Scaffold containing graphene oxide aerogel and biological glue and preparation method
By injecting bioglue with gelatin and dextran oxide into graphene oxide aerogel stents, the bioglue-graphene oxide aerogel composite scaffold was formed, and the problem of difficulty in retaining graphene oxide aerogel in the maxillofacial area was solved, better retention and mechanical properties were achieved, and controllable temperatures were generated locally, enhancing the tumor clearance and bone regeneration ability.
Patent Information
- Application Number
- CN202411872626.1
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-12-18
- Publication Date
- 2025-02-28
AI Technical Summary
Graphene oxide aerogel is difficult to relocate in complex and humid environments of the oral and maxillofacial area, and its brittle quality and poor mechanical properties, making it difficult to effectively repair tissue defects and prevent tumor recurrence in the treatment of maxillofacial osteosarcoma.
Graphene oxide aerogel grid-like scaffolds were prepared by 3D printing technology, and bioglue with gelatin and dextran oxide was injected into it. The bioglue was formed by cross-linking to enhance the bonding strength and mechanical properties of the scaffolds.
The retention ability and mechanical properties of graphene oxide aerogel in the maxillofacial area are improved, and the photothermal grading response is achieved locally generated by controlled temperatures, enhancing the tumor clearance and bone regeneration capabilities.
Smart Images
Figure CN119524198A_ABST
Abstract
Description
Technical Field
[0001] The invention belongs to the technical field of biomedicine and relates to a stent comprising graphene oxide aerogel and biological glue and a preparation method thereof. Background Art
[0002] Maxillofacial osteosarcoma is a malignant tumor derived from mesenchymal tissue. It is highly invasive and often causes significant bone destruction and functional impairment, such as affecting pronunciation, swallowing, and chewing. Treatment usually requires extensive surgical resection based on the biological characteristics of the tumor, as well as preoperative and postoperative chemotherapy to reduce the risk of recurrence. However, these treatments often lead to significant tissue defects that exceed the body's natural repair capacity. Therefore, how to effectively repair these defects and prevent tumor recurrence has become a key issue in the treatment of maxillofacial osteosarcoma.
[0003] Photothermal therapy (PTT) is a new research direction for tumor treatment that has emerged in recent years. Photothermal agents convert the radiation energy in near-infrared light (NIR) into thermal energy through energy conversion, intervene in surrounding tissues, and achieve therapeutic effects. Compared with traditional radiotherapy and chemotherapy, it has the advantages of less invasiveness, high specificity, and less systemic toxicity and side effects. Graphene oxide aerogel is a material with high photothermal conversion efficiency. Its excellent photothermal conversion performance, good biocompatibility and biodegradability make it an ideal photothermal material for constructing tissue engineering scaffolds. On the one hand, graphene oxide aerogel is difficult to adapt to the complex and humid mechanical environment of the oral and maxillofacial region, and retention becomes one of the factors affecting the effect; on the other hand, graphene oxide aerogel is brittle and has poor mechanical properties, and appropriate methods need to be adopted to enhance the strength of the material itself. Summary of the invention
[0004] The purpose of the present invention is to further improve the retention of graphene oxide aerogel in the complex humid environment of the oral and maxillofacial region in a simple and low-cost manner, and to improve the precise control of local temperature by graphene oxide aerogel to achieve photothermal graded response. To this end, the present invention provides a scaffold comprising graphene oxide aerogel and biological glue and a preparation method to solve this need in the art.
[0005] In one aspect, the present invention relates to a scaffold comprising graphene oxide aerogel and biological glue, wherein the scaffold comprises graphene oxide aerogel and biological glue;
[0006] The graphene oxide aerogel is in a grid shape;
[0007] The biological glue is a mixed liquid containing gelatin and oxidized dextran, which is arranged in any area inside the graphene oxide aerogel, and then the gelatin and oxidized dextran are cross-linked to form a gel.
[0008] Furthermore, in the scaffold comprising graphene oxide aerogel and biological glue provided by the present invention, the ratio of gelatin to oxidized dextran is 10:1 to 100:1 by mass ratio.
[0009] Further, in the scaffold comprising graphene oxide aerogel and biological glue provided by the present invention, the biological glue accounts for at least 5% of the graphene oxide aerogel by volume percentage;
[0010] The volume of the biological glue is not less than 0.5 mL.
[0011] Generally, the bioglue accounts for 5-10% of the graphene oxide aerogel in terms of volume percentage.
[0012] Preferably, the volume ratio of the biological glue to the graphene oxide aerogel is 1:9-20.
[0013] On the other hand, the present invention relates to a method for preparing the scaffold comprising graphene oxide aerogel and biological glue, comprising:
[0014] The graphene oxide aerogel is made into a grid shape as a main body;
[0015] After the gelatin solution and the oxidized dextran solution are mixed, they are injected into any area inside the main body before gelation, and the biological glue is formed after the gelatin and the oxidized dextran are cross-linked to form gel.
[0016] The present invention does not particularly limit the preparation of graphene oxide aerogel and the method of 3D printing different properties, and those skilled in the art can easily obtain them from existing preparation methods (for example: Y Jiang, Z Xu, T Huang, et al. Direct 3D printing of ultralight graphene oxide aerogel microlattices [J]. Advanced Functional Materials, 2018, 28, 1707024.).
[0017] The present invention does not specifically limit the position, quantity and dosage of BG in the main body, and those skilled in the art can easily make arbitrary adjustments based on the position of the maxillofacial osteosarcoma relative to the bracket.
[0018] Furthermore, in the method for preparing a scaffold comprising graphene oxide aerogel and biological glue provided by the present invention, the graphene oxide aerogel is manufactured into the main body by 3D printing.
[0019] On the other hand, the present invention relates to the use of the stent comprising graphene oxide aerogel and biological glue in the preparation of an oral and maxillofacial thermal therapy retention stent.
[0020] Furthermore, in the application provided by the present invention, the laser power used for the thermal therapy is 0.5 to 2 W / cm 2 .
[0021] Furthermore, in the application provided by the present invention, the temperature of the thermal therapy is not lower than 43°C.
[0022] Furthermore, in the application provided by the present invention, the biological glue is subjected to cyclic irradiation of laser to produce periodic temperature changes locally.
[0023] Furthermore, in the application provided by the present invention, the biological glue improves the bonding strength of the oral and maxillofacial thermal therapy retention bracket.
[0024] Compared with the prior art, the technical solution provided by the present invention has at least the following beneficial effects or advantages:
[0025] The present invention uses 3D printing technology to prepare a graphene oxide aerogel scaffold (GOAs) with photothermal conversion efficiency as the main body, and coats it with low swelling and fracture-resistant gelatin-oxidized dextran bioglue (Biomedical Glue, BG) to give it adhesion ability in the moist and dynamic mechanical environment of the oral cavity, and constructs a biomedical glue-graphene oxide aerogel scaffold (BG-GOAs) with tumor removal and bone regeneration capabilities. It has better mechanical strength, uses the photothermal effect to accurately control the local temperature, and realizes tumor removal and bone tissue regeneration with photothermal graded response, which provides more ideas for improving the thermal therapy effect of osteosarcoma. BRIEF DESCRIPTION OF THE DRAWINGS
[0026] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.
[0027] Figure 1 Schematic diagram of the GOAs bracket.
[0028] Figure 2 Schematic diagram of BG hydrogel formation.
[0029] Figure 3 Schematic diagram of BG-GOAs composite scaffold.
[0030] Figure 4 This is the photothermal efficiency test results of the BG-GOAs composite bracket.
[0031] Figure 5 This is a graph showing the experimental results of the effect of heat stimulation on the proliferation activity of MNNG / HOS cells.
[0032] Figure 6 This is the result of the scratch test.
[0033] Figure 7 This is the result of ALP staining.
[0034] Figure 8 This is the ARS staining result.
[0035] Fig. 9 Figure 2 shows the bonding strength test results of BG-GOAs. DETAILED DESCRIPTION
[0036] The technical scheme of the present invention is described below in conjunction with the embodiments, but the present invention is not limited to the following embodiments. The experimental methods and detection methods described in each embodiment are conventional methods unless otherwise specified; the reagents and materials described are available on the market unless otherwise specified. The % in the following embodiments are all mass percentages unless otherwise specified. The ratios in the following embodiments are all mass ratios unless otherwise specified.
[0037] Example 1
[0038] This example provides the preparation and performance testing of the BG-GOAs composite scaffold.
[0039] Refer to Y Jiang, Z Xu, T Huang, et al. Direct 3D printing of ultralightgraphene oxide aerogel microlattices [J]. Advanced Functional Materials, 2018, 28, 1707024. GOAs scaffolds were successfully prepared by 3D printing and freeze drying. Figure 1 As shown in the figure, the GOAs are black porous grid-like stents, arranged in 90° vertical and horizontal layers. Their shape can be designed by computer to print a personalized stent that matches the exact match.
[0040] The 20% gelatin solution and the 20% oxidized dextran solution were quickly taken out of the 45°C oven and subjected to a hydrogel crosslinking gelation test at room temperature of 23°C. The hydrogels with a gelatin: oxidized dextran mass ratio of 10:1, 20:1, 50:1, and 100:1 were able to gel, and the gelation time was 14s, 37s, 78s, and 122s, respectively. When the gelatin: oxidized dextran mass ratio was 200:1, the hydrogel could not gel stably. Figure 2 Schematic diagram of the gelation of hydrogel with a gelatin:oxidized dextran ratio of 20:1 (left: before gelation, right: after gelation).
[0041] like Figure 3 As shown, before the gelatin-oxidized dextran hydrogel is gelled, 0.5 mL is injected into the GOAs scaffold (BG accounts for 5% of the total volume of GOAs) through a syringe to obtain a composite scaffold with excellent mechanical properties and bonding properties.
[0042] like Figure 4 As shown in the figure, different temperature changes can be produced locally by irradiating the stent with lasers of different intensities. 2 The laser can heat the local area to 35℃, 1.5w / cm 2 The laser can raise the temperature locally to 58°C. 2 The near-infrared laser cyclically irradiates the stent, which can produce periodic temperature changes locally, and the photothermal stability of the material is reliable. Figure 4 It can be seen that with the introduction of BG, BG-GOAs can generate controllable temperature locally rather than overall when irradiated by laser.
[0043] Pigskin was used for simulated bonding. 2×2cm pigskin was bonded to an acrylic plate using 502. The GOAs bracket was placed in the center of the soaked pigskin. BG (gelatin: oxidized dextran mass 20:1) was injected on the surface and inside of the bracket. A silicone pad was used to prevent BG from overflowing. After complete gelation, 502 was used to bond the bottom surface of BG-GOAs to another acrylic plate. A 180° peel test was then used to test the bonding strength of BG-GOAs. The results are shown in the figure. Fig. 9 The present invention tests that any amount of BG can improve the bonding strength of BG-GOAs in a wet environment, and the bonding strength increases with the increase in the amount. Fig. 9 It can be seen that the bonding strength of the test group with a volume ratio of 20:1 is 15.30 kPa, and the bonding strength of the test group with a volume ratio of 10:1 is 12.68 kPa; in contrast, the bonding strength of GOAs in a wet environment is 0 kPa.
[0044] The composite of BG and GOAs can enhance the mechanical properties of the GOAs scaffold, which has poor mechanical properties; on the other hand, it can give the GOAs scaffold the ability to bond and retain. The strength of the material was tested using the uniaxial tensile method, and the material strengths of GOAs, 20:1 group BG, and 20:1 group composite BG-GOAs were tested respectively. The results showed that the strength of GOAs was only 13.7 kPa, the strength of 20:1 group BG was 116.44 kPa, and the strength of BG-GOAs scaffolds was 137.77 kPa.
[0045] Example 2
[0046] This example provides the response of MNNG / HOS cells to hyperthermia.
[0047] (1) Effect of thermal stimulation on the proliferation activity of MNNG / HOS cells
[0048] The cells were heated at 37, 40, 43, 46, and 49°C for 15 minutes, and the CCK8 kit was used to detect changes in cell proliferation activity. Compared with the 37°C control group, the proliferation activity of cells in the 43°C, 46°C, and 49°C groups decreased significantly (p<0.001), while there was no statistically significant change in the proliferation activity of cells in the 40°C group (p>0.05). Figure 5 It is shown that heat stimulation at 40℃ or below will not affect the proliferation of MNNG / HOS cells.
[0049] (2) Effect of hyperthermia on the invasiveness of MNNG / HOS cells
[0050] After scratching, the suspended cells were washed away and heated to 37, 40, 43, 46, and 49°C, respectively. Then, the cells were cultured and photographed by a cell photography system at 24 and 48 hours (e.g. Figure 6 ). The changes in scratch width were measured and the changes in the invasion rate of osteosarcoma cells were calculated. Compared with the 37℃ control group, the tumor invasion rate in the 40℃ group decreased slightly (p<0.05), and the tumor invasion rate in the 43℃ group decreased significantly (p<0.001). Due to the massive cell death in the 46℃ and 49℃ groups, the changes in scratch width could no longer be measured, and the changes in invasion rate could not be calculated.
[0051] (3) Alkaline phosphatase activity in BMSCs cells under different temperature stimulation
[0052] BMSCs cells were heated at 37°C, 40°C, and 43°C for 15 minutes every day. After 7 days, samples were collected and cells were stained with ALP staining reagent. Cell photography system was used to record the cells. The active site of alkaline phosphatase was blue-purple and located in the cytoplasm. The results of each group are shown in Figure 2. Figure 7 The cells in the 43℃ group were the heaviest stained, followed by the 40℃ group, and the 37℃ group were the lightest stained.
[0053] (4) Formation of mineralized nodules under different temperature stimulations
[0054] BMSCs cells were heated at 37°C, 40°C, and 43°C for 15 minutes every day. After 14 days, samples were collected and stained with ARS staining reagent. Cell photography system was used to record the mineralized nodules, which were red-orange and located outside the cells. Figure 8 The 43℃ group produced the most mineralized nodules, followed by the 40℃ group, and the 37℃ group produced the least mineralized nodules.
[0055] It can be seen from the above test results that during the thermal treatment of maxillofacial osteosarcoma, the BG-GOAs composite scaffold provided by this application is easy to use in surgical implantation scenarios. The graphene oxide scaffold can be placed in the bone defect area first, and then the oxidized dextran-gelatin hydrogel can be injected into the defect area, which can enhance the performance of the graphene scaffold, fill the tissue defect area and promote the retention of the graphene oxide scaffold. In addition, the BG-GOAs composite scaffold provided by this application has a laser power of 0.5 to 2 W / cm 2 The corresponding adaptive temperature range is suitable for the fixed-point controllable photothermal treatment of maxillofacial osteosarcoma.
[0056] As described above, the basic principle, main features and advantages of the present invention are well described. The above embodiments and descriptions are only descriptions of the preferred implementation modes of the present invention, and the present invention is not limited by the above embodiments. Without departing from the spirit and scope of the present invention, various changes and improvements made by ordinary technicians in this field to the technical solution of the present invention should fall within the protection scope determined by the present invention.
Claims
1. A scaffold comprising graphene oxide aerogel and biological glue, characterized in that: The scaffold comprises graphene oxide aerogel and biological glue; The graphene oxide aerogel is in a grid shape; The biological glue is a mixed liquid containing gelatin and oxidized dextran, which is arranged in any area inside the graphene oxide aerogel, and then the gelatin and oxidized dextran are cross-linked to form a gel.
2. The scaffold comprising graphene oxide aerogel and biological glue according to claim 1, characterized in that: In terms of mass ratio, the ratio of gelatin to oxidized dextran is 10:1 to 100:
1.
3. The stent comprising graphene oxide aerogel and biological glue according to claim 1, characterized in that: The bioglue accounts for at least 5% of the graphene oxide aerogel by volume percentage; The volume of the biological glue is not less than 0.5 mL.
4. A method for preparing a scaffold comprising graphene oxide aerogel and biological glue as claimed in any one of claims 1 to 3, characterized in that: include: The graphene oxide aerogel is made into a grid shape as a main body; After the gelatin solution and the oxidized dextran solution are mixed, they are injected into any area inside the main body before gelation, and the biological glue is formed after the gelatin and the oxidized dextran are cross-linked to form gel.
5. The method for preparing a scaffold comprising graphene oxide aerogel and biological glue according to claim 4, characterized in that: The graphene oxide aerogel is manufactured into the main body through 3D printing.
6. Use of the stent comprising graphene oxide aerogel and biological glue as claimed in any one of claims 1 to 3 in the preparation of oral and maxillofacial thermal therapy retention stents.
7. The application according to claim 6, characterized in that: The laser power used for the thermal therapy is 0.5-2 W / cm 2 .
8. The application according to claim 6, characterized in that: The temperature of the thermal therapy is not lower than 43°C.
9. The application according to claim 6, characterized in that: The biological glue is irradiated with laser in a cycle, thereby generating periodic temperature changes locally.
10. The application according to claim 6, characterized in that: The biological glue improves the bonding strength of the oral and maxillofacial thermal therapy retention bracket.