CN108392674B - Preparation method of high-bioactivity glass nanofiber scaffold - Google Patents

Abstract

The invention belongs to the field of biological functional materials, and discloses a preparation method of a high-bioactivity glass nanofiber scaffold. The method comprises the following steps: chemically reacting a pure bacterial cellulose film in a solution of ceric ammonium nitrate and ethylenediamine respectively, The amino group is grafted onto the hydroxyl group of bacterial cellulose to obtain aminated modified bacterial cellulose, which is freeze-dried to obtain aminated bacterial cellulose bulk. Then, using the aminated bacterial cellulose as a template, the precursors containing calcium and silicon elements were deposited on the surface of the bacterial cellulose by ultrasonic method, and then calcined to obtain a nano-biological glass fiber scaffold. The nanofiber glass scaffold can rapidly induce the formation of hydroxyapatite in body fluids due to its ultrafine nanoscale network structure and huge specific surface area, and has very high biological activity. The invention has the advantages of simple process, easy operation, low cost and the like, and has a good application prospect.

Description

A kind of preparation method of high bioactive glass nanofiber scaffold

technical field

The invention relates to the field of biological functional materials, in particular to a preparation method of a high biological activity glass nanofiber scaffold.

Background technique

Today, with the continuous development of science and technology, people pay more and more attention to the research of life science. Biomaterials, as the frontier field of life science and materials science, plays an immeasurable role in human rehabilitation engineering. As a representative of biomaterials, bioactive glass has attracted more and more attention as a kind of glass material that can repair, replace and regenerate body tissues, and can bond with tissues (especially bone tissue). One of the hot topics in the field of materials research. So far, a large number of studies have reported that bioglass with high specific surface area and porosity has higher bioactivity and good biomimetic mineralization performance, and can be immersed in simulated body fluid (SBF) for a period of time. The formation of bone-like hydroxycarbonated apatite crystals, which is also one of the most important criteria for measuring whether the material has biological activity. At present, the scale of bioglass scaffolds prepared at home and abroad is mainly concentrated before the micropores and macropores. The prepared biomaterials have a small specific surface area and low biological activity, which is difficult to meet the great demand for bioglass materials in human tissue engineering.

Because of its ultra-fine three-dimensional network structure, bacterial cellulose has a huge three-dimensional space outside the nanofibers, and it is easy to accommodate solutions or sol particles such as water, alcohols, inorganic substances, organic precursors, etc., which are the targets on the surface of bacterial cellulose. The deposition of bacterial cellulose provides conditions for the nanoscale pore size distribution and large specific surface area contained in bacterial cellulose, which is widely used in the field of biological functional nanomaterials. However, due to the low chemical activity of hydroxyl groups on the surface of bacterial cellulose, it is generally difficult to combine with cations and other components. Therefore, the method of amination modification of bacterial cellulose is used to increase the chemically active sites of bacterial cellulose. Greatly improved the binding capacity of cellulose to cations. In the present invention, the superfine network structure of bacterial cellulose is used as an organic template. After depositing the bioglass precursor components containing calcium and silicon, the bioglass nanofiber scaffold with the same superfine nano network structure is obtained by calcination. , so as to improve the biological activity of this glass scaffold. This method provides a new route for the preparation of inorganic bioglass nanofiber scaffolds, and has certain practical significance.

SUMMARY OF THE INVENTION

The technical problem to be solved by the present invention is to provide a method for preparing a nano-glass fiber scaffold with simple operation process, three-dimensional ultra-fine network structure, high specific surface area and high biological activity, which can realize mass production and is green and environmentally friendly. Amination modification of bacterial cellulose improves the induction and binding ability of cellulose nanofibers to the components of bioglass precursors, thereby preparing nanoscale bioglass fiber scaffolds with nano-network structure.

The technical scheme of the present invention is as follows:

(1) Cut the bacterial cellulose film into blocks, soak it in a 0.5-2 mol/L NaOH solution at 90 °C for 2-5 h, then wash it with a large amount of deionized water until it becomes neutral, and freeze it. Freeze-drying in a dryer to obtain bacterial cellulose bulk;

(2) The bacterial cellulose block was put into constant temperature water at 35 °C and nitrogen was continuously introduced to remove the oxygen dissolved in the water, and 0.1 mol/L cerium ammonium nitrate solution was added to react for 15 min, and then continued for 30 Drop into polyglycidyl methacrylate GMA for 2 h, wash it with deionized water after 2 h, and wash it with absolute ethanol again, so as to repeatedly wash to neutrality to obtain a white block, which is then freeze-dried; wherein nitric acid The ceric ammonium solution is dissolved with 1 mol/L HNO ₃ ; wherein the mass ratio of bacterial cellulose, ceric ammonium nitrate, and GMA is 1:20:2;

(3) Put the white block obtained in step (2) into 125 mL of mixed solution prepared by mass ratio of ethylenediamine:water=3:2, and stir at 80 ℃ for 2~5 h, and finally Wash with deionized water once, and then wash with absolute ethanol again, and repeat this for many times until the washing is neutral to obtain aminoated bacterial cellulose, which is then freeze-dried;

(4) Put the aminated cellulose obtained in step (3) into an ethanol solution of 0.1-1 mol/L Ca(NO ₃ ) ₂ ·4H ₂ O and sonicated for 3 h to obtain precalcified aminated bacterial fibers element, wherein the calcium-containing solution is changed every 1 h; wherein the ratio of the mass of the aminated bacterial cellulose block to the mass of the Ca(NO ₃ ) ₂ solution is 1:2000;

(6) (5) After taking out the pre-calcified aminated bacterial cellulose block obtained in step (4), put it directly into the silicon-containing solution and continue to sonicate for 3 h, and replace the silicon-containing solution every 1 h to maintain the silicon-containing solution. The concentration of ions, continue to be cleaned with deionized water after ultrasonication, and then freeze-dried; wherein the ratio of the mass of the precalcified aminated bacterial cellulose block to the mass of the silicon-containing solution is 1:2000; The aminated bacterial cellulose block was heated and sintered at 600-800 °C for 1-3 h, and the obtained bioactive nanoglass scaffold was obtained.

The silicon-containing solution in step (5) is specifically: by volume ratio, tetraethyl orthosilicate TEOS: anhydrous ethanol=1:40 mixed.

The solvent used in the freeze-drying step in the step is all tert-butanol, water or a mixed solution thereof.

The heating rate in the step (6) is 5-20 °C/min.

Compared with the prior art, the present invention has the following advantages:

1. The present invention has the advantages of simple process, easy operation, low cost and low time consumption, and has a good industrialization prospect.

2. The present invention improves the induction and binding ability of the bioglass precursor components on the bacterial cellulose nanofibers through the sol-gel method of amination modification of bacterial cellulose combined with ultrasonic treatment, and a 3D network structure is prepared. of nanobioglass fiber scaffolds. The prepared bioactive glass scaffold has high porosity, high specific surface area and high bioactivity.

Description of drawings

FIG. 1 is a schematic diagram of the process of preparing a highly bioactive glass nanofiber scaffold.

FIG. 2 is the SEM image of the aminated bacterial cellulose of the invention of Example 1 after ultrasonication.

Figure 3 is the FTIR image of the aminated bacterial cellulose NBC and bacterial cellulose BC prepared in the Example.

Specific implementation method

The technical solutions of the present invention will be described in detail below through examples, but the content protected by the present invention is not limited thereto.

Example 1

A preparation method of a highly bioactive glass nanofiber scaffold, comprising the following steps:

(1) Cut the bacterial cellulose film into blocks, soak it in a 1 mol/L NaOH solution at 90 °C for 2 h, then wash it with a large amount of deionized water until it becomes neutral, and freeze-dry it in a freeze dryer , to obtain bacterial cellulose bulk;

(2) Put the bacterial cellulose block into water at 35 °C with nitrogen gas, add 0.1 mol/L cerium ammonium nitrate solution (dissolved in 1 mol/L HNO ₃ ) to react for 15 min, and then continue for 30 min Polyglycidyl methacrylate (GMA) was added dropwise, and after 2 h of reaction, it was washed with deionized water and alcohol to neutrality for several times to obtain white blocks, which were then freeze-dried; the ceric ammonium nitrate solution was dissolved in 1 mol/ L HNO ₃ ; wherein the mass ratio of bacterial cellulose, ceric ammonium nitrate, GMA is 1:20:2; (3) the white block obtained in step (2) is put into ethylenediamine by mass ratio: water= 3:2 in 125 mL of mixed solution, and stirred at 80 °C for 2-5 h, and finally washed with deionized water, and then washed with absolute ethanol. To obtain aminoated bacterial cellulose, and then freeze-drying;

(4) Put the aminocellulose into the ethanol solution of 0.1 mol/L Ca(NO ₃ ) ₂ ·4H ₂ O and ultrasonically for 3 h, wherein the calcium-containing solution is changed every 1 h; The ratio of the mass of the cellulose block to the mass of the Ca(NO ₃ ) ₂ solution is 1:2000;

(5) After taking out the pre-calcified aminated bacterial cellulose block obtained in step (4), put it directly into the silicon-containing solution and continue to sonicate for 3 h, and replace the silicon-containing solution every 1 h to maintain the concentration of silicon ions , continue to wash with deionized water after ultrasonication, and then freeze-dry; wherein the ratio of the mass of the precalcified aminated bacterial cellulose block to the mass of the silicon-containing solution is 1:2000;

(6) The aminated bacterial cellulose bulk (CaSi/NBC) obtained after the final freeze-drying was calcined at 700° C. for 1 h to obtain a bioactive nanoglass scaffold (NBG).

Example 2

A preparation method of a highly bioactive glass nanofiber scaffold, comprising the following steps:

(1) Cut the bacterial cellulose film into blocks, soak it in a 1 mol/L NaOH solution at 90 °C for 2 h, then wash it with a large amount of deionized water until it becomes neutral, and freeze-dry it in a freeze dryer , to obtain bacterial cellulose bulk;

(2) Put the bacterial cellulose block into water at 35 °C with nitrogen gas, add 0.1 mol/L cerium ammonium nitrate solution (dissolved in 1 mol/L HNO ₃ ) to react for 15 min, and then continue for 30 min Polyglycidyl methacrylate (GMA) was added dropwise, and after 2 h of reaction, it was washed with deionized water and alcohol to neutrality for several times to obtain white blocks, which were then freeze-dried; the ceric ammonium nitrate solution was dissolved in 1 mol/ The HNO ₃ of L; wherein the mass ratio of bacterial cellulose, ceric ammonium nitrate, GMA is 1:20:2;

(3) Put the white block obtained in step (2) into 125 mL of mixed solution prepared by mass ratio of ethylenediamine:water=3:2, and stir at 80 ℃ for 2~5 h, and finally Wash with deionized water once, and then wash with absolute ethanol again, and repeat this for many times until the washing is neutral to obtain aminoated bacterial cellulose, which is then freeze-dried;

(4) Put the aminocellulose into the ethanol solution of 0.2 mol/L Ca(NO ₃ ) ₂ ·4H ₂ O and ultrasonically for 3 h, wherein the calcium-containing solution is changed every 1 h; The ratio of the mass of the cellulose block to the mass of the Ca(NO ₃ ) ₂ solution is 1:2000;

(5) After taking out the pre-calcified aminated bacterial cellulose block obtained in step (4), put it directly into the silicon-containing solution and continue to sonicate for 3 h, and replace the silicon-containing solution every 1 h to maintain the concentration of silicon ions , continue to wash with deionized water after ultrasonication, and then freeze-dry; wherein the ratio of the mass of the precalcified aminated bacterial cellulose block to the mass of the silicon-containing solution is 1:2000;

(6) The aminated bacterial cellulose block obtained after the final freeze-drying was calcined at 700 °C for 1 h, and the obtained bioactive nanoglass scaffold was obtained.

Example 3

A preparation method of a highly bioactive glass nanofiber scaffold, comprising the following steps:

(1) Cut the bacterial cellulose film into blocks, soak it in a 1 mol/L NaOH solution at 90 °C for 2 h, then wash it with a large amount of deionized water until it becomes neutral, and freeze-dry it in a freeze dryer , to obtain bacterial cellulose bulk;

(2) Put the bacterial cellulose block into water at 35 °C with nitrogen gas, add 0.1 mol/L cerium ammonium nitrate solution (dissolved in 1 mol/L HNO ₃ ) to react for 15 min, and then continue for 30 min Polyglycidyl methacrylate (GMA) was added dropwise, and after 2 h of reaction, it was washed with deionized water and alcohol to neutrality for several times to obtain white blocks, which were then freeze-dried; the ceric ammonium nitrate solution was dissolved in 1 mol/ The HNO ₃ of L; wherein the mass ratio of bacterial cellulose, ceric ammonium nitrate, GMA is 1:20:2;

(3) Put the white block obtained in step (2) into 125 mL of mixed solution prepared by mass ratio of ethylenediamine:water=3:2, and stir at 80 ℃ for 2~5 h, and finally Wash with deionized water once, and then wash with absolute ethanol again, and repeat this for many times until the washing is neutral to obtain aminoated bacterial cellulose, which is then freeze-dried;

(4) Put the aminocellulose into the ethanol solution of 0.5 mol/L Ca(NO ₃ ) ₂ ·4H ₂ O and ultrasonically for 3 h, wherein the calcium-containing solution is changed every 1 h; The ratio of the mass of the cellulose block to the mass of the Ca(NO ₃ ) ₂ solution is 1:2000;

(5) After taking out the pre-calcified aminated bacterial cellulose block obtained in step (4), put it directly into the silicon-containing solution and continue to sonicate for 3 h, and replace the silicon-containing solution every 1 h to maintain the concentration of silicon ions , continue to wash with deionized water after ultrasonication, and then freeze-dry; wherein the ratio of the mass of the precalcified aminated bacterial cellulose block to the mass of the silicon-containing solution is 1:2000;

(6) The aminated bacterial cellulose block obtained after the final freeze-drying was calcined at 700 °C for 1 h, and the obtained bioactive nanoglass scaffold was obtained.

Claims (4)

1. A preparation method of a high-bioactivity glass nanofiber scaffold is characterized by comprising the following steps: the method comprises the following steps:

(1) shearing a bacterial cellulose film into blocks, soaking the blocks in 0.5-2 mol/L NaOH solution at 90 ℃ for 2-5 h, then washing the blocks to be neutral by using a large amount of deionized water, and freeze-drying the blocks in a freeze dryer to obtain bacterial cellulose blocks;

(2) putting the bacterial cellulose block into constant-temperature water of 35 ℃, continuously introducing nitrogen to remove oxygen dissolved in the water, adding a ceric ammonium nitrate solution with the concentration of 0.1 mol/L for reaction for 15 min, then dripping polyglycidyl methacrylate GMA for 30 min, washing the bacterial cellulose block with deionized water after reacting for 2h, washing the bacterial cellulose block with absolute ethyl alcohol, repeatedly washing the bacterial cellulose block to be neutral to obtain a white block, and then freeze-drying the white block; wherein 1 mol/L HNO is dissolved in the ammonium ceric nitrate solution₃(ii) a Wherein the mass ratio of the bacterial cellulose to the ammonium ceric nitrate to the GMA is 1:20: 2;

(3) putting the white block obtained in the step (2) into ethylene diamine according to the mass ratio: the method comprises the following steps of (1) preparing 125 mL of mixed solution with water =3:2, stirring for 2-5 h at 80 ℃, finally washing with deionized water and ethanol to be neutral, and freeze-drying to obtain an aminated bacterial cellulose template;

(4) adding 0.1-1 mol/L Ca (NO) into the aminated bacterial cellulose₃)₂·4H₂Performing ultrasonic treatment in ethanol solution of O for 3 h, and replacing Ca (NO) every 1h₃)₂The solution is used for ensuring the concentration of calcium ions; wherein the mass of the aminated bacterial cellulose block and Ca (NO)₃)₂The mass ratio of the solution is 1: 2000;

(5) placing the pre-calcified aminated bacterial cellulose block into a silicon-containing solution, continuing to perform ultrasonic treatment for 3 hours, replacing the silicon-containing solution every 1 hour to maintain the concentration of silicon ions, continuing to perform deionized water cleaning after the ultrasonic treatment is completed, and then performing freeze drying;

(6) and (3) calcining the block obtained in the step (5) at 600-800 ℃ for 1-6 h, and removing the bacterial cellulose template to obtain the nano glass fiber support.

2. The method for preparing the glass nano fiber scaffold with high bioactivity according to claim 1, wherein the method comprises the following steps: the silicon-containing solution in the step (5) is specifically: tetraethyl orthosilicate TEOS: absolute ethanol = 1: 40 and mixing.

3. The method for preparing the glass nano fiber scaffold with high bioactivity according to claim 1, wherein the method comprises the following steps: the mass ratio of the pre-calcified aminated bacterial cellulose to the siliceous solution in the step (5) is 1: 2000.

4. The method for preparing the glass nano fiber scaffold with high bioactivity according to claim 1, wherein the method comprises the following steps: and (4) in the step (6), the heating rate of the calcination is 5-20 ℃/min.

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