby Example 1 shifted upward, indicating that after adding an appropriate concentration of TEOS, an ordered structural distribution of a certain size appeared inside the aggregate adhesive.

[0047] The results obtained through further simulation calculations using the software are shown in Table 3. Combined with Comparative Example 2, the adhesive of Example 1 was simulated and analyzed. The ordered structure size of the "mesoscopic phase" reinforced body prepared by this method is 18.17±0.28nm.

[0048] Table 3 Simulation results of Example 1

[0049] Note: N can represent the number of particles per unit volume, and σ can be used as the standard deviation of the size; the larger the value, the wider the size distribution. R0 represents the size of an ordered structure.

[0050] To further verify the synthesized condensed material enhanced by the silicide-mediated "mesoscopic phase," Fourier transform infrared spectroscopy was used to analyze TA, PF127, the material synthesized in Example 1, and the material synthesized in Comparative Example 2. Samples were prepared by pressing freeze-dried powder samples into potassium bromide (KBr) pellets and then tested using a Bruker Invenio infrared spectrometer in transmission mode, with a scan wavenumber range of 400-4000 cm⁻¹. -1 The scan was performed 4 times, with a resolution of 4 cm. -1 The result is as follows Figure 5 As shown.

[0051] It can be seen that Example 1 not only retains TA 1614 cm -1 The C=C stretching vibration peak on the aromatic ring, 1536 cm⁻¹ -1 The C-C stretching vibration peak on the aromatic ring, 1448 cm⁻¹ -1 CH bending vibration peak at 758 cm -1 The CH torsional vibration peak on the aromatic ring is also preserved at 2886 cm⁻¹. -1 The stretching vibration peak of nearby alkanes (CH). Additionally, a peak located at 1714 cm⁻¹. -1 The absorption peak is due to the stretching vibration of the carbonyl group (-C=O) in the intramolecular ester bond, shifted to 1724 cm⁻¹. -1 The energy of the C=O bond is increased, located at 2886 cm⁻¹. -1 The absorption peak is the stretching vibration peak of alkane (CH), shifted to 2921 cm⁻¹. -1 This indicates that it is influenced by the interaction with hydrogen donors (-OH), forming intramolecular hydrogen bonds, meaning that the material is mainly formed by multiple hydrogen bonds. Compared to Comparative Example 2, Example 1 at 474 cm⁻¹... -1 A peak appeared at the point, which corresponds to the bending vibration absorption peak of Si-O-Si, indicating the successful synthesis of condensed materials enhanced by the "mesoscopic phase" mediated by the silicide network.

[0052] Performance Testing—Biocompatibility, Antibacterial and Antioxidant Performance Testing: To further verify the biocompatibility, antibacterial properties, and antioxidant properties of the adhesive, the following tests were performed on the optimal group obtained in Example 1.

[0053] Cytotoxicity: Lyophilized adhesive material (1 mg) was extracted with 10 mL DMEM medium for 12 h to obtain a 100% extract, which was then serially diluted to obtain 50%, 25%, 12.5%, and 6.25% extracts. HUVEC and L929 cells were seeded at 8000 cells per well in 96-well plates and incubated with different concentrations of extract for 24 h. Cell viability was then assessed using the CCK8 assay. Figure 6 This is a graph showing the cell viability assay in Example 1 of the present invention.

[0054] HaCAT and L929 cells were treated with different concentrations of material extracts, respectively. Figure 6 The results of the assay showed that the adhesive had good biocompatibility and no obvious cytotoxicity.

[0055] Blood compatibility assay: The hemolytic properties of the adhesive were assessed by a hemolysis test. Briefly, 300 μL of 5% erythrocyte (RBC) suspension was incubated with 10 mg of adhesive powder at 37°C for 2 hours. Subsequently, 1 mL of PBS solution was added, and the mixture was centrifuged at 2000 rpm for 10 minutes. The absorbance (A) of the supernatant was then measured at 540 nm. gel For comparison, 300 μL of 5% RBC suspension was mixed with 1 mL of deionized water (A). DW A positive control was prepared by mixing 300 μL of 5% RBC suspension with 1 mL of PBS (A). PBS A mixture was used to prepare a negative control. The percentage of hemolysis was calculated using the following formula.

[0056] Hemolysis rate (%) = [(A gel - A PBS ) / (A DW - A PBS )] ×100%. Figure 7 This is a graph showing the hemolysis rate determination in Example 1 of the present invention.

[0057] Depend on Figure 7 As shown in the measurement chart, the hemolysis rate measured in Example 1 was less than 5%, indicating that the results meet the requirements for material biosafety.

[0058] Antibacterial performance test: Take bacterial suspension (OD 600) nm= 0.5) Dilute 10-fold with physiological saline. Then, mix 10 μL of the diluted bacterial suspension with 100 μL of adhesive extract solutions at different concentrations (1, 5, 10 mg / mL) and incubate at 37°C for 12 hours. After incubation, add 890 μL of PBS to the treated bacterial suspension and mix thoroughly. Then, further dilute the solution 10-fold with PBS. Next, spread 10 μL of the diluted bacterial suspension onto LB agar plates. Incubate the plates at 37°C for 24 hours, then photograph and count the colonies. The experiment was repeated 3 times. Figure 8 This is a graph showing the antibacterial performance test results of Example 1 of the present invention.

[0059] Depend on Figure 8 The test results show that when the concentration of the extract reaches 1 mg / mL, the antibacterial activity against Escherichia coli and Staphylococcus aureus is over 98%, indicating that the material has strong antibacterial properties.

[0060] Antioxidant performance test: HaCat cells were seeded at a density of 10,000 cells per well in 96-well plates and incubated in a cell culture incubator for 24 hours. Subsequently, the original medium was replaced with an adhesive extract (prepared by extracting the material in DMEM medium at a concentration of 50 μg / mL and adding 500 μM H2O2). DMEM medium without H2O2 served as a positive control, and DMEM medium containing 500 μM H2O2 served as a negative control. After an additional 24 hours of incubation, ROS probes were loaded in situ. DAPI was added after ROS, and the plates were incubated at room temperature for 5 minutes. DAPI and DCHF-DA staining solution were aspirated, and the plates were washed 2-3 times with PBS for 3-5 minutes each time, and examined under a fluorescence microscope. Figure 9 This is a graph showing the antioxidant performance measurement of Example 1 of the present invention.

[0061] Depend on Figure 9 The results showed that HaCat cells experienced increased reactive oxygen species (ROS) after H2O2 treatment, while treatment with a 50 μg / mL adhesive material extract significantly reduced ROS production, indicating that the material can effectively help cells alleviate oxidative stress.

[0062] The burst pressure of medical tissue adhesives is a core method for evaluating their resistance to rupture and sealing performance under physiological loads. It directly determines the structural stability of the adhesive layer under the action of body fluid pressure and tissue tension. Furthermore, the materials of Example 1 and Comparative Example 1 were tested on small intestinal tissue using a burst pressure testing device. The test procedure was as follows: A 100 mm diameter, 1 mm thick pig small intestine was cleaned. A hole approximately 2 mm in diameter was drilled in the outer casing using a syringe. The intestine was then cleaned with deionized water. One end of the small intestine was tied with a rope, and the other end was connected to a tube connected to a pneumatic pump and fixed. The pressure value in the entire closed system was measured using a digital pressure gauge. Next, the tissue adhesive was applied to the surface of the moistened intestinal casing's air outlet. Subsequently, a certain flow rate of gas was continuously introduced into the closed system until the casing ruptured. The maximum reading on the digital pressure gauge before the pressure drop was considered the maximum burst pressure. The results are as follows: Figure 10 As shown.

[0063] Depend on Figure 10 The test results show that, since normal human blood pressure generally does not exceed 120 mmHg (16 kPa), the burst pressure of the adhesive in Comparative Example 2 is about 18.7 kPa, only slightly higher than the safe pressure (16 kPa), indicating a certain risk of leakage. In contrast, the burst pressure of the adhesive in Example 1 is about 30.2 kPa, far exceeding the safe pressure. This demonstrates that the mesophase-reinforced aggregate adhesive synthesized in this invention better meets the requirements for tissue hemostasis and sealing. Due to the presence of the "mesophase" reinforcing structure in the TP-Si group, the cohesive effect of the adhesive is enhanced, and the maximum burst pressure it resists also increases accordingly.

[0064] The above description is merely a specific embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Any person skilled in the art can easily conceive of various equivalent modifications or substitutions within the technical scope disclosed in the present invention, and these modifications or substitutions should all be covered within the scope of protection of the present invention. Therefore, the scope of protection of the present invention should be determined by the scope of the claims.

Claims

1. A method for preparing a mesoscopically reinforced polyphenolic aggregate adhesive, characterized in that, Includes the following steps: Step 1: Prepare a solution of the polyphenol compound, stir it evenly, and set it aside for later use. This solution is called Solution A. The concentration of the polyphenol compound in the solution is 100-600 mg / mL; Step 2: Prepare a solution of the epoxy-based polymer material for later use, referred to as solution B; the epoxy-based polymer material is selected from Pluronic F127, and the concentration of the prepared solution is 100-400 mg / mL; Step 3: Dilute the silanizing reagent with acidic deionized water and set aside for later use; this solution is called solution C. The pH of the acidic deionized water is 3-4. Step 4: Mix the B solution prepared in Step 2 and the C solution prepared in Step 3 and react for 1 hour. Then add the mixture to the A solution prepared in Step 1 to obtain a mixed solution. After shaking and centrifugation, discard the supernatant to obtain a mesoscopic phase-enhanced polyphenolic aggregate adhesive. The volume ratio of the B solution to the A solution is 1:(0.1-5), and the concentration of the silanizing agent in the mixed solution is 0.1-10 mM.

2. The method for preparing the mesoscopic phase-reinforced polyphenolic aggregate adhesive as described in claim 1, characterized in that, In step one, the polyphenolic compound is selected from at least one of catechol, dopamine, 1-(3,4-dihydroxyphenyl)-2-aminoethanol, flavonol, ellagic acid, salicumin, hesperidin, kaempferol, morin, catechin, luteolin, quercetin, pyrogallol, gallic acid, baicalin, 1,2,4-phenylpyrogallol, tannic acid, epigallocatechin gallate, epicatechin gallate, and epigallocatechin.

3. The method for preparing the mesoscopic phase-reinforced polyphenolic aggregate adhesive as described in claim 1, characterized in that, In step three, the silanizing agent is selected from at least one of methyltrimethoxysilane, methyldiethoxysilane, ethyltriethoxysilane, dimethyldiethoxysilane, methyltriethoxysilane, tetraethyl orthosilicate, tetramethyl orthosilicate, bis(trimethylsilyl)acetamide, 3-aminopropyltriethoxysilane, 3-mercaptopropyltrimethoxysilane, trimethylchlorosilane, diethylenetriaminopropyltrimethoxysilane, chlorotriethoxysilane, N-aminoethyl-γ-aminopropyltrimethoxysilane, octyltrimethoxysilane, cyclohexylmethyldimethoxysilane, trimethoxysilane, triethoxysilane, propyltriethoxysilane, benzyltriethoxysilane, vinyltrimethoxysilane, and isobutyltriethoxysilane.

4. A mesoscopic phase-reinforced polyphenol-based aggregate adhesive, characterized in that, It is prepared by the method for preparing the mesoscopic phase-reinforced polyphenolic aggregate adhesive according to any one of claims 1-3.

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