CN112169007A - Preparation method of antibacterial fiber membrane based on electrostatic spinning - Google Patents

Abstract

The invention relates to a preparation method of an antibacterial fiber membrane based on electrostatic spinning, in particular to a method for preparing a fiber membrane containing Ag NPs @ MOFs antibacterial materials by adopting an electrostatic spinning technology. The nano silver is uniformly dispersed on the surface, the framework and the pores of the ZIF-8 in an in-situ growth mode, so that the spontaneous agglomeration of silver particles is avoided, the stability and the monodispersity of antibacterial particles are improved, silk fibroin which is good in biocompatibility and can be controllably degraded is used as a base material, and the adhesion, growth and proliferation of cells can be effectively promoted; meanwhile, the antibacterial agent is combined with antibacterial performance, and can be applied to wound repair, inflammation diminishing and the like. The antibacterial particles are further formed into a film by adopting an electrostatic spinning mode, the operation is simple, and the cost is low.

Description

Preparation method of antibacterial fiber membrane based on electrostatic spinning

Technical Field

The invention relates to preparation and application of a novel antibacterial fiber membrane, relates to a preparation method of an antibacterial fiber membrane based on electrostatic spinning, and particularly relates to a method for preparing a fiber membrane containing Ag NPs @ MOFs antibacterial materials by adopting an electrostatic spinning technology. The invention belongs to the field of nano antibacterial materials.

Background

The increase in living standards has prompted more people to pay attention to the health field, and biological contamination (such as air, water, soil and food contamination) caused by pathogenic parasites, bacteria and viruses seriously threatens human health. Among them, infection caused by microorganisms and their metabolites causes great pain to the wound healing of patients. Meanwhile, the main problems encountered in the treatment of bacterial infections by the rapid evolution of drug-resistant bacteria and biofilms. The traditional antibacterial methods, such as antibacterial adsorption, bacterial surface contact sterilization, antibacterial agent release and the like, cannot achieve satisfactory antibacterial effects. Therefore, the development of novel antibacterial materials is urgently required. The literature reports that the inorganic nanoparticles can inhibit the formation of a biological membrane and have a good antibacterial effect.

The currently commonly used nano antibacterial materials mainly comprise: a silver-based antibacterial material (Chinese patent: a preparation method of a silver titanium dioxide composite antibacterial material, publication No. CN 104472543A); copper ion antibacterial material (Chinese patent: a manufacturing method of antibacterial material, publication No. CN 105381460A.); zinc oxide (a preparation method and application of a nano-zinc composite antibacterial material, and the publication number is CN 107163654A) and the like. The silver-based antibacterial material has the advantages of strong antibacterial activity, wide antibacterial spectrum, low tendency of inducing bacteria to resist drugs and the like, and is an ideal antibacterial material. However, nano silver particles (Ag NPs) tend to spontaneously aggregate to form larger particles and even precipitate, and the antibacterial performance of the nano silver particles is seriously influenced. The Metal Organic Frameworks (MOFs) MOFs material has the advantages of large specific surface area, good biocompatibility, stable chemical and physical properties and biodegradability. The MOFs is adopted to wrap the Ag NPs, so that spontaneous agglomeration of Ag NPs particles can be solved, and a stable antibacterial material is formed, and the antibacterial performance of the Ag NPs is effectively exerted.

The electrostatic spinning can utilize an electric field to produce the jetted high molecular fluid into nano-scale polymer filaments, so as to prepare the superfine fiber membrane with higher specific surface area and porosity, thereby achieving the purpose of different functionalization. Because the antibacterial fiber has very important application prospect in various fields, the novel antibacterial fiber is prepared by adopting the electrostatic spinning technology and combining the nano-grade antibacterial material, and has extremely good research and practical values.

Disclosure of Invention

Aiming at the defects in the prior art, the invention aims to provide a preparation method of an antibacterial fiber membrane based on electrostatic spinning.

The purpose of the invention is realized by the following scheme: a preparation method of an antibacterial fiber membrane based on electrostatic spinning is characterized by firstly synthesizing Ag NPs @ MOFs antibacterial material, further adjusting the size of the Ag NPs and the particle size and porosity of an MOF structure by adjusting the use amounts of silver ammonia solution, 2-methylimidazole and zinc acetate, and simultaneously obtaining Ag NPs @ MOFs structures with different shapes; then, Ag NPs @ MOFs are dissolved in silk fibroin solution with certain concentration to obtain Ag NPs @ MOFs/SF spinning solution with different mass ratios, and the fiber membrane with antibacterial performance is prepared by utilizing an electrostatic spinning technology, and the method comprises the following steps:

step (1), preparing Ag NPs @ MOFs:

1) 0.34g of silver nitrate (Ag NO) is weighed₃) Preparing a fresh 20 mM silver ammonia solution by using 0.12 g of sodium hydroxide (Na OH) and a proper amount of ammonia water, transferring the silver ammonia solution to a brown bottle, and storing the silver ammonia solution in the shade;

2) dissolving 70-80 g of 2-methylimidazole in 90 mL of aqueous solution, uniformly stirring at 37 ℃, adding 10 mL of 2.5-6 mM zinc salt solution, uniformly stirring, adding 0.5-1 mL of 20 mM silver ammonia solution into the reaction solution, and reacting the mixed solution at 37 ℃ and 400 rpm in a dark place for 2 hours;

3) adding 5mL of PVP (50-100 mM) solution into the reaction solution obtained in the step 2), uniformly mixing, putting into a photochemical reactor, and irradiating for 2-4 hr by using a 300W mercury lamp until the reaction is finished; then, the user can use the device to perform the operation,

4) centrifuging at 8000 rpm/min for 20 min to remove supernatant, washing for three times, and drying at 80 ℃ to obtain final products Ag NPs @ MOFs particles;

spinning in step (2):

1) dissolving Ag NPs @ MOFs particles into a Silk Fibroin (SF) solution to ensure that the mass ratio of the Ag NPs @ MOFs to the SF is 0.01-0.2 mg/mg, and uniformly mixing at 60 ℃ to obtain a concentrated Ag NPs @ MOFs/SF spinning solution;

2) and injecting the uniformly mixed spinning solution into a 10 mL injector, setting the electrospinning pressure to be 20 kv, and carrying out electrostatic spinning at the flow speed of 0.1-1 mL/h within the acceptance distance of 10-20 cm to obtain the Ag NPs @ MOFs/SF fiber membrane with the thickness of 0.05-0.2 mm.

Wherein the zinc salt in the step (1) is zinc nitrate or zinc acetate, and the concentration of the zinc salt is 2.5-6 mM.

The SF concentration in the step (2) is 30% -40%.

The method for preparing the antibacterial electrospun fiber has the advantages of simple process, easy operation and good biocompatibility, and can be used for further preparing the antibacterial film material or the textile material on the biological surface.

And (3) testing antibacterial performance: escherichia coli was used as the test strain. Taking Escherichia coli cultured for more than three generations, and diluting with PBS buffer solution to about 10⁵ cfu/mL; taking 3X 3 cm²The prepared Ag NPs @ MOFs/SF fiber membrane is placed in a triangular flask, 45 mL of PBS containing 0.1% Tween-80 is added and mixed, and then 2.5 mL of the prefabricated bacterial suspension is added. The control group was prepared as described above for bacterial suspension of the microfilm without antimicrobial particles. The test group and the control group were incubated at 37 ℃ for 2 hr with shaking in a constant temperature shaker at 100 rpm. After the completion of shaking, the test group and the control group were appropriately diluted, inoculated on plates containing agar medium, 2 replicates were set for each concentration, the plates were cultured in an incubator at 37 ℃ for 24 hr, and the bacterial colonies were counted to calculate the antibacterial rate.

The invention has the advantages that:

(1) the nano silver is uniformly dispersed on the surface, the framework and the pores of the ZIF-8 in an in-situ growth mode, so that the spontaneous agglomeration of silver particles is avoided, and the stability and the monodispersity of antibacterial particles are improved;

(2) the silk fibroin which has good biocompatibility and controllable degradation is adopted as a matrix material, so that the adhesion, growth and proliferation of cells can be effectively promoted; meanwhile, the antibacterial agent is combined with antibacterial performance, and can be applied to wound repair, inflammation diminishing and the like.

(3) The antibacterial particles are further formed into a film by adopting an electrostatic spinning mode, the operation is simple, and the cost is low.

Detailed Description

The technical solution of the present invention is further described below by specific examples. The following examples are further illustrative of the present invention and do not limit the scope of the present invention.

Example 1

An antibacterial fiber membrane based on electrostatic spinning is characterized in that Ag NPs @ MOFs antibacterial materials are synthesized, the size of the Ag NPs and the particle size and porosity of an MOF structure are further adjusted by adjusting the using amounts of silver ammonia solution, 2-methylimidazole and zinc acetate, and meanwhile, Ag NPs @ MOFs structures with different shapes are obtained; then, dissolving Ag NPs @ MOFs in silk fibroin solution with certain concentration to obtain Ag NPs @ MOFs/SF spinning solution with different mass ratios, preparing a fiber membrane with antibacterial property by using an electrostatic spinning technology, and preparing the fiber membrane by the following steps:

1. preparation of Ag NPs @ MOFs:

1) 0.34g of silver nitrate (Ag NO) is weighed₃) Preparing a fresh 20 mM silver ammonia solution by using 0.12 g of sodium hydroxide (Na OH) and a proper amount of ammonia water, transferring the silver ammonia solution to a brown bottle, and storing the silver ammonia solution in the shade;

2) dissolving 75 g 2-methylimidazole in 90 mL of aqueous solution, stirring uniformly at 37 ℃, adding 10 mL of 5 mM zinc nitrate solution, stirring uniformly, adding 0.5mL of 20 mM silver ammonia solution into the reaction solution, and reacting the mixed solution at 37 ℃ and 400 rpm in a dark place for 2 hours;

3) adding 5mL of PVP 100 mM solution into the reaction solution obtained in the step 2), uniformly mixing, putting into a photochemical reactor, irradiating by using a 300W mercury lamp for 4 hr, and finishing the reaction; then, the user can use the device to perform the operation,

4) centrifuging at 8000 rpm/min for 20 min to remove supernatant, washing for three times, and drying at 80 ℃ to obtain final products Ag NPs @ MOFs particles;

2. spinning:

1) dissolving 350 mg of Ag NPs @ MOFs particles into 10 mL of Silk Fibroin (SF) solution with the mass concentration of 35%, and uniformly mixing at 60 ℃ to obtain concentrated Ag NPs @ MOFs/SF spinning solution;

2) and injecting the uniformly mixed spinning solution into a 10 mL injector, setting the electrospinning pressure to be 20 kv, and carrying out electrostatic spinning at the flow speed of 0.5mL/h within the acceptance distance of 15 cm to obtain the Ag NPs @ MOFs/SF fiber membrane with the thickness of 0.05-0.2 mm.

3. And (3) testing antibacterial performance: escherichia coli is used as a test strain. Taking Escherichia coli cultured for more than three generations, and diluting with PBS buffer solution to about 10⁵ cfu/mL; taking 3X 3 cm²The prepared Ag NPs @ MOFs/SF fiber membrane is placed in a triangular flask, 45 mL of PBS containing 0.1% Tween-80 is added and mixed, and then 2.5 mL of the prefabricated bacterial suspension is added. The control group was prepared as described above for bacterial suspension of the microfilm without antimicrobial particles. The test group and the control group were incubated at 37 ℃ for 2 hr with shaking in a constant temperature shaker at 100 rpm. After the completion of shaking, the test group and the control group were appropriately diluted, inoculated on plates containing agar medium, 2 replicates were set for each concentration, the plates were cultured in an incubator at 37 ℃ for 24 hr, and the bacterial colonies were counted to calculate the antibacterial rate.

Example 2

An antibacterial fiber membrane based on electrostatic spinning, similar to example 1, is prepared by the following steps:

1. preparation of Ag NPs @ MOFs:

1) 0.34g of silver nitrate (Ag NO) is weighed₃) Preparing a fresh 20 mM silver ammonia solution by using 0.12 g of sodium hydroxide (Na OH) and a proper amount of ammonia water, transferring the silver ammonia solution to a brown bottle, and storing the silver ammonia solution in the shade;

2) dissolving 80 g of 2-methylimidazole in 90 mL of aqueous solution, uniformly stirring at 37 ℃, adding 10 mL of 6 mM zinc nitrate solution, uniformly stirring, adding 0.75 mL of 20 mM silver ammonia solution into the reaction solution, and reacting the mixed solution at 37 ℃ and 400 rpm in a dark place for 2 hr;

3) adding 5mL of 100 mM PVP solution into the reaction solution, uniformly mixing, putting into a photochemical reactor, and irradiating for 3 hr by using a 300W mercury lamp;

4) after the reaction is finished, centrifuging at 8000 rpm/min for 20 min, separating and removing supernatant, washing for three times, and drying at 80 ℃ to obtain the final product Ag NPs @ MOFs particles.

2. Spinning:

1) dissolving 500 mg of Ag NPs @ MOFs particles in 10 mL of Silk Fibroin (SF) solution with the mass concentration of 35%, and uniformly mixing at 60 ℃ to obtain concentrated Ag NPs @ MOFs/SF spinning solution;

2) and injecting the uniformly mixed spinning solution into a 10 mL injector, setting the electrospinning pressure to be 20 kv, and carrying out electrostatic spinning at the flow speed of 0.5mL/h within an acceptance distance of about 15 cm to obtain the Ag NPs @ MOFs/SF fiber membrane of 0.05-0.2 mm.

3. And (3) testing antibacterial performance: escherichia coli was used as the test strain. Taking Escherichia coli cultured for more than three generations, and diluting with PBS buffer solution to about 10⁵ cfu/mL; taking 3X 3 cm²The prepared Ag NPs @ MOFs/SF fiber membrane is placed in a triangular flask, 45 mL of PBS containing 0.1% Tween-80 is added and mixed, and then 2.5 mL of the prefabricated bacterial suspension is added. The control group was prepared as described above for bacterial suspension of the microfilm without antimicrobial particles. The test group and the control group were incubated at 37 ℃ for 2 hr with shaking in a constant temperature shaker at 100 rpm. After the completion of shaking, the test group and the control group were appropriately diluted, inoculated on plates containing agar medium, 2 replicates were set for each concentration, the plates were cultured in an incubator at 37 ℃ for 24 hr, and the bacterial colonies were counted to calculate the antibacterial rate.

Example 3

An antibacterial fiber membrane based on electrostatic spinning, similar to example 1, is prepared by the following steps:

1. preparation of Ag NPs @ MOFs:

1) 0.34g of silver nitrate (Ag NO) is weighed₃) Preparing a fresh 20 mM silver ammonia solution by using 0.12 g of sodium hydroxide (Na OH) and a proper amount of ammonia water, transferring the silver ammonia solution to a brown bottle, and storing the silver ammonia solution in the shade;

2) dissolving 70 g of 2-methylimidazole in 90 mL of aqueous solution, stirring uniformly at 37 ℃, adding 10 mL of 5 mM zinc acetate solution, stirring uniformly, adding 0.5mL of 20 mM silver ammonia solution into the reaction solution, and reacting the mixed solution at 37 ℃ and 400 rpm in a dark place for 2 hr;

3) adding 5mL PVP 100 mM) solution into the reaction solution, uniformly mixing, putting into a photochemical reactor, and irradiating for 4 hr by using a 300W mercury lamp;

4) after the reaction is finished, centrifuging at 8000 rpm/min for 20 min, separating and removing supernatant, washing for three times, and drying at 80 ℃ to obtain the final product Ag NPs @ MOFs particles.

2. Spinning:

1) dissolving 400 mg of Ag NPs @ MOFs particles in 10 mL of Silk Fibroin (SF) solution with the mass fraction of 35%, and uniformly mixing at 60 ℃ to obtain concentrated Ag NPs @ MOFs/SF spinning solution;

2) and injecting the uniformly mixed spinning solution into a 10 mL injector, setting the electrospinning pressure to be 20 kv, and carrying out electrostatic spinning at the flow rate of 0.5mL/h within an acceptance distance of about 15 cm to obtain a fiber membrane of 0.05-0.2 mm.

3. And (3) testing antibacterial performance: escherichia coli was used as the test strain. Taking Escherichia coli cultured for more than three generations, and diluting with PBS buffer solution to about 10⁵ cfu/mL; taking 3X 3 cm²The prepared Ag NPs @ MOFs/SF fiber membrane is placed in a triangular flask, 45 mL of PBS containing 0.1% Tween-80 is added and mixed, and then 2.5 mL of the prefabricated bacterial suspension is added. The control group was prepared as described above for bacterial suspension of the microfilm without antimicrobial particles. The test group and the control group were incubated at 37 ℃ for 2 hr with shaking in a constant temperature shaker at 100 rpm. After the completion of shaking, the test group and the control group were appropriately diluted, inoculated on plates containing agar medium, 2 replicates were set for each concentration, the plates were cultured in an incubator at 37 ℃ for 24 hr, and the bacterial colonies were counted to calculate the antibacterial rate.

Table 1 shows the results of the antibacterial effect of the Ag NPs @ MOFs/SF fiber membranes prepared in the embodiments 1-3 of the present invention on Escherichia coli, and the results show that the killing efficiency of the prepared Ag NPs @ MOFs/SF fiber membranes on Escherichia coli is above 99.9%:

。

Claims (3)

1. a preparation method of an antibacterial fiber membrane based on electrostatic spinning is characterized by firstly synthesizing Ag NPs @ MOFs antibacterial material, further adjusting the size of the Ag NPs and the particle size and porosity of an MOF structure by adjusting the use amounts of silver ammonia solution, 2-methylimidazole and zinc acetate, and simultaneously obtaining Ag NPs @ MOFs structures with different shapes; then, Ag NPs @ MOFs are dissolved in silk fibroin solution with certain concentration to obtain Ag NPs @ MOFs/SF spinning solution with different mass ratios, and the fiber membrane with antibacterial performance is prepared by utilizing an electrostatic spinning technology, and the method comprises the following steps:

step (1), preparing Ag NPs @ MOFs:

1) 0.34g of silver nitrate (Ag NO) is weighed₃) Preparing a fresh 20 mM silver ammonia solution by using 0.12 g of sodium hydroxide (Na OH) and a proper amount of ammonia water, transferring the silver ammonia solution to a brown bottle, and storing the silver ammonia solution in the shade;

2) dissolving 70-80 g of 2-methylimidazole in 90 mL of aqueous solution, uniformly stirring at 37 ℃, adding 10 mL of 2.5-6 mM zinc salt solution, uniformly stirring, adding 0.5-1 mL of 20 mM silver ammonia solution into the reaction solution, and reacting the mixed solution at 37 ℃ and 400 rpm in a dark place for 2 hours;

3) adding 5mL of PVP (50-100 mM) solution into the reaction solution obtained in the step 2), uniformly mixing, putting into a photochemical reactor, and irradiating for 2-4 hr by using a 300W mercury lamp until the reaction is finished; then, the user can use the device to perform the operation,

4) centrifuging at 8000 rpm/min for 20 min to remove supernatant, washing for three times, and drying at 80 ℃ to obtain final products Ag NPs @ MOFs particles;

spinning in step (2):

1) dissolving Ag NPs @ MOFs particles into a Silk Fibroin (SF) solution, enabling the mass ratio of the Ag NPs @ MOFs to the SF to be 0.01-0.2, and uniformly mixing at 60 ℃ to obtain a concentrated Ag NPs @ MOFs/SF spinning solution;

2) and injecting the uniformly mixed spinning solution into a 10 mL injector, setting the electrospinning pressure to be 20 kv, and carrying out electrostatic spinning at the flow speed of 0.1-1 mL/h within the acceptance distance of 10-20 cm to obtain the Ag NPs @ MOFs/SF fiber membrane with the thickness of 0.05-0.2 mm.

2. The electrospun-based antibacterial fibrous membrane of claim 1, characterized in that the zinc salt in step (1) is zinc nitrate or zinc acetate.

3. The electrospinning-based antibacterial fiber membrane of claim 1, wherein the SF concentration in step (2) is 30% to 40%.