CN102051702A - Mesoporous silicon oxide particle/degradable polymer nano composite fiber, preparation method and application thereof - Google Patents

Abstract

The invention discloses a mesoporous silica particle/degradable polymer nanocomposite fiber, a preparation method and application thereof. Firstly, the mesoporous silica particles are evenly dispersed in the degradable polymer solution, and then the nanocomposite fibers are prepared by using an electrospinning process, wherein the mesoporous silica particles are evenly distributed in the degradable polymer fibers. Compared with the traditional polymer fiber, the nanocomposite fiber of the invention can more effectively control the release of drugs, has faster biodegradability and higher mechanical strength, and the material has better cell compatibility. The nanocomposite fibers of the present invention can be used as tissue engineering scaffolds, drug release carriers, wound covering materials or functional membranes.

Description

Mesoporous silica particle/degradable polymer nanocomposite fiber and its preparation method and application

technical field

The invention relates to a mesoporous material/polymer nanocomposite fiber and its preparation method and application, in particular to a mesoporous silicon oxide particle/degradable polymer nanocomposite fiber and its preparation method and application.

Background technique

With a structure similar to that of extracellular matrix (ECM) in vivo, high specific surface area/volume ratio and length/diameter ratio, high porosity and pore connectivity, and good biocompatibility and biodegradability, nanofibers, In particular, nanofibers based on biodegradable polymer materials have been shown to significantly promote cell adhesion, spreading, and directional growth, and exhibit excellent biological activity (Jin HJ, Fridrikh SV, Rutledge GC and Kaplan DL. Biomacromolecules, 2002, 3:1233; Li M, Mondrinos MJ, Gandhi MR, KoFK, Weiss AS and Lelkes PI. Biomaterials, 2005, 26:5999). Therefore, it is widely used as tissue engineering scaffold, controlled drug release, wound coating and functional diaphragm, and has attracted much attention (Matthews JA, WnekGE, Simpson DG and Bowlin GL Biomacromolecules, 2002, 3: 232; Yoshimoto H, Shin YM , Terai H and Vacanti JP Biomaterials, 2003, 24:2077).

In recent years, in order to further improve and optimize the performance of nanofibers, researchers have proposed and developed nanocomposite fibers. Among them, functional inorganic particles/polymer nanofibers are an important one. Existing studies have shown that the introduction of nanoparticles with special functions can greatly improve the mechanical properties and biological activities of nanofibers. For example, the introduction of nano-hydroxyapatite into nanofibrous materials for bone tissue engineering can significantly improve the adhesion, proliferation and differentiation of osteoblast-related cells, as well as the mineralization and deposition of bone (Fujihara K, Kotaki M and Ramakrishna S. Biomaterials , 2005, 26: 4139; Chen F, Tang QL, Zhu YJ, et al.Acta Biomaterialia 2010, 6: 3013); nanofibers composited with silver nanoparticles exhibited excellent antibacterial properties (Naddaf Sichani G, Morshed M, Amirnasr M, J Appl Polym Sci 2010, 116: 1021, ), etc. At the same time, nano functional particles can also improve the controllable release of drugs from nanofibers.

Mesoporous silica nanoparticles (MSNs) have excellent characteristics such as uniform pores, large pore volume and specific surface area, silanol bonds, easily chemically modified pore surfaces, and unique cell transport functions. Porous materials MSNs have shown great application potential in the storage, transport and controlled release of poorly soluble drugs and macromolecular drugs, and even protein molecules. Moreover, its ordered pores can be used as "microreactors" to realize the rapid circulation, diffusion and reaction of substances. In recent years, there have been many research reports on the controllable preparation, surface modification and application of MSNs, but few reports on their composites with nanofibers.

Contents of the invention

The purpose of the present invention is to provide a mesoporous silica particle/degradable polymer nanocomposite fiber, which can improve the high activity loading and controllable release of nanofibers to drugs/proteins through the compounding of mesoporous silica particles, and improve the efficiency of traditional polymerization. The biodegradability and mechanical strength of the nanofibers are improved without affecting the cytocompatibility of the nanofibers.

Another object of the present invention is to provide a method for preparing the mesoporous silica particle/degradable polymer nanocomposite fiber.

The third object of the present invention is to provide the application of the mesoporous silica particle/degradable polymer nanocomposite fiber.

The mesoporous silica particles/degradable polymer nanocomposite fibers of the present invention include mesoporous silica particles and degradable polymer fibers, wherein the mesoporous silica particles are distributed in the degradable polymer fibers.

According to the present invention, the mass ratio of the mesoporous silica particles to the degradable polymer is 1:5-500.

According to the present invention, the diameter of the nanocomposite fiber is 50nm-2μm, and the length is 20-800um.

According to the present invention, the pore diameter of the mesoporous silicon oxide particles is 1-50 nm, and the specific surface area is 200-1400 m ² /g. The mesoporous silicon oxide particle of the present invention has a typical MCM-41 structure material and is well dispersed.

According to the present invention, the biodegradable polymer includes natural polymer materials such as gelatin, collagen, and chitosan, and biodegradable polymer materials such as polylactic acid, polycaprolactone, and polylactic acid-glycolide.

According to the present invention, the mesoporous silicon oxide particles are drug-loaded mesoporous silicon oxide particles.

The preparation method of the mesoporous silica particle/degradable polymer nanocomposite fiber provided by the invention comprises the following steps:

(a), dissolving the degradable polymer material in a solvent to form a polymer solution;

(b), uniformly dispersing mesoporous silica particles in the polymer solution to form a mixed solution;

(c), using electrospinning technology to spin the mixed solution to obtain nanofibers, and collect them;

(d) Cross-linking the prepared nanofibers with 1% glutaraldehyde to obtain the mesoporous silica particles/degradable polymer nanocomposite fibers.

Wherein, the solvent includes: acetic acid, hexafluoroisopropanol, chloroform, acetone, dichloroethane, water, or a mixed solvent of the two or the three.

According to the present invention, the voltage used in the electrospinning is 5-40KV, the spinning rate is 1-3mL/h, the distance between the solution injection port and the collector is 5-20 cm, and the needle of the solution injection port The specifications are 18-27.

The mesoporous silicon oxide particle/degradable polymer nanocomposite fiber of the present invention can be used to prepare tissue engineering scaffolds, drug release carriers, wound covering materials, functional diaphragms and the like.

The invention introduces mesoporous silicon oxide nanoparticles with regular, uniform and adjustable channels, large pore volume and specific surface area into polymer nanofibers, and improves its mechanical strength and biodegradability while retaining the properties of traditional nanofibers , while realizing high-throughput, high-activity loading and controlled release of drugs/proteins.

Description of drawings

Fig. 1 is (a) XRD pattern and (b) TEM pattern of mesoporous silica nanoparticles.

Fig. 2 is (a) nitrogen isothermal adsorption-desorption diagram and (b) pore size distribution diagram of mesoporous silica nanoparticles.

Fig. 3 is the SEM picture of 0.25% MSN/Gel-AcOH fiber.

Fig. 4 is the SEM picture of 1% MSN/Gel-AcOH fiber.

Fig. 5 is the SEM picture of 10% MSN/Gel-HFIP-27 fiber.

Fig. 6 is a SEM image of 10% MSN/Gel-HFI-22 fiber.

Figure 7 shows the distribution of nanoparticles in 0.25% FITC-MSN/Gel-AcOH nanofibers: (a) under bright field, (b) under fluorescence.

Figure 8 is the distribution of nanoparticles in 10% FITC-MSN/Gel-HFIP nanofibers: (a) under bright field, (b) under fluorescence.

Figure 9 is a comparison of the degradability of composite nanofibers and ordinary nanofibers.

Figure 10 is a comparison of the release of dexamethasone from nanofibers.

Figure 11 is a comparison of the release of alendronate sodium from nanofibers.

Fig. 12 is the cytocompatibility of the nanocomposite fibers: where (a) is cell viability; (b) is cell morphology.

Detailed ways

The unique composition and structure make degradable polymer nanofibers widely used in biomedical fields such as tissue engineering and skin repair. As a porous material with uniform and adjustable channels, large pore volume and specific surface area, mesoporous silica nanoparticles have outstanding advantages in drug/protein molecule storage and controlled release. On this basis, the present invention uses mesoporous silica nanoparticles as a carrier to load drugs/proteins, then mixes them with polymers, and uses electrospinning technology to prepare nanocomposite fibers, completing the present invention.

The present invention will be further described below in conjunction with specific embodiments. It should be understood that the following examples are only used to illustrate the present invention but not to limit the scope of the present invention.

Evaluation method

(1) Characterization of materials

X-Ray diffraction analysis (D/max 2550VB/PC polycrystalline diffractometer) was used to analyze the crystalline state of the material at 0-10°, and a transmission electron microscope (TEM 2100F type) was used to observe the microstructure of the material. The microporous structure of the material was measured by nitrogen isothermal adsorption-desorption, and the specific surface area and pore volume of the material were calculated by BET, and the average pore diameter was calculated according to the Barrett-Joyner-Helen (BJH) formula. A scanning electron microscope (JSM-6360LV type) was used to observe the surface morphology and microstructure of the prepared materials. The distribution and cytocompatibility of the mesoporous silica nanoparticles labeled with fluorescent probes in the nanocomposite fibers were observed with a fluorescence microscope (TE2000-U).

(2) Degradability of materials

A certain amount of nanofibers and nanocomposite fibers were packaged in 20ml simulated body fluid bottles, and oscillated in a constant temperature vibration box at a constant temperature (37°C) at a frequency of 68r/min. Regularly and quantitatively replace the SBF solution, regularly sample, take out, weigh, and record changes in the quality of the nanofibers. Three parallel experiments were done for each sample, and the results were averaged.

(3) Mechanical performance test of materials

The fiber membrane is made into a tensile sample strip of 10mm×50mm, and then the tensile test is carried out on an Instron3365 tensile tester, wherein the thickness of the membrane is 0.25mm, the clamping length is 30mm, and the tensile speed is 15mm/min .

(4) In vitro drug release evaluation

Lipid-soluble and water-soluble drugs were respectively selected as drug release targets, and dexamethasone (fat-soluble) and or alendronate sodium (water-soluble) were used as model drugs to evaluate the drug release properties of the synthesized nanocomposite fibers. A certain amount of nanofibers loaded with dexamethasone and or alendronate sodium were placed in a dialysis bag, and then the dialysis bag was placed in a sealed plastic bottle of 20 mL phosphate buffer saline or saline, at a constant temperature of 37°C at 120rpm Velocity oscillates. In addition, the same amount of drug-loaded nanocomposite fibers and mesoporous silica nanoparticles packed in dialysis bags were used as the control group. Take out 4mL of the solution outside the dialysis bag at regular intervals to measure the concentration of the drug, and add 4ml of deionized water. Samples were obtained at each time point for measurement by UV-Vis spectrophotometer. The graph is plotted with time as the X-axis and cumulative release as the Y-axis.

(5) In vitro cell culture

上，置于24孔培养板中，在37℃、5％CO₂环境中，接种L929细胞。培养时间为3天后，将配制好的钙黄绿素AM和乙锭二聚体-1试剂加入样品中，在培养箱中孵育30分钟后，通过荧光倒置显微镜(TE2000-U)观察细胞的成活情况。Using L929 fibroblasts as a model, the cytotoxicity of the prepared nanofibers was tested using tetramethylazozolium salt and live/dead cytotoxicity assay kits, respectively. Soak the nanofibers in the anti-anti double antibody solution, and change the solution at least once to ensure complete sterilization. The treated fiber film is placed in fixed and embedded in a 24-well culture plate. L929 cells were inoculated at a certain cell concentration on the surface of nanocomposite fibers placed in a 24-well culture plate, cultured at a constant temperature of 37° C. and 5% CO ₂ for 1 to 3 days, and samples were taken regularly. After the culture, transplant the sample into a new well plate, add the culture medium, and then add 20 μL of tetramethyl azolium salt reagent to each well, continue to incubate at 37°C for 4 hours, discard the supernatant, and add 150 μL of DMSO , gently shaken for 20 min to dissolve the crystals, and after centrifugation, use a continuous spectrum microplate reader to measure the light absorption value of the solution at 490 nm. fix the nanocomposite fibers in the

Put it in a 24-well culture plate, and inoculate L929 cells in an environment of 37°C and 5% CO ₂ . After 3 days of incubation, the prepared calcein AM and ethidium dimer-1 reagents were added to the samples, incubated in the incubator for 30 minutes, and the viability of the cells was observed through a fluorescent inverted microscope (TE2000-U).

Example 1 Preparation of Mesoporous Silica Nanoparticles MSN-1

Weigh 1.116g of cetyl dimethyl sodium bromide, add 880 mL of deionized water, raise the temperature to 50°C, and keep the temperature for 2 hours until the cetyl dimethyl sodium bromide is completely dissolved. Then 52.8mL of 27% concentrated ammonia water was added, and after 10min, 5.6mL of ethyl orthosilicate was slowly added dropwise. Stir at constant temperature for 2h to stop heating, the reaction is terminated, and stand for aging for 2h. Then, pour off the supernatant, centrifuge and wash with ultrapure water and absolute ethanol twice respectively. Finally, it was extracted and refluxed in methanol/hydrochloric acid system for 24 hours, and then washed twice with ethanol. Finally dried, ground and collected. The obtained mesoporous material (referred to as MSN-1) was measured by X-Ray diffraction analysis, transmission electron microscope, nitrogen isothermal adsorption-desorption, etc. to measure the microstructure and morphology of the material, and the specific surface area and pore volume of the material were calculated by BET . The results are shown in Figure 1 and Figure 2. It can be seen from the XRD in Figure 1(a) that the synthesized material has obvious diffraction peaks between 2θ of 1 and 5, indicating that the material has a typical MCM-41 mesoporous structure. TEM shows that the mesoporous structure is clear and there are obvious hexagonal channels. Figure 2. Nitrogen isotherm adsorption-desorption shows that the material shows an obvious jump in the range of 0.2<p/p ₀ <0.4, and the desorption line is always above the adsorption line to form a _H2 hysteresis ring, which also shows that the There are mesoporous channels in the sample. At the same time, the pore size of the material is 3.8nm. The specific surface area of the material calculated by BET is 1400m ² /g.

Example 2 Preparation of Mesoporous Silica Nanoparticles MSN-2

Weighing 12g (EO ₂₀ PO ₇₀ EO ₂₀ ; average molecular weight 5800, Aldrich Company) was added to 312 mL of deionized water and 60 mL of 37% concentrated hydrochloric acid. After the surfactant was completely dissolved, 27.5 mL of ethyl orthosilicate was added. After stirring at room temperature for 24 hours, it was then poured into a special airtight tetrafluoroethylene container and hydrothermally crystallized at 100°C for 24 hours. The obtained suspension was centrifuged, washed twice with water and ethanol, calcined at 600°C for 6 hours, and then ground. The obtained mesoporous material (referred to as MSN-2) has a pore size of 9.8 nm. BET calculation results show that the specific surface area of the material is 520m ² /g.

Example 3 Preparation of Mesoporous Silica Nanoparticles MSN-3

Weigh 0.98g of hexadecyldimethyl sodium bromide, add it into 380mL of deionized water, raise the temperature to 30°C, and keep the temperature for 2 hours until the hexadecyldimethyl sodium bromide is completely dissolved. Then 52.8mL of 27% concentrated ammonia water was added, and after 10min, 5.6mL of ethyl orthosilicate was slowly added dropwise. Stir at constant temperature for 2h to stop heating, the reaction is terminated, and stand for aging for 2h. Then, pour off the supernatant, centrifuge and wash with ultrapure water and absolute ethanol twice respectively. Finally, it was extracted and refluxed in methanol/hydrochloric acid system for 24 hours, and then washed twice with ethanol. Finally dried, ground and collected. The obtained mesoporous material (referred to as MSN-3) has a pore size of 3.6 nm. The specific surface area of the material calculated by BET is 200m ² /g.

Example 3 Preparation of mesoporous silica nanoparticles/gelatin nanocomposite fibers

Dissolve 10 grams of gelatin in 50 mL of acetic acid/water, where the volume ratio of acetic acid and water is 70/30, and then add 25 mg of mesoporous silica nanoparticles (MSN-1) to it, and ultrasonically vibrate for 30 minutes to ensure that the nano The particles are uniformly dispersed in the solution.

Under the conditions of room temperature (25°C) and relative air humidity of about 50%, the above-mentioned mixed solution is sucked into a 10ml syringe, and the syringe is equipped with a No. 20 stainless steel needle (its inner diameter is 0.6mm, and the front end is ground flat), and then the above-mentioned syringe is Install it on the electrostatic spinning device and prepare the composite fiber by electrostatic spinning technology. Under the action of high-voltage static electricity, the mixed solution is sprayed from the stainless steel needle to the collecting device to form composite fibers. Among them, the stainless steel needle is connected to the positive pole of the high-voltage power supply, and aluminum foil is used to collect the composite fiber. The aluminum foil is connected to the negative pole of the high-voltage power supply. The distance between the stainless steel needle and the aluminum foil is 15 cm, and the power supply voltage is 20 kV. The as-prepared nanocomposite fibers were then crosslinked with 1% glutaraldehyde. The morphology of the prepared mesoporous silica nanoparticles/gelatin nanofibers (denoted as 0.25% MSN/Gel-AcOH) is shown in FIG. 3 . It can be seen that the prepared mesoporous silica nanoparticles/gelatin nanofibers are continuous and smooth, with a diameter of 20 nm-0.5 μm and a length of 20-50 μm.

Example 4 Preparation of mesoporous silica nanoparticles/gelatin nanocomposite fibers

Dissolve 10 grams of gelatin in 50 mL of acetic acid/water, where the volume ratio of acetic acid and water is 70/30, and then add 100 mg of mesoporous silica nanoparticles MSN-2 to it to ensure that the nanoparticles are evenly dispersed in the solution.

Under the conditions of room temperature (25°C) and relative air humidity of about 50%, the above-mentioned mixed solution is sucked into a 10ml syringe equipped with a No. 18 stainless steel needle (its inner diameter is 0.84mm, and the front end is ground flat), and then the above-mentioned syringe is Install it on the electrostatic spinning device and prepare the composite fiber by electrostatic spinning technology. Under the action of high-voltage static electricity, the mixed solution is sprayed from the stainless steel needle to the collecting device to form composite fibers. Among them, the stainless steel needle is connected to the positive pole of the high-voltage power supply, and aluminum foil is used to collect the composite fiber. The aluminum foil is connected to the negative pole of the high-voltage power supply. The distance between the stainless steel needle and the aluminum foil is 15 cm, and the power supply voltage is 20 kV. The as-prepared nanocomposite fibers were then crosslinked with 1% glutaraldehyde. The morphology of the prepared mesoporous silica nanoparticles/gelatin nanofibers (referred to as 1% MSN/Gel-AcOH) is shown in FIG. 4 . It can be seen from the figure that the prepared mesoporous silica nanoparticles/gelatin nanofibers are continuous and smooth, with a diameter of 5 nm-0.5 μm and a length of 40-300 μm.

Example 5 Preparation of mesoporous silica nanoparticles/gelatin nanocomposite fibers

Dissolve 2 grams of gelatin in 50 mL of hexafluoroisopropanol, and then add 200 mg of mesoporous silica nanoparticles MSN-2 to it to ensure that the nanoparticles are evenly dispersed in the solution.

Under the conditions of room temperature (25°C) and relative air humidity of about 50%, the above-mentioned mixed solution is sucked into a 10ml syringe, and the syringe is equipped with a No. 27 stainless steel needle (its inner diameter is 0.21mm, and the front end is ground flat), and then the above-mentioned syringe is Install it on the electrostatic spinning device and prepare the composite fiber by electrostatic spinning technology. Under the action of high-voltage static electricity, the mixed solution is sprayed from the stainless steel needle to the collecting device to form composite fibers. Among them, the stainless steel needle is connected to the positive pole of the high-voltage power supply, and aluminum foil is used to collect the composite fiber. The aluminum foil is connected to the negative pole of the high-voltage power supply. The distance between the stainless steel needle and the aluminum foil is 15 cm, and the power supply voltage is 20 kV. The as-prepared nanocomposite fibers were then crosslinked with 1% glutaraldehyde. The morphology of the prepared mesoporous silica nanoparticles/gelatin nanofibers (referred to as 10% MSN/Gel-HFIP-27) is shown in FIG. 5 . It can be seen from the figure that the prepared mesoporous silica nanoparticles/gelatin nanofibers are continuous and smooth, with a diameter of 20 nm-0.5 μm and a length of 50-600 μm.

Example 6 Preparation of mesoporous silica nanoparticles/gelatin nanocomposite fibers

Dissolve 2 grams of gelatin in 50 mL of hexafluoroisopropanol, and then add 200 mg of mesoporous silica nanoparticles MSN-1 to it to ensure that the nanoparticles are evenly dispersed in the solution.

Under the conditions of room temperature (25°C) and relative air humidity of about 50%, the above-mentioned mixed solution is sucked into a 10ml syringe, and the syringe is equipped with a No. 22 stainless steel needle (its inner diameter is 0.41mm, and the front end is ground flat), and then the above-mentioned syringe is Install it on the electrostatic spinning device and prepare the composite fiber by electrostatic spinning technology. Under the action of high-voltage static electricity, the mixed solution is sprayed from the stainless steel needle to the collecting device to form composite fibers. Among them, the stainless steel needle is connected to the positive pole of the high-voltage power supply, and aluminum foil is used to collect the composite fiber. The aluminum foil is connected to the negative pole of the high-voltage power supply. The distance between the stainless steel needle and the aluminum foil is 15 cm, and the power supply voltage is 20 kV. The as-prepared nanocomposite fibers were then crosslinked with 1% glutaraldehyde. The morphology of the prepared mesoporous silica nanoparticles/gelatin nanofibers (referred to as 10% MSN/Gel-HFIP-22) is shown in FIG. 6 . It can be seen from the figure that the prepared mesoporous silica nanoparticles/gelatin nanofibers are continuous and smooth, with a diameter of 20 nm-0.5 μm and a length of 100-800 μm.

Example 7

Dissolve 10 grams of polylactic acid in 50 mL of dichloromethane, and then add 100 mg of mesoporous silica nanoparticles MSN-2 to it to ensure that the nanoparticles are evenly dispersed in the solution.

Under the conditions of room temperature (25°C) and relative air humidity of about 50%, the above-mentioned mixed solution is sucked into a 10ml syringe equipped with a No. 18 stainless steel needle (its inner diameter is 0.84mm, and the front end is ground flat), and then the above-mentioned syringe is Install it on the electrostatic spinning device and prepare the composite fiber by electrostatic spinning technology. Under the action of high-voltage static electricity, the mixed solution is sprayed from the stainless steel needle to the collecting device to form composite fibers. Among them, the stainless steel needle is connected to the positive pole of the high-voltage power supply, and aluminum foil is used to collect the composite fiber. The aluminum foil is connected to the negative pole of the high-voltage power supply. The distance between the stainless steel needle and the aluminum foil is 15 cm, and the power supply voltage is 20 kV. The as-prepared nanocomposite fibers were then crosslinked with 1% glutaraldehyde. The prepared mesoporous silica nanoparticle/polylactic acid nanofiber (recorded as 1% MSN/PLA) is continuous and smooth, with a diameter of 5nm-0.5μm and a length of 40-300μm.

Example 8

Dissolve 2 grams of collagen in 50 mL of water, and then add 200 mg of mesoporous silica nanoparticles MSN-2 to it to ensure that the nanoparticles are evenly dispersed in the solution.

Under the conditions of room temperature (25°C) and relative air humidity of about 50%, the above-mentioned mixed solution is sucked into a 10ml syringe, and the syringe is equipped with a No. 22 stainless steel needle (its inner diameter is 0.41mm, and the front end is ground flat), and then the above-mentioned syringe is Install it on the electrostatic spinning device and prepare the composite fiber by electrostatic spinning technology. Under the action of high-voltage static electricity, the mixed solution is sprayed from the stainless steel needle to the collecting device to form composite fibers. Among them, the stainless steel needle is connected to the positive pole of the high-voltage power supply, and aluminum foil is used to collect the composite fiber. The aluminum foil is connected to the negative pole of the high-voltage power supply. The distance between the stainless steel needle and the aluminum foil is 15 cm, and the power supply voltage is 20 kV. The as-prepared nanocomposite fibers were then crosslinked with 1% glutaraldehyde. The prepared mesoporous silica nanoparticle/collagen nanofiber (referred to as 10% MSN/Col) is continuous and smooth, with a diameter of 20nm-0.5μm and a length of 100-800μm.

Example 9 Preparation of FITC-labeled mesoporous silica nanoparticles/gelatin nanocomposite fibers

In order to further understand the distribution of mesoporous silica nanoparticles in nanofibers, the experiment used fluorescein isothiocyanate (FITC)-labeled mesoporous silica material (MSN-1) to prepare nanocomposite fibers. The preparation of FITC-labeled mesoporous silica material is as follows: disperse 2 g of MSN-1 in absolute ethanol, add 0.2 g of γ-aminopropyltriethoxysilane, reflux at 74 ° C for 1 hour, centrifuge, and Dry the powder for later use; dissolve 0.05 g of FITC in 5 mL of carbonate buffer, then add the MSN powder pretreated by the above-mentioned coupling agent, stir at room temperature for 16 hours in the dark, centrifuge, and then wash off the surface of the powder with carbonate buffer FITC, dry it.

Dissolve 10 g of gelatin in 50 mL of acetic acid/water, where the volume ratio of acetic acid and water is 70/30, and then add 25 mg of FITC-labeled mesoporous silica nanoparticles FITC-MSN-1 to it to ensure that the nanoparticles are uniform dispersed in solution.

Under the conditions of room temperature (25°C) and relative air humidity of about 50%, the above-mentioned mixed solution is sucked into a 10ml syringe, and the syringe is equipped with a No. 23 stainless steel needle (its inner diameter is 0.6mm, and the front end is ground flat), and then the above-mentioned syringe is Install it on the electrostatic spinning device and prepare the composite fiber by electrostatic spinning technology. Under the action of high-voltage static electricity, the mixed solution is sprayed from the stainless steel needle to the collecting device to form composite fibers. Among them, the stainless steel needle is connected to the positive pole of the high-voltage power supply, and aluminum foil is used to collect the composite fiber. The aluminum foil is connected to the negative pole of the high-voltage power supply. The distance between the stainless steel needle and the aluminum foil is 15 cm, and the power supply voltage is 20 kV. The as-prepared nanocomposite fibers were then crosslinked with 1% glutaraldehyde. The prepared mesoporous silica nanoparticles/gelatin nanofibers (denoted as 0.25% FITC-MSN/Gel-AcOH). Further, the prepared nanocomposite fibers were observed with a fluorescence microscope, and the results are shown in FIG. 7 . It can be seen from the figure that the prepared nanofibers are smooth and uniform, and the mesoporous silica nanoparticles are evenly distributed in the fibers.

Preparation of Example 10 FITC-labeled mesoporous silica nanoparticles/gelatin nanocomposite fibers

Dissolve 2 g of gelatin in 50 mL of hexafluoroisopropanol, and then add 200 mg of FITC-labeled mesoporous silica nanoparticles (FITC-MSN-1) to it to ensure that the nanoparticles are evenly dispersed in the solution.

Under the conditions of room temperature (25°C) and relative air humidity of about 50%, the above-mentioned mixed solution is sucked into a 10ml syringe, and the syringe is equipped with a No. 22 stainless steel needle (its inner diameter is 0.41mm, and the front end is ground flat), and then the above-mentioned syringe is Install it on the electrostatic spinning device and prepare the composite fiber by electrostatic spinning technology. Under the action of high-voltage static electricity, the mixed solution is sprayed from the stainless steel needle to the collecting device to form composite fibers. Among them, the stainless steel needle is connected to the positive pole of the high-voltage power supply, and aluminum foil is used to collect the composite fiber. The aluminum foil is connected to the negative pole of the high-voltage power supply. The distance between the stainless steel needle and the aluminum foil is 15 cm, and the power supply voltage is 20 kV. The as-prepared nanocomposite fibers were then crosslinked with 1% glutaraldehyde. The morphology of the prepared mesoporous silica nanoparticles/gelatin nanofibers (referred to as 10% FITC-MSN/Gel-HFIP) is shown in FIG. 8 . It can be seen from the figure that the prepared mesoporous silica nanoparticles/gelatin nanofibers are continuous and smooth, and the mesoporous silica nanoparticles are evenly distributed in the fibers.

The preparation of embodiment 11 gelatin nanofibers

For comparison, conventional polymer nanofibers without mesoporous silica particles were prepared by the same method.

Dissolve 10 grams of gelatin in 50 mL of acetic acid/water (wherein the volume ratio of acetic acid and water is 70/30), and oscillate evenly with ultrasound. Under the conditions of room temperature (25°C) and relative air humidity of about 50%, the above-mentioned mixed solution is sucked into a 10ml syringe, and the syringe is equipped with a No. 20 stainless steel needle (its inner diameter is 0.6mm, and the front end is ground flat), and then the above-mentioned syringe is Install it on the electrostatic spinning device and prepare the composite fiber by electrostatic spinning technology. Under the action of high-voltage static electricity, the mixed solution is sprayed from the stainless steel needle to the collecting device to form composite fibers. Among them, the stainless steel needle is connected to the positive pole of the high-voltage power supply, and aluminum foil is used to collect the composite fiber. The aluminum foil is connected to the negative pole of the high-voltage power supply. The distance between the stainless steel needle and the aluminum foil is 15 cm, and the power supply voltage is 20 kV. The as-prepared nanocomposite fibers were then crosslinked with 1% glutaraldehyde. The prepared gelatin nanofibers (referred to as Gel-AcOH) have a morphology similar to that of Example 1.

The comparison of the degradability of embodiment 12 nanofibers

A certain amount of mesoporous silica nanocomposite fibers (Example 4 and Example 5) and gelatin nanofibers (Example 11) were packaged in 20ml SBF simulated body fluid bottles respectively, and kept at a constant temperature (37°C) in a constant temperature vibration box. ) oscillation, the oscillation frequency is 68r/min. Regularly and quantitatively replace the SBF solution, regularly sample, take out, weigh, and record changes in the quality of the nanofibers. Three parallel experiments were done for each sample, and the results were averaged. The result is shown in Figure 9. It can be seen that the addition of mesoporous silica particles can significantly promote the degradation of gelatin nanofibers.

The mechanical properties of embodiment 13 nanocomposite fibers

The prepared mesoporous silica nanocomposite fiber (embodiment 4 and embodiment 5) and gelatin nanofiber (embodiment 11) are respectively made into 10mm * 50mm tensile spline, then on the Instron3365 type strength elongation tester Tensile tests were performed in which the thickness of the film was 0.25 mm, the gripping length was 30 mm, and the tensile speed was 15 mm/min. The results are shown in Table 1. It can be seen from the table that the introduction of mesoporous silica particles can significantly improve the mechanical strength and elongation at break of nanofibers.

The mechanical strength comparison of table 1 nanofiber

samples Tensile stress (Mpa) Elongation at break (%) Gel-AcOH 1.58±0.32 3.9±0.72 1% MSN-Gel-AcOH 2.51±0.45 7.6±0.37 10% MSN-Gel-HFIP 2.28±0.37 6.7±0.52

Embodiment 14 dexamethasone release performance in vitro

Using dexamethasone as a hydrophobic model drug, the drug release properties of the synthesized nanocomposite fibers were evaluated.

Dissolve 2 grams of gelatin in 50 mL of hexafluoroisopropanol, then add 200 mg of dexamethasone-loaded mesoporous silica nanoparticles to it, and ultrasonically 10 minutes to ensure that the nanoparticles are evenly dispersed in the solution, wherein the The mesoporous silicon oxide nanoparticle loaded with dexamethasone adopts the physical adsorption method to immobilize dexamethasone in the pores of the mesoporous silicon oxide nanoparticle. Then, under the conditions of room temperature (25°C) and relative air humidity of about 50%, the above-mentioned mixed solution is sucked into a 10ml syringe, and the syringe is equipped with a No. 27 stainless steel needle (its inner diameter is 0.21mm, and the front end is ground flat), and then the above-mentioned The syringe is installed on the electrospinning device and the composite fiber is prepared by electrospinning technology. Under the action of high-voltage static electricity, the mixed solution is sprayed from the stainless steel needle to the collecting device to form composite fibers. Among them, the stainless steel needle is connected to the positive pole of the high-voltage power supply, and aluminum foil is used to collect the composite fiber. The aluminum foil is connected to the negative pole of the high-voltage power supply. The distance between the stainless steel needle and the aluminum foil is 15 cm, and the power supply voltage is 20 kV. Then, the prepared nanocomposite fibers were cross-linked with 1% glutaraldehyde to obtain drug-loaded mesoporous silica nanoparticles/gelatin nanofibers, labeled as 10% Dex-MSN/Gel-HFIP.

Dissolve 2 grams of gelatin and 2 mg of dexamethasone in 50 mL of hexafluoroisopropanol, and suck the above mixed solution into a 10 ml syringe at room temperature (25°C) and a relative air humidity of about 50%. No. 27 stainless steel needle (its inner diameter is 0.21 mm, the front end is ground flat), and then the above-mentioned syringe is installed on the electrospinning device and the composite fiber is prepared by electrospinning technology. Under the action of high-voltage static electricity, the mixed solution is sprayed from the stainless steel needle to the collecting device to form composite fibers. Among them, the stainless steel needle is connected to the positive pole of the high-voltage power supply, and aluminum foil is used to collect the composite fiber. The aluminum foil is connected to the negative pole of the high-voltage power supply. The distance between the stainless steel needle and the aluminum foil is 15 cm, and the power supply voltage is 20 kV. The prepared nanocomposite fibers were then cross-linked with 1% glutaraldehyde to obtain dexamethasone-loaded gelatin nanofibers marked as Dex-Gel-HFIP.

Place equal amounts of dexamethasone nanocomposite fibers (10% Dex-MSN/Gel-HFIP) and gelatin fibers (Dex-Gel-HFIP) in dialysis bags, and then place the dialysis bags in 20 mL of phosphate buffered saline Liquid in a plastic airtight bottle, 37 ℃ constant temperature stirring. In addition, the same amount of drug-loaded nanofibers and mesoporous silica nanoparticles packed in dialysis bags were used as the control group. Take out 4ml of the solution outside the dialysis bag at regular intervals to measure the concentration of the drug, and add 4ml of phosphate buffer. Taking time as the X-axis and cumulative release as the Y-axis, the results are shown in Figure 10. As shown, mesoporous silica nanoparticles/gelatin nanofibers are more effective in controlling the release of dexamethasone than drug-loaded gelatin nanofibers.

Embodiment 15 alendronate sodium release performance in vitro

The drug release properties of the synthesized nanocomposite fibers were evaluated with alendronate sodium (Ale) as the hydrophilic model drug.

Dissolve 2 grams of gelatin in 50 mL of hexafluoroisopropanol, then add 200 mg of mesoporous silica nanoparticles loaded with alendronate sodium, and ultrasonically vibrate for 10 minutes to ensure that the nanoparticles are evenly dispersed in the solution, wherein The mesoporous silicon oxide nanoparticles loaded with alendronate sodium is that sodium alendronate is immobilized in the pores of the mesoporous silicon oxide nanoparticles by physical adsorption. Then, under the conditions of room temperature (25°C) and relative air humidity of about 50%, the above-mentioned mixed solution is sucked into a 10ml syringe, and the syringe is equipped with a No. 23 stainless steel needle (its inner diameter is 0.34mm, and the front end is ground flat), and then the above-mentioned The syringe is installed on the electrospinning device and the composite fiber is prepared by electrospinning technology. Under the action of high-voltage static electricity, the mixed solution is sprayed from the stainless steel needle to the collecting device to form composite fibers. Among them, the stainless steel needle is connected to the positive pole of the high-voltage power supply, and aluminum foil is used to collect the composite fiber. The aluminum foil is connected to the negative pole of the high-voltage power supply. The distance between the stainless steel needle and the aluminum foil is 15 cm, and the power supply voltage is 20 kV. Then adopt 1% glutaraldehyde to carry out cross-linking to the prepared nanocomposite fiber, the prepared alendronate-loaded mesoporous silica nanoparticle/gelatin nanofiber (recorded as 10% Ale-MSN/Gel-HFIP ).

Dissolve 2 grams of gelatin and 2 mg of alendronate sodium in 50 mL of hexafluoroisopropanol, and suck the above mixed solution into a 10 ml syringe at room temperature (25° C.) and with a relative air humidity of about 50%. The syringe is equipped with a No. 23 stainless steel needle (its inner diameter is 0.34 mm, and the front end is ground flat), and then the above-mentioned syringe is installed on an electrospinning device to prepare composite fibers by electrospinning technology. Under the action of high-voltage static electricity, the mixed solution is sprayed from the stainless steel needle to the collecting device to form composite fibers. Among them, the stainless steel needle is connected to the positive pole of the high-voltage power supply, and aluminum foil is used to collect the composite fiber. The aluminum foil is connected to the negative pole of the high-voltage power supply. The distance between the stainless steel needle and the aluminum foil is 15 cm, and the power supply voltage is 20 kV. Then, 1% glutaraldehyde was used to cross-link the prepared nanocomposite fibers, and the prepared alendronate-loaded mesoporous silica nanoparticles/gelatin nanofibers were designated as Ale-Gel-HFIP.

Place equal amounts of nanocomposite fibers loaded with alendronate sodium (10% Ale-MSN/Gel-HFIP) and gelatin fibers (Ale-Gel-HFIP) in dialysis bags, and then place the dialysis bags in 100 mL of water In a beaker, stir at 37°C. In addition, the same amount of drug-loaded nanofibers and mesoporous silica nanoparticles packed in dialysis bags were used as the control group. Take out 4ml of the solution outside the dialysis bag at regular intervals to measure the concentration of the drug, and add 4ml of deionized water. Taking time as the X-axis and cumulative release as the Y-axis, the results are shown in Figure 11. As shown, mesoporous silica nanoparticles/gelatin nanofibers are more effective in controlling the release of alendronate than drug-loaded gelatin nanofibers.

Example 16 In Vitro Cell Characterization

固定)，于37℃恒温、5％CO₂中培养1～3天，定时取样。培养结束后，将支架移至新孔中，弃液并置换新培养基。配制MTS用试剂，每孔加入800μL该试剂，37℃继续孵育4h后，用连续光谱酶标仪在490nm处测定溶液的光吸收值。结果如图12(a)所示。由图可见，与对照组(24孔板，不加入任何材料)相比，纳米复合纤维对细胞的活性几乎无影响。Using L929 cells as a model, the cytotoxicity of the nanocomposite fibers prepared in Example 4 and Example 5 was tested using tetramethylazolate (MTT) and live/dead cytotoxicity assay kits, respectively. L929 cells were seeded on the surface of nanocomposite fibers placed in 24-well culture plates (firstly with

fixed), cultured at a constant temperature of 37° C. and 5% CO ₂ for 1 to 3 days, and samples were taken regularly. After incubation, the scaffolds were moved to new wells, discarded and replaced with new medium. Prepare the reagent for MTS, add 800 μL of the reagent to each well, and continue to incubate at 37° C. for 4 h, then measure the light absorption value of the solution at 490 nm with a continuous spectrum microplate reader. The result is shown in Fig. 12(a). It can be seen from the figure that, compared with the control group (24-well plate, without adding any material), the nanocomposite fiber has almost no effect on the activity of the cells.

上，置于24孔培养板中，在37℃、5％CO₂环境中，接种L929细胞。培养时间为3天后，将配制好的钙黄绿素AM和乙锭二聚体-1试剂加入样品中，在培养箱中孵育30分钟后，通过荧光倒置显微镜(TE2000-U)观察细胞的成活情况。从图12(b)中可以看出，材料表面的细胞呈现绿色荧光，说明该纳米复合纤维具有非常优异的生物相容性，细胞呈现纤维状伸展，表明细胞粘附、增殖状态良好。The prepared nanocomposite fibers were fixed in

Put it in a 24-well culture plate, and inoculate L929 cells in an environment of 37°C and 5% CO ₂ . After 3 days of incubation, the prepared calcein AM and ethidium dimer-1 reagents were added to the samples, incubated in the incubator for 30 minutes, and the viability of the cells was observed through a fluorescent inverted microscope (TE2000-U). It can be seen from Figure 12(b) that the cells on the surface of the material show green fluorescence, indicating that the nanocomposite fibers have excellent biocompatibility, and the cells show fibrous extension, indicating that the cells adhere and proliferate well.

The nanocomposite fiber of the present invention not only has the characteristics of higher specific surface area/volume ratio, length/diameter ratio, and high porosity of traditional polymer nanofibers, but also can more effectively control the release of drugs, and has faster biological Degradability and higher mechanical strength, while the material has better cell compatibility, used in tissue engineering scaffolds, drug release carriers, wound coating materials and functional diaphragms, etc., can greatly improve biological activity and repair Effect.

Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than limiting the protection scope of the present invention, although the present invention has been described in detail with reference to the preferred embodiments, those of ordinary skill in the art should understand , the technical solution of the present invention can be modified or equivalently replaced: for example, natural polymer materials such as collagen and chitosan, and synthetic polymers such as polylactic acid, polycaprolactone, and polylactic acid-glycolide are used to replace the present invention. gelatin, these equivalent forms do not depart from the spirit and scope of the technical solution of the present invention.

Claims (10)

1. mesopore silicon oxide particle/degradable polymer nano composite fibre is characterized in that, comprises mesopore silicon oxide particle and biodegradable polymer fiber, and wherein, described mesopore silicon oxide distribution of particles is in described biodegradable polymer fiber.

2. nano-composite fiber as claimed in claim 1 is characterized in that, the mass ratio of described mesopore silicon oxide particle and described degradable polymer 1: 10～500.

3. nano-composite fiber as claimed in claim 1 is characterized in that, the diameter of described nano-composite fiber is 50nm～2 μ m, and length is 20～800 μ m.

4. composite fibre as claimed in claim 1 is characterized in that, the aperture 2～10nm of described mesopore silicon oxide particle, specific area 200～1400m ²/ g.

5. composite fibre as claimed in claim 1 is characterized in that, described biodegradable polymers is gelatin, collagen, shitosan, PLA, polycaprolactone or PLA-glycolide.

6. composite fibre as claimed in claim 1 is characterized in that, described mesopore silicon oxide particle is the mesopore silicon oxide particle that is mounted with medicine.

7. the preparation method as each described mesopore silicon oxide particle/degradable polymer nano composite fibre in the claim 1～5 comprises the steps:

(a), the degradable polymer material dissolves is formed polymer solution in solvent;

(b), the mesopore silicon oxide particle is dispersed in the described polymer solution forms mixed solution;

(c), adopting electrostatic spinning technique that described mixed solution is carried out spinning obtains nanofiber, collects;

(d), to adopt 1% glutaraldehyde that prepared nanofiber is carried out crosslinked, promptly obtains described mesopore silicon oxide particle/degradable polymer nano composite fibre.

8. preparation method as claimed in claim 7 is characterized in that, described solvent is selected from a kind of in acetic acid, hexafluoroisopropanol, chloroform, acetone, carrene, the water or their mixture.

9. preparation method as claimed in claim 7, it is characterized in that the voltage that described electrostatic spinning adopts is 5～40KV, spinning speed is 1～3mL/h, the solution jet is 5～20 centimetres to the distance between the gatherer, and the specification of described solution jet syringe needle is 18～No. 27.

10. as the application of each described mesopore silicon oxide particle/degradable polymer nano composite fibre in the claim 1～5 in preparation tissue engineering bracket, medicine controlled release carrier, wound clad material or functional barrier film.

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