CN108815519A - A kind of Preparation method and use of silver nanoparticle fiber light thermit powder - Google Patents
A kind of Preparation method and use of silver nanoparticle fiber light thermit powder Download PDFInfo
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Abstract
The present invention provides a kind of preparation methods of silver nanoparticle fiber light thermit powder, belong to technical field of material；Specific steps include：Polypeptide solution is prepared first；Then silver nano-grain light thermit powder is prepared；Photo-thermal finally, which is carried out, using silver nano-grain light thermit powder kills drug-fast bacteria experiment；Light thermit powder prepared by the present invention irradiates 20-360 min under the near infrared light that the wavelength that power is 5W is 808nm, kills drug-fast bacteria efficiency 90% or more；The present invention not only promotes the biocompatibility of material using biomaterial as template, preferably the treatment for diseases such as human tumors, and reduces the toxicity of nano silver itself, and discharging silver ion for sterilization by photo-thermal can be realized controllable sterilization.
Description
Technical field
The invention belongs to technical field of material, and in particular to a kind of based on Human islet amyloid polypeptide is template
The preparation method of silver nanoparticle fiber light thermit powder.
Background technique
Partial size is exactly accomplished nanoscale metallic silver simple substance by nano silver, and nano silver particle size is generally 25 nanometers, right
The tens of kinds of pathogenic microorganisms such as Escherichia coli, gonococcus, chlamydia trachomatis have strong inhibition and killing effect, and will not
Generate drug resistance.But there is also inevitable disadvantages simultaneously：Itself is very easy to oxidation, is not sufficiently stable, and toxicity is too strong,
It is incompatible with biology, it cannot distinguish between beneficial bacterium or harmful bacteria, killing effect all had to mushroom in human body, toxicity is too strong.And
Reducing silver concentration makes it not have silver will be made to lose sterilizing ability while toxicity to human body.
There are human bodies itself for Human islet's amyloid polypeptide (hIAPP), by islet β cell under normal physiological conditions.
Overall length includes the small peptide of 37 amino acid, sequence KCNTATCATQRLANFLVHSSNNFGAILSSTNVGSNTY.Wherein the 20th
To 29 amino acid sequences, i.e. SNNFGAILSS, it is considered to be the most sequence of fiber ability.Photo-thermal therapy, which refers to, to be had
It is logical to convert light energy into thermal energy under near infrared light (NIR) irradiation for easily penetrating tissue for the light thermit powder of high light thermal conversion efficiency
Cell heating ablation approach is crossed to realize a kind for the treatment of method of irreversible cytoclasis.In recent years, not using precious metal
Light thermit powder with form production is widely used in treating tumour and sterilization field.Current main light thermit powder has：Gold nanorods,
Platinum modifies nano dot, multiple twin nanogold and nano silica；But the method preparation process of traditional preparation light thermit powder
It is cumbersome, and there are extreme reaction conditions, such as：Mechanical lapping, high temperature and pressure and high-power ultrasonic.Using biocompatible material as
For the silver nano-grain of templated synthesis as light thermit powder, preparation condition is mild, highly-safe, and it is anti-to be applied to photo-thermal release silver ion
Drug-fast bacteria has a good application prospect.
Summary of the invention
In order to make up the shortcomings of the prior art, the present invention provides a kind of preparation method of silver nanoparticle fiber light thermit powder,
Specific technical solution is as follows：
A kind of preparation method of silver nanoparticle fiber light thermit powder, carries out according to the following steps：
Hexafluoroisopropanol solution is added in the test tube containing Human islet's amyloid polypeptide 20-29 powder step 1, sets
Isothermal reaction is carried out in shaking table, the polypeptide solution of hexafluoroisopropanol dispersion is obtained, is placed in refrigerator cold-storage；
Step 2, the polypeptide solution for taking the hexafluoroisopropanol in step 1 to disperse, are placed in centrifuge tube, are sealed with sealed membrane,
Several apertures are pricked on sealed membrane, and polypeptide films are obtained after being dried；
Deionized water is added in step 3, the centrifuge tube containing polypeptide films into step 2 after drying, ultrasound, oscillation, and
It is repeated 3 times, is placed in constant-temperature table incubation, obtains polypeptide solution after incubation；
Step 4, by silver nitrate (AgNO3) be dissolved in deionized water, obtain silver nitrate solution；Then, by silver nitrate solution
The polypeptide solution being added in step 3 obtains mixed solution A after stirring；
Step 5, by sodium borohydride (NaBH4) be dissolved in the deionized water of ice, sodium borohydride solution is obtained, by sodium borohydride
Solution is added in step 4 in obtained mixed liquor A, then, deionized water is added to dilute to obtain solution B to get silver nanoparticle fiber
Photo-thermal agent solution；；
Further, in step 1, the hexafluoroisopropanol and polypeptide powder amount ratio are 1mL：2mg；The isothermal reaction
Temperature be 25 DEG C, time 12h.
Further, in step 2, the volumetric usage of the polypeptide solution is 10~40 μ L.
Further, in step 3, the deionized water dosage is 200 μ L；The temperature of incubation is 37 DEG C, time 5-
12h。
Further, in step 4, the concentration of the silver nitrate solution is 10mM, in the mixed liquor A silver nitrate solution with
The molar ratio of polypeptide solution is 100：1.
Further, in step 5, the concentration of the sodium borohydride solution is 10mM, and sodium borohydride is molten in the mixed liquid B
The molar ratio of liquid and silver nitrate solution is 2：1.
Further, the concentration of silver nanoparticle fiber is 1 μ g/mL in mixed liquid B described in step 5.
The present invention utilizes simple, mild synthesis condition, system using Human islet's amyloid polypeptide 20-29 fiber as template
Standby pattern uniform, controllable out has preferable photo-thermal effect silver nanoparticle fiber light thermit powder, under near infrared light effect, effectively kills
Dead drug-fast bacteria.
It tests and tests for drug-fast bacteria bactericidal effect：Solution prepared by 180 μ L is taken to mix with 20 μ L bacterial suspensions, it is quiet
Placement location 10min, near infrared light, wavelength 808nm, power 5W, irradiation time is 20~360min；Then, 2 are taken
The mixed solution of μ L light irradiation is diluted in the phosphate buffer solution of 2ml, and the solution after 100 μ L dilution is taken to be coated in solid fine jade
On rouge culture medium, 37 DEG C of incubator cultures for 24 hours, calculate final Survival probability of bacteria.
Bacterium used is methicillin-resistant staphylococcus aureus (MRSA), vancomycin resistance staphylococcus aureus
(VRSA) and one of vancomycin-resistant enterococcus (VRE).
The hexafluoroisopropanol acts on dispersing agent.
The Human islet amyloid polypeptide 20-29, functions as template.
The silver nitrate solution, Ag+ ion needed for effect is to provide preparation light thermit powder silver nanoparticle fiber sterilization.
The sodium borohydride solution, effect is reducing agent.
Beneficial effect：
(1) present invention synthesize using Human islet's amyloid polypeptide as the silver nanoparticle fiber light thermit powder of template, pattern can benefit
Realize that uniform, controllable realizes better photothermal conversion effect to regulate and control light thermal property with polypeptide fibers length.
(2) reaction condition of the invention is room temperature, normal atmosphere pressure, compared to the photo-thermal prepared under conventional high-temperature high pressure
Agent, preparation condition milder of the invention.
(3) present invention not only promotes the biocompatibility of material using biomaterial as template, preferably swollen for human body
The treatment of the diseases such as tumor, and the toxicity of nano silver itself is reduced, it can be realized by photo-thermal release silver ion for sterilization and controllably kill
Bacterium.
(4) raw material used in the present invention is simple and easy to get, and production method is easy.
(5) present invention is for the first time that templated synthesis silver nanoparticle is fine using Human islet's amyloid polypeptide 20-29 segment aggregation
Light thermit powder is tieed up, realizes higher photo-thermal effect.
(6) present invention is silver nanoparticle fiber photo-thermal of the template preparation with photo-thermal effect using Human islet's amyloid polypeptide
Agent, when high concentration, have a superior photo-thermal effect, and when low concentration does not have bio-toxicity, can be used for discharging silver ion under illumination condition
It sterilizes also, bactericidal property is much better than simple substance silver powder.
Specific embodiment
Silver nanoparticle fiber light thermit powder prepared by above-mentioned technical proposal is applied to photo-thermal and kills drug-fast bacteria, below with reference to specific reality
Applying example, the present invention will be further described.
Embodiment 1：
(1) preparation of polypeptide solution：
The hexafluoroisopropanol solution of 1mL is added in the test tube containing 2mg Human islet's amyloid polypeptide powder, is placed
Reaction 12h is rocked on 25 DEG C of constant-temperature table, obtains being scattered in the polypeptide solution in hexafluoroisopropanol, 10 μ L is taken to be scattered in six
Polypeptide solution in fluorine isopropanol is in the centrifuge tube of 1.5mL, with the mouth of ParafilmTM centrifuge tube, and with syringe needle in sealed membrane
It is upper to prick several apertures, drying at room temperature 2h in a vacuum drying oven is placed, 200 μ L of deionized water is then added into centrifuge tube, makes
Peptide concentration is 100 μM, then ultrasound, oscillation, it in triplicate, is placed in 37 DEG C of constant-temperature tables and is incubated for, control incubation time is
5h finally obtains polypeptide solution.
(2) preparation of silver nanoparticle fiber light thermit powder：
Taking 200 μ L concentration is the silver nitrate solution of 10mM, is added in step (1) obtained polypeptide solution, stirs 5 points
Clock obtains the molar ratio of silver nitrate and polypeptide solution as 100 mixed solution, and the ice water that 400 μ L concentration are 10mM is taken to prepare
Sodium borohydride solution, be added drop-wise in the mixed solution of silver nitrate and polypeptide, then be added deionized water be diluted to silver nanoparticle fibre
The concentration of dimension is 1 μ g/mL, is vibrated 5 seconds, stands 1h, finally obtains silver nanoparticle fiber light thermit powder solution.
(3) silver nanoparticle fiber light thermit powder photo-thermal kills drug-fast bacteria experiment
It takes light thermit powder prepared by 180 μ L and the MRSA bacterial suspension of 20 μ L to mix, stands and place 1h, be with power
5W, after wavelength is 808nm near infrared light 360min, the mixed solution for taking 2 μ L light to irradiate is diluted in the phosphate-buffered of 2ml
Solution in solution, and after taking 100 μ L to dilute is coated on solid agar medium, 37 DEG C of incubator cultures for 24 hours, calculate final
Survival probability of bacteria.Gained Survival probability of bacteria is shown in Table 1.
Embodiment 2：
The preparation method is the same as that of Example 1, and only changing bacterium used in 1 step of embodiment (3) is VRSA and VRE, gained bacterium
Survival rate is shown in Table 1.The result shows that under the same conditions, silver nanoparticle fiber light thermit powder prepared by the present invention is acted in near-infrared
Under can kill MRSA, VRSA and VRE drug-fast bacteria completely.
Germicidal efficiency of the 1 silver nanoparticle fiber of table to different bacterium type
| Bacteria types | Survival probability of bacteria (%) |
| MRSA | 0 |
| VRSA | 0 |
| VRE | 0 |
Embodiment 3：
The preparation method is the same as that of Example 1, and only changing light application time in 1 step of embodiment (3) is 0min, 20min, 240min.
Gained Survival probability of bacteria is shown in Table 2.MRSA is killed the result shows that reducing light application time and can reduce silver nanoparticle fiber light thermit powder simultaneously
Bacterium efficiency；When light application time is 360min, bactericidal effect 100%.
The different light application times of table 2 influence the sterilization of prepared light thermit powder
| Light application time (min) | Survival probability of bacteria (%) |
| 0 | 100 |
| 20 | 95 |
| 240 | 37 |
| 360 | 0 |
Embodiment 4：
The preparation method is the same as that of Example 1, and only changing the polypeptide solution taken in 1 step of embodiment (1) is 20 μ L, 40 μ L.Institute
It obtains Survival probability of bacteria and is shown in Table 3.The result shows that changing sterilization of the amount of polypeptide solution volume to silver nanoparticle fiber light thermit powder to MRSA
Rate does not influence, and bactericidal effect is 100%.
The different silver nanoparticle fiber germicidal efficiency of peptide concentration used in table 3
| Peptide concentration (μM) | Survival probability of bacteria (%) |
| 25 | 0 |
| 50 | 0 |
| 100 | 0 |
Embodiment 5：
The preparation method is the same as that of Example 1, only changes polypeptide incubation time 7h, 12h, gained bacterium in 1 step of embodiment (2) and deposits
Motility rate is shown in Table 4.The result shows that polypeptide incubation time does not influence the efficiency that silver nanoparticle fiber light thermit powder kills MRSA, change polypeptide
Incubation time, bactericidal effect are 100%.
The not homopolypeptide incubation time germicidal efficiency of table 4
| Polypeptide incubation time (h) | Survival probability of bacteria (%) |
| 5 | 0 |
| 7 | 0 |
| 12 | 0 |
| 360 | 0 |
Embodiment 6：
(1) preparation of elemental silver solution
Appropriate amount of deionized water is taken to dissolve simple substance silver powder, until concentration is 1 μ g/mL
(2) sterilization test
Elemental silver solution prepared by 180 μ L is taken to mix respectively with MRSA, VRSA and the VRE bacterial suspension of 20 μ L, it is quiet
It takes 2 μ L silver and bacterium mixed solution to be diluted in the phosphate buffer solution of 2ml after setting 1h, and takes the solution after 100 μ L dilution
Be coated on solid agar medium, 37 DEG C of incubator cultures for 24 hours, calculate final Survival probability of bacteria.Gained Survival probability of bacteria is shown in
Table 5.Description of test concentration is that the silver-colored solution of 1 μ g/mL does not have sterilizing ability.
Germicidal efficiency of the 5 silver nanoparticle fiber of table to different bacterium type
| Bacteria types | Survival probability of bacteria (%) |
| MRSA | 99 |
| VRSA | 100 |
| VRE | 98 |
Claims (10)
1. a kind of preparation method of silver nanoparticle fiber light thermit powder, which is characterized in that carry out according to the following steps：
Hexafluoroisopropanol solution is added in the test tube containing Human islet's amyloid polypeptide 20-29 powder step 1, is placed in perseverance
Warm shaking table is reacted, and is obtained the polypeptide solution of hexafluoroisopropanol dispersion, is placed in refrigerator cold-storage；
Step 2, the polypeptide solution for taking the hexafluoroisopropanol in step 1 to disperse, are placed in centrifuge tube, are sealed with sealed membrane, sealed
Several apertures are pricked on membrana oralis, and polypeptide films are obtained after being dried；
Deionized water is added in step 3, the centrifuge tube containing polypeptide films into step 2 after drying, ultrasound, oscillation, and repeat 3
It is secondary, it is placed in constant-temperature table incubation, obtains polypeptide solution after incubation；
Silver nitrate is dissolved in deionized water by step 4, obtains silver nitrate solution；Then, silver nitrate solution is added to step 3
In polypeptide solution, obtain mixed solution A after stirring；
Sodium borohydride is dissolved in the deionized water of ice by step 5, obtains sodium borohydride solution, and sodium borohydride solution is added and is walked
In rapid 4 in obtained mixed liquor A, then, deionized water is added to dilute to obtain solution B, as silver nanoparticle fiber light thermit powder solution.
2. a kind of preparation method of silver nanoparticle fiber light thermit powder according to claim 1, which is characterized in that institute in step 1
It states hexafluoroisopropanol and polypeptide powder amount ratio is 1mL：2mg.
3. a kind of preparation method of silver nanoparticle fiber light thermit powder according to claim 1, which is characterized in that institute in step 1
The temperature for the isothermal reaction stated is 25 DEG C, time 12h.
4. a kind of preparation method of silver nanoparticle fiber light thermit powder according to claim 1, which is characterized in that institute in step 2
The polypeptide solution for stating hexafluoroisopropanol dispersion is 10~40 μ L.
5. a kind of preparation method of silver nanoparticle fiber light thermit powder according to claim 1, which is characterized in that described in step 3
The dosage of middle deionized water is 200 μ L；The temperature of the incubation is 37 DEG C, time 5-12h.
6. a kind of preparation method of silver nanoparticle fiber light thermit powder according to claim 1, which is characterized in that institute in step 4
The concentration for stating silver nitrate solution is 10mM；The molar ratio of silver nitrate solution and polypeptide solution is 100 in the mixed liquor A：1.
7. a kind of preparation method of silver nanoparticle fiber light thermit powder according to claim 1, which is characterized in that institute in step 5
The concentration for stating sodium borohydride solution is 10mM；The molar ratio of sodium borohydride solution and silver nitrate solution is 2 in the mixed liquid B：
1。
8. a kind of preparation method of silver nanoparticle fiber light thermit powder according to claim 1, which is characterized in that described in step 5
The concentration of silver nanoparticle fiber is 1 μ g/mL in mixed liquid B.
9. a kind of preparation method system of silver nanoparticle fiber light thermit powder described in any one claim according to claim 1~8
Standby silver nanoparticle fiber light thermit powder is applied to photo-thermal and kills drug-fast bacteria.
10. silver nanoparticle fiber photo-thermal prepared by a kind of preparation method of silver nanoparticle fiber light thermit powder according to claim 9
Agent be applied particularly to photo-thermal kill methicillin-resistant staphylococcus aureus, vancomycin resistance staphylococcus aureus or it is resistance to through the ages
Mycin enterococcus.
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