Mesoporous bioactive enhanced cartilage-adhesive hydrogel and method of making the same

Abstract

This invention relates to the field of biomaterials technology, specifically to a mesoporous bioactive enhanced cartilage adhesion hydrogel and its preparation method. The preparation method is as follows: Ethyl acetate, ammonia solution, tetraethyl orthosilicate, and calcium nitrate are added to a hexadecyltrimethylammonium bromide dispersion, and the mixture is stirred at 55-65°C for 5-8 hours to obtain mesoporous bioactive glass nanoparticles (MBGNs). Highly reactive AG is prepared using gelatin, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride, and N-hydroxysuccinimide as raw materials. 2-8% (w / w) of MBGNs is added to a DAS solution to obtain a DAS / MBGNs mixed solution, and then the AG solution is added and stirred thoroughly to obtain the mesoporous bioactive enhanced cartilage adhesion hydrogel (ADGM). The prepared ADGM hydrogel provides stable support and stress dispersion, and also achieves covalent adhesion at the cartilage tissue interface and regulation of the biological microenvironment, realizing a synergistic unity of structural stability and functional repair.

Description

Technical Field

[0001] This invention relates to the field of biomaterials technology, and more specifically, to a mesoporous bioactive enhanced cartilage adhesion hydrogel and its preparation method. Background Technology

[0002] The growth plate is a hyaline cartilage tissue located between the epiphysis and metaphysis, composed of a quiescent layer, a proliferative layer, and a hypertrophic layer. The cell division activity of the proliferative layer directly determines the longitudinal growth of the bone, and its closure status can be determined by measuring the ratio of sound velocity using ultrasound. After puberty, the depletion of proliferative capacity leads to the closure of the growth plate, and after calcification into bone, longitudinal bone growth ceases. This cartilage tissue is regulated by both genes and the endocrine system; abnormal activity can lead to skeletal dysplasia or deformities.

[0003] However, in daily life, growth plates are often damaged due to traumatic fractures, iatrogenic injuries, or infections. The growth plate (epithelial plate) is a longitudinally arranged columnar cartilage structure that primarily bears longitudinal compressive stress. If abnormal osteogenesis occurs during the repair process, bone bridges can easily form, leading to growth disorders and limb deformities. Current clinical treatments mainly rely on Kirschner wire fixation, autologous fat grafting, or artificial material isolation repair. Kirschner wire fixation achieves mechanical fixation and structural stability by inserting rigid metal needles into the damaged area, but it is a purely mechanical support method, lacking the ability to biointegrate with cartilage tissue, requiring secondary surgery for removal, and unable to regulate the local microenvironment. Fat grafting only provides physical filling, but it suffers from a series of key drawbacks, including weak mechanical strength, unstable bone bridge inhibition, and a tendency to induce inflammation and complications, making it difficult to maintain long-term stability of the damaged area under dynamic load conditions. Artificial materials, such as traditional bone cement and cyanoacrylate, have high bonding strength, but suffer from problems such as thermal damage, poor biocompatibility, and non-degradability. While existing hydrogel materials have a certain degree of biocompatibility, they generally suffer from defects such as insufficient mechanical strength, poor interfacial stability in humid environments, and inability to withstand periodic compressive stress in joint and growth plate areas, making it difficult to replace Kirschner wires in providing stable support.

[0004] Therefore, it is of great significance to develop a biodegradable repair material that can provide Kirschner needle-like stable support and stress dispersion, as well as achieve covalent adhesion at the cartilage tissue interface and regulation of the biological microenvironment. Summary of the Invention

[0005] To address the aforementioned technical problems, the present invention aims to provide a mesoporous bioactive enhanced cartilage adhesion hydrogel. By constructing a composite hydrogel structure that combines a dynamic covalent cross-linked network with a mesoporous bioactive enhancement system, it can provide stable support and stress dispersion in the damaged area, and also achieve covalent adhesion and biological microenvironment regulation at the cartilage tissue interface. Thus, it achieves a synergistic unity of structural stability and functional repair without the need for internal metal fixation.

[0006] The objective of this invention is achieved through the following technical solution: A method for preparing a mesoporous bioactive enhanced cartilage adhesion hydrogel includes the following steps: (1) Disperse hexadecyltrimethylammonium bromide in deionized water, then add ethyl acetate, ammonia solution, tetraethyl orthosilicate and calcium nitrate in sequence, and stir the reaction at 55-65℃ for 5-8 hours; after centrifuging, washing and drying the obtained product, calcine at 600-700℃ to remove impurities, and obtain mesoporous bioactive glass nanoparticles MBGNs. (2) Dissolve gelatin in MES buffer, add 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and N-hydroxysuccinimide, stir for 1-2 hours; add ethylenediamine dropwise, adjust the reaction system to neutral, and continue stirring for 3.5-4.5 hours; after the reaction is complete, dialyze the mixture for 60-80 hours and freeze dry to obtain highly reactive AG; (3) Dissolve AG and DAS separately in phosphate-buffered saline (PBS) to prepare AG solution with a mass fraction of 30-35% and DAS solution with a mass fraction of 10-15%; add MBGNs with a mass fraction of 2-8% to the DAS solution to obtain DAS / MBGNs mixed solution, then add AG solution and stir thoroughly, freeze-dry to obtain mesoporous bioactive enhanced cartilage adhesion hydrogel ADGM. In addition to internal cross-linking during the formation process, the residual aldehyde groups can covalently bind with the amino groups of proteins at the tissue interface, and synergistically interact with hydrogen bonds to achieve mechanical intercalation with nanoparticles, thereby achieving robust adhesion and fixation of cartilage and bone tissue.

[0007] According to a preferred embodiment of the present invention, the mass ratio of tetraethyl orthosilicate to calcium nitrate in step (1) is (1.5-2.5):1.

[0008] According to a preferred embodiment of the present invention, the mass percentage concentration of ammonia in step (1) is 0.5-2.0%.

[0009] According to a preferred embodiment of the present invention, the calcination time in step (1) is 3-5 hours.

[0010] According to a preferred embodiment of the present invention, in step (2), the mass ratio of gelatin, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and N-hydroxysuccinimide is (0.5-1.5):(0.15-0.3):(0.1-0.3).

[0011] According to a preferred embodiment of the present invention, the reaction temperature before adding ethylenediamine in step (2) is 40-50°C, and the reaction temperature after adding ethylenediamine is 20-30°C.

[0012] According to a preferred embodiment of the present invention, in step (3), the preparation method of DAS is as follows: corn amylopectin is dissolved in deionized water, sodium periodate solution is added, and the mixture is stirred and reacted under light-protected conditions. The resulting solution is centrifuged, washed, and then freeze-dried. This component can undergo a Schiff base reaction with the amino groups on the AG molecular chain to form dynamic C=N covalent bonds, thereby constructing a three-dimensional cross-linked network. This dynamic cross-linked structure has a certain degree of reversibility under physiological conditions, enabling the material to have buffering and self-adaptive capabilities under external stress.

[0013] According to a preferred embodiment of the present invention, the reaction is carried out at 35-40°C for 10-12 hours; the freeze-drying time is 72-80 hours.

[0014] According to a preferred embodiment of the present invention, the volume ratio of AG solution to DAS / MBGNs mixed solution in step (3) is 1:1.

[0015] Compared with the prior art, the beneficial effects of the present invention are as follows: The preparation method of the mesoporous bioactive enhanced cartilage adhesion hydrogel of the present invention forms a nanoscale reinforced support framework by uniformly distributing mesoporous bioactive glass nanoparticles in a three-dimensional network. This enables the material to effectively transfer and disperse axial stress under compression, significantly improve the compressive modulus, and maintain a stable linear elastic response within the physiological strain range. Thus, it plays a structural stabilizing and supporting role similar to Kirschner wires in the damaged areas of cartilage and growth plate.

[0016] Unlike the rigid, concentrated support of traditional metal Kirschner wires, the mesoporous, bioactive, enhanced cartilage adhesion hydrogel of this invention forms a distributed micro-support network, providing stability while avoiding stress concentration damage. Simultaneously, the dynamic Schiff base cross-linking network endows the material with excellent ductility and fatigue resistance, enabling it to adapt to the dynamic deformation environment of cartilage tissue. Furthermore, during gelation, the material forms a covalent anchor with the tissue interface, combining hydrogen bonds and mechanical interlocking to achieve robust fixation and biointegration. Thus, it achieves a unified "Kirschner wire-like" support function and tissue repair function without the need for metal internal fixation. Attached Figure Description

[0017] The technical solution of the present invention will be further described in detail below with reference to the accompanying drawings, but this does not constitute any limitation on the present invention.

[0018] Figure 1Characterization of mesoporous bioactive glass nanoparticles (MBGNs) according to embodiments of the present invention. A is a SEM image of MBGNs; B is the particle size distribution curve of MBGNs (DLS test results); C is a statistical graph of the average particle size of MBGNs.

[0019] Figure 2 This is a SEM image of the AGDM hydrogel in an embodiment of the present invention.

[0020] Figure 3 The dynamic mechanical properties of the AGDM hydrogel in this embodiment of the invention are shown. A is the frequency-scan rheological curve of the AGDM hydrogel; B is the strain-scan rheological curve of the AGDM hydrogel.

[0021] Figure 4 The following are the uniaxial compression test results of the AGDM hydrogel according to an embodiment of the present invention. A is the compression curve of the AGDM hydrogel; B is the quantitative analysis result of the compressive modulus of the AGDM hydrogel.

[0022] Figure 5 This diagram illustrates the adhesion properties of the AGDM hydrogel in an embodiment of the present invention. A represents the overlap shear adhesion curve of the AGDM hydrogel on the bone fragment; B represents the average adhesion strength of the AGDM hydrogel to the bone fragment.

[0023] Figure 6 This image shows the recovery of growth plate damage 8 weeks post-surgery in an embodiment of the present invention. Image A shows an X-ray image of growth plate damage treated with Kirschner wires 8 weeks post-surgery; image B shows an X-ray image of growth plate damage treated with AGDM hydrogel 8 weeks post-surgery. Detailed Implementation

[0024] The technical solution of the present invention will be further described in detail below with reference to specific embodiments, but this does not constitute any limitation on the present invention. Example 1

[0025] (1) 1.40 g of hexadecyltrimethylammonium bromide (CTAB) was dispersed in 66 mL of deionized water. Then, 20 mL of ethyl acetate (EA), 28 mL of 1 mol / L ammonia solution, 14.40 mL of tetraethyl orthosilicate (TEOS), and 6.52 g of calcium nitrate were added slowly in sequence. The mixture was stirred at 60 °C for 5 hours. The resulting product was centrifuged at 8000-10000 r / min for 12 min. The lower layer was washed alternately with anhydrous ethanol and deionized water 3-5 times. The precipitate was collected and dried under vacuum at 70 °C for 10 hours, followed by calcination at 600 °C for 4 hours to remove organic matter and nitrates, yielding mesoporous bioactive glass nanoparticles (MBGNs). The characterization of the mesoporous bioactive glass nanoparticles (MBGNs) is as follows: Figure 1 As shown, Figure 1-A shows that the particles exhibit a uniformly dispersed, near-spherical nanostructure with a relatively uniform particle size distribution. Figure 1 -B indicates that the particle size is mainly concentrated in the range of approximately 100-200 nm. Figure 1 -C results show that its average particle size is approximately 150 nm, exhibiting good size uniformity and stability. Therefore, MBGNs possess a regular mesoporous structure, high specific surface area, and can be uniformly embedded in organic networks. Through physical filling and interfacial interactions, they enhance network rigidity and simultaneously act as ion release carriers to regulate the local microenvironment.

[0026] (2) Dissolve 1 g of gelatin in 20 mL of 0.1 mol / L, pH 5.5 MES buffer solution, and control the temperature at 40℃. Then add 0.25 g of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC) and 0.15 g of N-hydroxysuccinimide (NHS), and stir for 1 hour. Then add 0.3 mL of ethylenediamine (EDA), and adjust the pH of the system to 7.0 with 1 mol / L sodium hydroxide solution. Stir the reaction at room temperature for 4 hours. Put the reaction mixture into a dialysis bag with a molecular weight cutoff of 12 kDa, dialyze in distilled water for 3 days, and change the distilled water every 12 hours. Freeze-dry the mixture to obtain highly reactive AG, which provides sufficient reaction sites for subsequent cross-linking and interfacial bonding.

[0027] (3) Dissolve 5 g of corn amylopectin in 50 mL of deionized water at 37 °C, and then slowly add 5 g of sodium periodate solution to the suspension to prepare oxidized starch. The reaction mixture was stirred at 37 °C in the dark for 12 hours. After the oxidation reaction was completed, the resulting solution was transferred to a centrifuge tube and centrifuged at 3500 rpm for 6 minutes. The resulting white precipitate was repeatedly washed with deionized water and centrifuged. After washing, it was freeze-dried for 3 days to obtain dialdehyde starch (DAS).

[0028] (4) Dissolve AG in 1 mL of phosphate-buffered saline (PBS) to prepare a 30% AG solution; dissolve dialdehyde starch (DAS) in 1 mL of PBS to prepare a 10% DAS solution in a centrifuge tube. Add 2% MBGNs to the DAS solution and stir thoroughly to dissolve it; mix the AG solution and the DAS / MBGNs mixed solution at a volume ratio of 1:1 and stir thoroughly to obtain the pore bioactivity enhanced cartilage adhesion hydrogel ADGM. Example 2

[0029] Add 4% MBGNs by mass to the DAS solution and stir thoroughly to dissolve it; mix the AG solution with the DAS / MBGNs mixed solution at a volume ratio of 1:1 and stir thoroughly to obtain the porous bioactive enhanced cartilage adhesion hydrogel ADGM.

[0030] The remaining steps are the same as in Example 1. Example 3

[0031] Add 6% MBGNs to the DAS solution and stir thoroughly to dissolve it; mix the AG solution with the DAS / MBGNs mixed solution at a volume ratio of 1:1 and stir thoroughly to obtain the porous bioactive enhanced cartilage adhesion hydrogel ADGM. Example 4

[0032] Add 8% MBGNs to the DAS solution and stir thoroughly to dissolve it; mix the AG solution with the DAS / MBGNs mixed solution at a volume ratio of 1:1 and stir thoroughly to obtain the porous bioactive enhanced cartilage adhesion hydrogel ADGM.

[0033] The porous bioactive enhanced cartilage adhesion hydrogels prepared in Examples 1-4 were designated ADGM-1, ADGM-2, ADGM-3, and ADGM-4, respectively. The microstructure of the AGDM hydrogels was characterized using scanning electron microscopy (SEM), and the morphology and pore size results are shown below. Figure 2 As shown, all AGDM hydrogel samples exhibited a uniform, interconnected three-dimensional porous network structure, a key structural feature for tissue adhesion and regeneration scaffold applications. With increasing MBGN content, the hydrogel's porous structure maintained good connectivity, while the pore wall roughness gradually increased. This confirms that MBGNs were successfully and uniformly dispersed in the hydrogel cross-linked network, without significant particle aggregation. The interconnected porous structure not only endows the hydrogel with excellent deformation adaptability to match the dynamic physiological activities of cartilage tissue but also provides a favorable microenvironment for nutrient transport and cell infiltration during cartilage regeneration.

[0034] To further verify the performance of the prepared porous bioactive enhanced cartilage adhesion hydrogel, the following experiments were conducted: Experimental Example 1: Mechanical Properties of AGDM Hydrogel The dynamic mechanical properties of AGDM hydrogels with different MBGNs contents were evaluated using a rotational rheometer. Frequency sweep test results (Figure 3A) showed that, across the entire frequency range of 0.1–10 Hz, the storage modulus (G′) of all AGDM hydrogel samples was consistently significantly higher than the loss modulus (G″), and no intersection between G′ and G″ was observed. This rheological behavior indicates that all AGDM hydrogels exhibit typical elastic solid properties and maintain a stable dynamic cross-linked network structure under physiologically relevant frequency conditions. Furthermore, the G′ of the hydrogel initially increased and then stabilized with increasing MBGNs content, with AGDM-3 and AGDM-4 hydrogels exhibiting the highest elastic modulus. These results demonstrate that the introduction of inorganic MBGNs can effectively enhance the rigidity of the hydrogel cross-linked network and improve its resistance to deformation through physical filling and interfacial interaction with the organic polymer network.

[0035] The structural stability and critical fracture strain of AGDM hydrogels were further investigated using strain scanning experiments, and the results are shown in Figure 3B. All hydrogel samples maintained stable G′ and G″ strains in the low strain range. As the oscillating strain increased, the hydrogel cross-linking network was gradually disrupted, leading to a sharp decrease in G′ and its intersection with G″. The strain corresponding to this intersection point was defined as the critical fracture strain of the hydrogel. The results show that all AGDM hydrogels can withstand significant deformation, with critical fracture strains exceeding 100%. Notably, the deformation resistance of the hydrogels further improved with increasing MBGN content, with AGDM-3 hydrogel exhibiting the highest critical fracture strain. This indicates that MBGN doping did not weaken the network extensibility of the hydrogel; instead, it further enhanced its structural robustness, enabling it to adapt to the dynamic deformation requirements of cartilage tissue during physiological activities.

[0036] Uniaxial compression test results ( Figure 4 The results show that the compressive mechanical properties of AGDM hydrogels are a key indicator for their application as cartilage adhesives in load-bearing joint environments. The compressive stress-strain curves (Figure 4A) indicate that all AGDM hydrogels exhibit typical compressive characteristics of porous elastic materials, demonstrating good linear elastic response within the 0-30% strain range without significant brittle fracture. Quantitative analysis of the compressive modulus (Figure 4B) shows that the compressive modulus of the hydrogels increases significantly with increasing MBGNs content, with AGDM-4 hydrogel exhibiting the highest compressive modulus, several times that of the undoped MBGNs hydrogel matrix. This improvement in mechanical properties is attributed to the strong interfacial interaction between MBGNs and the hydrogel's organic network: the inorganic nanoparticles effectively transfer and disperse compressive stress, thereby significantly enhancing the hydrogel's compressive strength and matching its mechanical strength to that of natural cartilage tissue, meeting the application requirements of load-bearing environments in joints.

[0037] Overlap shear curve ( Figure 5 A) indicates that all hydrogel samples exhibit typical tough adhesion characteristics, with the adhesion force initially increasing and then decreasing with increasing displacement, and no sudden debonding at the interface. Quantitative statistical results ( Figure 5 B) shows that the adhesion strength of AGDM hydrogel to bone fragments first increases and then slightly decreases with increasing MBGNs content, with AGDM-3 hydrogel exhibiting the highest adhesion strength, significantly higher than AGDM-1 and AGDM-2 groups. The excellent cartilage tissue adhesion properties of AGDM hydrogel mainly stem from the synergistic effect of multiple interfacial interactions: the abundant amino groups on the AG molecular chain can form hydrogen bonds with the carboxyl groups of proteins on the bone / cartilage tissue surface, and simultaneously interact with the residual aldehyde groups on DAS to form dynamic Schiff base bonds with the amino groups on the tissue surface, achieving covalent cross-linking with the tissue interface; the introduction of MBGNs further enhances the mechanical interlocking between the hydrogel and the tissue interface, and strengthens the adhesion effect through interfacial hydrogen bond interactions, ultimately achieving strong adhesion between the hydrogel and cartilage / bone tissue. Excessively high MBGNs content (8 wt%, AGDM-4) leads to a slight decrease in hydrogel adhesion strength, possibly because the excessive inorganic nanoparticles partially shield the reactive functional groups in the hydrogel network, weakening the chemical interaction with the tissue interface.

[0038] To verify the structural stability and repair effect of the hydrogel prepared in this invention in the repair of growth plate damage, the following experiments were conducted: Experiment Example 2 Immature experimental pigs (approximately 8-12 weeks old) were selected, routinely anesthetized, and fixed in a supine position. The knee joint area was shaved, disinfected, and draped with sterile towels. A 2-3 cm incision was made on the medial side of the proximal tibia to expose the proximal metaphysis of the tibia layer by layer. The growth plate was located by X-ray fluoroscopy. After establishing a cortical bone window in the metaphysis, a low-speed bone drill with a diameter of approximately 1.5-2 mm was used to drill a hole from the metaphysis toward the growth plate, forming a drilled hole-type defect spanning the metaphysis and the growth plate in the central region of the growth plate (usually occupying approximately 20%-30% of the cross-sectional area of ​​the growth plate). The drilling depth was approximately 3-5 mm to disrupt the local growth plate structure but avoid damaging the articular cartilage surface. The drilled hole was then flushed with sterile saline to remove bone fragments, and hemostasis was achieved by gentle pressure, thereby establishing a stable and repeatable growth plate injury model.

[0039] The treatment of growth plate damage was divided into two groups: the Kirschner wire fixation group and the hydrogel treatment group. In the hydrogel treatment group, a pre-prepared AGDM composite hydrogel precursor solution was loaded into a sterile syringe and slowly injected into the damaged area through a fine needle along the drilled channel, so that the material could fully fill the growth plate defect and part of the drilled channel. The injection volume for a single defect was generally controlled at about 50-200 μL, and the injection needle was kept stable for several tens of seconds to allow the hydrogel to complete in-situ gelation in vivo, thereby forming a stable three-dimensional support structure. Subsequently, the muscle layer and skin were sutured layer by layer and routine postoperative care was performed.

[0040] Kirschner wire fixation group ( Figure 6 A) and the AGDM composite hydrogel treatment group of the present invention ( Figure 6 B) Imaging evaluation. Eight weeks post-surgery, X-ray examination was used to observe the recovery of bone structure and growth plate morphology in the injured area. Results showed that while the Kirschner wire fixation group maintained local structural stability to some extent, incomplete recovery of growth plate continuity was still observed. In contrast, the AGDM composite hydrogel treatment group of this invention formed a stable supporting structure in the injured area, with a more regular morphology in the growth plate region, significantly reduced bone bridge formation, and overall structural recovery approaching that of a normal growth plate. These results indicate that the hydrogel material of this invention can provide structural support similar to Kirschner wires in the injured area, while simultaneously promoting orderly repair of growth plate tissue through biomaterial interface integration, thereby achieving a synergistic effect of structural stability and tissue regeneration.

[0041] The above embodiments are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above embodiments. Any changes, modifications, substitutions, combinations, or simplifications made without departing from the spirit and principle of the present invention shall be considered equivalent substitutions and shall be included within the protection scope of the present invention.

Claims

1. A method for preparing a mesoporous bioactive enhanced cartilage adhesion hydrogel, characterized in that, Includes the following steps: (1) Disperse hexadecyltrimethylammonium bromide in deionized water, then add ethyl acetate, ammonia solution, tetraethyl orthosilicate and calcium nitrate in sequence, and stir the reaction at 55-65℃ for 5-8 hours; after centrifuging, washing and drying the obtained product, calcine at 600-700℃ to remove impurities, and obtain mesoporous bioactive glass nanoparticles MBGNs. (2) Dissolve gelatin in MES buffer, add 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and N-hydroxysuccinimide, stir for 1-2 hours; add ethylenediamine dropwise, adjust the reaction system to neutral, and continue stirring for 3.5-4.5 hours; after the reaction is complete, dialyze the mixture for 60-80 hours and freeze dry to obtain highly reactive AG; (3) Dissolve AG and DAS in phosphate buffer PBS to prepare AG solution with a mass fraction of 30-35% and DAS solution with a mass fraction of 10-15%; add MBGNs with a mass fraction of 2-8% to DAS solution to obtain DAS / MBGNs mixed solution, then add AG solution and stir thoroughly to obtain mesoporous bioactive enhanced cartilage adhesion hydrogel ADGM.

2. The method for preparing a mesoporous bioactive enhanced cartilage adhesion hydrogel according to claim 1, characterized in that, In step (1), the mass ratio of tetraethyl orthosilicate to calcium nitrate is (1.5-2.5):

1.

3. The method for preparing a mesoporous bioactive enhanced cartilage adhesion hydrogel according to claim 1, characterized in that, The mass percentage concentration of ammonia in step (1) is 0.5-2.0%.

4. The method for preparing a mesoporous bioactive enhanced cartilage adhesion hydrogel according to claim 1, characterized in that, The calcination time in step (1) is 3-5 hours.

5. The method for preparing a mesoporous bioactive enhanced cartilage adhesion hydrogel according to claim 1, characterized in that, In step (2), the mass ratio of gelatin, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and N-hydroxysuccinimide is (0.5-1.5):(0.15-0.3):(0.1-0.3).

6. The method for preparing a mesoporous bioactive enhanced cartilage adhesion hydrogel according to claim 1, characterized in that, In step (2), the reaction temperature is 40-50℃ before adding ethylenediamine and 20-30℃ after adding it.

7. The method for preparing a mesoporous bioactive enhanced cartilage adhesion hydrogel according to claim 1, characterized in that, In step (3), the preparation method of DAS is as follows: dissolve corn amylopectin in deionized water, add sodium periodate solution and stir the reaction under light-protected conditions, centrifuge and wash the resulting solution, and then freeze dry it.

8. The method for preparing a mesoporous bioactive enhanced cartilage adhesion hydrogel according to claim 7, characterized in that, The reaction is carried out at 35-40℃ for 10-12 hours; the freeze-drying time is 72-80 hours.

9. The method for preparing a mesoporous bioactive enhanced cartilage adhesion hydrogel according to claim 1, characterized in that, In step (3), the volume ratio of AG solution to DAS / MBGNs mixed solution is 1:

1.

10. The mesoporous bioactive enhanced cartilage adhesion hydrogel obtained by the preparation method according to any one of claims 1-9.

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