CN114344544B - Light activated antibiotic dressing and preparation method thereof - Google Patents
Light activated antibiotic dressing and preparation method thereof Download PDFInfo
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Abstract
The invention provides a light-activated antibacterial dressing and a preparation method thereof, wherein the preparation method comprises the following steps: s1, etching MAX ceramic into MXene nanosheets; s2, preparing MXene/Ag 3 PO 4 A heterojunction; s3, modifying MXene/Ag by polydopamine 3 PO 4 A heterojunction; and S4, modifying MXene/Ag by polycaprolactone particles and polydopamine 3 PO 4 And dissolving the heterojunction, uniformly mixing and forming a film to obtain the nanofiber film. The wound dressing for treating the infected wound can quickly sterilize in a short term, inhibit bacteria in a long term and promote the wound healing.
Description
Technical Field
The invention belongs to the technical field of medical materials, and particularly relates to a light-activated antibacterial dressing and a preparation method thereof.
Background
The skin, as the largest organ and first line of defense of the human body, plays a vital role in protecting the internal environment of the body from external damage. Once the skin is damaged and loses its original defense function, the subcutaneous tissue is attacked by microorganisms, resulting in infection, physical and mental suffering to the patient, and even more, amputation or death. In addition, if the patient is accompanied by basic diseases such as diabetes, chronic wounds are easy to generate, and repeated treatment also easily causes the appearance of drug-resistant bacteria.
At present, for the treatment of infected wounds, a debridement dressing method is mostly adopted. However, the traditional wound dressing only has the function of absorbing wound exudate/forming a physical barrier, so that bacteria remained on the local wound can continuously proliferate to generate stubborn wounds. In order to obtain antibacterial performance, antibiotics, nano-silver and other antibacterial agents are added into some dressings. However, overuse of these antimicrobials is often associated with some biotoxicity, and abuse of antibiotics has led to the prevalence of drug-resistant bacteria.
Disclosure of Invention
In view of the many deficiencies in the prior art, one of the objects of the present invention is to address one or more of the problems in the prior art. For example, it is an object of the present invention to provide a light-activated antimicrobial dressing that solves the problem of existing light treatment methods that the antimicrobial effect is lost when the light source is removed.
The invention provides a preparation method of a light-activated antibacterial dressing, which comprises the following steps: s1, etching MAX ceramic into MXene nanosheets; s2, preparing MXene/Ag 3 PO 4 A heterojunction; s3, modifying MXene/Ag by polydopamine 3 PO 4 A heterojunction; and S4, modifying MXene/Ag by polycaprolactone particles and polydopamine 3 PO 4 And dissolving the heterojunction, uniformly mixing and forming a film to obtain the nanofiber film.
The invention also provides a preparation method of the light-activated antibacterial dressing, which comprises the following steps: s1, etching MAX ceramic into MXene nanosheets; s2, preparing an MXene/AgS heterojunction; s3, modifying the MXene/AgS heterojunction by using polydopamine; and S4, dissolving the polycaprolactone particles and the polydopamine modified MXene/AgS heterojunction, uniformly mixing, and forming a film to obtain the nanofiber film.
In an embodiment, the step S1 may include: s11, soaking the MXene phase raw material in the etching solution, and continuously stirring for 12-24 hours at the constant temperature of 37-45 ℃.
In an embodiment, the etching solution may be a mixture of a lithium fluoride and hydrochloric acid solution.
In an embodiment, the step S1 may further include: s12, washing the precipitate with deionized water, and centrifuging at 3500-4500 r/min; the product was lyophilized in a vacuum desiccator to yield etched MXene.
In an embodiment, step S2 may comprise: s21, dispersing MXene obtained in the step S1 in distilled water, and adding Ag + Solution of Ag + The product is dialyzed until there is no Ag in the dialysate + Residue is left; s22, strongly stirring the dialysate, and gradually adding a first precipitate aqueous solution, wherein the first precipitate aqueous solution is one or more of disodium hydrogen phosphate aqueous solution, silver phosphite aqueous solution and silver phosphate aqueous solution; and S23, re-freeze-drying the precipitate to obtain purified MXene/Ag 3 PO 4 A heterojunction.
In an embodiment, step S2 may comprise: s21', dispersing MXene obtained in the step S1 in distilled water, and adding Ag + Solution of Ag + The product is dialyzed until there is no Ag in the dialysate + Residue is left; s22', stirring the dialyzate intensely, and gradually adding a second precipitation aqueous solution, wherein the second precipitation aqueous solution is a sulfur ion aqueous solution; and S23', re-lyophilizing the precipitate to obtain the purified MXene/AgS heterojunction.
In embodiments, the Ag + The solution can be silver nitrate solution, and the concentration can be 0.1-0.15M.
In the embodiment, S4, polycaprolactone particles and polydopamine modified MXene/AgS heterojunction are distributed in 1, 3-hexafluoro-2-propanol, the mixture is uniformly mixed, and an electrostatic spinning method is adopted to form a film to obtain the nanofiber film.
In another aspect, the invention provides a light-activated antibacterial dressing, which is prepared by the preparation method.
In an embodiment, the light-activated antimicrobial dressing may include MXene/Ag 3 PO 4 Heterojunction, polydopamine coating and polycaprolactone nanofiber membrane.
In an embodiment, the light-activated antimicrobial dressing may include an MXene/AgS heterojunction, a polydopamine coating, and a polycaprolactone nanofiber film.
In an embodiment, the light-activated antimicrobial dressing exerts a phototherapeutic effect under near-infrared light irradiation.
In an embodiment, the wavelength of the near infrared light is 808nm and/or 1064nm.
Compared with the prior art, the invention has the beneficial effects that:
(1) The invention discloses a wound dressing for treating infected wounds, which can quickly sterilize in a short term, inhibit bacteria in a long term and promote wound healing.
(2) The invention can be used repeatedly, has low cost for single treatment and has great advantages for treating chronic repeated wounds.
(3) The invention has simple operation, high feasibility and excellent antibacterial effect.
Drawings
The accompanying drawings, which are included to provide a further understanding of the inventive concepts, are incorporated in and constitute a part of this specification, illustrate exemplary embodiments of the inventive concepts and together with the description serve to explain the principles of the inventive concepts.
FIG. 1 is a scanning electron microscope image of MX @ AgP heterojunction and MX @ AgP-polycaprolactone nanofiber membrane prepared in example 1 of the present invention.
Fig. 2 is a scanning electron microscope image of an MXene nanosheet prepared in example 2 of the present invention.
Fig. 3 is a scanning electron microscope image of MXene nanosheets prepared in comparative example 1 of the present invention.
Fig. 4 is a schematic diagram of recording and evaluating photothermal effects of different nanofiber membranes by a thermal imaging instrument in application example 1 of the present invention, where fig. 4a shows real-time temperature changes of mx @ agp-PCL nanofiber membrane under near-infrared light irradiation, and fig. 4b shows photothermal stability of mx 8 @ agp-polycaprolactone nanofiber membrane.
Fig. 5 is a graph for evaluating the effect of the photodynamic of different nanofiber membranes according to application example 2 of the present invention, wherein fig. 5a shows a Methylene Blue (MB) reduction mechanism, fig. 5b and 5c show absorption spectra, fig. 5d shows a diphenyl isobenzofuran (DPBF) consumption mechanism, and fig. 5e and 5f show absorption spectra.
FIG. 6 is a graph showing the results of in vitro evaluation of antibacterial ability according to application example 3 of the present invention, and FIG. 6a is a graph showing the results against Staphylococcus aureus; FIG. 6b shows a graph of the results against E.coli.
Fig. 7 shows a scanning electron microscope image of the antibacterial mechanism of different nanofiber membranes in application example 4 of the present invention. Wherein figure 7a shows a scanning electron micrograph of anti-staphylococcus aureus (s. Aureus); fig. 7b shows a scanning electron micrograph of anti-e.coli (e.coil).
Fig. 8 is a graph showing the results of evaluating biocompatibility of different nanofiber membranes according to application example 5 of the present invention, wherein fig. 8a shows the morphology of cells cultured on different membranes recorded by confocal microscopy; FIG. 8b shows cell nodules captured by a scanning electron microscope.
Detailed Description
Hereinafter, a light-activated antimicrobial dressing and a method for manufacturing the same according to the present invention will be described in detail with reference to the accompanying drawings and exemplary embodiments.
In recent years, phototherapy has gained much attention from researchers because it can combat bacterial infections without the risk of antibiotic resistance.
The light therapy is mainly divided into photodynamic therapy and photothermal therapy. Under irradiation with light of an acceptable wavelength, the active oxygen species (ROS) generated by the photosensitizer and the photothermal agent convert the light energy into heat for antimicrobial therapy. The phototherapy can quickly kill most bacteria without generating drug-resistant bacteria; however, in the treatment, the bacteria remained on the local part of the wound can continue to proliferate, so that repeated infection is caused.
Cu 2+ ，Ag + The plasma metal ions have remarkable antibacterial effect, but the long-term use of the metal ions by a human body can cause organ toxicity; and as the ions are released, the antimicrobial effect of the ion source is diminished.
The invention provides a reusable antibacterial nano platform which can efficiently sterilize in a short term and persistently inhibit bacteria in a long term. In an exemplary embodiment, the present invention provides a wound dressing for treatment of infected wounds comprising MXene/Ag 3 PO 4 (MX @ AgP) heterojunction, polydopamine coating and polycaprolactone nanofiber membrane.
Wherein the MX @ AgP heterojunction exerts phototherapeutic effect under 808 nanometer near-infrared irradiation, generates ROS and converts light energy into heat energy, and can quickly sterilize in a short period; and after the light treatment, the silver ions can be continuously released on the part of the wound for bacteriostasis. However, the invention is not limited to this, and light in the near infrared band is applicable to other wavelengths besides 808nm and 1064nm, and has stronger tissue penetrability compared with 808 nm.
The polydopamine coating is positioned on the surface of the MX @ AgP heterojunction to play a role in recovering silver ions, and the released silver ions can be reduced to silver simple substances in situ and adhered to the surface of the polydopamine, namely the silver ion recovery process can be used for the next treatment.
The structure of the polycaprolactone nanofiber membrane is similar to that of extracellular matrix, so that the polycaprolactone nanofiber membrane is beneficial to cell crawling and wound healing, and can provide an anchoring platform for nanoparticles.
Alternatively, the wound dressing according to the invention may also be used with 0.1M Ag 2 S replacement of 0.1MAG 3 PO 4 After the heterojunction is synthesized, the photodynamic and antibacterial ability of silver ion release can be obtained.
The invention also provides a preparation method of the wound dressing for treating the infected wound, which comprises the following steps:
s1, etching the MAX ceramic into MXene nanosheets.
Specifically, S11, firstly, the MAX ceramic raw material is soaked in the etching solution and is continuously stirred for 12 to 24 hours at a constant temperature of 37 to 45 ℃.
Wherein the etching solution may be a mixture of lithium fluoride and hydrochloric acid solution. However, the present invention is not limited thereto, and the direct finished hydrofluoric acid may be used. Compared with the use of finished hydrofluoric acid, the present embodiment uses the mixture of lithium fluoride and hydrochloric acid solution as the etching solution, which can avoid the processes of transferring and pouring hydrofluoric acid, and is safer and less dangerous.
MXene material is a new two-dimensional layered transition metal carbide crystal material with chemical formula M n+1 X n (n =1, 2 or 3, M is a transition metal element, and X is carbon or nitrogen element.) MAX ceramic starting Material (MAX) may include Ti 3 AlC 2 、Ti 2 C、V 2 C and Mo 3 C 2 One or more of (a). More preferably Ti 3 AlC 2 . The source of the MXene material is not particularly limited in the present invention, and commercially available products known to those skilled in the art may be used. In the invention, the MXene material is used as a carrier raw material of the catalyst. MXene was obtained after MAX etching.
The amount of MAX ceramic raw materials may be 0.8-1.2 g. If the MAX amount is too small, the hydrofluoric acid is excessive; MAX excess does not react adequately.
The stirring is continued for 12 to 24 hours at a constant temperature of 37 to 45 ℃ in order to sufficiently etch. If the temperature is too low, the MAX aluminum layer is insufficiently etched due to too short time; the product is easy to oxidize when the temperature is too high, and the shape of the product can be influenced by the time course.
S12, after etching, preferably, the method further comprises the steps of carrying out solid-liquid separation on the etched system, washing the obtained solid component with water, and drying to obtain MXene.
The solid-liquid separation mode is not particularly limited, and a solid-liquid separation mode known to those skilled in the art can be adopted, specifically, filtration or centrifugal separation; the conditions for the centrifugation in the present invention are not particularly limited, and the centrifugation conditions known to those skilled in the art may be used. In the invention, the washing with water is preferably washing with deionized water, the frequency of the washing with water is not particularly limited, and the acid on the surface of the solid component can be removed completely. In the present invention, the drying is preferably freeze-drying, more preferably, lyophilization in a vacuum dryer.
In the present embodiment, the precipitate is preferably washed with deionized water, centrifuged at 3500 to 4500r/min for 5 to 7 times, and the centrifugation time is respectively 5min,15min,30min,1h and 1h. The sixth and seventh repeated centrifugation times were 1h. The purpose of centrifugation is as follows: removing impurities, e.g. Al, from the product by stepwise centrifugation 3+ 、F - (pour off with supernatant). Here, too low a centrifugation speed results in incomplete deposition of the product, and too high a centrifugation speed results in deposition of impurities.
The product was lyophilized in a vacuum desiccator to yield etched MXene.
S2, preparing MXene/Ag 3 PO 4 A heterojunction. The method specifically comprises the following steps:
s21, dispersing MXene obtained in the step S1 in distilled water, and adding Ag + Solution of Ag + The product is dialyzed until there is no Ag in the dialysate + And (4) remaining.
Wherein, ag + The solution can be silver nitrate solution, and the concentration can be 0.1-0.15M.
In this example, 0.1-0.2 g of MXene obtained in step S1 was added with 0.1-0.15M silver nitrate (AgNO) 3 ) 100-200 mL of solution, making Ag + The static electricity is adsorbed on the MXene surface. The product was dialyzed overnight until no Ag was present in the dialysate + And (4) remaining. Here, the presence or absence of Ag can be checked with sodium chloride + And (4) remaining.
And S22, intensively stirring the dialyzate, and gradually adding a first precipitation aqueous solution, wherein the first precipitation aqueous solution is one or more of disodium hydrogen phosphate aqueous solution, silver phosphite aqueous solution and silver phosphate aqueous solution.
In this example, the dialysate was vigorously stirred, and 0.01 to 0.02M disodium hydrogen phosphate (Na) was gradually added 2 HPO 4 ) An aqueous solution.
S23, precipitating againSub-lyophilizing to obtain purified MXene/Ag 3 PO 4 (MX @ AgP) heterojunction.
Here, the MAX ceramic is etched into MXene nanosheets, so that a higher specific surface area is obtained, more silver phosphate can grow in situ, and the light absorption capacity is better. The MXene nanosheets are loaded with silver phosphate and synthesized into MX @ AgP heterojunction, so that the nanoparticles not only have a photothermal antibacterial effect, but also can obtain a photodynamic and anion releasing antibacterial effect.
S3, modifying MXene/Ag by polydopamine 3 PO 4 A heterojunction.
And then modifying MX @ AgP by polydopamine (polydopamine, 2mg/mL in 10mM Tris-HCl, pH = 8.0-8.4), wherein the dopamine monomer can be polymerized into polydopamine within the pH range more quickly to obtain the polydopamine modified MX @ AgP heterojunction.
Polydopamine can be obtained by dissolving Dopamine (DA) in Tris-HCl (Tris-HCl) solution for a short reaction.
In this example, 2mg/mL in 10mM Tris-HCl means that 2mg/mL Dopamine (DA) is dissolved in 10mM Tris-HCl solution, and poly dopamine solution can be obtained by short reaction.
The outer coating of MX @ AgP is coated with polydopamine coating, so that the nano particles can obtain the capability of recovering and re-releasing silver ions.
S4, modifying MXene/Ag by polycaprolactone Particles (PCL) and polydopamine 3 PO 4 The heterojunction is distributed in 1, 3-hexafluoro-2-propanol, and the MX @ AgP-PCL nanofiber membrane is obtained by uniformly mixing and forming a membrane.
Here, polycaprolactone particles and polydopamine are modified to MXene/Ag 3 PO 4 The purpose of the distribution of the heterojunctions in 1,1,1,3, 3-hexafluoro-2-propanol is to be able to dissolve polycaprolactone to form an ester solution, alternatively, 1, 3-hexafluoro-2-propanol may be used as a substitute such as acetone, ethylene glycol.
In this embodiment, the blending method is stirring overnight, and the film may be formed by an electrostatic spinning method. The invention is not limited to the method, and the 3D printing can be adopted to form the film, so that the gap and the thickness can be accurately controlled, but compared with the electrostatic spinning film, the electrostatic spinning film has higher efficiency and lower price.
Specifically, three different concentrations of polydopamine modified mx @ agp (2 wt.%, 5wt.% and 8 wt.%) and polycaprolactone particles (1g, mw = 80kda) were distributed in 1,1,1,3, 3-hexafluoro-2-propanol (HFIP, 10 mL) and stirred overnight. Here, the selection of three different concentrations can be used to evaluate in vitro the performance of the drug at three concentrations, such as biosafety, photothermal power, and photodynamic power. Finally, the concentration for in vivo selection is considered comprehensively.
The electrospinning mixed solution was supplied at room temperature at a rate of 1 mm/min. The working voltage is 15.0kV, and the distance between the spinning device and the metal roller collector is 20cm. The electrospinning films were randomly collected on a metal drum trap at a speed of 150 rpm, and the aforementioned three different concentrations of polydopamine-modified mx @ agp (2 wt.%, 5wt.% and 8 wt.%) were named 2mx @ agp-PCL,5mx @ agp-PCL and 8mx @ agp-PCL. The pure polycaprolactone electrospun membrane is prepared by adopting the same electrostatic spinning method. The microstructure and morphology of the mx @ agp heterojunction and nanofiber membrane were observed using a scanning electron microscope.
The working principle of the antibacterial nano platform is as follows: under the irradiation of 808 nanometer near-infrared light, MX @ AgP heterojunction in the nanofiber membrane shows good photo-thermal/photodynamic performance, and silver ions are released for antibiosis, so that photo-thermal/photodynamic/silver ion synergetic antibiosis is realized. When the near infrared light is removed, the nanofiber membrane can still continuously release silver ions, meanwhile, the polydopamine serves as a 'water storage tank', the released silver ions are reduced into silver simple substances and adhered to the surface of the polydopamine, the 'self-charging' function is realized, the silver ions are recovered, the potential toxicity is reduced, and the polydopamine is used for next phototherapy. Then, the light leaks in near infrared light again, the photo-thermal and photo-dynamic effects are generated again, active oxygen generated by the photo-dynamic effect can create an easily oxidized environment, silver simple substances which are recovered and adhered to the surface of the polydopamine are oxidized into silver ions and released, and the effect of ion treatment is not weakened.
However, the invention is not so limited and in another embodiment, 0.1MAG may be used 2 S replacement for 0.1MAG 3 PO 4 After the heterojunction is synthesized, the photodynamic and antibacterial ability of silver ion release can be obtained.
Specifically, the preparation method thereof and the preparation method of the above-mentioned light-activated antimicrobial dressing in this embodiment may include:
s1' is the same as step S1 above.
S2', preparing the MXene/AgS heterojunction, which specifically comprises the following steps:
s21', dispersing MXene obtained in the step S1 in distilled water, and adding Ag + Solution of Ag + The product is dialyzed until there is no Ag in the dialysate + And (4) remaining.
Wherein, ag + The solution can be silver nitrate solution, and the concentration can be 0.1-0.15M.
In this example, 0.1 to 0.2g of MXene obtained in step S1 may be added with 0.1 to 0.15M silver nitrate (AgNO) 3 ) 100-200 mL of solution, make Ag + The static electricity is adsorbed on the MXene surface. The product was dialyzed overnight until no Ag was present in the dialysate + And (4) remaining. Here, the presence or absence of Ag can be checked by using sodium chloride + And (4) remaining.
S22', vigorously stirring the dialysate, and gradually adding a second precipitating aqueous solution, which is an aqueous solution of sulfide ions.
In this example, the dialysate was vigorously stirred, and 0.01 to 0.02M aqueous solution of sulfide ions was gradually added.
S23', the precipitate is lyophilized again to obtain a purified MXene/AgS heterojunction.
The MAX ceramic is etched into MXene nanosheets, so that a higher specific surface area is obtained, more silver sulfate can grow in situ, and the light absorption capacity is better. The MXene/AgS heterojunction is synthesized by loading silver sulfate on the MXene nanosheets, so that the nanoparticles not only have the effect of photothermal antibiosis, but also can obtain the effects of photodynamic and anion release antibiosis.
S3', modifying MXene/AgS heterojunction by polydopamine.
And then MX @ AgS is modified by polydopamine (polydopamine, 2mg/mL in 10mM Tris-HCl, pH = 8.0-8.4), and the dopamine monomer can be polymerized into polydopamine within the pH range more quickly to obtain the polydopamine modified MX @ AgS heterojunction.
Polydopamine can be obtained by dissolving Dopamine (DA) in Tris-HCl (Tris-HCl) solution for a short reaction.
In this example, 2mg/mL in 10mM Tris-HCl means that 2mg/mL Dopamine (DA) is dissolved in 10mM Tris-HCl (Tris-HCl) solution, and poly-dopamine solution can be obtained by short reaction.
And a polydopamine coating is coated outside the MX @ AgS, so that the nano particles obtain the capability of recovering and re-releasing silver ions.
S4', dissolving the polycaprolactone particles and the poly MXene/AgS heterojunction, uniformly mixing and forming a film to obtain the MX @ AgS-polycaprolactone nanofiber film.
Specifically, polycaprolactone particles and poly MXene/AgS heterojunction are distributed in 1, 3-hexafluoro-2-propanol, and the mixture is uniformly mixed and filmed to obtain the MX @ AgS-polycaprolactone nanofiber membrane.
Here, the polycaprolactone particles and polydopamine modified MXene/AgS heterojunctions are distributed in 1, 3-hexafluoro-2-propanol in order to be able to dissolve polycaprolactone to form an ester solution, alternatively, 1, 3-hexafluoro-2-propanol may be used such as acetone, ethylene glycol, as an alternative.
In this example, the blending was performed overnight by stirring, and the film was formed by electrospinning. The invention is not limited to the method, and the 3D printing can be adopted to form the film, so that the gap and the thickness can be accurately controlled, but compared with the electrostatic spinning film, the electrostatic spinning film has higher efficiency and lower price.
In one exemplary embodiment, a method of making a wound dressing for treatment of an infected wound of the present invention may comprise:
0.8 to 1.2gMAX (Ti) 3 AlC 2 ) Immersed in an etching solution which was a mixture of lithium fluoride (LiF, 1 g) and hydrochloric acid (HCl) solution (9M, 20mL), andcontinuously stirring for 12-24 hours at the constant temperature of 37-45 ℃. Then, the precipitate is washed with deionized water and centrifuged several times at 3500-4500 r/min. The product was lyophilized in a vacuum desiccator to yield etched MXene.
MXene (0.1 g) obtained above was dispersed in distilled water, and 0.1M silver nitrate (AgNO) was added 3 ) Solution 100mL, make Ag + The static electricity is adsorbed on the MXene surface. The resulting product was dialyzed overnight until no Ag was present in the dialysate + Residual (checked with sodium chloride). The dialysate was then vigorously stirred and 0.01M disodium hydrogen phosphate (Na) was gradually added 2 HPO 4 ) An aqueous solution. The precipitate was lyophilized again to give a purified MX @ AgP heterojunction.
And 3, etching the MAX ceramic into MXene nanosheets to obtain a higher specific surface area, so that more silver phosphate can grow in situ and the MXene nanosheets have better light absorption capacity. The MXene nanosheets are loaded with silver phosphate and synthesized into MX @ AgP heterojunction, so that the nanoparticles not only have the effect of photothermal antibiosis, but also can obtain the effects of photodynamic and anion release antibiosis. The outer coating of MX @ AgP is coated with polydopamine coating, so that the nano particles can obtain the capability of recovering and re-releasing silver ions.
And 4, preparing the pure polycaprolactone electrospun membrane by adopting the same electrostatic spinning method. In addition, 3D printing is adopted to form a film, but in comparison, the electrostatic spinning film is higher in efficiency and lower in price, but the gap and the thickness cannot be accurately controlled.
In order that the above-described exemplary embodiments of the invention may be better understood, they are further described below in connection with specific examples.
Example 1
1gMAX (Ti) 3 AlC 2 ) The substrate was immersed in an etching solution which was a mixture of lithium fluoride (LiF, 1 g) and hydrochloric acid (HCl) (9M, 20mL), and stirred at a constant temperature of 40 ℃ for 12 hours. Then, the sediment is washed by deionized water, and is centrifuged for 7 times at 3500r/min, wherein the centrifugation time is respectively 5min,15min,30min,1h and 1h. The sixth and seventh repeated centrifugation for 1h. The product was lyophilized at-20 c,etched MXene is obtained.
MXene (0.1 g) obtained above was dispersed in distilled water, and 0.1M silver nitrate (AgNO) was added 3 ) 100mL of solution, make Ag + The static electricity is adsorbed on the MXene surface. The resulting product was dialyzed overnight until no Ag was present in the dialysate + The residue was checked with sodium chloride. Then, the dialyzate was vigorously stirred, and 0.05M silver phosphite (Na) was gradually added 2 HPO 4 ) An aqueous solution. The precipitate was lyophilized again to give a purified MX @ AgP heterojunction.
Subsequently, MX @ AgP is modified by polydopamine (polydopamine, 2mg/mL in 10mM Tris-HCl, pH = 8.0) to obtain polydopamine modified MX @ AgP heterojunction.
Three different concentrations of polydopamine modified mx @ agp (2,5 and 8 wt.%) and polycaprolactone particles (1g, mw = 80kda) were distributed in 1,1,1,3,3,3-hexafluoro-2-propanol (HFIP, 10 mL) and stirred overnight. The electrospinning mixed solution was supplied at room temperature at a speed of 1 mm/min. The working voltage is 15.0kV, and the distance between the spinning device and the metal roller collector is 20cm. The electrospinning films are randomly collected on a metal roller trap at a speed of 120 r/min, and the electrospinning films with different concentrations of polydopamine modified MX @ AgP are named as 2MX @ AgP-PCL, MX 5 @ AgP-PCL and 8MX @ AgP-PCL. The pure polycaprolactone electrospun membrane is prepared by adopting the same electrostatic spinning method.
The microstructure and morphology of the mx @ agp heterojunction and nanofiber membrane were observed using a scanning electron microscope. Specifically, as shown in fig. 1, the MX @ AgP heterojunction and the MX @ AgP-polycaprolactone nanofiber membrane are characterized, and the appearances of the MX @ AgP heterojunction and the MX @ AgP-polycaprolactone nanofiber membrane are characterized through a scanning electron microscope. Wherein the morphology of the (a) MX @ AgP heterojunction is characterized by scanning electron microscopy in FIG. 1 a. In FIG. 1b, the morphology of MX @ AgP-PCL nanofiber membranes was characterized by scanning electron microscopy. The result shows that the MX @ AgP heterojunction and the MX @ AgP-PCL nanofiber membrane are successfully synthesized.
Example 2
1gMAX (Ti) 3 AlC 2 ) Immersed in an etching solution, which was a mixture of lithium fluoride (LiF, 1 g) and hydrochloric acid (HCl) (9M, 20mL), and continuously stirred at a constant temperature of 40 ℃ for 1For 2 hours. Then, washing the sediment with deionized water, centrifuging for 7 times at 4000r/min, wherein the centrifuging time is respectively 5min,15min,30min,1h and 1h; the sixth and seventh repeated centrifugation times were 1h. The product was lyophilized in a vacuum lyophilizer to yield etched MXene.
MXene (0.15 g) obtained above was dispersed in distilled water, and 0.15M silver nitrate (AgNO) was added 3 ) 100mL of solution, make Ag + The static electricity is adsorbed on the MXene surface. The resulting product was dialyzed overnight until no Ag was present in the dialysate + The residue was checked with sodium chloride. Then, the dialyzate was vigorously stirred, and 0.075M silver phosphite (Na) was gradually added 2 HPO 4 ) An aqueous solution. The precipitate was lyophilized again to give a purified MX @ AgP heterojunction.
And then modifying MX @ AgP by polydopamine (polydopamine, 2mg/mL in 10mM Tris-HCl, pH = 8.0) to obtain the polydopamine modified MX @ AgP heterojunction.
Three different concentrations of polydopamine modified mx @ agp (2 wt.%, 5wt.% and 8 wt.%) and polycaprolactone particles (1g, mw = 80kda) were distributed in 1,1,1,3, 3-hexafluoro-2-propanol (HFIP, 10 mL) and stirred overnight.
The electrospinning mixed solution was supplied at room temperature at a speed of 1 mm/min. The working voltage is 15.0kV, and the distance between the spinning device and the metal roller collector is 20cm. The electrospun membranes were randomly collected on a metal drum trap at a rate of 150 rpm, and the electrospun membranes of polydopamine-modified MX @ AgP of different concentrations were named 2MX @ AgP-PCL,5MX @ AgP-PCL and 8MX @ AgP-PCL. The pure polycaprolactone electrospun membrane is prepared by adopting the same electrostatic spinning method. And observing the microstructure and the morphology of the MX @ AgP heterojunction and the nanofiber membrane by using a scanning electron microscope.
Fig. 2 shows an example 2 of the present invention, wherein MXene nanosheets were prepared under different reaction conditions from example 1, and the morphology was characterized by scanning electron microscopy.
Comparative example 1
1gMAX (Ti) 3 AlC 2 ) The substrate was immersed in an etching solution which was a mixture of lithium fluoride (LiF, 1 g) and hydrochloric acid (HCl) solution (9M, 20mL), and stirred at a constant temperature of 25 ℃ for 10 hours. Then, the polymer is deionizedRinsing the precipitate with seed water, and centrifuging at 3500r/min for several times. The product was lyophilized in a vacuum lyophilizer to yield etched MXene. MXene nanoplatelets and mx @ agp heterojunctions were observed using a scanning electron microscope.
Fig. 3 shows comparative example 1 of the present invention, wherein MXene nanosheets were prepared under different reaction conditions from examples 1 and 2, and morphology characterization was performed by scanning electron microscopy. As can be seen from fig. 3, if the conditions for etching MXene are not within the range given in the present invention, MXene nanosheets cannot be obtained, and the aluminum layer therein cannot be etched away.
Application example 1
The thermal infrared camera is used for collecting different nanofiber membranes at 1.5W/cm 2 And (3) a real-time thermal imaging graph under near infrared light, which is used for evaluating the photo-thermal performance of the photo-thermal imaging graph. At 1.5W/cm 2 And testing the photo-thermal effect stability of the nanofiber membrane through three heating-cooling cycles under near-infrared light illumination, wherein each cycle comprises illumination heating for 10 minutes and natural cooling for 15 minutes.
The real-time temperature change of the MX @ AgP-PCL nanofiber membrane under near infrared light irradiation (as shown in FIG. 4 a) and the photo-thermal stability of the 8MX @ AgP-PCL nanofiber membrane (as shown in FIG. 4 b) were recorded by a thermal imaging instrument. The result shows that the nanofiber membrane has good photo-thermal effect and photo-thermal stability.
Application example 2
The formation of OH is detected spectrophotometrically. 200 μ LMB solution (100 mg) was added to 200 μ L of the sample suspension. After centrifugation, the absorption spectrum of MB was measured after irradiation with or without near-infrared light for 10 min. Deionized water was used as a control. The absorbance was measured for three predetermined times (0, 5, 10 minutes) using ultraviolet visible light spectroscopy (UV 1800 PC) in the wavelength range of 450 nm to 750 nm. Detection Using the same method 1 O 2 And O 2 - The sample was suspended in 200. Mu.l of ethanol, added to the well plate, and 3mL of ethanol containing 30. Mu.M DPBF was added. 808nm laser is used for illumination, and an ultraviolet visible spectrophotometer is used for detecting the absorption of the DPBF solution within the wavelength range of 300-500 nm.
To examine the photodynamic effect of nanofiber membranes: evaluating different nanofiber film lightKinetic effects, fig. 5a shows the Methylene Blue (MB) reduction mechanism, fig. 5b and 5c show the absorption spectra, fig. 5d shows the Diphenylisobenzofuran (DPBF) consumption mechanism, and fig. 5e and 5f show the absorption spectra. The result shows that the 8MX @ AgP-PCL nanofiber membrane can generate hydroxyl radical (.OH) and superoxide radical (.O) under illumination 2 -), singlet oxygen (c) 1 O 2 ) Three active oxygen species.
Application example 3
The in-vitro antibacterial effect of PCL,2MX @ AgP-PCL,5MX @ AgP-PCL and 8MX @ AgP-PCL on staphylococcus aureus and escherichia coli under 808-nanometer near-infrared irradiation/non-irradiation is measured by adopting a plate counting method. In summary, each nanofiber membrane was incubated with bacterial suspension (200 ml, 1X 10) 5 CFU/mL) were co-placed in 48-well plates and subjected to different treatments (808 nm near-infrared illumination for 10 min; dark groups were incubated in the dark for 10 minutes). 50. Mu.L of the treated bacterial suspension was spread evenly on an agar plate, incubated at 37 ℃ for 24 hours, and a typical image of the plate was recorded using a digital camera.
The antibacterial ability of different nanofiber membranes was evaluated by the plate coating method. Fig. 6 is a graph showing the results of in vitro antibacterial ability evaluation according to application example 3 of the present invention, wherein fig. 6a is a graph showing the results against staphylococcus aureus (s. Aureus); fig. 6b shows a graph of the results against e.coli (e.coil). The result shows that the nanofiber membrane has excellent antibacterial effect on staphylococcus aureus and escherichia coli under illumination, and the antibacterial effect is gradually enhanced along with the increase of the concentration of the MX @ AgP heterojunction contained in the nanofiber membrane.
Application example 4
The morphology of the bacteria treated by different nanofiber membranes was studied by scanning electron microscopy. Bacterial suspension (200. Mu.l, staphylococcus aureus 1X 10) 7 CFU/mL, E.coli 1X 10 7 CFU/mL) was mixed with the sample and placed in 48-well plates. Near infrared light (808 nm, 1.5W/cm) 2 ) The irradiation was carried out for 10 minutes. Subsequently, excess bacterial suspension was eliminated, 2.5% (v/v) glutaraldehyde was used to immobilize the bacteria, and then the samples were dehydrated with ethanol at various concentrations. Finally, after the nanofiber membrane was dried and gold-coated, the nanofiber membrane was observed by SEMThe morphology of (2).
Fig. 7 shows a scanning electron microscope image of the antibacterial mechanism of different nanofiber membranes in application example 4 of the present invention. In order to explore the antibacterial mechanism, the antibacterial mechanism of different nanofiber membranes was explored by scanning electron microscopy. Wherein fig. 7a shows a scanning electron micrograph of anti-staphylococcus aureus (s. Aureus); fig. 7b shows a scanning electron micrograph of anti-e.coli (e.coil). The results show that the cell membrane of the bacteria is damaged and the integrity of the bacteria is damaged through the nanofiber membrane to kill the bacteria.
Application example 5
Mouse fibroblasts L929 were evaluated for morphology on PCL,2MX @ AgP-PCL,5MX @ AgP-PCL,8MX @ AgP-PCL samples. The nanofiber membrane and 24-well cell slide were placed in a 24-well culture plate and incubated with L929 cells (1X 10) 4 one/mL) was cultured at 37 ℃ for 3 days. The nanofiber membranes were rinsed twice with PBS and fixed with 4% formaldehyde tissue fixative for 2 hours. Then, the inside of the cells was penetrated with Triton X-100 (0.1% v/v) for 20 minutes, and then cytoskeleton and nuclei were counterstained with FITC-Coprin and 4', 6-diamino-2-phenylindole (DAPI), and fluorescence images were recorded with CLSM. In addition, the morphology of the L929 cells growing on the surface of different samples was observed using a scanning electron microscope. Placing the nanofiber membrane in a 24-well culture plate, and culturing with L929 cells (1X 10) 4 one/mL) was cultured at 37 ℃ for 3 days. After fixation with 4% formaldehyde tissue fixative, dehydration was continued for 10min with ethanol solution (30, 50, 70, 80, 90, 100%), followed by observation of cells on the fiber membrane with a scanning electron microscope.
Fig. 8 is an application example 5 of the present invention to evaluate biocompatibility of different nanofiber membranes, wherein fig. 8a shows the morphology of cells cultured on different membranes recorded by confocal microscopy (CLSM) (arrows indicate cellular pseudopodia); fig. 8b shows cell nodules captured by a Scanning Electron Microscope (SEM) (arrows indicate cellular pseudopodia). The result shows that the nanofiber membrane has excellent biocompatibility, cells can grow on the nanofiber membrane, and the shape is good.
The light-activated antibacterial dressing can be repeatedly used, is mainly used for treating infected wounds, is applied to the wounds, can achieve high-efficiency antibacterial effect after being locally applied by near infrared light for 10 minutes, and slowly releases silver ions for a long time after the near infrared light treatment because the silver ions are contained in the dressing, so that the dressing can continuously inhibit bacteria. The effect can be repeatedly realized through the self-charging function, and the wound healing agent can be used for treating chronic infection wounds.
According to the invention, the problem that the antibacterial effect is not existed again when the light source is removed in the existing light treatment method can be solved; the problem that the ion source can release along with ions in ion antibiosis and the antibiosis effect is weakened can be solved.
Although certain exemplary embodiments and implementations have been described herein, other embodiments and modifications will be apparent from the description. Accordingly, the inventive concept is not limited to such exemplary embodiments, but is to be defined by the appended claims along with their full scope of equivalents.
Claims (10)
1. A preparation method of a light-activated antibacterial dressing is characterized in that the light-activated antibacterial dressing comprises a heterojunction, a polydopamine coating and a polycaprolactone nanofiber membrane, wherein the heterojunction is MXene/Ag3PO4, and the preparation method comprises the following steps:
s1, etching MAX ceramic into MXene nanosheets;
s2, preparing an MXene/Ag3PO4 heterojunction;
s3, modifying the MXene/Ag3PO4 heterojunction with polydopamine to coat a polydopamine coating outside the MXene/Ag3PO4 heterojunction, wherein the polydopamine coating can reduce released silver ions into a silver simple substance in situ and adhere to the surface of the polydopamine; and
and S4, dissolving the polycaprolactone particles and the polydopamine modified MXene/Ag3PO4 heterojunction, uniformly mixing, and forming a film to obtain the nanofiber film.
2. A preparation method of a light-activated antibacterial dressing is characterized in that the light-activated antibacterial dressing comprises a heterojunction, a polydopamine coating and a polycaprolactone nanofiber membrane, wherein the heterojunction is an MXene/AgS heterojunction, and the preparation method comprises the following steps:
s1, etching MAX ceramic into MXene nanosheets;
s2, preparing an MXene/AgS heterojunction;
s3, modifying the MXene/AgS heterojunction with polydopamine to coat a polydopamine coating outside the MXene/AgS heterojunction, wherein the polydopamine coating can reduce released silver ions into silver simple substances in situ and adhere to the surface of the polydopamine; and
and S4, dissolving the polycaprolactone particles and the polydopamine modified MXene/AgS heterojunction, uniformly mixing, and forming a film to obtain the nanofiber film.
3. The method for preparing a light-activated antimicrobial dressing according to claim 1 or 2, wherein the step S1 comprises:
s11, soaking the MAX ceramic raw material in an etching solution, and continuously stirring for 12-24 hours at a constant temperature of 37-45 ℃.
4. The method for preparing a light-activated antimicrobial dressing according to claim 3, wherein the etching solution is a mixture of lithium fluoride and hydrochloric acid solution.
5. The method for preparing a light-activated antimicrobial dressing according to claim 1 or 2, wherein the step S1 further comprises: s12, washing the precipitate with deionized water, and centrifuging at 3500-4500 r/min; the product was lyophilized in a vacuum desiccator to yield etched MXene.
6. The method for preparing a light-activated antimicrobial dressing according to claim 1, wherein the step S2 comprises:
s21, dispersing MXene obtained in the step S1 in distilled water, adding an Ag + solution to enable Ag + to be adsorbed on the MXene surface in a static manner, and dialyzing the obtained product until no Ag + residue exists in a dialyzate;
s22, strongly stirring the dialysate, and gradually adding a first precipitation aqueous solution, wherein the first precipitation aqueous solution is one or more of a disodium hydrogen phosphate aqueous solution, a silver phosphite aqueous solution and a silver phosphate aqueous solution; and
s23, the precipitate is freeze-dried again to obtain the purified MXene/Ag3PO4 heterojunction.
7. The method for preparing a light-activated antimicrobial dressing according to claim 2, wherein the step S2 comprises:
s21', dispersing the MXene obtained in the step S1 in distilled water, adding an Ag + solution to enable Ag + to be electrostatically adsorbed on the surface of the MXene, and dialyzing the obtained product until no Ag + remains in a dialyzate;
s22', stirring the dialysate vigorously, and gradually adding a second precipitate aqueous solution, wherein the second precipitate aqueous solution is a sulfur ion aqueous solution; and
s23', the precipitate is lyophilized again to obtain a purified MXene/AgS heterojunction.
8. The method for preparing a light-activated antibacterial dressing according to claim 6 or 7, wherein the Ag + solution is silver nitrate solution with a concentration of 0.1-0.15M.
9. The preparation method of the light-activated antibacterial dressing according to claim 2, wherein S4, the polycaprolactone particles and the polydopamine modified MXene/AgS heterojunction are distributed in 1, 3-hexafluoro-2-propanol, and are uniformly mixed, and the nanofiber membrane is obtained by forming a membrane by an electrostatic spinning method.
10. A light-activated antibacterial dressing prepared by the preparation method of any one of claims 1 to 9, wherein the light-activated antibacterial dressing comprises a heterojunction, a polydopamine coating and a polycaprolactone nanofiber membrane, and the heterojunction is an MXene/Ag3PO4 or MXene/AgS heterojunction.
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