SA-GO / SiO2 composite injectable hydrogel as well as preparation method and application thereof

Abstract

The invention discloses SA-GO / SiO2 composite injectable hydrogel and a preparation method and application thereof.The preparation method comprises the steps that graphene oxide (GO) powder is added into a sodium alginate (SA) solution, and the mixture is stirred to be uniform through a magnetic stirrer till the powder is completely dispersed in the solution; adding nano silicon dioxide (nSiO2) powder into the solution, and uniformly stirring by using a magnetic stirrer until the powder is completely dispersed in the solution; a CaCl2 solution is slowly dropwise added into the solution obtained in the previous step at the dropwise adding speed of 20 drops per minute, and SA-GO / SiO2 composite hydrogel containing SiO2 with different concentrations is formed through Ca < 2 + > crosslinking and can be used as an injectable biological scaffold material for bone tissue repair, so that proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells are promoted, and the bone tissue repair efficiency is improved.

Description

Technical Field

[0001] The present invention belongs to the field of material technology, and specifically relates to a sodium alginate@graphene oxide-silicon dioxide (SA@GO / SiO 2 )Composite injectable hydrogel and its preparation method and application. Background Art

[0002] Sodium alginate (SA) is a biodegradable anionic polymer with good tissue compatibility. 2+ The interaction of divalent cations such as SA and SA forms a stable gel. This gelation ability makes SA widely used as a hydrogel scaffold material. SA hydrogel is a porous three-dimensional network structure, similar to the structure of the extracellular matrix, which is conducive to material exchange with the surrounding tissue and is a very suitable scaffold material. However, SA alone is difficult to apply to bone tissue engineering because of its weak mechanical strength. Therefore, it is necessary to combine SA with other materials for research on bone tissue engineering.

[0003] Graphene oxide (GO) is an important derivative of graphene-based materials and an emerging carbon material for bone tissue engineering. It has the characteristics of high mechanical strength, easy functionalization, large surface area and high biocompatibility. Adding GO to various types of hydrogels can significantly improve their mechanical properties. GO composites have been used to manufacture various bioactive scaffolds and have become a research hotspot in the field of bone tissue engineering.

[0004] Si is mostly introduced into various biomaterials in the form of nano-silicon dioxide, and exhibits good bone integration, bone induction and bone conduction abilities. Studies have shown that silicon ions (Si 4+ ) can upregulate bone activity, promote osteogenic differentiation and bone mineralization, stimulate angiogenesis, and accelerate bone regeneration. 2 The surface of nanoparticles is rich in hydroxyl groups, which can form stable chemical bonds with other biomaterials, thereby improving the overall performance of the composite material. Summary of the invention

[0005] The purpose of the present invention is to provide a SA@GO / SiO 2 Composite injectable hydrogel and preparation method and application thereof.

[0006] The present invention is achieved through the following technical solutions:

[0007] SA@GO / SiO 2 The preparation method of the composite injectable hydrogel comprises the following steps:

[0008] 1) preparing a SA solution with a concentration of 1-5 wt.%;

[0009] 2) Prepare CaCl with a concentration of 0.5-1wt.% 2 Solution;

[0010] 3) Add GO powder to the SA solution and stir evenly with a magnetic stirrer until the powder is completely dispersed in the solution; the mass ratio of GO powder to SA is 1:20-25;

[0011] 4) Add nSiO to the solution in step 3) 2 The powder was stirred evenly with a magnetic stirrer until the powder was completely dissolved in the solution. 2 The powder mass ratio is 1:1-4;

[0012] 5) Slowly add CaCl to the solution in step 4) 2 The solution was added at a rate of 20 drops per minute and mixed to obtain a solution containing SiO 2 SA@GO / SiO 2 Composite hydrogel, CaCl 2 The amount of solution added is 1 / 5-1 / 4 of the volume of the SA solution.

[0013] The preparation method described herein comprises sodium alginate: molecular weight 20,000-50,000, viscosity 15-60 mPa.s; CaCl 2 : Dihydrate, analytical grade AR; nano-SiO 2 Commercially available silicon dioxide, 99.5%, particle size 50±5nm; graphene oxide: multilayer, 98%.

[0014] In the preparation method, the SA solution is 5wt.%.

[0015] The preparation method, CaCl 2 The solution is 1 wt.%.

[0016] The preparation method, nSiO 2 The powder addition amount is 10-40mg.

[0017] In the preparation method, the rotation speed of the magnetic stirring magnet is 300-600 r / min.

[0018] SA@GO / SiO prepared by any of the preparation methods 2 Composite injectable hydrogel.

[0019] The SA@GO / SiO 2 The application of composite injectable hydrogels in the preparation of injectable biological scaffold materials for promoting bone tissue repair.

[0020] Compared with the prior art, the present invention has the following beneficial effects:

[0021] 1. According to this preparation method, SA, GO, SiO 2 The three have good compatibility in the composite scaffold, forming a three-dimensional network with high pore connectivity, which is beneficial to the transport and release of substances.

[0022] 2. The composite hydrogels prepared by the present invention all exhibit good cell compatibility and will not produce significant toxic effects on cells.

[0023] 3. The present invention loads graphene oxide (GO) and nano-silicon dioxide (nSiO) on sodium alginate (SA) hydrogel cross-linked with calcium ions. 2 ), synthesize SA@GO / SiO 2 The composite hydrogel can be used as an injectable biological scaffold material for bone tissue repair to promote the proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells and improve the efficiency of bone tissue repair. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 The SEM images of the composite hydrogels prepared by the method of the present invention are (A) SA hydrogel, (B) SA@GO / SiO 2 Hydrogel;

[0025] Figure 2 SA@GO / SiO prepared by the method of the present invention 2 XPS spectrum of composite hydrogel;

[0026] Figure 3 SA@GO / SiO prepared by the method of the present invention 2 Thermogravimetric analysis curve of composite hydrogel;

[0027] Figure 4 Rheological test of the composite hydrogel prepared by the method of the present invention, (A) frequency scanning result, (B) strain scanning result;

[0028] Figure 5 The swelling properties of the composite hydrogel prepared by the method of the present invention;

[0029] Figure 6 The biocompatibility evaluation results of the composite hydrogel prepared by the method of the present invention. (AC) CCK8 method was used to detect the cell viability of rat bone marrow mesenchymal stem cells (BMSCs) after co-culture with the hydrogel extract for 24, 48, and 72 hours; (DF) live / dead cell staining results after BMSCs were co-cultured with the hydrogel extract for 24 hours. DETAILED DESCRIPTION

[0030] The present invention is described in detail below in conjunction with specific embodiments.

[0031] Example 1

[0032] The present invention discloses a method for preparing a graphene oxide / nano-silicon dioxide composite sodium alginate injectable hydrogel, and the method steps are as follows:

[0033] 1) preparing a SA solution with a concentration of 5 wt.%;

[0034] 2) Prepare CaCl with a concentration of 1wt.% 2 Solution;

[0035] 3) Add 10 mg of GO powder to 4 mL of SA solution (5 wt.%) and stir evenly using a magnetic stirrer until the powder is completely dispersed in the solution;

[0036] 4) Add 10 mg nSiO to the solution in step 3) 2 Powder, use a magnetic stirrer to stir evenly until the powder is completely dispersed in the solution;

[0037] 5) Slowly add 1 mL of CaCl to the solution in step 4) 2 Solution (1wt.%), dripping speed is 20 drops per minute, after mixing, SA@GO / SiO 2 Composite hydrogel.

[0038] Example 2

[0039] The present invention discloses a method for preparing a graphene oxide / nano-silicon dioxide composite sodium alginate injectable hydrogel, and the method steps are as follows:

[0040] 1) preparing a SA solution with a concentration of 5 wt.%;

[0041] 2) Prepare CaCl with a concentration of 1wt.% 2 Solution;

[0042] 3) Add 10 mg of GO powder to 4 mL of SA solution (5 wt.%) and stir evenly using a magnetic stirrer until the powder is completely dispersed in the solution;

[0043] 4) Add 20 mg nSiO to the solution in step 3) 2 Powder, use a magnetic stirrer to stir evenly until the powder is completely dispersed in the solution;

[0044] 5) Slowly add 1 mL of CaCl to the solution in step 4) 2 Solution (1wt.%), dripping speed is 20 drops per minute, after mixing, SA@GO / SiO 2 Composite hydrogel.

[0045] Example 3

[0046] The present invention discloses a method for preparing a graphene oxide / nano-silicon dioxide composite sodium alginate injectable hydrogel, and the method steps are as follows:

[0047] 1) preparing a SA solution with a concentration of 5 wt.%;

[0048] 2) Prepare CaCl with a concentration of 1wt.% 2 Solution;

[0049] 3) Add 10 mg of GO powder to 4 mL of SA solution (5 wt.%) and stir evenly using a magnetic stirrer until the powder is completely dispersed in the solution;

[0050] 4) Add 40 mg nSiO to the solution in step 3) 2 Powder, use a magnetic stirrer to stir evenly until the powder is completely dispersed in the solution;

[0051] 5) Slowly add 1 mL of CaCl to the solution in step 4) 2 Solution (1wt.%), dripping speed is 20 drops per minute, after mixing, SA@GO / SiO 2 Composite hydrogel.

[0052] Example 4

[0053] SA@GO / SiO 2 SEM test of composite hydrogel

[0054] After freeze drying of Examples 1-3, the microscopic morphology of the cross section was studied by SEM. The results are as follows: Figure 1 . It can be observed that SA, GO, SiO 2 The three have good compatibility in the composite hydrogel, and a large number of pores inside are interconnected in a three-dimensional network, forming a highly porous interconnected structure, which is conducive to the transport and release of substances.

[0055] SA@GO / SiO 2 Structural analysis of composite hydrogels

[0056] In order to more clearly elucidate the structure of the composite hydrogel material, SA@GO / SiO 2 XPS and TGA analysis were performed. Figure 2 and Figure 3 ,The analysis results indicated the successful incorporation of graphene oxide and nano-silica in the composite hydrogel.

[0057] SA@GO / SiO 2 Rheological testing of composite hydrogels

[0058] In order to examine the rheological properties of the composite hydrogels, e.g. Figure 4 As shown, Figure 4 A is the frequency scanning result, Figure 4 B is the strain scanning result. SA@GO / SiO 2 The storage modulus (G') and loss modulus (G") of the hydrogel were significantly higher than those of the SA hydrogel, indicating that GO and SiO 2 The addition of increased the mechanical strength of the hydrogel, making SA@GO / SiO 2 The hydrogel has better stability.

[0059] SA@GO / SiO 2 Swelling rate detection of composite hydrogel

[0060] like Figure 5 As shown in the figure, the composite hydrogel absorbed water quickly in the first 3 h and then gradually stabilized. 2 The final swelling ratios of the hydrogels were 49.9% and 24.6%, respectively. 2 The doping of GO and SiO reduced the swelling capacity of the hydrogel. This may be due to the 2 The formation of hydrogen bonds between the functional groups (hydroxyl and carboxyl) on the surface and those on the alginate polymer network resulted in a tighter pore structure of the composite hydrogel, which to some extent hindered the entry of water molecules.

[0061] SA@GO / SiO 2 Biocompatibility evaluation of composite hydrogels

[0062] like Figure 6 As shown, the biocompatibility of the injectable hydrogels was evaluated using CCK-8 cell viability assay and cell live / dead staining. Figure 6 A shows that the cell viability of rat bone marrow mesenchymal stem cells (BMSCs) was detected by CCK-8 method after co-culturing the extracts of different hydrogels with rat bone marrow mesenchymal stem cells (BMSCs) for 24h, 48h, and 72h. As shown in the figure, there was no significant difference in the cell activity of different groups after 24h of co-culture with the control group. After 48h, the cell viability of SA@GO / SiO 2 The cell activity of the SA@GO / SiO group was significantly increased compared with that of the Control and SA groups. 2 The SA@GO / SiO 2 Hydrogel can significantly promote the proliferation of BMSCs and can be used as an injectable biological scaffold material for bone tissue repair to improve the efficiency of bone tissue repair. Figure 6As shown in B, live / dead staining was used to evaluate the effects of different hydrogel extracts on BMSCs after 24 h of co-culture. As shown in the figure, the cell morphology of each group was good, and almost no dead cells were found in all groups, indicating that the hydrogel had no toxic effect on the cells. The above results show that SA@GO / SiO 2 The hydrogel has good biocompatibility.

[0063] It should be understood that those skilled in the art can make improvements or changes based on the above description, and all these improvements and changes should fall within the scope of protection of the appended claims of the present invention.

Claims

1. A method for preparing a SA@GO / SiO2 composite injectable hydrogel, characterized in that: The preparation method comprises the following steps: 1) preparing a SA solution with a concentration of 1-5 wt.%; 2) preparing a CaCl2 solution having a concentration of 0.5-1wt.%; 3) Add GO powder to the SA solution and stir evenly with a magnetic stirrer until the powder is completely dispersed in the solution; the mass ratio of GO powder to SA is 1:20-25; 4) Add nSiO2 powder to the solution in step 3) and stir evenly using a magnetic stirrer until the powder is completely dispersed in the solution, and the mass ratio of GO powder to nSiO2 powder is 1:1-4; 5) Slowly add CaCl2 solution to the solution in step 4) and mix well to obtain SA@GO / SiO2 composite hydrogel containing SiO2. The amount of CaCl2 solution added is 1 / 5-1 / 4 of the volume of the SA solution.

2. The preparation method according to claim 1, characterized in that: Sodium alginate: molecular weight 20,000-50,000, viscosity 15-60mPa.s; CaCl2: dihydrate, analytical grade AR; nano-SiO2 is commercially available silicon dioxide, 99.5%, particle size 50±5nm; graphene oxide: multilayer, 98%.

3. The preparation method according to claim 1, characterized in that: The SA solution was 5 wt.%.

4. The preparation method according to claim 1, characterized in that: The CaCl2 solution was 1 wt.%, and the dropping speed was 20 drops per minute.

5. The preparation method according to claim 1, characterized in that: The amount of nSiO2 powder added is 10-40mg.

6. The preparation method according to claim 1, characterized in that: The magnetic stirring magnet has a rotation speed of 300-600 r / min.

7. SA@GO / SiO2 composite injectable hydrogel prepared according to any one of the preparation methods described in claims 1-6.

8. Use of the SA@GO / SiO2 composite injectable hydrogel according to claim 7 in the preparation of injectable biological scaffold materials for promoting bone tissue repair.

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