CN108210990A - It is a kind of using graphene oxide as the new liquid adhesive bandage of antibacterial components - Google Patents
It is a kind of using graphene oxide as the new liquid adhesive bandage of antibacterial components Download PDFInfo
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- CN108210990A CN108210990A CN201611194777.1A CN201611194777A CN108210990A CN 108210990 A CN108210990 A CN 108210990A CN 201611194777 A CN201611194777 A CN 201611194777A CN 108210990 A CN108210990 A CN 108210990A
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L26/00—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form
- A61L26/0004—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form containing inorganic materials
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A61L26/00—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form
- A61L26/0009—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form containing macromolecular materials
- A61L26/0014—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form containing macromolecular materials obtained by reactions only involving carbon-to-carbon unsaturated bonds
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A61L26/00—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form
- A61L26/0061—Use of materials characterised by their function or physical properties
- A61L26/0066—Medicaments; Biocides
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- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/10—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices containing or releasing inorganic materials
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/40—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
- A61L2300/404—Biocides, antimicrobial agents, antiseptic agents
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Abstract
The present invention provides a kind of using graphene oxide as the new liquid adhesive bandage of antibacterial components.The present invention utilizes graphene oxide wettability and surface-active, give full play to the physical antibacterial characteristic of graphene oxide, using graphene oxide as the antipathogenic composition of liquid adhesive bandage, while the series of advantages for keeping liquid adhesive bandage, its long-term efficient antibacterial action is given full play to, enhances the fungistatic effect of liquid adhesive bandage.New liquid adhesive bandage disclosed by the invention preferably using graphene oxide as antibacterial components contains graphene 2mg/ml, ethyl alcohol, 5% polyvinyl butyral, 10% ethyl acetate, 5% glycerine also comprising weight percent 75%.
Description
Technical field
The present invention relates to medical instruments field, more particularly it relates to which one kind is using graphene oxide as antibacterial components
New liquid adhesive bandage.
Background technology
Graphene (Graphene) is the only one layer of atomic thickness for being stripped out from graphite material, being made of carbon atom
Two dimensional crystal.2004, Univ Manchester UK physicist An Deliegaimu and Constantine's Nuo Woxiao loves,
Graphene is isolated in success from graphite, it was demonstrated that it can with individualism (Geim A K, Nature Materials, 2007
(6)：183-191), therefore two people also obtain Nobel Prize in physics in 2010 jointly.
Graphene oxide (graphene oxide, GO) is one of representative derivative that graphene is modified, graphene oxide
It is essentially identical with the structure of graphene, a large amount of oxygen-containing group is only connected on the basal plane of single layer of carbon atom structure.Such as Fig. 1
Shown in the structure diagram of graphene oxide, containing-OH and C-O-C in plane, and contain C=O and COOH at its lamella edge.
Compared with graphene, graphene oxide has good wettability and surface-active.
The antibacterial action research of graphene oxide starts from 2010, the yellow celebrating subject of Shanghai Applied Physics institute of the Chinese Academy of Sciences
Group first reported research (Hu W, et al.Graphene-based of the graphene in terms of anti-biotic material
Antibacterial paper.ACS Nano, 2010,4 (7)：4317-4323).They find, graphene oxide suspension with
Inhibiting rate reaches more than 90% after Escherichia coli are incubated 2h, and antibiotic property is from graphene oxide to Bacillus coli cells film
Machine cuts are destroyed.(Zhao J M, the et al.Graphene oxidebased antibacterial cotton such as Zhao
Fabrics.Advanced Health care Materials, 2013 (2)：1259-1266.) use radiation grafted method
Graphene oxide is connected on cotton and is prepared for graphite oxide alkenyl antibacterial cotton fabric, and measures its antibiotic property.Result of study
Show that graphite oxide alkenyl antibacterial cotton fabric shows very strong antibiotic property, bacteriostasis rate is more than 98%, and this cotton fabric is in quilt
Bacteriostasis rate is still maintained at more than 90% after washing 100 times.Qu little Gang (Nurunnabi M1, In vivo
biodistribution and toxicology of carboxylated graphene quantum dots.ACS
Nano.2013Aug 27；7(8)：Hydrogen peroxide 6858-67.) et al. is catalytically decomposed using graphene quantum dot, make its generate it is strong and
Strong sterilization hydroxyl radical free radical.On the bandage similar to adhesive bandage, infiltration contains graphene and hydrogen peroxide, you can significantly
Reduce the quantity of the bacterium of mouse site of injury.(Akhavan O, the Toxicity of graphene and such as Akhavan
Grapheme oxide nanowalls against bacteria.ACS Nano, 2010,4 (10)：5731-5736) respectively
Using gram-positive bacteria (staphylococcus aureus) and Gram-negative bacteria (Escherichia coli) to being deposited on stainless steel matrix
Stannic oxide/graphene nano wall and the antibiotic property of graphene nano wall studied, find graphene oxide and graphene to leather
Lan Shi negative bacteriums and gram-positive bacteria all have good anti-microbial property.Liu et al. (Liu S, et al.Antibacterial
Activity of graphite, graphite oxide, graphene oxide and reduced graphene
oxide：Membrane and oxidative stress.ACS Nano, 2011,5 (9)：6971-6980) have studied oxidation stone
The influence of black alkene and redox graphene (rGO) to Escherichia coli vigor finds that the antibacterial ability of graphene oxide GO is higher than
Redox graphene rGO, meanwhile, the study found that graphene oxide and redox graphene can be in oxidizing bacteria bodies
Reduced glutathione.Therefore, it is considered that the antibiotic property of graphene oxide GO is other than from the destruction to cell membrane, it is also possible to
From the GO oxygen radicals caused and then the oxidative damage induced.Its antibiotic property may be：Bacterial adhesion first is on material, so
Graphene stress eventually lead to bacterial death by sharp corner incising cell film, then superoxide ion mediated oxidative afterwards.
(Tu Y, the et al.Destructive extraction of phospholipids from such as Tu in 2013
Escherichia coli membranes by graphene nanosheets.Nat Nano, 2013,8 (8)：594-601)
A kind of new Graphene antibiosis mechanism is proposed, as shown in Fig. 2, they are observed directly by transmission electron microscope experiment
The cavity structure generated after bacterial cell membrane and graphene oxide effect.Thus propose that graphene can not only be cut by contacting
Effect destroys bacterial cell membrane, can also be by the phospholipid molecule on large-scale direct extraction cell membrane, to destroy
Cell membrane simultaneously kills bacterium.
With the research that deepens continuously to Graphene antibiosis mechanism, the application about Graphene antibiosis effect has also been obtained into one
The development of step.(Zhu Z J, the et al.Preparation of grapheme oxide-silver such as Zhu
Nanoparticle nanohybrids with highly an tibacterial capability.Talanta, 2013
(117)：449-455.) nano-Ag particles are loaded to using electrostatic self-assembled technology, GO-Ag has been prepared on graphene oxide
Composite material, antibacterial experiment is the result shows that nanometer GO-Ag composite materials have good antibacterial property.Lim etc. (Lim H N,
Facile preparation of grapheme-based chitosan films：Enhanced thermal,
Mechanical and antibacterial properties.Joural of Non-Crystalline Solids, 2012
(358)：525-530) GO, rGO are carried out respectively with chitosan it is compound after measure the antibiotic property of its compound, the results showed that GO or
There is synergetic antibacterial effect after rGO and chitosan are compound.This illustrates that grapheme material still maintains its antibacterial in recombination process
Characteristic has broad application prospects in coating, packaging, antibiotic fabric and medical material.
Adhesive bandage also known as " hemostasis plaster ", are most common a kind of surgical medicines in people's life.General adhesive bandage is
By an elongated adhesive plaster, the sliver that centre is accompanied by one piece of dipped drug is formed.In currently available technology, patent
CN201510783493.5 has been disclosed for a kind of " adhesive bandage of the ingredient containing graphene oxide and preparation method thereof ", which is
The active component of the compositions such as graphene oxide and quaternary ammonium salt, sodium carboxymethylcellulose and polyacrylamide is coated on normal
The stromal surface of adhesive bandage is advised, achievees the purpose that anti-inflammation hemostasia analgesic, convergence myogenic.But even if there is graphene as advanced
Antimicrobial component, but the invention can not still avoid the shared shortcoming of conventional solid-state adhesive bandage：(1) since its structure limits, wound
The wound emergence therapeutic for being only used for fritter can be pasted, the protection and healing of the large area surface of a wound can not be met；(2) sliver or matrix
Water absorbing capacity is stronger, easy adhesion wound, and when replacement causes mechanicalness to be damaged again, dressing bleeding pain；(3) waterproof effect is very
Difference, bacterium easily penetrate and cause secondary infection, make wound deterioration；(4) incrustation is easily generated, hinders contraction of wound.
For conventional solid-state adhesive bandage, there are drawbacks, now have been developed for liquid adhesive bandage.Liquid adhesive bandage not only has
There are hemostasis, the protection wound face that conventional adhesive bandage has, also with good biocompatibility, do not limited by wound face size
System, using liquid adhesive bandage, wound face is afraid of water, can bathing, joint part uses, and activity is freely unrestricted, does not have to skin
There is stimulation, it is safe to use without sensitization.Liquid adhesive bandage is a kind of biological film forming disinfectant, is smeared (or spraying) and existed
During wound surface, it can volatilize rapidly, form one layer very thin of soft and flexible glued membrane, and playing prevents wound from polluting and protecting
The effect of shield.Importantly, this glued membrane has waterproof action.On the one hand external water pollution wound is prevented, on the other hand
The moistening of wound is kept, incrustation is not generated and promotes wound healing.
The antipathogenic composition of presently commercially available liquid adhesive bandage mainly divides 2 classes：One kind be chemical bacteriostatic agent (such as bromogeramine,
Povidone iodine, parachlorometaxylenol, benzoic acid, glyceryl monolaurate etc.) it is another kind of be the traditional Chinese medicine extraction for having bacteriostasis
Object (such as callicarpa nudiflora).The antimicrobial mechanism of Chinese medical extract is unclear, and it is slow to work；The mechanism of chemical bacteriostatic agent mainly has following
Several aspects：1st, film mechanism：By inhibiting the mobility of film, change membrane passage；2nd, mechanism of acidification：It is thin by entering
Cause intracellular acidification inside bacterium, influence ATP metabolism；3rd, ion transport system is influenced：The function of bacterium inner ion pump is influenced,
Influence ionic equilibrium；4th, it influences signal to transmit, interferes the expression of gene.These effects can ensure chemical bacteriostatic agent than relatively low
The growth that can effectively inhibit bacteria of concentration, still, however it remains the defects of term of validity is shorter, bacterial resistance.
What is utilized in this part patent is graphene oxide wettability and surface-active, plays its physical antibacterial characteristic, will
Antipathogenic composition of the graphene oxide as liquid adhesive bandage while the series of advantages for keeping liquid adhesive bandage, is fully sent out
Its long-term efficient antibacterial action is waved, enhances the fungistatic effect of liquid adhesive bandage.
Invention content
The present invention provides a kind of liquid adhesive bandages containing graphene oxide, are resisted using the stronger physics of graphene oxide
Bacterium advantage enhances the fungistatic effect of liquid adhesive bandage.
The invention discloses：
It is a kind of using graphene oxide as the new liquid adhesive bandage of antibacterial components, comprising film forming component, solvent, feature exists
In the antimicrobial component in the liquid adhesive bandage is graphene oxide.
It is as described above a kind of using graphene oxide as the new liquid adhesive bandage of antibacterial components, which is characterized in that described
Antimicrobial component graphene oxide content is 0.006mg/ml-4mg/ml.
A kind of new liquid adhesive bandage using graphene oxide as antibacterial components as described above pastes, which is characterized in that institute
Graphene oxide content is stated as 0.5mg/ml or 1mg/ml or 2mg/ml.
It is as described above a kind of using graphene oxide as the new liquid adhesive bandage of antibacterial components, which is characterized in that described
Graphene oxide content is 2mg/ml.
It is as described above a kind of using graphene oxide as the new liquid adhesive bandage of antibacterial components, which is characterized in that described
Film forming component is polyvinyl butyral.
It is as described above a kind of using graphene oxide as the new liquid adhesive bandage of antibacterial components, which is characterized in that described
Solvent is ethyl alcohol.
It is as described above a kind of using graphene oxide as the new liquid adhesive bandage of antibacterial components, which is characterized in that described
Liquid adhesive bandage also includes plasticizer.
It is as described above a kind of using graphene oxide as the new liquid adhesive bandage of antibacterial components, which is characterized in that described
Plasticizer is ethyl acetate.
It is as described above a kind of using graphene oxide as the new liquid adhesive bandage of antibacterial components, which is characterized in that described
Liquid adhesive bandage also includes glycerine.
It is as described above a kind of using graphene oxide as the new liquid adhesive bandage of antibacterial components, which is characterized in that include
The ethyl alcohol of weight percent 75%, 5% polyvinyl butyral, 10% ethyl acetate, 5% glycerine.
The present invention provides a kind of using graphene oxide as the new liquid adhesive bandage of antibacterial components, contain graphene
0.06mg/ml, 0.125mg/ml, 0.25mg/ml, 0.5mg/ml, lmg/ml, 2mg/ml, 4mg/ml also include weight percent
75% ethyl alcohol, 5% polyvinyl butyral, 10% ethyl acetate, 5% glycerine, wherein preferred embodiment be containing
There is the liquid adhesive bandage of graphene oxide 2mg/ml.The property of its physical sterilization using graphene oxide, overcomes existing liquid
In body adhesive bandage the drawbacks of antimicrobial component, using graphene oxide as the antimicrobial component of liquid adhesive bandage, more traditional Chinese medicine
Antimicrobial component is rapid-action, the phenomenon that also avoiding the term of validity of chemical bacteriostatic agent in the prior art short, is more not in that bacterium is resistance to
The drawbacks of medicine.
The present invention first confirms that free graphene oxide can effectively inhibit Gram-negative bacteria, gram-positive bacteria and true
Proliferation of the bacterium in rich medium, and Gram-negative bacteria, gram-positive bacteria can be effectively killed in physiological saline, but
It is limited to the killing ability of fungi；It is green with tetracyclin resistance and ammonia benzyl to be further discovered that free graphene oxide can be killed effectively
Mycin and the Double Escherichia coli of kanamycins illustrate that graphene oxide has the bacteriostasis of wide spectrum.
Secondly, it has been found that the liquid adhesive bandage containing graphene oxide is after film forming, to Gram-negative bacteria, gram
Positive bacteria also has significant lethal effect, but to the lethal effect unobvious of fungi.Its fungistatic effect is better than certain of commercialization
Inlet liquid adhesive bandage, but it is inferior to certain domestic liquid adhesive bandage.
Third, we establish epidermis holostrome wound and infection model in mouse body surface, on the mold we have found that containing
The liquid adhesive bandage of graphene oxide can not only effectively inhibit surface of a wound bacterial multiplication, and can wound healing well.
Its fungistatic effect is not only far superior to the film forming matrix of not graphene-containing, but also can better than the commercial liquid wound of current commercialization
Patch.
The invention demonstrates that graphene oxide has the potential as the antibacterial component of liquid adhesive bandage, it is hopeful to develop into one
The novel liquid adhesive bandage of kind.
Description of the drawings
The structure diagram of Fig. 1, graphene oxide；
Fig. 2, graphene oxide act on the electron micrograph after bacterium；
Fig. 3, graphene oxide inhibit bacteria the effect of growth；
Fig. 4, graphene oxide are to the lethal effects of all kinds of bacteriums；
Fig. 5, the lethal effect of graphene oxide antagonism Escherichia coli；
Fig. 6, using graphene oxide it is the liquid adhesive bandage of antibacterial components to the lethal effects of Escherichia coli；
Fig. 7, using graphene oxide it is the liquid adhesive bandage of antibacterial components to the lethal effect of staphylococcus aureus；
Fig. 8, using graphene oxide it is the liquid adhesive bandage of antibacterial components to the lethal effect of Candida albicans；
Fig. 9, NIH3T3 cell and liquid adhesive bandage are incubated result figure jointly；
Figure 10, postoperative 4th day wounds in mice bacterial population summarize data；
Figure 11, postoperative 10th day wounds in mice healing state comparison (first group)；
Figure 12, postoperative 10th day wounds in mice healing state comparison (second group)；
Figure 13, postoperative 10th day wounds in mice healing state comparison (third group)；
Figure 14, postoperative 10th day wounds in mice healing state comparison (the 4th group)；
Figure 15, postoperative 10th day wounds in mice healing state comparison (the 5th group).
Specific embodiment
The present invention is further detailed in following embodiment, experimental example, should not be construed as limiting the invention.It is real
It applies example and does not include detailed descriptions of conventional methods.
The effectively antibacterial dosage of experimental example 1, graphene oxide determines
(1) preparation of graphene oxide suspension：
5g graphene oxides are weighed, add in 20ml absolute ethyl alcohols, are ultrasonically treated 30min, 85 DEG C of drying add in 100ml
DdH2O is ultrasonically treated 60min, graphene oxide suspension (50mg/ml) is made.
(2) culture medium is prepared：
LB fluid nutrient mediums (Escherichia coli, staphylococcus aureus)：Peptone 10g, dusty yeast 5g, sodium chloride (NaCl)
10g is settled in 1000ml sterile waters, autoclave sterilization processing.LB agar mediums：Peptone 10g, dusty yeast 5g, chlorine
Change sodium (NaCl) 10g, agar powder 15g is settled in 1000ml sterile waters, autoclave sterilization processing, paved tablet.Pancreas junket is big
Beans peptone fluid nutrient medium (staphylococcus aureus)：Trypticase 15.0g, soybean papain hydrolysis object 5.0g, sodium chloride
5.0g is settled to 1000ml, mixing, and it is 7.3 ± 0.2 to adjust pH, packing, autoclave sterilization.Pancreas junket soya peptone agar culture
Base (staphylococcus aureus)：Trypticase 15.0g, soybean papain hydrolysis object 5.0g, sodium chloride 5.0g, agar 15.0g,
It is settled to 1000ml, autoclave sterilization, paved tablet.Sabouraud dextrose fluid nutrient medium (Candida albicans)：Animal tissue
Pepsin hydrolysate and trypticase equal amount of mixture 10.0g, glucose 20.0g water 1000ml, in addition to glucose, take it is above-mentioned into
Point, it mixes, dissolving, it is 5.6 ± 0.2 to adjust pH, adds human glocose, shakes up, and is dispensed, sterilizing.Sabouraud glucose agar
(Candida albicans)：Animal tissue's pepsin hydrolysate and trypticase equal amount of mixture 10.0g, glucose 40.0g agar
15.0g, water 1000ml in addition to glucose, take mentioned component, mix, and dissolving, it is 5.6 ± 0.2 to adjust pH, adds human glocose, shakes
It is even, it dispenses, sterilizing.Paved tablet.
(3) Bacteria Culture：
Escherichia coli：
Monoclonal escherichia coli DH5a is inoculated with to 3ml LB culture mediums, 37 DEG C of shake cultures are stayed overnight, and next day connects by 1: 100
It plants into 50ml LB culture mediums, 37 DEG C of shake culture 2h, obtains the bacterium solution of exponential phase.
Staphylococcus aureus：
Monoclonal staphylococcus aureus is inoculated with to 3mlLB fluid nutrient mediums, 30-35 DEG C of shake culture is stayed overnight, and next day presses
Be seeded in 50ml pancreas junket soya peptone fluid nutrient medium at 1: 100,30-35 DEG C of shake culture 2h, obtains the bacterium solution of exponential phase.
Candida albicans：
Monoclonal Candida albicans is inoculated with to 3ml Sabouraud dextrose fluid nutrient mediums, 20-25 DEG C of shake culture 24-48h,
It is seeded in 50ml Sabouraud dextrose fluid nutrient mediums by 1: 100,20-25 DEG C of shake culture 4-5h obtains exponential phase
Bacterium solution.
(4) bacteriostasis of graphene oxide
CCK8 methods detect：The bacterium solution of exponential phase is diluted to OD600=0.2 with respective liquid culture medium；By graphite
Alkene suspension is diluted to 40 with ddH2O, 20,10,5,2.5,12.5mg/ml, following solution (μ l) is added in corresponding each group hole：Such as
Liquid feeding (every group of 4 multiple holes) in each group hole shown in table 1
Table 1, each group hole liquid feeding table of comparisons
Escherichia coli and staphylococcus aureus, in 37 DEG C of shake culture 1h, Candida albicans is in 25 DEG C of shake culture 3h.
Microplate reader detects 450nm light absorptions, and 650nm is reference wavelength, detects graphene fungistatic effect.The results are shown in Figure 3, with training
The raising of graphene concentration in base is supported, the activity of bacterium is continuously decreasing, having significant dose-effect relationship, illustrate free graphite alkene pair
The growth of bacterium has inhibiting effect.Fungistatic effect of the graphene concentration more than 2mg/ml is close to platform.Graphene is to Escherichia coli
It is preferable with the growth inhibitory effect of staphylococcus aureus, can be more than 60%；It is general to the inhibition of Candida albicans, only
Close to 35%.The inhibition that graphene is better than to fungi the inhibition of Gram-negative bacteria and gram-positive bacteria is prompted to imitate
Fruit.
The bactericidal effect of experimental example 2, graphene oxide to different type bacterium
Escherichia coli, staphylococcus aureus and the Candida albicans being incubated overnight meet bacterium 3ml, and 37 DEG C of shaken cultivations 2 are small
When；Bacterium solution is collected by centrifugation, is washed 3 times with physiology salt；It is resuspended with physiological saline, adjusts bacterial concentration to 106/ml (according to pre- before
The concentration dilution of experiment is to suitable bacterial concentration), it is dispensed into 48 orifice plates：0.3ml/ holes * 3；It is separately added into various concentration graphite
Alkene 4,2,1mg/ml；20ul coated plates (12 hole LB tablets) were taken at 3,6 hours respectively, are inverted, 37 DEG C of overnight incubations；Secondary number of days gram
It is grand.
2 free graphite alkene of table is to the killing effect of different type bacterium
Graphene oxide to the lethal effect result of all kinds of bacteriums as shown in table 2 and Fig. 4, as a result data from table 2 and Fig. 4
It can be seen that free graphite alkene has apparent lethal effect, and fragmentation effect to Escherichia coli and staphylococcus aureus
The year raising of graphene concentration significantly increases, and the graphene of 2-4mg/ml can kill both bacteriums completely substantially.With altogether
With the time lengthening being incubated, fragmentation effect further enhances, and is incubated 6 hours common, the graphene of 1mg/ml can also reach non-
Often good fragmentation effect.However it is very limited to the fragmentation effect of Candida albicans, only as a concentration of 4mg/ml of graphene,
The killing of 10-20% just can be observed, remaining 2 concentration is without fragmentation effect.
The result illustrates graphene oxide to Gram-negative bacteria (Escherichia coli) and gram-positive bacteria (golden yellow grape
Coccus) there is significant killing ability, and it is very limited to the fragmentation effect of fungi (Candida albicans).
Experimental example 3, graphene oxide are to the bactericidal effects of the Escherichia coli of different resistances
The tetracyclin resistance bacterium that is incubated overnight, ammonia benzyl the dual anti-bacterium of kanamycins meet bacterium 3ml, 37 DEG C of shaken cultivations 2 hours；
Bacterium solution is collected by centrifugation, is washed 3 times with physiology salt；It is resuspended with physiological saline, adjusts bacterial concentration (can be according to pre- reality before to 107/ml
The concentration dilution tested is to suitable bacterial concentration), it is dispensed into 48 orifice plates：0.3ml/ holes * 3；It is separately added into various concentration graphene
0.5、0.25、0.125mg/ml；20ul coated plates (12 hole LB tablets) were taken at 4,24 hours respectively, are inverted, 37 DEG C of overnight incubations；It is secondary
Number of days is cloned.
Clone's number (4 hours) of the different resistant E. colis of 3 graphene of table processing
Clone's number (24 hours) of the different resistant E. colis of 4 graphene of table processing
The lethal effect result of graphene oxide antagonism Escherichia coli as shown in Fig. 5 and table 4, the results show that
The graphene of 0.125mg/ml can kill only 50% tetracyclin resistance Escherichia coli and more than 90% when handling 4 hours
The dual anti-Escherichia coli of ampicillin/kanamycins, when treated between up to 24 hours when, all differences can be killed substantially
The Escherichia coli of resistance.The graphene of 0.25mg/ml substantially completely killed the effect of resistant E. coli at 4 hours with regard to that can reach
Fruit.
The In Vitro Bacteriostasis effect assessment of experimental example 4, liquid adhesive bandage containing graphene oxide
1), the preparation of the liquid adhesive bandage containing graphene oxide：
The preparation of liquid adhesive bandage membrane matrix containing graphene oxide：
Formula is as follows：75% ethyl alcohol, 5% polyvinyl butyral, 10% ethyl acetate, 5% glycerine and distilled water,
1hr (800 revs/min) is stirred at room temperature, liquid adhesive bandage film forming matrix is made.
2), the preparation of various concentration graphene oxide liquid adhesive bandage：
Different volumes graphene oxide is called in liquid adhesive bandage film forming matrix, preparing becomes oxygen-containing graphite alkene difference
The liquid adhesive bandage of concentration (2,1,0.5mg/ml).
3) graphene adhesive bandage is to Escherichia coli Bacteriostatic Effect on solid medium after, forming a film：
By 104Bacterium (or fungi) 20ul of/100ul exponential phases is uniformly coated on 12 hole agar plates, coating
The liquid adhesive bandage of various concentration graphene, liquid adhesive bandage film forming matrix of the coating without graphene oxide, as negative right
According to coating commercial liquid adhesive bandage is not coated with any film forming matrix, as blank control as positive control.It is inverted in 37 DEG C
Constant incubator overnight incubation, next day bacterium colony count.Each diluted concentration is coated with 3 holes, is averaged in triplicate.
4) influence of the graphene oxide to Bacterium and to eukaryocyte after, forming a film：
The preparation of homalographic membranization liquid adhesive bandage：
The liquid adhesive bandage of 2mg/ml graphene oxides is coated on repeatedly in 96 orifice bores, after 5-10min film forming, uses tweezer
Son connects down spare.Film forming matrix without graphene oxide is negative control, and commercial liquid adhesive bandage is positive control.
The bacterium solution 10ul of exponential phase is inoculated in 24 orifice plates of the corresponding culture mediums of 1ml, by various concentration graphene
Membranization adhesive bandage add in culture medium, 37 DEG C of oscillation incubations 2,6,20hr.
20ul is taken to be coated on LB solid cultures primary surface (12 orifice plate), is inverted, secondary number of days clone compares the oxidation after film forming
Graphene is to the killing effect of bacterium.
Graphene adhesive bandage is to Escherichia coli Bacteriostatic Effect on solid medium after film forming：
All solid culture primary surface (liquid adhesive bandages including film forming matrix, graphene-containing for smearing liquid adhesive bandage
With commercial liquid adhesive bandage) not long bacterium, all blank control wells for not smearing liquid adhesive bandage, bacterium can normal growth.
Supposition reason may be that the growth of this three classes bacterium is required to oxygen, and after smearing liquid adhesive bandage, block oxygen and bacterium
Contact, cause bacterium that can not grow.
To further look at the graphene oxide after comparing film forming to Bacterium, we are employed containing graphite oxide
The film forming adhesive bandage of alkene is incubated altogether with variety classes bacterium, then by way of colony count, observation graphene oxide into
The bactericidal effect in adhesive bandage after film.As a result it is as follows：
To the lethal effect of all kinds of bacteriums after the liquid adhesive bandage film forming of 5 graphene-containing of table
It is the liquid adhesive bandage of antibacterial components to lethal effect result such as Fig. 6 and table 5 of Escherichia coli using graphene oxide
Shown, it can be seen that, negative control film forming matrix is to all kinds of bacteriums at each time point without lethal effect from Fig. 6 and table 5；With
Graphene oxide for antibacterial components liquid adhesive bandage to the lethal effect result of staphylococcus aureus as shown in Fig. 7 and table 5,
There is a degree of effect for promoting growth to Candida albicans.The liquid adhesive bandage film forming of the graphene oxide containing 2mg/ml
Afterwards, there is very significant killing effect to Escherichia coli, Escherichia coli, bactericidal effect can be killed completely after being incubated 6 hours altogether
It is consistent with the liquid adhesive bandage of the domestic and imported of commercialization.
Also there is preferable killing effect to staphylococcus aureus, the germicidal efficiency nearly 90% of 6 hours is incubated altogether, with commodity
The liquid adhesive bandage of change is compared, and better than import adhesive bandage, is inferior to domestic adhesive bandage.
It is the liquid adhesive bandage of antibacterial components to lethal effect result such as Fig. 8 and table of Candida albicans using graphene oxide
Shown in 5, only have short-term killing activity to Candida albicans, only in common be incubated 2-4 hours, there is certain fragmentation effect；When altogether
When being incubated 20 hours, the facilitation effect of fragmentation effect and film forming matrix is offset, and does not observe effective killing.With commercialization
Liquid adhesive bandage is compared, and better than import adhesive bandage, is inferior to domestic adhesive bandage.
The result is consistent to the fragmentation effect of all kinds of bacteriums with free graphite alkene：Graphene oxide after forming a film is blue to leather
Family name's negative bacterium and gram-positive bacteria have notable lethal effect, and limited to the fragmentation effect of fungi.
Influence of the graphene liquid adhesive bandage to eukaryocyte after experimental example 5, film forming：
By the liquid containing graphene 0.06mg/ml, 0.125mg/ml, 0.25mg/ml, 0.5mg/ml, 1mg/ml, 2mg/ml
Body adhesive bandage is incubated jointly with NIH3T3 (mouse embryonic fibroblasts system cell), observes the liquid adhesive bandage containing graphene
Influence to NIH3T3 cell growths, with identical experiment condition to blank control group, import adhesive bandage and domestic adhesive bandage into
Row comparative experiments, is as a result shown in Fig. 9.
The results are shown in Figure 9, from the common numerical value for being incubated 1 hour and 4 hours as it can be seen that compared with blank control group, liquid wound
The film forming that can be attached in 96 holes significantly affects the adherent of NIH3T3 cells and growth, and cell activity is created significantly lower than not Covering Liguid
The blank control wells that can be pasted；The graphene of 2mg/ml has certain facilitation to the adherent of cell, and cell activity is significantly higher than
Remaining each concentration.After the liquid adhesive bandage film forming of commercialization, it appears that have certain toxicity to cell, cell viability is significantly lower than the moon
Property control film forming matrix.
When common incubation time was up to 16 hours, in holes of the NIH3T3 after the graphene liquid adhesive bandage film forming of various concentration
Well-grown, the graphene concentration indifference in cell activity and liquid adhesive bandage.Illustrate life of the graphene to eukaryocyte
Length does not influence.Therewith formed sharp contrast be commercialization liquid adhesive bandage film forming after, have notable shadow to the activity of cell
It rings, import adhesive bandage is smaller, and domestic adhesive bandage impact effect is especially apparent.Prompting has inhibition very in commercial liquid adhesive bandage
The adherent certain ingredients with growth of nucleus.It may influence wound healing.
Liquid adhesive bandage containing graphene oxide is in short term having an impact the adherent of eukaryocyte, and for a long time to eukaryocyte
Growth do not influence, effect be better than commercialization liquid adhesive bandage.
The internal fungistatic effect evaluation of experimental example 6, liquid adhesive bandage containing graphene oxide
The surface of a wound is manufactured in mouse body surface, using liquid adhesive bandage covering treatment, by detecting bacterial population under covering surface after 3 days
Amount observes Wound healing situation after 10 days, evaluate the internal fungistatic effect of the liquid adhesive bandage containing graphene oxide.
30 Balb/c mouse, half male and half female are divided into 5 groups, every group 6.Mouse back makes full thickness skin trauma model,
Surface smear Escherichia coli and staphylococcus aureus mixed liquor manufacture infected wound model.Using backbone as boundary, according to the form below carries out
It handles respectively：
Table 6, experiment mice grouping situation
After 4 days, cervical dislocation puts to death mouse (3/group), 75% ethyl alcohol cleaning mouse back, and removal surface miscellaneous bacteria influences,
Wound film is opened, with the surgical instrument clip wound and tissue around after high pressure steam sterilization；Tissue is shredded, immerses 0.5ml
PBS is fully vibrated, Aspirate supernatant 10ul with vortex oscillator, is added in 90ul PBS after ten times of dilutions, absorption 20ul tissues
Leachate is added in LB solid mediums, and coating is uniform.It is inverted, 37 DEG C of incubators, overnight incubation.Next day carries out colony count.Its
Remaining 15, the 10th day after surgery, wound healing situation was compared in observation.
To polluting the inhibiting effect of surface of a wound bacterium, result figure 10 and table 7 show postoperative graphene oxide liquid adhesive bandage
4th day wound tissue count of bacteria result.
The bacterial population initial data of the postoperative 4th day each group wound tissue of table 7
From result it can be seen that：The fungistatic effect (3L, 4L) of liquid adhesive bandage without graphene oxide film forming matrix with
(the 1L, 2L) wound not handled is compared, and bacterial number increases；Illustrate the liquid adhesive bandage film forming matrix without antipathogenic composition to wound
The bacterial growth of mouth does not act on not only always facilitation instead.
The fungistatic effect highly significant of liquid adhesive bandage containing graphene oxide better than the wound that does not handle and film forming base
Matter；The graphene liquid adhesive bandage of 0.5mg/ml contents can reach more than 90% bacteriostasis rate, and the liquid wound containing 2mg/ml can
The internal fungistatic effect of patch is up to 95%.
Compared with the liquid adhesive bandage of commercialization, the fungistatic effect of the liquid adhesive bandage containing graphene oxide is better than commercialization
Liquid adhesive bandage.The fungistatic effect of domestic liquid adhesive bandage is better than import adhesive bandage again.The trend is found with experiment in vitro
Trend is consistent.
Influence of the graphene oxide liquid adhesive bandage to Wound Contaminated：Postoperative 10th day, adhesive bandage film is thrown off, is compared
Each group wound healing situation, as a result five groups of postoperative 10th day wounds in mice healing state comparisons as shown in figs. 11 and 15.
As a result as it can be seen that wound has different degrees of cracking and infection performance on the left of 1-4 groups, healing situation is general；It is and right
The linear complete healing of side wound, no infection performance.Its healing state and the postoperative 4th day wound tissue bacterial population observed
It measures basically identical.5th group of commercialization adhesive bandage, the healing state of domestic adhesive bandage better than import adhesive bandage, this experiment it is oxygen-containing
The healing state of the liquid adhesive bandage of graphite alkene is better than commercialization adhesive bandage.
Claims (10)
1. a kind of using graphene oxide as the new liquid adhesive bandage of antibacterial components, comprising film forming component, solvent, feature exists
In the antimicrobial component in the liquid adhesive bandage is graphene oxide.
2. according to claim 1 a kind of using graphene oxide as the new liquid adhesive bandage of antibacterial components, feature exists
In the antimicrobial component graphene oxide content is 0.06mg/ml-4mg/ml.
3. according to claim 2 a kind of using graphene oxide as the new liquid adhesive bandage of antibacterial components, feature exists
In the graphene oxide content is 0.5mg/ml or 1mg/ml or 2mg/ml.
4. according to claim 3 a kind of using graphene oxide as the new liquid adhesive bandage of antibacterial components, feature exists
In the graphene oxide content is 2mg/ml.
5. according to claim 1 a kind of using graphene oxide as the new liquid adhesive bandage of antibacterial components, feature exists
In the film forming component is polyvinyl butyral.
6. according to claim 1 a kind of using graphene oxide as the new liquid adhesive bandage of antibacterial components, feature exists
In the solvent is ethyl alcohol.
7. according to claim 1 a kind of using graphene oxide as the new liquid adhesive bandage of antibacterial components, feature exists
In the liquid adhesive bandage also includes plasticizer.
8. according to claim 7 a kind of using graphene oxide as the new liquid adhesive bandage of antibacterial components, feature exists
In the plasticizer is ethyl acetate.
9. according to claim 1 a kind of using graphene oxide as the new liquid adhesive bandage of antibacterial components, feature exists
In the liquid adhesive bandage also includes glycerine.
10. according to claim 1 a kind of using graphene oxide as the new liquid adhesive bandage of antibacterial components, feature exists
In the ethyl alcohol comprising weight percent 75%, 5% polyvinyl butyral, 10% ethyl acetate, 5% glycerine.
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| CN109276738A (en) * | 2018-11-12 | 2019-01-29 | 张小伏 | A kind of high ventilative, antiphlogistic antibacterial bandage preparation method |
| CN109395152A (en) * | 2018-09-21 | 2019-03-01 | 南通强生石墨烯科技有限公司 | A kind of compound hydrocolloid medical dressing of graphene and preparation method thereof |
| CN110143588A (en) * | 2019-04-18 | 2019-08-20 | 浙江大学 | A method of graphene is shifted based on liquid adhesive bandage protective layer |
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