CN114848819B - Preparation method and application of photosensitive silver nano chitosan microsphere - Google Patents
Preparation method and application of photosensitive silver nano chitosan microsphere Download PDFInfo
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- 229920001661 Chitosan Polymers 0.000 title claims abstract description 125
- 239000004005 microsphere Substances 0.000 title claims abstract description 124
- 229910052709 silver Inorganic materials 0.000 title claims abstract description 52
- 239000004332 silver Substances 0.000 title claims abstract description 52
- BQCADISMDOOEFD-UHFFFAOYSA-N Silver Chemical compound [Ag] BQCADISMDOOEFD-UHFFFAOYSA-N 0.000 title claims abstract description 48
- 238000002360 preparation method Methods 0.000 title claims abstract description 26
- 239000000463 material Substances 0.000 claims abstract description 35
- SQGYOTSLMSWVJD-UHFFFAOYSA-N silver(1+) nitrate Chemical compound [Ag+].[O-]N(=O)=O SQGYOTSLMSWVJD-UHFFFAOYSA-N 0.000 claims abstract description 28
- 108091003079 Bovine Serum Albumin Proteins 0.000 claims abstract description 25
- 229940098773 bovine serum albumin Drugs 0.000 claims abstract description 25
- RBTBFTRPCNLSDE-UHFFFAOYSA-N 3,7-bis(dimethylamino)phenothiazin-5-ium Chemical compound C1=CC(N(C)C)=CC2=[S+]C3=CC(N(C)C)=CC=C3N=C21 RBTBFTRPCNLSDE-UHFFFAOYSA-N 0.000 claims abstract description 20
- 229960000907 methylthioninium chloride Drugs 0.000 claims abstract description 20
- 230000000844 anti-bacterial effect Effects 0.000 claims abstract description 17
- 229910001961 silver nitrate Inorganic materials 0.000 claims abstract description 14
- 241000894006 Bacteria Species 0.000 claims abstract description 10
- PLKATZNSTYDYJW-UHFFFAOYSA-N azane silver Chemical compound N.[Ag] PLKATZNSTYDYJW-UHFFFAOYSA-N 0.000 claims abstract description 10
- 241000192125 Firmicutes Species 0.000 claims abstract description 6
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- 241000588724 Escherichia coli Species 0.000 claims description 25
- 230000001580 bacterial effect Effects 0.000 claims description 18
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 claims description 18
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- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 claims description 5
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- 229910052724 xenon Inorganic materials 0.000 claims description 5
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- 238000005286 illumination Methods 0.000 abstract description 18
- 239000003504 photosensitizing agent Substances 0.000 abstract description 18
- FOIXSVOLVBLSDH-UHFFFAOYSA-N Silver ion Chemical compound [Ag+] FOIXSVOLVBLSDH-UHFFFAOYSA-N 0.000 abstract description 12
- 239000000126 substance Substances 0.000 abstract description 5
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- -1 oxygen radical Chemical class 0.000 description 12
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- 230000000694 effects Effects 0.000 description 9
- ZKSVYBRJSMBDMV-UHFFFAOYSA-N 1,3-diphenyl-2-benzofuran Chemical compound C1=CC=CC=C1C1=C2C=CC=CC2=C(C=2C=CC=CC=2)O1 ZKSVYBRJSMBDMV-UHFFFAOYSA-N 0.000 description 8
- 230000007935 neutral effect Effects 0.000 description 8
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- MPMSMUBQXQALQI-UHFFFAOYSA-N cobalt phthalocyanine Chemical compound [Co+2].C12=CC=CC=C2C(N=C2[N-]C(C3=CC=CC=C32)=N2)=NC1=NC([C]1C=CC=CC1=1)=NC=1N=C1[C]3C=CC=CC3=C2[N-]1 MPMSMUBQXQALQI-UHFFFAOYSA-N 0.000 description 3
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Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K41/00—Medicinal preparations obtained by treating materials with wave energy or particle radiation ; Therapies using these preparations
- A61K41/0057—Photodynamic therapy with a photosensitizer, i.e. agent able to produce reactive oxygen species upon exposure to light or radiation, e.g. UV or visible light; photocleavage of nucleic acids with an agent
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K33/00—Medicinal preparations containing inorganic active ingredients
- A61K33/24—Heavy metals; Compounds thereof
- A61K33/38—Silver; Compounds thereof
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K41/00—Medicinal preparations obtained by treating materials with wave energy or particle radiation ; Therapies using these preparations
- A61K41/0057—Photodynamic therapy with a photosensitizer, i.e. agent able to produce reactive oxygen species upon exposure to light or radiation, e.g. UV or visible light; photocleavage of nucleic acids with an agent
- A61K41/0071—PDT with porphyrins having exactly 20 ring atoms, i.e. based on the non-expanded tetrapyrrolic ring system, e.g. bacteriochlorin, chlorin-e6, or phthalocyanines
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
- A61K9/16—Agglomerates; Granulates; Microbeadlets ; Microspheres; Pellets; Solid products obtained by spray drying, spray freeze drying, spray congealing,(multiple) emulsion solvent evaporation or extraction
- A61K9/1605—Excipients; Inactive ingredients
- A61K9/1629—Organic macromolecular compounds
- A61K9/1652—Polysaccharides, e.g. alginate, cellulose derivatives; Cyclodextrin
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- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/04—Antibacterial agents
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B22—CASTING; POWDER METALLURGY
- B22F—WORKING METALLIC POWDER; MANUFACTURE OF ARTICLES FROM METALLIC POWDER; MAKING METALLIC POWDER; APPARATUS OR DEVICES SPECIALLY ADAPTED FOR METALLIC POWDER
- B22F1/00—Metallic powder; Treatment of metallic powder, e.g. to facilitate working or to improve properties
- B22F1/05—Metallic powder characterised by the size or surface area of the particles
- B22F1/054—Nanosized particles
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- B22F9/00—Making metallic powder or suspensions thereof
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- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
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Abstract
The invention belongs to the technical field of material preparation, and discloses a preparation method and application of photosensitive silver nano chitosan microspheres. The novel environment-friendly reaction system is provided, bovine serum albumin is used for preparing silver nano particles through the green reduction of silver nitrate by silver ammonia reaction, the silver nano particles are mixed with chitosan solution to prepare microspheres, and the microspheres are placed in photosensitizer solution (methylene blue and phthalocyanines) to obtain silver nano chitosan microspheres loaded with photosensitive substances. The microsphere has strong antibacterial performance on gram-negative bacteria, and also has strong antibacterial performance on gram-positive bacteria under illumination conditions.
Description
Technical Field
The invention belongs to the technical field of material preparation, and relates to a preparation method and application of photosensitive silver nano chitosan microspheres.
Background
In order to solve the problems as soon as possible, saving resources and developing new energy sources become a great premise for sustainable development of countries around the world. The metal nanomaterial combined with biology is more upstairs in terms of environmental protection and green, and attracts attention. In the continuous development of the nano technology, after innovative combination with the biological field, the metal nano material is simpler and more convenient in synthesis, more green and environment-friendly in application and more economical and feasible.
Antibiotic antibacterial agents have made great progress in the treatment of some diseases over the last decades, greatly promoting the development of the medical field. However, in recent years, the number of antibiotic-resistant strains has been increasing, which is a serious problem for headache. The continuous evolution of drug-resistant strains will have a tremendous impact on the medical field today, and will also make current medical modalities weak, even ineffective. There is therefore a need for innovative and effective methods to combat these diseases caused by drug resistance.
Photodynamic therapy (PDT) is a promising treatment modality, and its advantages of spatial controllability, low-density light irradiation, adjustable toxicity to specific tissues, and minimal trauma have attracted a large number of researchers. Antibacterial photodynamic therapy is photodynamic therapy directed to drug-resistant strains of bacteria for killing or eliminating pathogens. In the prior researches, the key point of the antibacterial photodynamic therapy is oxygen free radicals generated by photosensitizers, and the strong oxidability of the oxygen free radicals can have a certain effect on the cell membrane of bacteria and even enter the bacteria to change the physiological activity of the bacteria. However, insufficient oxygen radical production has been a major factor in impeding the popularization of antibacterial photodynamic therapy.
Disclosure of Invention
In order to overcome the defects of the prior art, the invention provides a preparation method and application of photosensitive silver nano chitosan microspheres, and provides a novel environment-friendly reaction system, wherein bovine serum albumin is used for preparing silver nano particles through silver ammonia reaction and green reduction of silver nitrate, the silver nano particles are mixed with chitosan solution to prepare microspheres, and the microspheres are placed in photosensitizer methylene blue solution to obtain the silver nano chitosan microspheres loaded with photosensitive substance methylene blue. The microsphere has strong antibacterial performance on gram-negative bacteria and strong antibacterial performance on gram-positive bacteria under illumination conditions.
The first aim of the invention is to claim a preparation method of photosensitive silver nano chitosan microsphere, which has the advantages of green and environment-friendly raw materials and simple and convenient operation, and the prepared photosensitive silver nano chitosan microsphere can obviously inhibit gram-negative bacteria represented by escherichia coli or gram-positive bacteria represented by staphylococcus aureus.
The second purpose of the invention is to claim the application of the photosensitive silver nano chitosan microsphere prepared by the preparation method in bacteriostasis.
The above object of the present invention is achieved by the following technical solutions:
a photosensitive silver nanometer chitosan microsphere preparation method, use silver nitrate as raw materials, regard bovine serum albumin as reducing agent end capping agent to prepare silver nanometer material; mixing silver nano material with chitosan to prepare microsphere, and loading in methylene blue solution; wherein the mass ratio of the silver nitrate to the bovine serum albumin is 1:1.27-2.35.
The specific preparation steps are as follows:
s1, weighing silver nitrate, dissolving the silver nitrate in trace ultrapure water, and dropwise adding 0.05mL of concentrated ammonia water, wherein the solution becomes turbid first and then becomes a clear solution to obtain 15mmol/L silver ammonia solution;
s2, weighing 0.05-0.15 g of bovine serum albumin, and dissolving the bovine serum albumin in ultrapure water to obtain 0.01-0.03 mg/L bovine serum albumin solution;
s3, adding the silver ammonia solution into the bovine serum albumin solution, and heating in a water bath at 40-60 ℃ for 90-120 min to obtain clear light yellow liquid, namely the silver nano material, namely AgNPs@BSA;
s4, weighing 2g of chitosan powder, dissolving in 5-10% (v/v) citric acid solution, and stirring until the solution is completely dissolved, wherein the solution is transparent; mixing and stirring the prepared silver nano material and chitosan solution to obtain a yellowish solution material;
s5, filling the solution material obtained in the step S4 into a syringe, slowly dripping the solution material into 2.0-3.0 mol/L NaOH at 1-1.5 mm/min, and continuously stirring to obtain light yellow microspheres; washing the microsphere for 5-10 times by deionized water until the pH of the microsphere reaches neutrality by using pH test paper, and placing the microsphere in a refrigerator for hardening at 4 ℃ for 24-48 hours to obtain AgNPs@BSA/chitosan microsphere; the concentration of the preparation is 1 multiplied by 10 -4 ～1×10 -3 Adding the prepared AgNPs@BSA/chitosan microspheres into the photosensitizer solution in mol/L, stirring for 3-6 hours after ultrasonic oscillation for 3-6 hours, putting the pellets into a baking oven for drying for 1-2 hours to obtain the AgNPs@BSA@photosensitizer/chitosan microspheres, and storing in a refrigerator at 4 ℃ for later use.
Further, the ultrapure water in the step S2 is the minister ultrapure water.
Further, the silver nano material in the step S4 is mixed and stirred with the chitosan solution for 2 to 4 hours.
Further, the concentration of the photosensitizer in step S5 should be lower than 500mg/L to avoid injury to human body, and should be higher than 1mg/L to achieve the minimum excitation concentration of the photosensitizer. The photosensitizer is methylene blue or phthalocyanine photosensitizer.
The AgNPs@BSA@photosensitizer/chitosan microsphere prepared by the preparation method is applied to bacteriostasis, and particularly applied to inhibiting gram-negative bacteria represented by escherichia coli or gram-positive bacteria represented by staphylococcus aureus.
The specific application is as follows: the AgNPs@BSA@photosensitizer/chitosan microsphere is placed into bacterial liquid, cultured for 1h in a constant temperature shaking box, and then placed under a xenon lamp (150 w) for irradiation for 20min. Among them, ag NPs@BSA and chitosan in a ratio of 2:1 are most preferable.
After the AgNPs@BSA@photosensitizer/chitosan microsphere is applied, detection of escherichia coli and staphylococcus aureus shows that compared with bacterial liquid without materials, the bacterial liquid growth effect of the materials is inhibited, the inhibition effect is obvious, and even the sterilization effect is achieved.
The silver nano material and the photosensitizer are interacted under the illumination condition, so that the problem of insufficient yield of active oxygen is solved, and the antibacterial performance of the silver nano material is further enhanced.
Compared with the prior art, the invention has the beneficial effects that:
(1) The invention uses bovine serum albumin as a template to prepare nano materials and uses chitosan to prepare microspheres, the materials are all green and environment-friendly biological materials, the concentration of chemical reagents used in the preparation process is almost trace, and the preparation has plasticity and can obtain the material with the required shape.
(2) The sulfhydryl carboxyl hydroxyl in the bovine serum albumin is a key for reducing silver ions into silver, and the three-dimensional structure of the protein can limit the size of the nano material, so that the size of the prepared silver nano particles reaches the nano level and cannot be agglomerated. Meanwhile, the chitosan microsphere also contains corresponding carboxyl hydroxyl sulfhydryl, so that unreduced silver ions of bovine serum albumin can be further reduced into silver nano particles.
(3) In the invention, in the process that the photosensitizer is excited by light and releases oxygen free radicals, the release of the oxygen free radicals is further promoted due to the existence of the silver nano-particles, and meanwhile, partial silver nano-particles are changed into silver ions due to the existence of the oxygen free radicals, so that interaction of the silver nano-particles and the silver ions is promoted, and the integral antibacterial performance of the silver nano-particles is enhanced. The yield of oxygen radicals of the photosensitive silver nano-chitosan microsphere prepared by the invention can be reflected by the reduction degree of ultraviolet absorption peak of an indirect indicator 3-diphenyl isobenzofuran (DPBF) at 410nm, as shown in figure 11.
Drawings
FIG. 1 is a schematic diagram of various microspheres prepared in example 2, example 3 and example 4, wherein a is chitosan microsphere, b is AgNPs@BSA chitosan microsphere, c is AgNPs@BSA@MB/chitosan microsphere, d is AgNPs@BSA@Co Pc/chitosan microsphere, and e is AgNPs@BSA@Zn Pc/chitosan microsphere.
FIG. 2 is a schematic diagram showing the growth curve of E.coli in the presence of AgNPs@BSA@MB/chitosan microspheres in application example 1.
FIG. 3 is a schematic diagram showing the growth curve of Staphylococcus aureus in the presence of AgNPs@BSA@MB/chitosan microspheres in application example 2.
FIG. 4 is a schematic diagram showing the growth curve of E.coli in the presence of AgNPs@BSA@Co Pc/chitosan microspheres in application example 1.
FIG. 5 is a schematic diagram showing the growth curve of Staphylococcus aureus in the presence of AgNPs@BSA@Co Pc/chitosan microspheres in application example 2.
FIG. 6 is a schematic diagram showing the growth curve of E.coli in the presence of AgNPs@BSA@Zn Pc/chitosan microspheres in application example 1.
FIG. 7 is a schematic diagram showing the growth curve of Staphylococcus aureus in the presence of AgNPs@BSA@Zn Pc/chitosan microspheres in application example 2.
FIG. 8 is a graph showing the colony survival rate of E.coli at different methylene blue concentrations and different material ratios in application example 3.
FIG. 9 is a graph showing the survival rate of Staphylococcus aureus colonies at different methylene blue concentrations and different material ratios in application example 4.
FIG. 10 is a graph showing the effect of different illumination times on the antibacterial effect of Ag NPs@BSA@MB/chitosan microspheres.
FIG. 11 is a graph showing the change in the yield of oxygen radicals of Ag NPs@BSA@MB/chitosan microspheres, wherein graph a shows the comparison of absorbance values before and after 18min of illumination of DPBH, DPBH+MB, DPBH+MB+Ag NPs, and graph b shows the change in ultraviolet absorbance at 410nm after DPBH+MB+Ag mixing with time.
Detailed Description
The present invention is described in detail below by way of specific examples, but the scope of the present invention is not limited thereto. Unless otherwise specified, the experimental methods used in the present invention are all conventional methods, and all experimental equipment, materials, reagents, etc. used can be obtained from commercial sources.
Example 1
The preparation method of the silver nano chitosan microsphere with the antibacterial effect comprises the following steps:
(1) Preparing silver ammonia solution. 0.0637g of silver nitrate is weighed and dissolved in trace ultrapure water, 0.05mL of concentrated ammonia water is added dropwise, the solution becomes turbid first and then becomes clear solution, and the solution is transferred to a 25mL volumetric flask for volume metering, so that 15mmol/L of silver ammonia solution is obtained.
(2) Bovine serum albumin was prepared. 0.05g of bovine serum albumin was weighed and dissolved in 5ml of ultrapure water to prepare a 0.01mg/L bovine serum albumin solution.
(3) Synthesizing silver nano particles. Adding silver nitrate into the bovine serum albumin solution, heating in a water bath at 50 ℃ for 90min to obtain clear pale yellow liquid, namely the silver nano material, namely Ag NPs@BSA.
(4) Preparing the silver nano chitosan material. 2g of chitosan powder was weighed and dissolved in 100mL of 10% (v/v) citric acid solution, and the solution was stirred until complete dissolution, and the solution was transparent. And pouring 20ml of the prepared silver nano material into 20ml of chitosan solution, and stirring for 3 hours to obtain the 1:1 Ag NPs@BSA/chitosan material.
(5) Preparing chitosan microsphere and silver nanometer chitosan microsphere.
And (3) filling the chitosan solution into a syringe, slowly dripping the chitosan solution into 2.0mol/L NaOH, and continuously stirring to obtain the milky microsphere. Washing the microsphere with deionized water for several times to make the pH of the microsphere neutral, placing in deionized water, and placing in refrigerator at 4deg.C for hardening for 24 hr to obtain chitosan microsphere as shown in figure 1a.
The Ag NPs@BSA/chitosan solution is filled into a syringe, slowly dripped into 2.0mol/L NaOH, and continuously stirred to obtain light yellow microspheres. Washing the microsphere with deionized water for several times to make the pH of the microsphere neutral, placing in deionized water, and placing in a refrigerator at 4deg.C for hardening for 24 hr to obtain Ag NPs@BSA/chitosan microsphere as shown in figure 1b.
Example 2
The preparation method of the photosensitive substance methylene blue loaded silver nano chitosan microsphere with the antibacterial effect comprises the following steps:
(1) Preparing silver ammonia solution. 0.0637g of silver nitrate is weighed and dissolved in trace ultrapure water, 0.05mL of concentrated ammonia water is added dropwise, the solution becomes turbid first and then becomes clear solution, and the solution is transferred to a 25mL volumetric flask for volume metering, so that 15mmol/L of silver ammonia solution is obtained.
(2) Bovine serum albumin was prepared. 0.05g of bovine serum albumin was weighed and dissolved in 5ml of ultrapure water to prepare a 0.01mg/L bovine serum albumin solution.
(3) Synthesizing silver nano particles. Adding silver nitrate into the bovine serum albumin solution, heating in a water bath at 50 ℃ for 90min to obtain clear pale yellow liquid, namely the silver nano material, namely Ag NPs@BSA.
(4) Preparing the silver nano chitosan material. 2g of chitosan powder was weighed and dissolved in 100mL of 5% (v/v) citric acid solution, and the solution was stirred until complete dissolution, and the solution was transparent. And pouring 20ml of the prepared silver nano material into 20ml of chitosan solution, and stirring for 1h to obtain the 1:1 Ag NPs@BSA/chitosan material.
(5) Preparing silver nano chitosan microsphere. The Ag NPs@BSA material is filled into a syringe, slowly dripped into 2.0mol/L NaOH, and continuously stirred to obtain the light yellow microsphere. Washing the microsphere for several times by deionized water to make the pH of the microsphere neutral, placing the microsphere in deionized water, placing the microsphere in a refrigerator and hardening the microsphere for 24 hours at 4 ℃ to obtain the Ag NPs@BSA/chitosan microsphere.
(6) Loading a photosensitizer. The concentration of the preparation is 1 multiplied by 10 -4 And (3) taking 20 pellets of the methylene blue solution with the mol/L, respectively placing the pellets into 20mL of the methylene blue solution with the concentration, carrying out ultrasonic vibration for 1h, and stirring for 12h to obtain yellow-green pellets. And then placing the mixture into an oven, and drying the mixture to obtain the dark green Ag NPs@BSA@MB/chitosan microspheres, as shown in figure 1c.
Example 3
And (3) loading a photosensitizer in the step (6). The concentration of the preparation is 1 multiplied by 10 -4 And (3) taking 20 pellets of a mol/L cobalt phthalocyanine (Co Pc) solution, respectively placing the 20 pellets into 20mL cobalt phthalocyanine (Co Pc) solution with the concentration, carrying out ultrasonic vibration for 1h, and stirring for 12h to obtain pellets. And then placing the mixture into an oven to finally obtain Ag NPs@BSA@Co Pc/chitosan microspheres, as shown in FIG. 1d, and the other steps are the same as those in example 2.
Example 4
And (3) loading a photosensitizer in the step (6). The concentration of the preparation is 1 multiplied by 10 -4 And (3) taking 20 pellets of a mol/L zinc phthalocyanine (Zn Pc) solution, respectively placing the 20 pellets into 20mL cobalt phthalocyanine (Co Pc) solution with the concentration, carrying out ultrasonic vibration for 1h, and stirring for 12h to obtain pellets. And then placing the mixture into an oven to finally obtain Ag NPs@BSA@Zn Pc/chitosan microspheres, as shown in FIG. 1e, and the other steps are the same as those in example 2.
Example 5
And (3) loading a photosensitizer in the step (6). The concentration of the preparation is 5 multiplied by 10 -4 Taking 20 pellets of the methylene blue solution with mol/L and respectively placing 20mL of the methylene blue solution in the concentrationIn the methylene blue solution, ultrasonic oscillation is carried out for 1h, and the pellets are yellow-green after stirring for 12 h. Then put into an oven, otherwise as in example 2.
Example 6
And (3) loading a photosensitizer in the step (6). The concentration of the preparation is 1 multiplied by 10 -3 And (3) taking 20 pellets of the methylene blue solution with the mol/L, respectively placing the pellets into 20mL of the methylene blue solution with the concentration, carrying out ultrasonic vibration for 1h, and stirring for 12h to obtain yellow-green pellets. Then put into an oven, otherwise as in example 2.
Example 7
And (3) preparing the silver nano chitosan material in the step (4). 2g of chitosan powder was weighed and dissolved in 100mL of 5% (v/v) citric acid solution, and the solution was stirred until complete dissolution, and the solution was transparent. And pouring 40ml of silver nano material into 20ml of chitosan solution, and stirring for 1h, wherein the solution is yellowish, so as to obtain the 1:2 Ag NPs@BSA/chitosan material. Otherwise, the same as in example 2 is carried out.
Example 8
And (3) preparing the silver nano chitosan material in the step (4). 2g of chitosan powder was weighed and dissolved in 100mL of 5% (v/v) citric acid solution, and the solution was stirred until complete dissolution, and the solution was transparent. 10ml of silver nano material is poured into 20ml of chitosan solution, and the solution is stirred for 1h, so that the solution is yellowish, and the Ag NPs@BSA/chitosan material with the ratio of 2:1 is obtained. Otherwise, the same as in example 2 is carried out.
Application example 1
The Ag NPs@BSA@MB/chitosan microspheres, the Ag NPs@BSA@Zn Pc/chitosan microspheres and the Ag NPs@BSA@Co Pc/chitosan microspheres prepared in the examples 2, 3 and 4 are selected to apply bacteriostasis in escherichia coli.
(1) LB medium was prepared. 1.0g of tryptone, 0.5g of yeast extract and 0.5g of NaCl are weighed into a 250ml conical flask, 100ml of deionized water is added, and 2.0mol/L of NaOH is added dropwise to adjust the pH, so that the solution becomes neutral. Sealing, and sterilizing in a sterilizing pot.
(2) E.coli bacterial liquid is cultivated. Taking the stock solution of the escherichia coli stored at 80 ℃, sucking 50 mu L of the stock solution of the escherichia coli, adding the stock solution into 100mL of liquid culture medium, placing the liquid culture medium into a constant-temperature shaking box at 37 ℃, culturing for 12 hours, taking out 2mL of the stock solution of the escherichia coli, diluting the stock solution to the concentration with the OD value of 0.6, and placing the stock solution at 4 ℃ for later use.
(3) 20ml of LB liquid medium is respectively taken and placed in 5 conical flasks, and 20 mu L of E.coli bacterial liquid with OD value of 0.6 concentration is respectively added. Selecting one group as a blank control, adding chitosan microspheres, respectively adding 5 MB/chitosan microspheres, 5 Ag NPs@BSA/chitosan microspheres, 5 Ag NPs@BSA@MB/chitosan microspheres and 5 Ag NPs@BSA@MB/chitosan microspheres, wherein the samples of the fourth group and the fifth group are the same, and controlling the illumination condition. Three replicates were used in each group. Five groups of samples are placed in a constant temperature shaking box at 37 ℃ for culture, and 2ml of liquid is taken every 6 hours to measure OD600.
The operations of Ag NPs@BSA@Zn Pc/chitosan microsphere and Ag NPs@BSA@Co Pc/chitosan microsphere are all synchronous (3).
As shown in the results of figures 2, 4 and 6, the Ag NPs@BSA@MB/chitosan microspheres, ag NPs@BSA@Zn Pc/chitosan microspheres and Ag NPs@BSA@Co Pc/chitosan microspheres have strong antibacterial performance on gram-negative bacteria represented by escherichia coli, and even have certain antibacterial performance under illumination.
Application example 2
The Ag NPs@BSA@MB/chitosan microspheres, the Ag NPs@BSA@Zn Pc/chitosan microspheres and the Ag NPs@BSA@Co Pc/chitosan microspheres prepared in the examples 2, 3 and 4 are selected to apply bacteriostasis to staphylococcus aureus.
(1) LB medium was prepared. 1.0g of tryptone, 0.5g of yeast extract and 0.5g of NaCl are weighed into a 250ml conical flask, 100ml of deionized water is added, and 2.0mol/L of NaOH is added dropwise to adjust the pH, so that the solution becomes neutral. Sealing, and sterilizing in a sterilizing pot.
(2) And culturing staphylococcus aureus bacterial liquid. Taking staphylococcus aureus stock solution stored at 80 ℃, sucking 50 mu L of the bacterial solution, adding the bacterial solution into 100mL of liquid culture medium, placing the liquid culture medium into a constant temperature shaking box at 37 ℃, culturing for 12 hours, taking out 2mL of liquid culture medium, diluting the liquid culture medium to a concentration with an OD value of 0.6, and placing the liquid culture medium at 4 ℃ for later use.
(3) 20ml of LB liquid medium are respectively taken and placed in 5 conical flasks, and 100 mu L of staphylococcus aureus bacterial liquid with OD value of 0.6 concentration is respectively added. Selecting one group as a blank control, adding chitosan microspheres, respectively adding 5 MB/chitosan microspheres, 5 Ag NPs@BSA/chitosan microspheres, 5 Ag NPs@BSA@MB/chitosan microspheres and 5 Ag NPs@BSA@MB/chitosan microspheres, wherein the samples of the fourth group and the fifth group are the same, and controlling the illumination condition. Three replicates were used in each group. Five groups of samples are placed in a constant temperature shaking box at 37 ℃ for culture, and 2ml of liquid is taken every 6 hours to measure OD600.
The operations of Ag NPs@BSA@Zn Pc/chitosan microsphere and Ag NPs@BSA@Co Pc/chitosan microsphere are all synchronous (3). As shown in fig. 3, 5 and 7, the antibacterial effect of the Ag nps@bsa@mb/chitosan microspheres, ag nps@bsa@zn Pc/chitosan microspheres and Ag nps@bsa@co Pc/chitosan microspheres on gram positive bacteria represented by staphylococcus aureus is not obvious or even not effective under the condition of no illumination, but has a certain antibacterial performance through interaction of a photosensitizer and a silver nanomaterial under the condition of illumination.
Application example 3
(1) LB medium was prepared. 1.0g of tryptone, 0.5g of yeast extract and 0.5g of NaCl are weighed into a 250ml conical flask, 100ml of deionized water is added, and 2.0mol/L of NaOH is added dropwise to adjust the pH, so that the solution becomes neutral. Sealing, and sterilizing in a sterilizing pot.
(2) E.coli bacterial liquid is cultivated. Taking stock solution of Escherichia coli stored at 80deg.C, sucking 50 μl of the stock solution, adding into 100mL of liquid culture medium, placing into a constant temperature shaking box at 37deg.C, culturing for 12 hr, taking out, and diluting to 10 -5 The concentration is kept at 4 ℃ for standby.
(3) 5ml of medium was added to each tube, 50. Mu.L was added and diluted to 10 -5 Concentration of E.coli. No. 1 is used as a blank control with chitosan microspheres, no. 2 is used as a control with MB/chitosan microspheres, and No. 3 is used as a control with Ag NPs@BSA/chitosan microspheres. No. 4 was added to the Ag NPs@BSA@MB/chitosan microspheres prepared in example 2. No. 5 was added to the Ag NPs@BSA@MB/chitosan microspheres prepared in example 5. No. 6 was added to the Ag NPs@BSA@MB/chitosan microspheres prepared in example 6. No. 7 was added to the Ag NPs@BSA@MB/chitosan microspheres prepared in example 7. No. 8 was added to the Ag NPs@BSA@MB/chitosan microspheres prepared in example 8.
Three replicates were run for each group. After all samples were cultured in a constant temperature shaking incubator for 1h, they were placed under a xenon lamp (150 w) and irradiated for 20min, 100. Mu.L each was smeared on agar medium and incubated in the incubator for 24h. The colony count was observed and recorded using a colony counter.
The results are shown in FIG. 8 and Table 1 below: under the condition of illumination, the higher the concentration of the photosensitive substance in the material, the stronger the growth inhibition effect on the escherichia coli bacterial liquid, but the influence of the content of the photosensitive agent on the water environment and the human body health is considered, and the photosensitive agent with high concentration is selected in the safety range. The higher the proportion of the silver nano material in the material, namely the concentration, the stronger the growth inhibition effect on bacterial liquid.
TABLE 1 survival of E.coli in different materials
Application example 4
(1) LB medium was prepared. 1.0g of tryptone, 0.5g of yeast extract and 0.5g of NaCl are weighed into a 250ml conical flask, 100ml of deionized water is added, and 2.0mol/L of NaOH is added dropwise to adjust the pH, so that the solution becomes neutral. Sealing, and sterilizing in a sterilizing pot.
(2) And culturing staphylococcus aureus bacterial liquid. Taking staphylococcus aureus stock solution stored at 80 ℃, sucking 50 mu L of the stock solution, adding the stock solution into 100mL of liquid culture medium, placing the liquid culture medium into a constant temperature shaking box at 37 ℃ for culturing for 12 hours, taking out and diluting to 10 -5 The concentration is kept at 4 ℃ for standby.
(3) 5ml of medium was added to each tube, 50. Mu.L was added and diluted to 10 -5 Concentration of staphylococcus aureus. Chitosan microspheres were added as a blank control, MB/chitosan microspheres were added as a control, and Ag nps@bsa/chitosan microspheres were added as a control, no. 3. No. 4 was added to the Ag NPs@BSA@MB/chitosan microspheres prepared in example 2. No. 5 was added to the Ag NPs@BSA@MB/chitosan microspheres prepared in example 5. No. 6 was added to the Ag NPs@BSA@MB/chitosan microspheres prepared in example 6. No. 7 was added to the Ag NPs@BSA@MB/chitosan microspheres prepared in example 7. No. 8 the Ag NPs@B prepared in example 8 was addedSA@MB/chitosan microspheres.
Three replicates were run for each group. After all samples were cultured in a constant temperature shaking incubator for 1h, they were placed under a xenon lamp (150 w) and irradiated for 20min, 100. Mu.L each was smeared on agar medium and incubated in the incubator for 24h. The colony count was observed and recorded using a colony counter.
Under the condition of illumination, the higher the concentration of the photosensitive substance in the material is, the stronger the growth inhibition effect on bacterial liquid is. The higher the proportion of silver nano material in the material, namely the concentration, the stronger the growth inhibition effect on bacterial liquid.
TABLE 2 survival of Staphylococcus aureus in different materials
Application example 5
(1) LB medium was prepared. 1.0g of tryptone, 0.5g of yeast extract and 0.5g of NaCl are weighed into a 250ml conical flask, 100ml of deionized water is added, and 2.0mol/L of NaOH is added dropwise to adjust the pH, so that the solution becomes neutral. Sealing, and sterilizing in a sterilizing pot.
(2) E.coli bacterial liquid is cultivated. Taking the stock solution of the escherichia coli stored at 80 ℃, sucking 50 mu L of the stock solution of the escherichia coli, adding the stock solution into 100mL of liquid culture medium, placing the liquid culture medium into a constant-temperature shaking box at 37 ℃, culturing for 12 hours, taking out 2mL of the stock solution of the escherichia coli, diluting the stock solution to the concentration with the OD value of 0.6, and placing the stock solution at 4 ℃ for later use.
(3) 5ml of LB liquid medium was placed in 9 conical flasks, respectively, and 50. Mu.L of E.coli bacterial liquid with an OD of 0.6 was added, respectively. 5 Ag NPs@BSA@MB/chitosan microspheres prepared in example 2 are respectively added, and different illumination conditions of 0, 10, 20, 30, 40, 50, 60, 90 and 120min are set. Three replicates were used in each group. 9 groups of samples are placed in a constant temperature shaking box at 37 ℃ for culture, and liquid is taken every 6 hours to measure OD600.
The data are plotted to form fig. 10, and as can be seen from fig. 10, the overall bacteriostatic effect of the material is good after 20-60 min of illumination, but as a certain temperature is generated by illumination, the material is placed in the constant temperature environment of the ice-water mixture during illumination, but in order to avoid the influence of the temperature on the material, the illumination condition is best after 20min of illumination.
Experimental mechanism research
In order to verify the interaction mechanism of the prepared silver nano material and the photosensitizer, 1, 3-diphenyl isobenzofuran (DPBF) is selected as an indirect indicator of oxygen free radicals for exploration. Since DPBF is very sensitive to oxygen radicals, the absorbance of DPBF at 410 decreases gradually after encountering oxygen radicals, and is therefore widely used as an indirect indicator of oxygen radicals. Preparing 1000ppm of DPBF in dimethyl sulfoxide (DMSO), sucking 2mL of DMSO, adding the mixture into an ultraviolet cuvette, then sucking 20 mu L of DPBF, adding the mixture into the cuvette, and measuring an ultraviolet absorption spectrum; based on the above operation, 100. Mu.L of methylene blue solution was added dropwise, and the ultraviolet absorption spectrum was measured; based on the above operation, 100. Mu.L of the prepared Ag NPs@BSA material was additionally added dropwise. Three sets of solutions were placed under a xenon lamp fitted with a 600nm filter and the absorption spectrum was measured every 2 min. From the measured spectra (i.e., FIG. 11), the absorbance decrease at 410 was most pronounced with the addition of DPBF, methylene blue solution and silver nanomaterial solution. From this, it can be determined that the silver nanomaterial and the photosensitizer such as methylene blue generate more oxygen radicals under the catalysis of light.
The above-described embodiments are only preferred embodiments of the invention, and not all embodiments of the invention are possible. Any obvious modifications thereof, which would be apparent to those skilled in the art without departing from the principles and spirit of the present invention, should be considered to be included within the scope of the appended claims.
Claims (3)
1. A preparation method of photosensitive silver nano chitosan microsphere with antibacterial performance is characterized in that silver nitrate is used as a raw material, bovine serum albumin is used as a reducing agent end capping agent to prepare silver nano material; mixing silver nano material with chitosan to prepare microsphere, and loading in methylene blue solution; wherein the mass ratio of the silver nitrate to the bovine serum albumin is 1:1.27-2.35;
the specific preparation steps are as follows:
s1, weighing silver nitrate, dissolving the silver nitrate in trace ultrapure water, dropwise adding two drops of concentrated ammonia water, firstly turning the solution into turbid, and stopping dropwise adding the concentrated ammonia water when the solution becomes a clear solution to obtain 15mmol/L silver ammonia solution;
s2, weighing 0.05-0.15 g of bovine serum albumin, and dissolving the bovine serum albumin into ultrapure water to obtain 0.01-0.03 mg/L bovine serum albumin solution;
s3, adding the silver ammonia solution into the bovine serum albumin solution, and heating in a water bath at 40-60 ℃ for 90-120 min to obtain clear light yellow liquid, namely the silver nano material Ag NPs@BSA;
s4, weighing 2g of chitosan powder, dissolving in 5-10 v/v% citric acid solution, and stirring until the solution is completely dissolved, wherein the solution is transparent; mixing and stirring the prepared silver nano material and chitosan solution according to the proportion of 2:1 to obtain a yellowish solution material;
s5, filling the solution material obtained in the step S4 into a syringe, slowly dripping the solution material into 2.0-3.0 mol/L NaOH at a speed of 1-1.5 mm/min, and continuously stirring to obtain light yellow microspheres; washing the microspheres with deionized water for 5-10 times until the pH of the microspheres reaches neutrality by using pH test paper, and placing the microspheres in a refrigerator for hardening at 4 ℃ for 24-48 hours to obtain AgNPs@BSA/chitosan microspheres; the concentration of the preparation is 1 multiplied by 10 -4 ~1×10 -3 Adding the prepared AgNPs@BSA/chitosan microspheres into the methylene blue solution, stirring for 3-6 hours after ultrasonic oscillation for 3-6 hours, putting the pellets into a baking oven to be dried for 1-2 hours to obtain AgNPs@BSA@MB/chitosan microspheres, and storing in a refrigerator at 4 ℃ for later use;
the ultrapure water in the step S2 is the ultra-pure water of the chen type.
2. The method for preparing photosensitive silver nano chitosan microspheres according to claim 1, wherein the silver nano material and the chitosan solution in the step S4 are mixed and stirred for 2-4 hours.
3. The application of the AgNPs@BSA@MB/chitosan microsphere prepared by the preparation method of the photosensitive silver nano chitosan microsphere according to any one of claims 1-2 in bacteriostasis, which is characterized by being specifically applied to gram-negative bacteria represented by escherichia coli and gram-positive bacteria represented by staphylococcus aureus; the specific application is as follows: the AgNPs@BSA@MB/chitosan microspheres are placed into bacterial liquid, cultured in a constant temperature shaking box for 1h, and then placed under a xenon lamp 150w for irradiation for 20min.
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| CN107812957A (en) * | 2017-10-26 | 2018-03-20 | 太原工业学院 | A kind of method that fluorescence silver nano-grain is prepared based on silver mirror reaction |
| CN110432320A (en) * | 2018-05-04 | 2019-11-12 | 天津科技大学 | A kind of preparation method of nano silver chitosan complex microsphere |
| CN113855844A (en) * | 2021-09-18 | 2021-12-31 | 军事科学院军事医学研究院环境医学与作业医学研究所 | Antibacterial material and preparation method and application thereof |
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| CN107812957A (en) * | 2017-10-26 | 2018-03-20 | 太原工业学院 | A kind of method that fluorescence silver nano-grain is prepared based on silver mirror reaction |
| CN110432320A (en) * | 2018-05-04 | 2019-11-12 | 天津科技大学 | A kind of preparation method of nano silver chitosan complex microsphere |
| CN113855844A (en) * | 2021-09-18 | 2021-12-31 | 军事科学院军事医学研究院环境医学与作业医学研究所 | Antibacterial material and preparation method and application thereof |
Non-Patent Citations (2)
| Title |
|---|
| Studies on the antimicrobial properties of colloidal silver nanoparticles stabilized by bovine serum albumin;Thomas V. Mathew等;《Colloids and Surfaces B: Biointerfaces》;第101卷;第14-18页，第2.5节，第3.4节第2段 * |
| synthesis and anti-fungal effect of silver nanoparticles–chitosan composite particles;Lung-shuo Wang等;《International Journal of Nanomedicine》;第10卷;第2685-2696页，第2686页右栏第1段、第2692页结论 * |
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