Graphene oxide hydrogel for treating rotator cuff injury and method of making the same
By combining graphene oxide hydrogel with microfracture technology, the problem of rapid loss of repair signals at the tendon-bone interface was solved, achieving effective healing and improvement of the tendon-bone interface and significantly improving the repair effect of rotator cuff injuries.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- AFFILIATED HOSPITAL OF NANTONG UNIV
- Filing Date
- 2026-04-09
- Publication Date
- 2026-05-29
Smart Images
Figure CN122097249A_ABST
Abstract
Description
Technical Field
[0001] This application relates to the field of medical materials technology, and more specifically, to a graphene oxide hydrogel for treating rotator cuff injuries and a method for preparing the same. Background Technology
[0002] Rotator cuff injuries are a common clinical condition that often results in tearing at the tendon-bone junction (tendon-bone insertion). Poor healing quality and a high rate of re-tearing after surgical repair are two major clinical challenges, primarily due to the inherently limited regenerative capacity of the insertion site.
[0003] To improve healing, microfracture techniques have been introduced as a biological enhancement method in rotator cuff repair. This technique involves drilling into the bone bed to release bone marrow mesenchymal stem cells (BMSCs) and various endogenous growth factors (such as TGF-β and FGF) to stimulate a local regenerative response. Studies have confirmed that this technique promotes the recruitment of BMSCs to the interface and has been observed to accelerate early healing in animal models.
[0004] However, existing microfracture techniques suffer from a key and common technical deficiency: the mobilized repair cells and bioactive factors lack an effective local retention mechanism. Due to the absence of a carrier microenvironment at the injury interface capable of accommodating and retaining these biological components, they rapidly dissipate from the target site, failing to form and maintain sufficient local effective concentrations. This loss results in short-lived and diffusely distributed regenerative signals, severely limiting the full realization of the potential of microfracture techniques and becoming a major bottleneck restricting further improvements in long-term healing outcomes and reductions in re-tear rates.
[0005] Therefore, overcoming the problem of rapid loss of repair signals locally is a key technical obstacle that urgently needs to be addressed in improving the quality of rotator cuff tendon-bone healing. Summary of the Invention
[0006] The purpose of this application is to provide a graphene oxide hydrogel and a method for preparing the same, which combines graphene oxide with polysaccharides to enhance the healing of the tendon-bone interface after rotator cuff injury, thereby improving the repair outcome.
[0007] To solve the above-mentioned technical problems, the technical solution adopted in this application is as follows: On one hand, this application provides a method for preparing a graphene oxide hydrogel for treating rotator cuff injuries, comprising the following steps: S1. Graphene oxide is uniformly dispersed in water to form a graphene oxide suspension; S2. Dissolve the water-soluble synthetic polymer framework material in water to obtain a framework material solution, then add a pore-forming agent to form a mixture after dissolution. S3. The graphene oxide suspension, anionic crosslinking agent and solution stabilizer are sequentially incorporated into the above mixture. After being mixed evenly, the mixture is cooled to room temperature, frozen at low temperature and freeze-dried to obtain the graphene oxide hydrogel.
[0008] On the other hand, this application provides a graphene oxide hydrogel prepared by the above method, and uses it to treat rotator cuff injuries. The preparation, use process, and working principle of the graphene oxide hydrogel of this application are illustrated in the diagram below. Figure 1 As shown.
[0009] Compared with the prior art, the embodiments of this application have at least the following advantages or beneficial effects: This application combines microfracture technology with graphene oxide hydrogel (GO-Gel). GO promotes the controlled release of growth factors through covalent, non-covalent, and electrostatic interactions, enhancing tendon-bone interface healing after rotator cuff injury and thus improving repair outcomes. Experimental results show that graphene oxide (GO-Gel) promotes microfracture treatment of rotator cuff injuries and significantly enhances tendon-bone healing in a rat model. We demonstrate that this combination delivers significant early biomechanical advantages, exhibiting significantly better failure load and stiffness at 3 weeks, and promoting enhanced histological maturation, including improved collagen fiber tissue and interfacial continuity. Material characterization confirms that the integration of GO enhances the structural complexity and stability of the scaffold. This work validates a transferable combinatorial strategy that directly addresses the scaffold-less limitations of microfracture, providing a promising approach for improving rotator cuff repair. Attached Figure Description
[0010] To more clearly illustrate the technical solutions of the embodiments of this application, the accompanying drawings used in the embodiments will be briefly introduced below. It should be understood that the following drawings only show some embodiments of this application and should not be regarded as a limitation of the scope. For those skilled in the art, other related drawings can be obtained based on these drawings without creative effort.
[0011] Figure 1 This diagram illustrates the preparation, use, and working principle of the graphene oxide hydrogel in this application.
[0012] Figure 2 The flowchart shows the research design for the experimental examples in this application.
[0013] Figure 3 Characterization images of the GO and GO-Gel composite materials in the experimental examples of this application are shown. 3A is the FTIR spectrum; 3B is the UV-Vis absorption spectrum; 3C is the XRD pattern; 3D is the Zeta potential measurement; and 3E is the TEM photomicrograph. Scale bar: 1 μm (TEM).
[0014] Figure 4 The GO-Gel composite material was characterized using SEM, Raman spectroscopy, and cell viability assays in the experimental examples of this application. Image 4A is an SEM image; image 4B is a Raman spectrum; and image 4C shows tendon cell viability at different concentrations (0.00625–0.05 mg / mL). SEM scale bar: 20 μm on the left, 2 μm on the right.
[0015] Figure 5 The biomechanical characteristics of the experimental cases in this application are shown at 3 and 9 weeks post-operation.
[0016] Figure 6 Comparison of hematoxylin-eosin (HE) staining and Masson trichrome (MT) staining at 3 and 9 weeks for the MF, MF+Gel, and MF+GO-Gel groups in the experimental examples of this application regarding the reattachment of tendon-bone interface.
[0017] Figure 7 This is a comparison of picric acid Sirius red staining at 3 and 9 weeks for the MF, MF+Gel, and MF+GO-Gel groups in the experimental examples of this application, showing the reattachment of tendon-bone interfaces. Detailed Implementation
[0018] To make the objectives, technical solutions, and advantages of the embodiments of this application clearer, the technical solutions in the embodiments of this application will be clearly and completely described below. Where specific conditions are not specified in the embodiments, conventional conditions or conditions recommended by the manufacturer shall apply. Reagents or instruments whose manufacturers are not specified are all conventional products that can be purchased commercially.
[0019] It should be noted that, unless otherwise specified, the embodiments and features described in this application can be combined with each other. The present application will now be described in detail with reference to specific embodiments.
[0020] A method for preparing a graphene oxide hydrogel for treating rotator cuff injuries includes the following steps: S1. Graphene oxide is uniformly dispersed in water to form a graphene oxide suspension; S2. Dissolve the water-soluble synthetic polymer framework material in water to obtain a framework material solution, then add a pore-forming agent to form a mixture after dissolution. S3. The graphene oxide suspension, anionic crosslinking agent and solution stabilizer are sequentially incorporated into the above mixture. After being mixed evenly, the mixture is cooled to room temperature, frozen at low temperature and freeze-dried to obtain the graphene oxide hydrogel.
[0021] In some embodiments of this application, the ratio of graphene oxide to water in the graphene oxide suspension in step S1 above is 1 g: (1-10) mL, and the dispersion specifically involves stirring for 30-60 min followed by ultrasonic treatment for 3-5 min. In this application, reduced graphene oxide or functionalized graphene can also be used as substitutes.
[0022] In some embodiments of this application, the preparation of the aforementioned framework material solution specifically involves: dissolving the water-soluble synthetic polymer framework material in water at a ratio of 1g:(20-30)mL, then autoclaving at 115-125°C for 5-20 minutes, cooling to 70-90°C and stirring, and finally diluting with water to obtain a framework material solution with a concentration of 1.5-3%. To improve the uniformity of dissolution and the accuracy of concentration of the framework material, this application adopts the method of first preparing a stock solution and then diluting it to the specified concentration. If zero water loss is ensured in a strictly sealed container, the specified concentration can also be prepared directly.
[0023] The water-soluble synthetic polymer backbone material is polyvinyl alcohol, polyvinylpyrrolidone, polyacrylamide, polyacrylic acid, or copolymers thereof.
[0024] In some embodiments of this application, the mass ratio of the water-soluble synthetic polymer framework material to the pore-forming agent in step S2 is 1:(0.25-2); after the pore-forming agent is added, it is stirred at 700-900 rpm at 50-70°C until completely dissolved; the pore-forming agent is D-lactose monohydrate, sucrose, trehalose, glucose, mannitol or maltose.
[0025] In some embodiments of this application, the final concentration of graphene oxide in the hydrogel during step S3 is 0.0125–0.2 mg / mL.
[0026] In some embodiments of this application, the anionic crosslinking agent is sodium alginate, chitosan, sodium hyaluronate, carboxymethyl cellulose, gelatin, or collagen. The solution stabilizer is glycerol, ethylene glycol, propylene glycol, polyethylene glycol-200, polyethylene glycol-400, or sorbitol.
[0027] In some embodiments of this application, the mass ratio of the above-mentioned anionic crosslinking agent to the water-soluble synthetic polymer framework material is 1:(1.5-2.5). After adding the anionic crosslinking agent in step S3, the mixture is stirred at 700-900 rpm at 60°C until completely dissolved, and then the solution stabilizer is added. The solution stabilizer accounts for 5-15% of the final mixture volume. After adding the solution stabilizer, the mixture is stirred at the same temperature and speed until it is homogeneous.
[0028] In some embodiments of this application, the low-temperature freezing temperature in step S3 is -70 to -90°C, and the time is 4 to 12 hours; the vacuum degree during freeze drying is 10 to 20 Pa, the temperature is ≤ -60°C, and the time is 24 to 48 hours.
[0029] A graphene oxide hydrogel for treating rotator cuff injuries was prepared using the method described above.
[0030] The features and performance of this application will be further described in detail below with reference to the embodiments. Example 1
[0031] This embodiment prepares a graphene oxide hydrogel (GO-Gel) for treating rotator cuff injuries, and the specific steps are as follows: S1. All laboratory instruments must be sterilized before use. Graphene oxide (GO) is a lightweight powder; for safety reasons, it is best used in a fume hood. Weigh 10 mg of GO using an electronic balance, disperse it in 10 mL of sterile water, stir thoroughly for a sufficient time, and sonicate for 3 min to obtain a homogeneous GO suspension.
[0032] S2. Dissolve 4g of polyvinyl alcohol (PVA) in 100mL of distilled water. Autoclave the solution at 121°C for 10min to promote dissolution. After autoclaving, remove the solution when the temperature drops to approximately 80°C and stir to ensure homogeneity. Then, dilute the solution by adding 100mL of sterile water to halve the concentration. Next, add 4g of D-lactose monohydrate to the PVA solution and stir at 800 rpm at 60°C until completely dissolved.
[0033] S3. Incorporate the GO suspension into the solution to adjust the ratio to 0.025 mg / mL. Then, add 2 g of sodium alginate to the mixture and continue stirring at 800 rpm at 60°C until completely dissolved. Next, add 20 mL of glycerol to stabilize the reaction and stir the mixture at the same speed and temperature until homogeneous. After the solution is prepared, dispense it into petri dishes, adjusting the thickness by adjusting the volume. After the solution cools to room temperature, transfer the petri dishes to -80°C and freeze for 6 hours. Finally, freeze-dry the frozen petri dishes under vacuum for 36 hours to obtain graphene oxide hydrogel (GO-Gel).
[0034] To conduct the following experiments, experimental groups with different GO concentrations were set up in step S3 of this embodiment, with concentrations of 0, 0.0125, 0.025, 0.05, 0.075, 0.1 and 0.2 mg / mL, respectively. Example 2
[0035] This embodiment prepares a graphene oxide hydrogel (GO-Gel) for treating rotator cuff injuries, and the specific steps are as follows: S1. All laboratory instruments are sterilized before use. Graphene oxide (GO) is a lightweight powder; for safety reasons, it is best used in a fume hood. Weigh 10 mg of GO using an electronic balance, disperse it in 8 mL of sterile water, stir thoroughly for a sufficient time, and sonicate for 5 min to obtain a homogeneous GO suspension.
[0036] S2. Dissolve 4g of polyvinylpyrrolidone in 120mL of distilled water. Autoclave the solution at 121°C for 15min to promote dissolution. After autoclaving, remove the solution when the temperature drops to approximately 80°C and stir to ensure homogeneity. Then, dilute the solution by half by adding 100mL of sterile water. Next, add 4g of trehalose to the polyvinylpyrrolidone solution and stir at 800rpm at 60°C until completely dissolved.
[0037] S3. Incorporate the GO suspension into the solution, adjusting the ratio to 0.025 mg / mL. Then, add 2 g of sodium hyaluronate to the mixture and continue stirring at 800 rpm at 60°C until completely dissolved. Next, add 20 mL of polyethylene glycol-200 to stabilize the reaction, and stir the mixture at the same speed and temperature until homogeneous. After the solution is prepared, dispense it into petri dishes, adjusting the thickness by changing the volume. After the solution cools to room temperature, transfer the petri dishes to -85°C and freeze for 10 h. Finally, freeze-dry the frozen petri dishes under vacuum for 40 h to obtain graphene oxide hydrogel (GO-Gel). Experimental Example
[0038] The graphene oxide hydrogel used in this experimental example is the product prepared in Example 1.
[0039] 1. Research Design Animal experiments were approved by the university's animal research ethics committee. A total of 42 male Sprague-Dawley rats, each weighing approximately 320g, were used. Bilateral supraspinatus tendon resection was performed on each rat at the greater tuberosity. Subjects were randomly assigned to three groups using a computer-generated random list: 14 rats were assigned to the microfracture (MF) group, another 14 to the microfracture combined with polysaccharide-based hydrogel (MF+Gel) group, and the remaining 14 to the microfracture treatment combined with graphene oxide gel (MF+GO-Gel) group. All animals underwent immediate surgical repair. At 3 and 9 weeks post-surgery, 7 rats from each group were sacrificed for subsequent biomechanical and histological analyses. The design flowchart is shown below. Figure 2 As shown, Figure 2 The data shows the number of rats in each group, the time point of sacrifice, and the assessment results. RCR indicates rotator cuff repair.
[0040] 2. Animal models All rats were anesthetized by intraperitoneal injection of 3% sodium pentobarbital. The skin was disinfected with povidone-iodine and then covered with a sterile surgical drape. A 1 cm longitudinal incision was made into the proximal humeral region, followed by fixation with mosquito forceps. The acromioclavicular joint was exposed according to previous studies. The supraspinatus tendon was carefully exposed and sharply severed at the greater tubercle, then retracted caudally. No bleeding was observed at the exposed site prior to the microfracture manipulation. Subsequently, six uniformly spaced burrs were created perpendicular to the bone surface and laterally towards the greater tubercle at the insertion site, approximately 2 mm deep and 0.2 mm wide, spaced 2 mm apart, until bleeding was observed at all burr sites. Two intramedullary tunnels, each 0.4 mm in diameter, were created using a small electric drill. The supraspinatus tendon was then reattached to its insertion point using a modified Mason-Allen suture and 5-0 Prolene sutures (Ethicon).
[0041] In the experimental group, after tendon repair, a lyophilized gel and graphene oxide gel (GO-Gel) scaffold was fixed to the retracted supraspinatus tendon surface using 5-0 absorbable sutures. Conversely, in the control group, the tendon was sutured directly to the bone without the use of a lyophilized scaffold. The acromioclavicular joint was repaired using mattress sutures with 4-0 Vicryl (Ethicon) sutures. Finally, the skin wound was sutured with 3-0 silk sutures. Postoperatively, all rats had free access to food and water.
[0042] 3. In vitro cell proliferation experiment.
[0043] When primary rat tendon cells reached 80% confluence, they were passaged and seeded into 96-well plates at a density of 20,000 cells per well. After 12 hours, the culture medium in these wells was replaced with a mixture containing gel suspensions of different concentrations of GO (0, 0.0125, 0.025, 0.05, 0.075, 0.1, and 0.2 mg / mL). Triple replicates of each experimental group were then incubated at 37°C and 5% CO2 for 24 hours and 72 hours. At each time point, tendon cell proliferation was quantitatively assessed using the CCK-8 assay.
[0044] 4. Biomechanical testing For biomechanical analysis, the complete tendon-bone complex (including the supraspinatus tendon, epiphysis, and humerus) was carefully dissected from the scapula and stored at -80°C until further processing. Prior to mechanical testing, surgical sutures were carefully removed, and the specimen was thawed at 4°C for 2 hours to ensure gradual rehydration while minimizing tissue degradation. The specimen was secured using large, non-slip serrated clamps, and uniaxial tension was applied using an Instron 3365 benchtop universal testing machine until failure, as described in previously published studies. Testing was terminated when the reattached tendon fractured at the tendon-bone repair site. Failure load data were automatically recorded using a synchronous data acquisition system, and structural stiffness was calculated as the slope of the linear elastic region of the load-displacement curve.
[0045] 5. Histological analysis Seven shoulder joints were collected from each group at 3 and 9 weeks post-surgery. Histological analysis was performed on the supraspinatus tendon-humeral head complex from rats. Tissues were fixed overnight in 10% neutral buffered formalin, decalcified with a rapid decalcification solution, and then embedded in paraffin. Specimens were roughly trimmed to expose the tendon-humeral head junction and then finely sectioned in the coronal plane to 5 μm thickness. Routine histological techniques were used, including hematoxylin and eosin, Masson's trichrome, and Sirius red staining. Tissue sections were examined immediately after picric acid Sirius red staining using an optical microscope (DM4B; Leica) equipped with a polarization module. Six normalized rectangular regions (300 × 150 μm²) were randomly selected from the repair area. Two independent observers unaware of the experimental conditions calculated histological scores, and semi-quantitative analysis of picric acid Sirius red staining was performed using ImageJ software (National Institutes of Health). Based on literature, we evaluated the continuity, parallel arrangement, density, vascular distribution, and cellular composition of collagen fibers at the tendon-bone interface. Histological findings were assessed using a semi-quantitative scoring system, with each parameter scored from 0 to 3. For the continuity and parallel alignment of collagen fibers, the scoring was based on percentage: 0 = 0%–25%; 1 = 26%–50%; 2 = 51%–75%; 3 = 76%–100%. Regarding collagen fiber density, the scoring system was defined as follows: 0 = very loose, 1 = loose, 2 = dense, 3 = very dense. Vascular distribution and cellular composition were also scored based on percentage presence: 0 = absent or very few, 1 = mildly present, 2 = moderately present, 3 = severe or significant. Each section was examined using an Olympus microscope and analyzed using ImageJ software.
[0046] 6. Statistical Analysis Based on preliminary data (mean failure loads: 22.02 N, 21.06 N, 29.97 N; SD = 3.54 N) and comparable studies, efficacy analysis was performed to determine that at least 5 shoulder joints were required per group to achieve 90% efficacy at a significance level of α = 0.05 (β = 0.1). The normality of the data was assessed using the Kolmogorov-Smirnov test to confirm a normal distribution. Data are expressed as mean ± standard deviation. One-way ANOVA was performed, followed by post-hoc Fisher's minimum significance analysis to determine p-values; p < 0.05 was considered statistically significant. Statistical analysis was performed using GraphPad Prism 10.1.2.
[0047] 7. Results 7.1 Characterization Analysis of GO-Gel Fourier transform infrared spectroscopy was used to analyze the chemical structural changes and potential interactions of graphene oxide (GO) after incorporation into polysaccharide-based gels. Figure 3 As shown in Figure A, the FTIR spectrum is in the range of 1000 to... Multiple characteristic absorption peaks of GO were observed within the range of 3600 to [missing value]. Within the wavenumber range, both GO and GO-Gel exhibit broad absorption bands, primarily attributed to the stretching vibrations of OH bonds, indicating a high hydroxyl content in the materials. Compared to GO, GO-Gel shows stronger absorption, likely due to the addition of hydroxyl-containing compounds such as sodium alginate and polyvinyl alcohol. and Two absorption peaks were observed at the GO-Gel, which are attributed to the stretching vibrations of the CH bonds in the organic compound. In GO-Gel, these peaks are stronger than in GO, possibly due to the CH bonds in the added glycerol and D-lactose monohydrate.
[0048] UV-Vis spectroscopy analysis showed that both GO and GO-Gel exhibited significant absorption peaks at approximately 230 nm. Figure 3 B), usually related to π-π in carbon materials Electronic transitions are involved. Furthermore, a small absorption peak is observed near 300 nm, which may be related to residual functional groups in the material. XRD analysis shows that the (002) crystal plane in the GO sample exhibits a strong diffraction peak at approximately 10°. Figure 3 C), indicating its typical layered structure. Compared to GO, the (001) peak in GO-Gel weakens and shifts to a higher angle, implying a decrease in interlayer spacing. The disappearance of the lower angle peak may indicate partial exfoliation or structural reorganization of the GO layers due to gelation. Zeta potential analysis showed that the Zeta potential of GO was -27 mV, while that of GO-Gel was -36 mV ( Figure 3The higher negative value (D) indicates enhanced stability of GO-Gel in aqueous media. TEM analysis shows that GO exhibits its typical thin-layer structure; in contrast, GO-Gel displays a more complex structure. Figure 3 E), including increased irregularity and interlayer connectivity, which may enhance the mechanical properties and functionality of the material.
[0049] SEM analysis showed that the gel had a uniform and continuous network structure with small and evenly distributed pores; in contrast, GO-Gel exhibited a more complex structure, especially the tight integration of the GO layer with the gel matrix. Figure 4 A) indicates that GO has been successfully incorporated into the gel. The porosity of the GO-Gel not only increases but also becomes irregular, reflecting the significant impact of GO addition on the gel network structure. Raman spectroscopy analysis showed that... and The main D and G peaks were identified at the location. Figure 4 B). The higher intensity of peak D compared to peak G indicates the presence of numerous defects in GO, suggesting its strong chemical reactivity. This comprehensive analysis highlights the significant changes in the properties of GO after integration into the gel matrix, providing insights into its enhanced functionality for various applications.
[0050] At 24 hours, tendon cell viability exhibited a concentration-dependent response: viability gradually increased with increasing GO-Gel concentration up to 0.025 mg / mL, then decreased at higher concentrations (0.05 mg / mL). A similar trend was observed at the 72-hour time point, with peak cell viability reached at 0.025 mg / mL GO-Gel. Figure 4 C). Based on these findings, 0.025 mg / mL GO-Gel was determined to be the optimal concentration for promoting tendon cell proliferation in vitro.
[0051] 7.2 Biomechanical Testing Biomechanical characteristics at 3 and 9 weeks post-surgery, such as Figure 5 As shown, values are expressed as mean ± standard deviation. MF, simple microfracture; MF+Gel, gel microfracture group; MF+GO-Gel, graphene oxide gel microfracture group; W, week. P < 0.05. From Figure 5As can be seen, at 3 weeks, the failure load of the MF+GO-Gel group (29.57±4.98 N) was significantly higher than that of the MF group (21.47±2.65 N; P = 0.0179), and the stiffness of the MF+GO-Gel group (9.24±1.48 N / mm) was significantly greater than that of the MF group (5.00±1.82 N / mm; P = 0.0039). At 9 weeks, the stiffness of the MF group (11.50±3.06 N / mm) was significantly lower than that of the MF+GO-Gel group (15.82±4.06 N / mm; P = 0.0342). There were no statistically significant differences in failure load and stiffness between the MF+Gel group and the MF+GO-Gel group and the MF group at 3 and 9 weeks postoperatively.
[0052] 7.3 Histological Analysis Figure 6 Hematoxylin-eosin (HE) staining and Masson trichrome (MT) staining of the tendon-bone interface at 3 and 9 weeks were performed in the MF, MF+Gel, and MF+GO-Gel groups. Values are expressed as mean ± standard deviation. W, weeks. P < 0.05; P <0.01. From Figure 6 As can be seen, after 3 weeks, compared with the MF group, the specimens in the MF+GO-Gel group showed improved collagen fiber continuity, more ordered and denser collagen fibers, fewer inflammatory cells, and increased vascular distribution at the tendon-bone interface (all P<0.05). Detailed histological scores are shown in Table 1. At 9 weeks, compared with the MF group, the MF+GO-Gel group showed improved collagen fiber continuity and fewer inflammatory cells (P < 0.01). Detailed histological scores are shown in Tables 1 and 2.
[0053] Table 1. Histological grading results at 3 weeks in the MF, MF+Gel, and MF+GO-Gel groups.
[0054] Table 2. Histological grading results of the MF, MF+Gel, and MF+GO-Gel groups at 9 weeks.
[0055] Collagen continuity / orientation: 0 (0–25%), 1 (26–50%), 2 (51–75%), 3 (76–100%); collagen fiber density: 0 (very loose) to 3 (very dense); blood vessel distribution / cellular composition: 0 (<5%), 1 (5–25%), 2 (26–50%), 3 (>50%).
[0056] Further quantitative analysis using picric acid-Sirius red staining showed that, as Figure 7 As shown, the semi-quantitative analysis of collagen content was based on light intensity. Values are expressed as mean ± standard deviation. W, weeks. P < .05; P < 0.001. From Figure 7 As can be seen, compared with the MF group, the MF+GO-Gel group had significantly higher brightness at both time points (both P < 0.001).
[0057] The results indicate that the synergistic combination of microfractures and graphene oxide-polysaccharide hydrogel (GO-Gel) significantly enhances tendon-bone healing, providing a promising therapeutic strategy for rotator cuff repair. The observed improvements in biomechanical strength and histological structure suggest that this approach may address key challenges in soft tissue-bone regeneration. Beyond rotator cuff injuries, this technology also has the potential for translational applications in other musculoskeletal fields, including articular cartilage repair and bone defect regeneration in the knee joint. The unique physicochemical properties of graphene oxide—including its high specific surface area, mechanical strength, and bioactivity—make it an ideal platform for next-generation regenerative therapies.
[0058] The embodiments described above are some, but not all, of the embodiments of this application. The detailed description of the embodiments of this application is not intended to limit the scope of the claimed application, but merely to illustrate selected embodiments. All other embodiments obtained by those skilled in the art based on the embodiments of this application without inventive effort are within the scope of protection of this application.
Claims
1. A method for preparing a graphene oxide hydrogel for treating rotator cuff injuries, characterized in that, Includes the following steps: S1. Graphene oxide is uniformly dispersed in water to form a graphene oxide suspension; S2. Dissolve the water-soluble synthetic polymer framework material in water to obtain a framework material solution, then add a pore-forming aid to form a mixture after dissolution. S3. The graphene oxide suspension, anionic crosslinking agent and solution stabilizer are sequentially incorporated into the above mixture. After being mixed evenly, the mixture is cooled to room temperature, frozen at low temperature and freeze-dried to obtain the graphene oxide hydrogel.
2. The method for preparing a graphene oxide hydrogel for treating rotator cuff injuries according to claim 1, characterized in that, In step S1, the ratio of graphene oxide to water in the graphene oxide suspension is 1 g: (1-10) mL. The dispersion is specifically achieved by stirring for 30-60 min and then ultrasonically treating for 3-5 min.
3. The method for preparing a graphene oxide hydrogel for treating rotator cuff injuries according to claim 1, characterized in that, The preparation of the framework material solution in step S2 is specifically as follows: a water-soluble synthetic polymer framework material is dissolved in water at a ratio of 1g:(20-30)mL, then autoclaved at 115-125℃ for 5-20min, cooled to 70-90℃ and stirred, and finally diluted with water to a framework material solution with a concentration of 1.5-3%; the water-soluble synthetic polymer framework material is polyvinyl alcohol, polyvinylpyrrolidone, polyacrylamide, polyacrylic acid or copolymers thereof.
4. The method for preparing a graphene oxide hydrogel for treating rotator cuff injuries according to claim 1, characterized in that, In step S2, the mass ratio of the water-soluble synthetic polymer framework material to the pore-forming agent is 1:(0.25-2); after the pore-forming agent is added, it is stirred at 700-900 rpm at 50-70°C until completely dissolved; the pore-forming agent is D-lactose monohydrate, sucrose, trehalose, glucose, mannitol, or maltose.
5. The method for preparing a graphene oxide hydrogel for treating rotator cuff injuries according to claim 1, characterized in that, In step S3, the final concentration of graphene oxide in the hydrogel is 0.0125–0.2 mg / mL.
6. The method for preparing a graphene oxide hydrogel for treating rotator cuff injuries according to claim 1, characterized in that, The anionic crosslinking agent is sodium alginate, chitosan, sodium hyaluronate, carboxymethyl cellulose, gelatin, or collagen.
7. The method for preparing a graphene oxide hydrogel for treating rotator cuff injuries according to claim 1, characterized in that, The solution stabilizer is glycerol, ethylene glycol, propylene glycol, polyethylene glycol-200, polyethylene glycol-400, or sorbitol.
8. The method for preparing a graphene oxide hydrogel for treating rotator cuff injuries according to claim 1, wherein the mass ratio of the anionic crosslinking agent to the water-soluble synthetic polymer framework material is 1:(1.5-2.5), in step S3, after adding the anionic crosslinking agent, the mixture is stirred at 700-900 rpm at 60°C until completely dissolved, and then a solution stabilizer is added, wherein the solution stabilizer accounts for 5-15% of the final mixture volume, and after addition, the mixture is stirred at the same temperature and speed until homogeneous.
9. The method for preparing graphene oxide hydrogel for treating rotator cuff injuries according to claim 1, wherein the low-temperature freezing temperature in step S3 is -70 to -90°C and the time is 4 to 12 hours; the vacuum degree during freeze-drying is 10 to 20 Pa, the temperature is ≤ -60°C, and the time is 24 to 48 hours.
10. A graphene oxide hydrogel for treating rotator cuff injuries, characterized in that, It is prepared by the method described in any one of claims 1 to 9.