Injectable-oxygen-carrying-antibacterial composite hydrogel as well as preparation method and application thereof

Abstract

The invention belongs to the technical field of biomedical materials, and particularly relates to injectable-oxygen-carrying-antibacterial composite hydrogel as well as a preparation method and application thereof. The method comprises the following steps: dissolving dopamine hydrochloride in a buffer solution, adding bioactive glass, carrying out ultrasonic treatment to obtain BGs (at) PDA, dispersing the BGs (at) PDA in water, dropwise adding perfluorooctanoyl chloride, and carrying out ultrasonic treatment to obtain fluorinated bioactive glass; the preparation method comprises the following steps: dissolving hyaluronic acid in water, adding sodium periodate, reacting in a dark place, adding ethylene glycol, continuously reacting, dialyzing and freeze-drying to obtain an oxidized hyaluronic acid solid; and dispersing the fluorinated bioactive glass into a gelatin solution, and then uniformly mixing with an oxidized hyaluronic acid solution, thereby obtaining the product. The hydrogel provided by the invention has injectability, excellent mechanical property and oxygen carrying effect, has good antibacterial effect and biocompatibility, can effectively promote bone defect repair, and has wide application prospect in the field of bone tissue engineering.

Description

Technical Field

[0001] This application belongs to the field of biomedical materials technology, specifically relating to an injectable-oxygen-carrying-antibacterial composite hydrogel, its preparation method, and its application. Background Technology

[0002] Bones are a vital component of the human body, undertaking crucial physiological functions such as protection, movement, and metabolism. However, due to traumatic fractures, osteonecrosis, and bone tumors, the number of patients with bone defects is rapidly increasing. Smaller bone defects can repair themselves; larger defects require surgical implantation of bone graft analogs. While autologous bone grafting is considered the "gold standard" in traditional treatment, it is limited by secondary complications, donor shortages, and immune rejection. Traditional implants suffer from poor stability and low biocompatibility, easily leading to rejection, degeneration, and infection. Furthermore, bone defect repair is often accompanied by local microvascular rupture; ischemia and hypoxia inhibit osteoblast proliferation and differentiation, bone formation, and angiogenesis, resulting in insufficient blood supply and affecting bone regeneration.

[0003] While emerging materials such as bioactive ceramics and metal implants can provide some mechanical support and bioactivity, they struggle to meet the complex needs of bone regeneration for cell growth, angiogenesis, and oxygen supply. Furthermore, their insufficient plasticity prevents them from precisely filling irregular bone defects. Hydrogel materials, on the other hand, address some of these shortcomings: they are highly plastic, capable of precisely filling irregular bone defects, and possess excellent biocompatibility. They can serve as scaffolds to facilitate osteoblast attachment and proliferation, and can also load growth factors to enhance osteoinduction and antibacterial activity. However, ordinary hydrogels still have limitations—their mechanical strength is far lower than that of bone tissue, making them prone to deformation; their degradation rate is difficult to match bone regeneration; and they lack vascularization channels and long-lasting antibacterial capabilities, thus limiting the repair effect. Summary of the Invention

[0004] The purpose of this invention is to overcome the shortcomings of the prior art and provide an injectable, oxygen-carrying, and antibacterial composite hydrogel, its preparation method, and its application. Specifically, the following technical solution is adopted: In a first aspect, the present invention provides a method for preparing an injectable, oxygen-carrying, and antibacterial composite hydrogel, comprising the following steps: S1. Dissolve dopamine hydrochloride in Tris-HCl buffer solution, then add bioactive glass and sonicate, stir evenly, centrifuge, wash, freeze dry to obtain BGs@PDA; S2. Disperse the BGs@PDA in water, add perfluorooctanoic acid dropwise, sonicate, stir evenly, centrifuge, wash, freeze dry to obtain fluorinated bioactive glass; S3. Dissolve hyaluronic acid in water, then add sodium periodate, react in the dark, add ethylene glycol to continue the reaction, dialyze after the reaction is complete, freeze dry to obtain solid oxidized hyaluronic acid; S4. Dissolve gelatin and oxidized hyaluronic acid solid in water to obtain gelatin solution and oxidized hyaluronic acid solution respectively; S5. Disperse the fluorinated bioactive glass in the gelatin solution, and then mix it evenly with the oxidized hyaluronic acid solution to obtain the injectable-oxygen-carrying-antibacterial composite hydrogel.

[0005] As a further preferred embodiment, the ratio of dopamine hydrochloride, Tris-HCl buffer solution and bioactive glass in S1 is 40 mg-80 mg: 20 mL-40 mL: 80 mg-160 mg.

[0006] As a further preferred embodiment, the pH of the Tris-HCl buffer solution is 8.5.

[0007] As a further preferred embodiment, the ratio of hyaluronic acid to sodium periodate in S3 is 1 g-5 g: 0.5 g-2.5 g.

[0008] As a further preferred embodiment, the ratio of gelatin, oxidized hyaluronic acid, and fluorinated bioactive glass in S4 and S5 is 2 g-3 g: 1.2 g-2 g: 0.4 g-2 g.

[0009] As a further preferred embodiment, the concentration of the gelatin solution in S4 is 10-15% w / v.

[0010] As a further preferred embodiment, the concentration of the oxidized hyaluronic acid solution in S4 is 6-10% w / v.

[0011] Secondly, the present invention provides an injectable-oxygen-carrying-antibacterial composite hydrogel, which is prepared by the above-described preparation method.

[0012] Thirdly, the present invention provides the application of the above-mentioned injectable-oxygen-carrying-antibacterial composite hydrogel in the preparation of bone defect repair materials.

[0013] Fourthly, the present invention also provides a bone defect repair material, including the above-mentioned injectable-oxygen-carrying-antibacterial composite hydrogel.

[0014] The beneficial effects of this invention are as follows: (1) The preparation method of the present invention prepares a hydrogel with both injectability and mechanical strength, which is suitable for irregular bone defect sites; wherein, oxidized hyaluronic acid and gelatin form a dynamic reversible cross-linked network through Schiff base reaction to achieve good injectability and adaptability, and the introduction of bioactive glass enhances the mechanical properties of the hydrogel.

[0015] (2) The injectable-oxygen-carrying-antibacterial composite hydrogel prepared by the present invention integrates antibacterial, oxygen-carrying and osteogenic functions. Perfluorooctanoyl chloride provides antibacterial and certain oxygen-carrying capacity to the hydrogel, allowing the hydrogel to resist bacterial infection and insufficient oxygen supply caused by bone defect environment. Bioactive glass mediates the directed differentiation of bone marrow mesenchymal stem cells into osteoblasts, thereby accelerating the regeneration and repair process of bone tissue. The efficient integration and synergistic enhancement of natural polymer and functional materials significantly improve the comprehensive therapeutic potential of hydrogel in bone defect repair. Attached Figure Description

[0016] To more clearly illustrate the technical solutions in the embodiments of the present invention, the accompanying drawings used in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0017] Figure 1 The diagram shows the preparation process of the injectable-oxygen-carrying-antibacterial composite hydrogel of Example 1 of the present invention.

[0018] Figure 2 The image shows an injectable, oxygen-carrying, and antibacterial composite hydrogel. Figure 3 The image shown is the 1H NMR spectrum of oxidized hyaluronic acid. Figure 4 The images shown are: (A) Comparison of the hydrogel prepared in Example 1 before and after gelation; (B) An image of the hydrogel prepared in Example 1 writing letters with a syringe; (C) An experimental image of the self-healing hydrogel prepared in Example 1. Figure 5 The image shown is a SEM image of the hydrogels prepared in Example 1 and Comparative Examples 1-2, respectively. Figure 6 The figures shown are: (A) a comparison of swelling of the hydrogels prepared in Example 1 and Comparative Examples 1-2; (B) a comparison of representative compressive stress-strain curves of the hydrogels prepared in Example 1 and Comparative Examples 1-2; and (C) a comparison of compressive elastic modulus of the hydrogels prepared in Example 1 and Comparative Examples 1-2. Figure 7 The figure shown is an experimental diagram of oxygen-carrying hydrogels prepared in Example 1 and Comparative Examples 1-2, respectively. Figure 8 The figure shows a comparison of the proliferation activity of MSCs cells after 1, 3, and 5 days of culture, as well as the hydrogel extracts prepared in Example 1 and Comparative Examples 1-2 and the blank control group. Figure 9The figures shown are: (A) Comparison of the antibacterial rates of the hydrogels prepared in Example 1 and Comparative Examples 1-2 with the blank control group against Escherichia coli and Staphylococcus aureus; (B) Colony images of the hydrogels prepared in Example 1 and Comparative Examples 1-2 with Escherichia coli and Staphylococcus aureus after co-culturing with the blank control group. Figure 10 The images show: ALP staining of hydrogel-induced osteogenic differentiation of bone MSCs (A); ARS staining of hydrogel-induced osteogenic differentiation of bone MSCs (B); ALP quantitative map of hydrogel-induced osteogenic differentiation of bone MSCs (C); and ARS quantitative map of hydrogel-induced osteogenic differentiation of bone MSCs (D). Figure 11 The following are in vivo evaluation results of hydrogel repair of bone defects in SD rats: A is a schematic diagram of the establishment of the rat bone defect model and the treatment process; B shows the skull repair status by micro-CT after 4 and 8 weeks of hydrogel treatment. C-F are the quantitative analysis results of bone parameters (BV / TV, Tb.Th, BMC, BMD). Detailed Implementation

[0019] The technical solutions of the embodiments of this application will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of this application, not all embodiments. Based on the embodiments of this application, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of this application.

[0020] Example 1 A method for preparing an injectable, oxygen-carrying, and antibacterial composite hydrogel (the flowchart is shown below) Figure 1 As shown), the specific steps include: (1) Weigh 40 mg of dopamine hydrochloride and dissolve it in 20 mL of Tris-HCl buffer solution at pH 8.5. Then add 80 mg of bioactive glass, sonicate for 15 min, and stir the solution vigorously at room temperature for 12 h to complete the encapsulation of bioactive glass with polydopamine. Then centrifuge the mixture (centrifuge speed is 8000 rpm, centrifuge time is 5 min) to remove the supernatant, wash with deionized water 3 times, freeze dry to obtain BGs@PDA; (2) Weigh 20 mg BGs@PDA and disperse it in 40 mL of deionized water. Add perfluorooctanoyl chloride (0.1 mM) and sonicate gently. Then stir vigorously at room temperature for 12 h. Centrifuge (centrifuge speed is 8000 rpm and centrifuge time is 5 min) to remove the supernatant. Wash with water 3 times and freeze dry to obtain fluorinated bioactive glass. (3) Weigh 1 g of HA (hyaluronic acid) and dissolve it in 100 mL of deionized water until the solution is homogeneous. Then add 0.54 g of sodium periodate to the hyaluronic acid solution and react in the dark for 2 h. Add ethylene glycol and react for half an hour to terminate the oxidation reaction. Put it into a dialysis bag and dialyze with pure water for 3 days, changing the water four times a day. Freeze dry and collect the oxidized hyaluronic acid solid and store it at -20 ℃. (4) Weigh 3 g of gelatin and dissolve it in 20 mL of deionized water to obtain a 15% w / v gelatin solution. Weigh 1.2 g of the oxidized hyaluronic acid solid prepared in step (3) and dissolve it in 20 mL of deionized water to obtain a 6% w / v oxidized hyaluronic acid solution. Finally, uniformly disperse 0.8 g of the fluorinated bioactive glass prepared in step (2) in the 15% w / v gelatin solution and mix it with the 6% w / v oxidized hyaluronic acid solution to finally obtain an injectable oxygen-carrying antibacterial composite hydrogel with Schiff base crosslinking.

[0021] The above-mentioned injectable-oxygen-carrying-antibacterial composite hydrogel's states before and after gelation are as follows: Figure 4 As shown in Figure A, the hydrogel changes from a flowing state to a solidified state.

[0022] The injectability of the prepared hydrogel was macroscopically evaluated. The stained hydrogel was placed in a syringe for injection testing. The hydrogel was squeezed out, and the word "GEL" was written on a whiteboard. Figure 4 As shown in B, the hydrogel has good fluidity, and the hydrogel retains a certain shape after extrusion.

[0023] The self-healing properties of the prepared hydrogels were evaluated. The stained and unstained hydrogels were cut separately, combined, and left to stand for a period of time; their state was then recorded.

[0024] The result is as follows Figure 4 As shown in Figure C, the two cut hydrogels can complete their self-repair function after being left for a period of time.

[0025] Comparative Example 1 A method for preparing a hydrogel is basically the same as that in Example 1. The difference between the two methods is that this comparative example only includes steps (3) and (4), and in step (4), fluorinated bioactive glass is not used. Instead, gelatin solution and oxidized hyaluronic acid solution are directly mixed to obtain the hydrogel.

[0026] Comparative Example 2 A method for preparing a hydrogel is basically the same as that in Example 1. The difference between Example 1 and Example 1 is that this comparative example only includes steps (3) and (4), and in step (4), fluorinated bioactive glass is not used, but bioactive glass is used instead. That is, bioactive glass is uniformly dispersed in a 15% w / v gelatin solution and then mixed with a 6% w / v oxidized hyaluronic acid solution to obtain a hydrogel.

[0027] Example 2 The materials prepared in Example 1 and Comparative Examples 1-2 were characterized and tested as follows: (1) Fourier transform infrared spectroscopy analysis was performed on the fluorinated bioactive glass synthesized in Example 1. A small amount of fluorinated bioactive glass powder, bioactive glass, and perfluorooctanoyl chloride were placed on a Fourier transform attenuated total reflectance infrared spectrometer for scanning. The number of scans was 32, and the wavenumber was 4000 cm⁻¹. -1 ~ 400 cm -1 .

[0028] The result is as follows Figure 2 As shown, the characteristic absorption of bioactive glass appears at 1037 cm⁻¹. -1 and 962 cm -1 Assigned to O-P tensile vibration. Compared with using bioactive glass alone, at 1322 cm⁻¹ -1 ~ 1376 cm -1 The C-F stretching absorption was observed in the fluorinated bioactive glass curve.

[0029] (2) The oxidized hyaluronic acid and hyaluronic acid prepared in Example 1 were subjected to 1H NMR spectroscopy (NMR spectroscopy). 1 For H NMR analysis, 6.0 mg of oxidized hyaluronic acid powder and lyophilized hyaluronic acid flocculent solid were weighed and dissolved in 0.6 mL of D2O. After complete dissolution, the solutions were transferred to NMR tubes for analysis. 1 1H NMR analysis (600 MHZ).

[0030] The result is as follows Figure 3 As shown in the oxidized hyaluronic acid spectrum, 4.8 × 10 -6 ~ 5.2×10 -6 The appearance of a new chemical shift indicates that the aldehyde group on the oxidized hyaluronic acid forms a hemiacetal proton with the adjacent hydroxyl group, indicating that the oxidized hyaluronic acid was successfully prepared.

[0031] (3) Morphological characterization was performed using a scanning electron microscope. Double-sided magnetic conductive adhesive was attached to a substrate. A small amount of the lyophilized hydrogel samples from Example 1 and Comparative Examples 1-2 were picked up and adhered to the magnetic conductive adhesive. The dust that was not adhered to the surface was blown away with a blower. Gold was sprayed for 15 seconds and observed under a scanning electron microscope.

[0032] The result is as follows Figure 5 As shown, the hydrogel exhibits a three-dimensional porous network structure with uniform pore size distribution and interconnected pores. The addition of bioactive glass and fluorinated bioactive glass makes the hydrogel structure more compact, with the group in Example 1 having the smallest pore size.

[0033] Example 3 The hydrogels prepared in Example 1 and Comparative Examples 1-2 were subjected to performance tests, as detailed below: (1) Swelling performance test of hydrogel: The hydrogels of Example 1 and Comparative Examples 1-2 were placed in PBS buffer and incubated at 37 °C for several hours to calculate the degree of swelling.

[0034] The result is as follows Figure 6 As shown in Figure A, the increase in fluorinated bioactive glass leads to a decrease in the swelling degree of the hydrogel. This indicates that with the increase in cross-linking degree, the three-dimensional pores of the hydrogel decrease, making it more compact and thus significantly inhibiting the expansion of the hydrogel.

[0035] (2) Mechanical property testing of the hydrogel: For the hydrogels of Example 1 and Comparative Examples 1-2, cylindrical hydrogel samples with a diameter of 10 mm and a height of 5 mm were fabricated using polytetrafluoroethylene plates of a specific shape. Three samples were prepared for each group. The samples were placed on the test bench, and the instrument parameters were set as follows: compression rate of 2 mm / min, preload force of 0.01 N, and termination at 40% deformation. Stress and strain data were collected. The value at 10% of the stress-strain curve was selected as the Young's modulus.

[0036] The result is as follows Figure 6 As shown in Figures B and C, the compressive stress-strain curves indicate that the addition of fluorinated bioactive glass increases the compressive strength of the hydrogel from 50 kPa to 240 kPa, with the hydrogel of Example 1 exhibiting the best compressive strength. The elastic modulus at 10% strain also shows a corresponding trend.

[0037] (3) Test the oxygen-carrying capacity of the hydrogel: 10 μL of 10 mg / L [Ru(dpp)3]Cl2 solution was added to each well of a 24-well plate containing 100 μL of hydrogel sample. After gently shaking to disperse the probe evenly, the plate was incubated at 37 ℃ for 15 min. Fluorescence imaging was then performed using the IVIS small animal fluorescence imaging system (excitation wavelength 488 nm, emission wavelength 620 nm). Finally, the region of interest in each well of hydrogel was selected using the system's built-in analysis software, and the average fluorescence intensity value was extracted and calculated.

[0038] The results are as follows Figure 7 As shown, in fluorescence imaging, the hydrogel in Comparative Example 1 exhibits a high-intensity fluorescence distribution dominated by yellow, indicating that its oxygen-carrying capacity is relatively weak. The fluorescence intensity of the hydrogel in Comparative Example 2 is significantly reduced and dominated by red tones, while the fluorescence signal of Example 1 is the weakest and is close to the dark area overall. The addition of fluorinated bioactive glass has a better effect on enhancing the oxygen-carrying capacity of the hydrogel, verifying that the hydrogel prepared in Example 1 enhances the oxygen-carrying performance of the hydrogel more efficiently.

[0039] Example 4 The hydrogels prepared in Example 1 and Comparative Examples 1-2 were subjected to cytotoxicity testing, as detailed below: Examples 1 and Comparative Examples 1-2 were sterilized with 75% alcohol and repeatedly washed with PBS to remove ethanol. The culture medium was then added at 60 mg / mL to α-MEM containing 10% fetal bovine serum and incubated at 37°C for 24 h in a shaker at 100 rpm / min. The supernatant was sterilized using a sterile syringe filter, and the filtered liquid was the hydrogel extract. Cells were seeded at a density of 5000 cells / well in 96-well plates and incubated overnight. Cells were then treated with the hydrogel extract for 1, 3, 5, and 7 days. After removing the supernatant and diluting with growth medium, CCK-8 reagent was applied to each well. The absorbance was measured at 450 nm after 1 hour of incubation using a microplate reader. The percentage of cell viability relative to the blank control group was determined.

[0040] The result is as follows Figure 8 As shown, toxicity assessments were performed at intervals of 1, 3, 5, and 7 days of culture. The absorbance values ​​of cells in each group increased significantly with the extension of time, and there was no significant difference in absorbance values ​​between groups at the same time point.

[0041] Example 5 The hydrogels prepared in Example 1 and Comparative Examples 1-2 were subjected to in vitro antibacterial performance tests, as detailed below: With Escherichia coli Escherichia coli (E. coli) Staphylococcus aureus Staphylococcus aureus (S .aureus)To evaluate the antibacterial properties of the hydrogels used in Example 1 and Comparative Examples 1-2, 15 g of soybean peptone was dissolved in 500 mL of ultrapure water and sterilized at high temperature. After cooling, a liquid bacterial culture medium was obtained. 15 g of soybean peptone and 8 g of agar were dissolved in 500 mL of ultrapure water and sterilized at high temperature. The solution was poured into a petri dish and cooled to obtain a solid culture medium. The bacterial suspension was added to the liquid culture medium and activated at 37 °C and 130 rpm for approximately 16 h. The solution was then diluted to the desired concentration before use. The bacterial suspension was added to well plates containing the hydrogels of Example 1 and Comparative Examples 1-2 and incubated for 24 h. The turbidity of the bacterial suspensions in different groups was observed to preliminarily determine the antibacterial effect of the hydrogel. The optical density (OD600) of the well plates was measured using a microplate reader. The bacterial suspension was then diluted 10,000 times, mixed thoroughly using a vortex mixer, and 100 μL was transferred to a culture plate and spread evenly. After incubating in a ventilated incubator at 37 ℃ for 18 h, the number of colonies on the culture plates was observed. Each group was tested in triplicate, and the results were expressed as antibacterial activity: antibacterial activity (%) = (Cc-Cs) / Cc×100, where Cc and Cs represent the colony counts of the control group and the sample group, respectively.

[0042] The result is as follows Figure 9 As shown, the hydrogel of Example 1 exhibits excellent broad-spectrum antibacterial effects. S. aureus The antibacterial rate was 84%, and it had a good antibacterial effect. E. coli The antibacterial rate was 79%. After co-culturing with the hydrogel, the number of bacterial colonies in the hydrogel of Example 1 was significantly reduced compared with that in Comparative Examples 1-2.

[0043] Example 6 The osteogenic induction properties of the hydrogels prepared in Example 1 and Comparative Examples 1-2 were tested as follows: Bone culture medium was prepared in advance: α-MEM medium + 10% FBS + 100 nmol / L dexamethasone + 50 μmol / L ascorbic acid + 10 mmol / L β-glycerophosphate sodium. Cell samples were fixed in 4% paraformaldehyde for 30 min on days 7 and 14 after osteogenic induction. ALP activity in BMSCs was qualitatively and quantitatively assessed using an alkaline phosphatase activity assay kit and a BCIP / NBT ALP staining kit. At day 14 of bone induction, cell samples were washed with PBS according to Alizarin Red (AR) staining instructions and then fixed in 4% paraformaldehyde for 30 min. The samples were then incubated with Alizarin Red staining agent for 30 min, and images were observed under an inverted fluorescence microscope. Calcium content was quantified using a calcium colorimetric kit.

[0044] The result is as follows Figure 10As shown, compared with the hydrogel of Comparative Example 1, the ALP staining intensity of the hydrogels of Example 1 and Comparative Example 2 was enhanced, with the highest intensity in Example 1. This indicates that the cells in these groups were more active in expressing early osteogenic markers. Further quantitative analysis of ALP activity also confirmed this trend. In addition, Alizarin Red staining was performed in this example to assess the osteogenic differentiation capacity of the cells in each group. According to the Alizarin Red staining results, significant mineral deposition was observed in the cells of Example 1 and Comparative Examples 1-2, indicating that the cells in these groups could effectively promote calcium salt deposition during osteoogenesis. Quantitative analysis also showed that calcium phosphate deposition was significantly increased in the Example 1 group compared with Comparative Examples 1-2.

[0045] Example 7 The hydrogels prepared in Example 1 and Comparative Examples 1-2 were used for in vivo evaluation of skull defects in SD rats, as detailed below: Rats were anesthetized by intraperitoneal injection of 3% sodium pentobarbital (0.2 mL / kg) before the experiment. An incision was made in the rat skull to expose the parietal bone surface, and a 5 mm diameter cranial drill was used to puncture the full-thickness bone defect area to create a skull defect model. All surgeries were performed under sterile conditions. 20 μL of different groups of hydrogel were injected into the bone defect area, and the wound was closed in layers. At 4 and 8 weeks post-surgery, rat samples were necropsies performed, and the skulls were removed to assess bone formation. Skull specimens were stored in 4% paraformaldehyde solution for at least 24 hours. Micro-CT images of the skull samples were acquired using NEMO. Bone volume / total volume (BV / TV), trabecular thickness (Tb.Th), bone mineral content (BMC), and bone mineral density (BMD) were analyzed using software provided by the Micro-CT manufacturer.

[0046] Figure 11 A is a schematic diagram of the establishment of a rat skull model. Figure 11 As shown in Figure B, at 4 weeks, no effective new bone tissue filling was observed in the defect area of ​​the Blank group. Although Comparative Examples 1 and 2 showed some tendency for bone repair, the defect outline was still clearly discernible. In Example 1, the distribution of new bone tissue at the defect edge was significantly wider, showing a more active bone regeneration trend. At 8 weeks, the Blank group only showed limited self-healing bone repair, and the defect was still obvious. The defect area of ​​Comparative Examples 1 and 2 decreased, and the repair effect gradually improved. In contrast, the defect area of ​​Example 1 was covered by a large amount of new bone tissue, further highlighting its bone regeneration advantage. Based on the quantitative analysis results of bone parameters, it is evident that... Figure 11As observed in the CF (Computed Tomography) results, the bone volume fraction (BV / TV), trabecular thickness (Tb.Th), bone mineral content (BMC), and trabecular number (Tb.N) of each experimental group showed an increasing trend over time. At week 8, the above indicators in the Example 1 group were significantly higher than those in other groups (***P<0.001), demonstrating superior efficacy in promoting bone formation, enhancing bone mineralization, and improving trabecular structure. In conclusion, the hydrogel in Example 1 significantly promoted the repair of skull defects in rats by regulating new bone formation and collagen fiber deposition, with the advantage becoming more pronounced over time, providing a basis for bone defect repair.

[0047] The embodiments of this application have been described above with reference to the accompanying drawings. Specific examples have been used to illustrate the principles and implementation methods of this application. The description of the above embodiments is only for the purpose of helping to understand the core ideas of this application. However, this application is not limited to the specific embodiments described above. The specific embodiments described above are merely illustrative and not restrictive. Those skilled in the art can make many other forms under the guidance of this application without departing from the spirit and scope of the claims, and all of these forms are within the protection scope of this application.

Claims

1. A method for preparing an injectable-oxygen-carrying-antibacterial composite hydrogel, characterized in that, Includes the following steps: S1. Dissolve dopamine hydrochloride in Tris-HCl buffer solution, then add bioactive glass and sonicate, stir evenly, centrifuge, wash, freeze dry to obtain BGs@PDA; S2. Disperse the BGs@PDA in water, add perfluorooctanoic acid dropwise, sonicate, stir evenly, centrifuge, wash, freeze dry to obtain fluorinated bioactive glass; S3. Dissolve hyaluronic acid in water, then add sodium periodate, react in the dark, add ethylene glycol to continue the reaction, dialyze after the reaction is complete, freeze dry to obtain solid oxidized hyaluronic acid; S4. Dissolve gelatin and oxidized hyaluronic acid solid in water to obtain gelatin solution and oxidized hyaluronic acid solution respectively; S5. Disperse the fluorinated bioactive glass in the gelatin solution, and then mix it evenly with the oxidized hyaluronic acid solution to obtain the injectable-oxygen-carrying-antibacterial composite hydrogel.

2. The preparation method according to claim 1, characterized in that, The ratio of dopamine hydrochloride, Tris-HCl buffer solution, and bioactive glass in S1 is 40 mg-80 mg: 20 mL-40 mL: 80 mg-160 mg.

3. The preparation method according to claim 2, characterized in that, The pH of the Tris-HCl buffer solution is 8.

5.

4. The preparation method according to claim 1, characterized in that, The ratio of hyaluronic acid to sodium periodate in S3 is 1g-5g: 0.5g-2.5g.

5. The preparation method according to claim 1, characterized in that, The ratio of gelatin, oxidized hyaluronic acid, and fluorinated bioactive glass used in S4 and S5 is 2 g-3 g : 1.2 g-2 g : 0.4 g-2 g.

6. The preparation method according to claim 5, characterized in that, The concentration of the gelatin solution described in S4 is 10-15% w / v.

7. The preparation method according to claim 6, characterized in that, The concentration of the oxidized hyaluronic acid solution described in S4 is 6-10% w / v.

8. An injectable, oxygen-carrying, antibacterial composite hydrogel, characterized in that, It is prepared by the preparation method according to any one of claims 1-7.

9. The application of the injectable-oxygen-carrying-antibacterial composite hydrogel according to claim 8 in the preparation of bone defect repair materials.

10. A bone defect repair material, characterized in that, Including the injectable-oxygen-carrying-antibacterial composite hydrogel as described in claim 8.

Images