CN113817181B - Carbon quantum dot modified double-network hydrogel and preparation method thereof - Google Patents
Carbon quantum dot modified double-network hydrogel and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a carbon quantum dot modified double-network hydrogel, which is prepared by introducing kanamycin carbon quantum dots into a double-network hydrogel system in a chemical crosslinking mode; the aperture of the double-network hydrogel modified by the carbon quantum dots is 10-20 mu m, and the size of the kanamycin carbon quantum dots is 1-5 nm. In addition, the invention also provides a preparation method of the hydrogel. The first layer of network of the double-network hydrogel modified by the carbon quantum dots is Schiff base bonds formed by the reaction of aldehyde groups and amino groups, the second layer of network is hydrogen bonds formed by polyvinyl alcohol through a circulating low-temperature freezing method, and the Schiff base bonds and the hydrogen bonds are reversible dynamic covalent bonds, so that the bonds can be broken under the inflammatory microenvironment of wounds, the carbon quantum dots are released, and the aims of sterilization and drug resistance are fulfilled.
Description
Technical Field
The invention belongs to the technical field of biomedical materials, and particularly relates to a carbon quantum dot modified double-network hydrogel and a preparation method thereof.
Background
The hydrogel is a material with a three-dimensional network structure formed by crosslinking water-soluble macromolecules, and the hydrogel can absorb a large amount of water but does not swell because of the crosslinked structure, so that the hydrogel has strong water absorbability and water retention property, can absorb dozens of times to thousands of times of water per se, and basically does not lose the water. In the case of wounds, most wounds are exposed after exposure to tissue damage, bacterial infection may occur during repair, and severe inflammatory reactions often slow the healing of wounds. To address the problem of bacterial infection, which is lost in thousands of people every year, hydrogels containing antibiotics are a common choice for surgical infection, and have significant advantages, such as topical administration and sustained effective load release, thereby improving biocompatibility and minimizing antibiotic toxicity, while adding antibiotics to hydrogels can effectively prevent infection, overuse of antibiotics can lead to bacterial resistance. Therefore, effective bacteriostasis is carried out, and bacteria can not generate drug resistance to treat wound surface bacterial infection, and the method is one effective way for reducing inflammation and even death caused by wound infection.
Due to the multiple antibacterial modes, nanotechnology brings a strategy to alleviate the drug resistance problem of antibacterial hydrogel, and metal oxide nanoparticles have been introduced into hydrogel for wound treatment, but the use of metal or metal oxide nanomaterials may cause harm to human health and environment, so the development of a material with good biocompatibility, high antibacterial activity and no drug resistance to bacteria is urgent.
Disclosure of Invention
The invention aims to overcome the problems in the prior art and provide a carbon quantum dot modified double-network hydrogel. The first layer network of the hydrogel is Schiff base bonds formed by the reaction of aldehyde groups and amino groups, the second layer network is hydrogen bonds formed by polyvinyl alcohol through a circulating low-temperature freezing method, and the Schiff base bonds and the hydrogen bonds are reversible dynamic covalent bonds, so that the bonds can be broken under the inflammatory microenvironment of wounds, and carbon quantum dots are released, thereby achieving the purposes of sterilization and drug resistance.
In order to solve the technical problems, the invention adopts the technical scheme that: the carbon quantum dot modified double-network hydrogel is characterized in that the carbon quantum dot modified double-network hydrogel is prepared by introducing kanamycin carbon quantum dots into a double-network hydrogel system in a chemical crosslinking mode; the aperture of the double-network hydrogel modified by the carbon quantum dots is 10-20 mu m, and the size of the kanamycin carbon quantum dots is 1-5 nm.
In addition, the invention also provides a method for preparing the double-network hydrogel modified by the carbon quantum dots, which is characterized by comprising the following steps:
mixing an oxidized glucan aqueous solution and a recombinant collagen aqueous solution to obtain an oxidized glucan-recombinant collagen mixed solution;
adding kanamycin carbon quantum dots into the oxidized dextran-recombinant collagen mixed solution obtained in the step one to obtain a carbon quantum dot-oxidized dextran-recombinant collagen solution mixed solution;
step three, adding a polyvinyl alcohol aqueous solution into the mixed solution of the carbon quantum dots, the oxidized dextran and the recombinant collagen solution in the step two to obtain a mixed solution of the polyvinyl alcohol, the carbon quantum dots, the oxidized dextran and the recombinant collagen solution;
and step four, circularly freezing the mixed solution of the polyvinyl alcohol, the carbon quantum dots, the oxidized dextran and the recombinant collagen solution in the step three at low temperature to obtain the carbon quantum dot modified double-network hydrogel.
The method is characterized in that in the step one, the mass percentage of the oxidized dextran aqueous solution is 6-12%, the mass percentage of the recombinant collagen in the recombinant collagen aqueous solution is 6-12%, and the volume ratio of the oxidized dextran aqueous solution to the recombinant collagen aqueous solution is 1 (1-2).
The method is characterized in that the preparation method of the kanamycin carbon quantum dot in the step two comprises the following steps: dissolving kanamycin in a reaction kettle, treating for 1-3 h at 160-185 ℃, centrifuging, dialyzing, filtering by using a filter membrane of 0.22 mu m, and freeze-drying to obtain kanamycin carbon quantum dots.
The method is characterized in that the rotating speed of the centrifugal machine is 10000-20000 rpm, the centrifugation time is 20-40 min, the interception amount of the dialysis bag is 1000Da, the dialysis time is 3d, and the freeze-drying time is 3d.
The method is characterized in that the concentration of the carbon quantum dots of the kanamycin in the mixed solution of the carbon quantum dots, the oxidized glucan and the recombinant collagen in the step two is not more than 4mg/mL.
The method is characterized in that the polyvinyl alcohol in the polyvinyl alcohol aqueous solution in the third step is 6-12% by mass.
The method is characterized in that the volume ratio of the carbon quantum dot-oxidized dextran-recombinant collagen solution mixed solution to the polyvinyl alcohol aqueous solution in the third step is 1 (1-2).
The method is characterized in that the number of times of low-temperature freezing in the fourth step is 2-3 times; each circulation low-temperature freezing comprises low-temperature freezing and unfreezing, the low-temperature freezing temperature is-10 ℃ to-40 ℃, the low-temperature freezing time is 2h to 30h, the unfreezing temperature is 20 ℃ to 40 ℃, and the unfreezing time is 2h to 10h.
The method is characterized by further comprising the step of placing the mixed solution of the polyvinyl alcohol-carbon quantum dots-oxidized dextran-recombinant collagen solution at room temperature for 2-40 min before circulating low-temperature freezing.
Compared with the prior art, the invention has the following advantages:
1. the pore diameter of the double-network hydrogel modified by the carbon quantum dots is 10-20 microns, the size of the carbon quantum dots is 1-5 nm, the first layer network of the double-network hydrogel is a Schiff base bond formed by the reaction of aldehyde groups and amino groups, the second layer network of the double-network hydrogel is a hydrogen bond formed by polyvinyl alcohol through a circulating low-temperature freezing method, the Schiff base bond and the hydrogen bond are reversible dynamic covalent bonds, the bond fracture can be realized under the inflammatory microenvironment of wounds, the carbon quantum dots are released, and the aims of sterilization and drug resistance are fulfilled.
2. The double-network hydrogel modified by the carbon quantum dots can effectively control water loss and keep the wound surface moist for a long time, so that the double-network hydrogel can resist bacteria and drug resistance, can preserve moisture, absorb seepage, block bacteria, adhere to the skin, prevent dressing from falling off and promote wound healing.
3. The carbon quantum dot modified double-network hydrogel can be directly attached to the skin, so that the treatment and nursing procedures are reduced, and the cell proliferation is promoted to accelerate the wound healing.
4. The invention adopts two crosslinking modes of chemical crosslinking and physical crosslinking to prepare the double-network hydrogel, and takes the oxidized glucan with good biocompatibility as a crosslinking agent, thereby having the characteristics of safety, no toxicity and convenience.
The technical solution of the present invention will be described in further detail with reference to the following detailed description and accompanying drawings.
Drawings
FIG. 1 is a high power transmission electron micrograph of a carbon quantum dot in example 1 of the present invention;
FIG. 2 is a graph showing a distribution of particle diameters of carbon quantum dots in example 1 of the present invention;
FIG. 3 is a plot of the Fourier infrared spectra of dextran and oxidized dextran;
FIG. 4 is a NMR spectrum of dextran and oxidized dextran;
FIG. 5 is a scanning electron micrograph of a double-network hydrogel according to example 1 of the present invention;
FIG. 6 is a comparison graph of the bacteriostatic properties of the double-network hydrogel in example 1 of the present invention;
FIG. 7 is a diagram showing a cell compatibility test of the double-network hydrogel in example 1 of the present invention.
Detailed Description
The recombinant collagen used in the invention is provided by Xianju biological gene technology GmbH; the oxidized dextran can be purchased directly or prepared by the following method:
4g of dextran and 3.4g of sodium periodate were dissolved in 50mL of ultrapure water with magnetic stirring. The mixture was reacted at room temperature in the dark for 24h and the solution was pale yellow. 1g of ethylene glycol was then added and the reaction was allowed to proceed for two hours to terminate further oxidation of the glucan. Continuously dialyzing the product with deionized water for 3 days (the cut-off molecular weight of the dialysis bag is 8000 Da-12000 Da), replacing dialysis water for 3 times every day, and finally freeze-drying the solution in the dialysis bag to obtain the oxidized dextran.
Example 1
The preparation method of the carbon quantum dot modified double-network hydrogel of the embodiment comprises the following steps:
mixing an oxidized glucan aqueous solution and a recombinant collagen aqueous solution to obtain an oxidized glucan-recombinant collagen mixed solution; the mass percentage content of the oxidized dextran aqueous solution is 6%, the number average molecular weight of the oxidized dextran is 40000Da, the mass percentage content of the recombinant collagen in the recombinant collagen aqueous solution is 6%, and the volume ratio of the oxidized dextran aqueous solution to the recombinant collagen aqueous solution is 1:1;
step two, dissolving kanamycin in a reaction kettle, treating for 2 hours at 160 ℃, then centrifuging for 30 minutes at 15000rpm, dialyzing the centrifugate for 3d by using a dialysis bag with the cut-off of 1000Da, filtering by using a filter membrane with the diameter of 0.22 mu m, and freeze-drying for 3d to obtain a kanamycin carbon quantum dot; adding kanamycin carbon quantum dots into the oxidized dextran-recombinant collagen mixed solution obtained in the step one, and preparing carbon quantum dot-oxidized dextran-recombinant collagen solution mixed solutions with different concentrations; the concentrations of the kanamycin carbon quantum dots in the carbon quantum dot-oxidized glucan-recombinant collagen solution mixed solution are respectively 0mg/mL, 1mg/mL, 2mg/mL, 3mg/mL and 4mg/mL;
dissolving polyvinyl alcohol with the number average molecular weight of 70000Da in water at the temperature of 90 ℃ to obtain a polyvinyl alcohol aqueous solution with the mass percentage of the polyvinyl alcohol of 6%; adding a polyvinyl alcohol aqueous solution into the mixed solution of the carbon quantum dots, the oxidized dextran and the recombinant collagen solution in the second step to obtain a mixed solution of the polyvinyl alcohol, the carbon quantum dots, the oxidized dextran and the recombinant collagen solution; the volume ratio of the mixed solution of the carbon quantum dots, the oxidized dextran and the recombinant collagen solution to the polyvinyl alcohol aqueous solution is 1:1;
step four, circularly freezing the mixed solution of the polyvinyl alcohol-carbon quantum dots-oxidized dextran-recombinant collagen solution in the step three for 3 times at a low temperature, wherein each circulation low temperature freezing comprises low temperature freezing and unfreezing, the low temperature freezing temperature is-20 ℃, the low temperature freezing time is 12 hours, the unfreezing temperature is 20 ℃, and the unfreezing time is 6 hours; obtaining the double-network hydrogel modified by the carbon quantum dots.
The aperture of the double-network hydrogel modified by the carbon quantum dots is 10-20 μm, and the size of the carbon quantum dots is 1-5 nm, and the double-network hydrogel modified by the carbon quantum dots in the embodiment can realize bond fracture and release of the carbon quantum dots under an inflammation microenvironment of a wound, so that the purposes of sterilizing and promoting wound healing are achieved.
Example 2
The preparation method of the carbon quantum dot modified double-network hydrogel of the embodiment comprises the following steps:
mixing an oxidized glucan aqueous solution and a recombinant collagen aqueous solution to obtain an oxidized glucan-recombinant collagen mixed solution; the mass percentage of the oxidized dextran aqueous solution is 10%, the number average molecular weight of the oxidized dextran is 40000Da, the mass percentage of the recombinant collagen in the recombinant collagen aqueous solution is 8%, and the volume ratio of the oxidized dextran aqueous solution to the recombinant collagen aqueous solution is 1.5;
step two, dissolving kanamycin in a reaction kettle, treating for 2 hours at 165 ℃, then centrifuging for 30 minutes at 15000rpm, dialyzing the centrifugate for 3d by using a dialysis bag with the cutoff of 1000Da, filtering by using a filter membrane of 0.22 mu m, and freeze-drying for 3d to obtain a kanamycin carbon quantum dot; adding kanamycin carbon quantum dots into the oxidized dextran-recombinant collagen mixed solution obtained in the step one, and preparing carbon quantum dot-oxidized dextran-recombinant collagen solution mixed solutions with different concentrations; the concentrations of the kanamycin carbon quantum dots in the carbon quantum dot-oxidized glucan-recombinant collagen solution mixed solution are respectively 0mg/mL, 1mg/mL, 2mg/mL, 3mg/mL and 4mg/mL;
dissolving polyvinyl alcohol with the number average molecular weight of 70000Da in water at the temperature of 90 ℃ to obtain a polyvinyl alcohol aqueous solution with the polyvinyl alcohol mass percentage of 8%; adding a polyvinyl alcohol aqueous solution into the mixed solution of the carbon quantum dots, the oxidized dextran and the recombinant collagen solution in the second step to obtain a mixed solution of the polyvinyl alcohol, the carbon quantum dots, the oxidized dextran and the recombinant collagen solution; the volume ratio of the mixed solution of the carbon quantum dots, the oxidized glucan and the recombinant collagen solution to the polyvinyl alcohol aqueous solution is 1.5;
placing the mixed solution of the polyvinyl alcohol-carbon quantum dots-oxidized dextran-recombinant collagen solution in the third step for 2min at room temperature, and then performing low-temperature freezing for 2 times, wherein the low-temperature freezing for each time comprises low-temperature freezing and unfreezing, the low-temperature freezing temperature is-20 ℃, the low-temperature freezing time is 12 hours, the unfreezing temperature is 20 ℃, and the unfreezing time is 6 hours; obtaining the double-network hydrogel modified by the carbon quantum dots.
The carbon quantum dot modified double-network hydrogel prepared in the embodiment has the pore size of 10-20 microns and the size of 1-5 nm, and can realize bond fracture and release of carbon quantum dots under the inflammatory microenvironment of wounds, so that the purposes of sterilizing and promoting wound healing are achieved.
Example 3
The carbon quantum dot modified double-network hydrogel is prepared by introducing kanamycin carbon quantum dots into a double-network hydrogel system in a chemical crosslinking manner; the aperture of the double-network hydrogel modified by the carbon quantum dots is 10-20 mu m, and the size of the kanamycin carbon quantum dots is 1-5 nm.
The preparation method of the carbon quantum dot modified double-network hydrogel of the embodiment comprises the following steps:
mixing an oxidized glucan aqueous solution and a recombinant collagen aqueous solution to obtain an oxidized glucan-recombinant collagen mixed solution; the mass percentage content of the oxidized dextran aqueous solution is 10%, the number average molecular weight of the oxidized dextran is 40000Da, the mass percentage content of the recombinant collagen in the recombinant collagen aqueous solution is 10%, and the volume ratio of the oxidized dextran aqueous solution to the recombinant collagen aqueous solution is 1:1;
step two, dissolving kanamycin in a reaction kettle, processing at 175 ℃ for 1h, centrifuging at 10000rpm for 40min, dialyzing the centrifugate for 3d by using a dialysis bag with the cut-off of 1000Da, filtering by using a filter membrane with the diameter of 0.22 mu m, and freeze-drying for 3d to obtain a kanamycin carbon quantum dot; adding kanamycin carbon quantum dots into the oxidized dextran-recombinant collagen mixed solution obtained in the step one, and preparing carbon quantum dot-oxidized dextran-recombinant collagen solution mixed solutions with different concentrations; the concentrations of the kanamycin carbon quantum dots in the carbon quantum dot-oxidized glucan-recombinant collagen solution mixed solution are respectively 0mg/mL, 1mg/mL, 2mg/mL, 3mg/mL and 4mg/mL;
dissolving polyvinyl alcohol with the number average molecular weight of 70000Da in water at the temperature of 90 ℃ to obtain a polyvinyl alcohol aqueous solution with the polyvinyl alcohol mass percentage of 10%; adding a polyvinyl alcohol aqueous solution into the mixed solution of the carbon quantum dots, the oxidized dextran and the recombinant collagen solution in the second step to obtain a mixed solution of the polyvinyl alcohol, the carbon quantum dots, the oxidized dextran and the recombinant collagen solution; the volume ratio of the mixed solution of the carbon quantum dots, the oxidized dextran and the recombinant collagen solution to the polyvinyl alcohol aqueous solution is 1:1;
step four, placing the mixed solution of the polyvinyl alcohol-carbon quantum dots-oxidized dextran-recombinant collagen solution in the step three for 10min at room temperature, and then performing 3 times of circulating low-temperature freezing, wherein each circulating low-temperature freezing comprises low-temperature freezing and unfreezing, the low-temperature freezing temperature is-10 ℃, the low-temperature freezing time is 2 hours, the unfreezing temperature is 20 ℃, and the unfreezing time is 2 hours; obtaining the double-network hydrogel modified by the carbon quantum dots.
The aperture of the double-network hydrogel modified by the carbon quantum dots is 10-20 μm, and the size of the carbon quantum dots is 1-5 nm, and the double-network hydrogel modified by the carbon quantum dots in the embodiment can realize bond fracture and release of the carbon quantum dots under an inflammation microenvironment of a wound, so that the purposes of sterilizing and promoting wound healing are achieved.
Example 4
The preparation method of the carbon quantum dot modified double-network hydrogel of the embodiment comprises the following steps:
mixing an oxidized glucan aqueous solution and a recombinant collagen aqueous solution to obtain an oxidized glucan-recombinant collagen mixed solution; the mass percentage of the oxidized dextran aqueous solution is 10%, the number average molecular weight of the oxidized dextran is 40000Da, the mass percentage of the recombinant collagen in the recombinant collagen aqueous solution is 12%, and the volume ratio of the oxidized dextran aqueous solution to the recombinant collagen aqueous solution is 1:2;
step two, dissolving kanamycin in a reaction kettle, treating for 3h at 175 ℃, centrifuging for 20min at 20000rpm, dialyzing the centrifugate for 3d by using a dialysis bag with the cut-off of 1000Da, filtering by using a filter membrane of 0.22 mu m, and freeze-drying for 3d to obtain a kanamycin carbon quantum dot; adding kanamycin carbon quantum dots into the oxidized dextran-recombinant collagen mixed solution obtained in the step one, and preparing carbon quantum dot-oxidized dextran-recombinant collagen solution mixed solutions with different concentrations; the concentrations of the kanamycin carbon quantum dots in the carbon quantum dot-oxidized glucan-recombinant collagen solution mixed solution are respectively 0mg/mL, 1mg/mL, 2mg/mL, 3mg/mL and 4mg/mL;
dissolving polyvinyl alcohol with the number average molecular weight of 70000Da in water at the temperature of 90 ℃ to obtain a polyvinyl alcohol aqueous solution with the polyvinyl alcohol mass percentage of 10%; adding a polyvinyl alcohol aqueous solution into the mixed solution of the carbon quantum dots, the oxidized dextran and the recombinant collagen solution in the second step to obtain a mixed solution of the polyvinyl alcohol, the carbon quantum dots, the oxidized dextran and the recombinant collagen solution; the volume ratio of the mixed solution of the carbon quantum dots, the oxidized glucan and the recombinant collagen solution to the polyvinyl alcohol aqueous solution is 1:2;
placing the mixed solution of the polyvinyl alcohol-carbon quantum dots-oxidized dextran-recombinant collagen solution in the third step for 40min at room temperature, and then performing 3 times of circulating low-temperature freezing, wherein each circulating low-temperature freezing comprises low-temperature freezing and unfreezing, the low-temperature freezing temperature is-40 ℃, the low-temperature freezing time is 2 hours, the unfreezing temperature is 40 ℃, and the unfreezing time is 2 hours; obtaining the double-network hydrogel modified by the carbon quantum dots.
The aperture of the double-network hydrogel modified by the carbon quantum dots is 10-20 μm, and the size of the carbon quantum dots is 1-5 nm, and the double-network hydrogel modified by the carbon quantum dots in the embodiment can realize bond fracture and release of the carbon quantum dots under an inflammation microenvironment of a wound, so that the purposes of sterilizing and promoting wound healing are achieved.
Example 5
The preparation method of the carbon quantum dot modified double-network hydrogel of the embodiment comprises the following steps:
step one, mixing an oxidized dextran aqueous solution and a recombinant collagen aqueous solution to obtain an oxidized dextran-recombinant collagen mixed solution; the mass percentage of the oxidized dextran aqueous solution is 10%, the number average molecular weight of the oxidized dextran is 40000Da, the mass percentage of the recombinant collagen in the recombinant collagen aqueous solution is 12%, and the volume ratio of the oxidized dextran aqueous solution to the recombinant collagen aqueous solution is 1:1;
step two, dissolving kanamycin in a reaction kettle, processing for 1h at 180 ℃, centrifuging for 40min at 10000rpm, dialyzing the centrifugate for 3d by using a dialysis bag with the cut-off of 1000Da, filtering by using a filter membrane with the diameter of 0.22 mu m, and freeze-drying for 3d to obtain a kanamycin carbon quantum dot; adding kanamycin carbon quantum dots into the oxidized dextran-recombinant collagen mixed solution obtained in the step one, and preparing carbon quantum dot-oxidized dextran-recombinant collagen solution mixed solutions with different concentrations; the concentrations of the kanamycin carbon quantum dots in the carbon quantum dot-oxidized glucan-recombinant collagen solution mixed solution are respectively 0mg/mL, 1mg/mL, 2mg/mL, 3mg/mL and 4mg/mL;
dissolving polyvinyl alcohol with the number average molecular weight of 70000Da in water at the temperature of 90 ℃ to obtain a polyvinyl alcohol aqueous solution with the polyvinyl alcohol mass percentage of 12%; adding a polyvinyl alcohol aqueous solution into the mixed solution of the carbon quantum dots, the oxidized dextran and the recombinant collagen solution in the second step to obtain a mixed solution of the polyvinyl alcohol, the carbon quantum dots, the oxidized dextran and the recombinant collagen solution; the volume ratio of the mixed solution of the carbon quantum dot-oxidized dextran-recombinant collagen solution to the polyvinyl alcohol aqueous solution is 1;
step four, placing the mixed solution of the polyvinyl alcohol-carbon quantum dot-oxidized dextran-recombinant collagen solution in the step three for 20min at room temperature, and then performing low-temperature freezing for 2 times, wherein each low-temperature freezing cycle comprises low-temperature freezing and unfreezing, the low-temperature freezing temperature is-10 ℃, the low-temperature freezing time is 30 hours, the unfreezing temperature is 20 ℃, and the unfreezing time is 10 hours; obtaining the double-network hydrogel modified by the carbon quantum dots.
The aperture of the double-network hydrogel modified by the carbon quantum dots is 10-20 μm, and the size of the carbon quantum dots is 1-5 nm, and the double-network hydrogel modified by the carbon quantum dots in the embodiment can realize bond fracture and release of the carbon quantum dots under an inflammation microenvironment of a wound, so that the purposes of sterilizing and promoting wound healing are achieved.
Example 6
The preparation method of the carbon quantum dot modified double-network hydrogel of the embodiment comprises the following steps:
mixing an oxidized glucan aqueous solution and a recombinant collagen aqueous solution to obtain an oxidized glucan-recombinant collagen mixed solution; the mass percentage content of the oxidized dextran aqueous solution is 12%, the number average molecular weight of the oxidized dextran is 40000Da, the mass percentage content of the recombinant collagen in the recombinant collagen aqueous solution is 12%, and the volume ratio of the oxidized dextran aqueous solution to the recombinant collagen aqueous solution is 1:1;
step two, dissolving kanamycin in a reaction kettle, processing for 2h at 185 ℃, centrifuging for 20min at 20000rpm, dialyzing the centrifugate for 3d by using a dialysis bag with the cutoff of 1000Da, filtering by using a filter membrane of 0.22 mu m, and freeze-drying for 3d to obtain a kanamycin carbon quantum dot; adding kanamycin carbon quantum dots into the oxidized dextran-recombinant collagen mixed solution obtained in the step one, and preparing carbon quantum dot-oxidized dextran-recombinant collagen solution mixed solutions with different concentrations; the concentrations of the kanamycin carbon quantum dots in the carbon quantum dot-oxidized glucan-recombinant collagen solution mixed solution are 0mg/mL, 1mg/mL, 2mg/mL, 3mg/mL and 4mg/mL respectively;
dissolving polyvinyl alcohol with the number average molecular weight of 70000Da in water at the temperature of 90 ℃ to obtain a polyvinyl alcohol aqueous solution with the polyvinyl alcohol mass percentage of 12%; adding a polyvinyl alcohol aqueous solution into the mixed solution of the carbon quantum dots, the oxidized dextran and the recombinant collagen solution in the second step to obtain a mixed solution of the polyvinyl alcohol, the carbon quantum dots, the oxidized dextran and the recombinant collagen solution; the volume ratio of the mixed solution of the carbon quantum dots, the oxidized glucan and the recombinant collagen solution to the polyvinyl alcohol aqueous solution is 1:1;
step four, placing the mixed solution of the polyvinyl alcohol-carbon quantum dots-oxidized dextran-recombinant collagen solution in the step three for 10min at room temperature, and then performing 3 times of circulating low-temperature freezing, wherein each circulating low-temperature freezing comprises low-temperature freezing and unfreezing, the low-temperature freezing temperature is-20 ℃, the low-temperature freezing time is 12 hours, the unfreezing temperature is 20 ℃, and the unfreezing time is 6 hours; obtaining the double-network hydrogel modified by the carbon quantum dots.
The aperture of the double-network hydrogel modified by the carbon quantum dots is 10-20 μm, and the size of the carbon quantum dots is 1-5 nm, and the double-network hydrogel modified by the carbon quantum dots in the embodiment can realize bond fracture and release of the carbon quantum dots under an inflammation microenvironment of a wound, so that the purposes of sterilizing and promoting wound healing are achieved.
And (4) performance testing:
taking the carbon quantum dots prepared in example 1 as an example, it can be seen from fig. 1 of a high-power scanning electron microscope that the prepared carbon quantum dots are in a quasi-spherical distribution shape with uniform distribution and obvious lattice fringes, and it can be seen from fig. 2 of a particle size distribution diagram that the average particle size of the prepared carbon quantum dots is 2.28 ± 0.028nm.
It can be seen from the Fourier infrared spectrum of FIG. 3 that the peak value is at 1727cm -1 The shrinkage vibration peak of the aldehyde group can be obviously seen; the NMR spectrum of FIG. 4 shows that the substance has significant chemical shifts at 8-9 pm, and FIGS. 3-4 show that dextran has been successfully modified to oxidized dextran.
Taking the carbon quantum dot modified double-network hydrogel prepared in example 1 as an example, as can be seen from fig. 5, the pore size of the double-network hydrogel of the invention is relatively uniform, and the pore size of the hydrogel is 10 to 20 μm.
The antibacterial performance evaluation method is a co-culture method, a gel sample to be tested and a bacterial liquid are co-cultured for 24 hours, the co-cultured bacterial suspension (escherichia coli and staphylococcus aureus) is uniformly coated on an agar culture medium, and the growth condition of bacteria is observed and photographed for 24 hours (figure 6). In the figure, the blank is a blank group without the gel sample to be tested, and one to five samples respectively correspond to the carbon quantum dot modified double-network hydrogel with the concentration of the kanamycin carbon quantum dot in the mixed solution of the carbon quantum dot-oxidized dextran-recombinant collagen solution in the example 1 being 0mg/mL, 1mg/mL, 2mg/mL, 3mg/mL and 4mg/mL respectively. As can be seen from fig. 6, the carbon quantum dot-doped double-network hydrogels of the present invention (samples two to five) have significant growth inhibition ability against both escherichia coli and staphylococcus aureus, relative to the blank group (blank) and the hydrogel having a carbon quantum dot content of 0mg (sample one).
The method for evaluating the cell compatibility (FIG. 7) was MTT method. In the figure, the blank is a blank group without the gel sample to be tested, and one to five samples respectively correspond to the carbon quantum dot modified double-network hydrogel with the concentration of the kanamycin carbon quantum dot in the mixed solution of the carbon quantum dot-oxidized dextran-recombinant collagen solution in the example 1 being 0mg/mL, 1mg/mL, 2mg/mL, 3mg/mL and 4mg/mL respectively. As can be seen from FIG. 7, the double-network hydrogel modified by the carbon quantum dots has good biocompatibility, and the cell compatibility of different samples shows that the double-network hydrogel sample has good cell compatibility, and the cell survival rate is basically over 85%.
The above description is only a preferred embodiment of the present invention, and is not intended to limit the present invention, and all simple modifications, changes and equivalent structural changes made to the above embodiment according to the technical spirit of the present invention still fall within the protection scope of the technical solution of the present invention.
Claims (9)
1. A preparation method of a carbon quantum dot modified double-network hydrogel is characterized in that the carbon quantum dot modified double-network hydrogel is prepared by introducing kanamycin carbon quantum dots into a double-network hydrogel system in a chemical crosslinking mode; the aperture of the double-network hydrogel modified by the carbon quantum dots is 10-20 mu m, and the size of the kanamycin carbon quantum dots is 1nm-5nm;
the preparation method comprises the following steps:
mixing an oxidized glucan aqueous solution and a recombinant collagen aqueous solution to obtain an oxidized glucan-recombinant collagen mixed solution;
adding kanamycin carbon quantum dots into the oxidized glucan-recombinant collagen mixed solution obtained in the step one to obtain a carbon quantum dot-oxidized glucan-recombinant collagen solution mixed solution;
step three, adding a polyvinyl alcohol aqueous solution into the carbon quantum dot-oxidized dextran-recombinant collagen solution mixed solution obtained in the step two to obtain a polyvinyl alcohol-carbon quantum dot-oxidized dextran-recombinant collagen solution mixed solution;
and step four, circularly freezing the mixed solution of the polyvinyl alcohol-carbon quantum dots-oxidized dextran-recombinant collagen solution in the step three at a low temperature to obtain the carbon quantum dot modified double-network hydrogel.
2. The preparation method of the carbon quantum dot modified double-network hydrogel as claimed in claim 1, wherein in the first step, the oxidized dextran aqueous solution contains oxidized dextran 6-12 wt%, the recombinant collagen aqueous solution contains recombinant collagen 6-12 wt%, and the volume ratio of the oxidized dextran aqueous solution to the recombinant collagen aqueous solution is 1 (1~2).
3. The method for preparing a carbon quantum dot modified double-network hydrogel according to claim 1, wherein the method for preparing the kanamycin carbon quantum dot in the second step comprises the following steps: dissolving kanamycin in a reaction kettle, treating at 160-185 ℃ for 1h-3h, centrifuging, dialyzing, filtering with a 0.22 mu m filter membrane, and freeze-drying to obtain the kanamycin quantum dot.
4. The method for preparing the carbon quantum dot modified two-network hydrogel according to claim 3, wherein the rotation speed of a centrifuge is 10000rpm to 20000rpm, the centrifugation time is 20min to 40min, the cut-off of a dialysis bag is 1000Da, the dialysis time is 3d, and the freeze-drying time is 3d.
5. The method for preparing the carbon quantum dot modified double-network hydrogel as claimed in claim 1, wherein the concentration of the carbon quantum dots of the kanamycin in the mixed solution of the carbon quantum dot, the oxidized dextran and the recombinant collagen in the step two is not more than 4mg/mL.
6. The method for preparing the carbon quantum dot modified double-network hydrogel according to claim 1, wherein the polyvinyl alcohol aqueous solution in the third step is 6-12% by mass of polyvinyl alcohol.
7. The method for preparing the carbon quantum dot modified double-network hydrogel according to claim 1, wherein the volume ratio of the carbon quantum dot-oxidized dextran-recombinant collagen solution mixed solution to the polyvinyl alcohol aqueous solution in the third step is 1 (1~2).
8. The method for preparing the double-network hydrogel modified by the carbon quantum dots according to claim 1, wherein the number of times of low-temperature freezing in the four steps is 2~3; each circulation low-temperature freezing comprises low-temperature freezing and unfreezing, wherein the low-temperature freezing temperature is minus 10 ℃ to minus 40 ℃, the low-temperature freezing time is 2h to 30h, the unfreezing temperature is 20 ℃ to 40 ℃, and the unfreezing time is 2h to 10h.
9. The method for preparing the carbon quantum dot modified double-network hydrogel according to claim 1, further comprising placing the mixed solution of the polyvinyl alcohol-carbon quantum dot-oxidized dextran-recombinant collagen solution at room temperature for 2min to 40min before circulating low-temperature freezing.
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