CN114767925A - Fluorescent pH-sensitive antibacterial hydrogel dressing and preparation method and application thereof - Google Patents
Fluorescent pH-sensitive antibacterial hydrogel dressing and preparation method and application thereof Download PDFInfo
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- CN114767925A CN114767925A CN202210484134.XA CN202210484134A CN114767925A CN 114767925 A CN114767925 A CN 114767925A CN 202210484134 A CN202210484134 A CN 202210484134A CN 114767925 A CN114767925 A CN 114767925A
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Abstract
The invention discloses a preparation method of a fluorescent pH-sensitive antibacterial hydrogel dressing, and belongs to the technical field of medical materials. The method comprises the steps of firstly obtaining chitosan hydrogel from chitosan and gelatin, then placing the chitosan hydrogel into a carbon quantum dot solution after freeze drying, enabling the carbon quantum dots to enter a gel network, wherein the gel has a three-dimensional network structure, and is beneficial to cell adhesion and tissue growth. The dressing not only promotes the repair of skin wounds, but also can be used for reflecting the pH state of the wounds, the hydrogel dressing is coated on the wounds, the fluorescence of the hydrogel dressing is gradually reduced along with the reduction of the pH value of the environment, and the fluorescence intensity is gradually enhanced along with the increase of the pH value. Based on the change of the pH value of the wound, the change of the pH value of the wound can be monitored in real time through ultraviolet light excitation, and simple visual monitoring is realized.
Description
Technical Field
The invention relates to the technical field of medical materials, in particular to a fluorescent pH-sensitive antibacterial hydrogel dressing, a preparation method thereof and application thereof in detecting wound infection.
Background
The skin serves as the first line of immunological defense of the human body, covering the whole body, and it protects various tissues and organs in the body from physical, mechanical, chemical and pathogenic microbial attacks. The pH value of the surface of normal skin is about 5.0-7.0; the pH of the skin averages about 5.8. Following skin injury, wound healing is a physiological process involved in the repair of the injury to living tissue, restoring its anatomical integrity and function to the injured site. If infection exists at the wound, the pathogenic bacteria excessively decompose extracellular matrix and generate ammonia, so that the wound is alkaline, and oxygen released by oxygenated hemoglobin in an alkaline environment is reduced, which is not beneficial to healing. Clinical researches show that the pH value of the chronic wound is 7.15-8.90, the range is an ideal environment for growth and proliferation of most germs, and for example, the pH value suitable for survival of staphylococcus aureus is 7.0-7.5. Thus, elevated pH can be used as an important indicator of wound infection. Detection of the pH of the wound surface in a non-healing or infectious state is critical to prevent disease progression and is also helpful for therapeutic intervention in wound care.
The concept of dressings has been around for a long time, but the dressings used by people for wounds for a long time are essentially gauze. Traditional dressings such as gauze, cotton pads and the like have good water absorption, are simple to manufacture, low in price and widely used, but in the healing process, the wound surface cannot be kept moist and cannot promote wound healing; granulation tissue formed on the surface of the wound is easy to adhere to the dressing, so that secondary wound is generated on the wound when the dressing is changed.
In recent years, hydrogel dressings have attracted wide attention, and the hydrogel dressings have the advantages of high water content, capability of keeping wound surfaces in a moist environment, convenience for granulation growth, good moisture retention, toughness and strength, capability of being tightly pasted on irregular wound surfaces, reduction in bacterial invasion, capability of absorbing a large amount of exudates, shortened dressing change time and alleviation of pain of patients.
Chitosan is a natural biological polysaccharide, consists of glucosamine and N-acetylglucosamine units, and has antioxidant, antibacterial, anticoagulant and antitumor activities. The chitosan polymer compound has many high-activity functional groups, has the characteristics similar to antibiotics, and can induce natural blood coagulation and stimulate collagen deposition and fibroblast proliferation.
Gelatin, a macromolecular hydrocolloid, is the product of partial hydrolysis of collagen. Because of high biocompatibility and biodegradability, no other byproducts are generated after in vivo degradation, no immunogenicity and blood compatibility, and the same components and biological properties as collagen, the collagen-based collagen is widely applied to tissue engineering and drug delivery systems.
As a new generation of fluorescent nano material, carbon quantum dots attract extensive attention due to the advantages of good water solubility, simple synthesis method, stable photoluminescence, low toxicity, environmental protection and the like.
In order to improve the life quality of patients, the development of materials for promoting wound healing is clinically significant. In view of this, the invention is particularly proposed.
Disclosure of Invention
In view of the above disadvantages, a first technical problem to be solved by the present invention is to provide a method for preparing a fluorescent pH-sensitive antibacterial hydrogel dressing, a second technical problem to be solved by the present invention is to provide a fluorescent pH-sensitive antibacterial hydrogel dressing prepared by the method, and a third technical problem to be solved by the present invention is to provide an application of the fluorescent pH-sensitive antibacterial hydrogel dressing in detecting wound infection.
In order to solve the technical problems, the technical scheme adopted by the invention is as follows:
a preparation method of a fluorescent pH-sensitive antibacterial hydrogel dressing comprises the following steps:
s1, dissolving citric acid and ciprofloxacin in ultrapure water, placing the ultrapure water in a reaction kettle, heating to 160-180 ℃, reacting for 2-3 hours, taking out, cooling, and dialyzing by using a dialysis bag to obtain a transparent carbon quantum dot solution; the molar ratio of the citric acid to the ciprofloxacin is 3-7: 1;
s2, dissolving chitosan in 1% acetic acid aqueous solution to prepare chitosan solution, adding gelatin aqueous solution, cross-linking agent glutaraldehyde aqueous solution and plasticizer glycerol to obtain mixed solution, reacting for 10min at 50-80 ℃ to obtain chitosan hydrogel, and freeze-drying the gel to obtain xerogel; the mass ratio of the chitosan to the gelatin is 1: 1-3, and the volume ratio of the chitosan solution to the cross-linking agent glutaraldehyde aqueous solution is 10: 1-2;
s3, putting the xerogel prepared in the S2 into a carbon quantum dot solution to obtain the fluorescent pH-sensitive antibacterial hydrogel dressing containing the carbon quantum dots.
Further, in step S2, in the mixed solution, the mass percentage concentration of chitosan is 1 to 3%, the mass percentage concentration of gelatin is 5 to 15%, and the mass percentage concentration of glutaraldehyde is 0.1 to 1%.
Further, in step S2, in the mixed solution, the mass percentage concentration of chitosan is 1.5-2.5%, and the mass percentage concentration of gelatin is 8-12%.
Preferably, in step S2, the mixed solution contains chitosan 2 wt%, gelatin 10 wt%, and glutaraldehyde 0.5 wt%.
Further, in step S2, the mass ratio of chitosan to gelatin is 1: 2.
Further, in step S2, the volume ratio of glycerin to water in the glycerin water solution is 0.01-0.02: 1.
The fluorescent pH-sensitive antibacterial hydrogel dressing prepared by the method.
The hydrogel dressing is coated on a wound, an ultraviolet lamp is used for irradiating, the change of fluorescence intensity is observed, the pH value of the environment where the hydrogel dressing is located is found to be in positive correlation with the fluorescence intensity, the fluorescence is gradually reduced along with the reduction of the pH value of the environment, and the fluorescence intensity is gradually enhanced along with the increase of the pH value.
The fluorescent pH-sensitive antibacterial hydrogel dressing is applied to promotion of skin wound repair and monitoring of change of pH value of a wound, and real-time monitoring of wound healing state can be achieved.
Has the beneficial effects that: compared with the prior art, the invention has the advantages that:
the invention provides a preparation method of a fluorescent pH-sensitive antibacterial hydrogel dressing, which comprises the steps of firstly obtaining chitosan hydrogel from chitosan and gelatin, then placing the chitosan hydrogel in a carbon quantum dot solution after freeze drying, and enabling carbon quantum dots to enter a gel network, wherein the gel has a three-dimensional network structure and is beneficial to cell adhesion and tissue growth. The dressing not only promotes the repair of skin wounds, but also can be used for reflecting the pH state of the wounds, the hydrogel dressing is coated on the wounds, the fluorescence of the hydrogel dressing is gradually reduced along with the reduction of the pH value of the environment, and the fluorescence intensity is gradually enhanced along with the increase of the pH value. Based on the change of the pH value of the wound, the change of the pH value of the wound can be monitored in real time through ultraviolet light excitation, and simple visual monitoring is realized.
Drawings
FIG. 1 is a reference diagram of a method of using a fluorescent hydrogel dressing made in accordance with the present invention;
FIG. 2 is a graph of fluorescence intensity as a function of pH for quantum dots prepared in accordance with the present invention;
FIG. 3 is a linear plot of quantum dots prepared according to the present invention at different pH;
FIG. 4 is a schematic view of UV irradiation at different pH for a fluorescent hydrogel prepared according to the present invention; the left part of the graph is simulated normal skin pH5.8; figure right is simulated infection wound ph 7.5;
FIG. 5 is a graph of the effect of the gel on a simulated wound according to the present invention;
FIG. 6 is a scatter plot of water loss versus time for the present invention;
FIG. 7 is a graph of water absorption as a function of time for the present invention.
Detailed Description
The invention is further described with reference to specific examples. These examples are intended to illustrate the invention and are not intended to limit the scope of the invention. Modifications or substitutions to methods, steps or conditions of the present invention may be made without departing from the spirit and scope of the invention. In the following examples, the technical means used in the examples are conventional means well known to those skilled in the art, unless otherwise specified.
Example 1
Weighing 2g of citric acid and 0.4g of ciprofloxacin, dissolving the citric acid and the ciprofloxacin in 5mL of water, transferring the mixture into a polytetrafluoroethylene reaction kettle, reacting for 3 hours at 180 ℃, and dialyzing for 24 hours by using a 3000KDa dialysis bag to obtain a transparent carbon quantum dot solution for later use.
0.5g of chitosan was dissolved in 20ml of a 1% aqueous acetic acid solution, 1g of gelatin was dissolved in 10ml of an aqueous solution, and 3ml of glutaraldehyde with a mass concentration of 0.25% and 0.5ml of glycerol were added. Reacting at 70 ℃ for 10min to obtain the chitosan hydrogel. And (3) freezing the chitosan hydrogel in a refrigerator at the temperature of-22 ℃ for 24h, and then freeze-drying the chitosan hydrogel in a freeze dryer for 72h to obtain the chitosan xerogel.
And soaking the dried gel in a carbon quantum dot solution, and fully reacting to make the carbon quantum dots enter a gel network to obtain the fluorescent hydrogel dressing. In the dressing, carbon quantum dots are present in the gel network by ionic interaction.
The prepared fluorescent pH-sensitive antibacterial hydrogel and each intermediate product are respectively detected, and the results are as follows:
FIG. 2 is a graph showing the change of fluorescence intensity of the prepared quantum dots with pH, and the detection method specifically comprises the following steps: the quantum dot solution is diluted 200 times, 1ml is placed in 5ml PBS buffer solutions with different pH values for 20min, and then the fluorescence intensity of the mixed solution is measured. As can be seen from fig. 2, the fluorescence intensity of the prepared carbon quantum dot solution gradually increased with increasing pH.
FIG. 3 is a fluorescence linear graph at different pH values, wherein 360nm is selected as the excitation wavelength, and the highest value of the fluorescence values at different pH values is taken at the same excitation wavelength. And drawing a pH linear graph by taking the pH as an abscissa and the fluorescence value as an ordinate.
FIG. 4 shows fluorescence intensities of the prepared fluorescent pH-sensitive antibacterial hydrogel under different pH values, and the detection method specifically comprises the following steps: the prepared chitosan xerogel is put into quantum dot solutions with different pH values for soaking, and after full reaction, redundant quantum dot solutions are wiped off. And then, the fluorescence intensity of the fluorescent hydrogel is obtained under a gel ultraviolet lamp. As can be seen, the fluorescence intensity of the fluorescent hydrogel gradually increased with increasing pH.
FIG. 5 is a view showing the effect of the gel on wound simulation, when in use, the fluorescent hydrogel dressing is covered on the wound, after a period of time (e.g. 30min), the ultraviolet lamp with the wavelength of 360nm is used for irradiation, the fluorescence intensity is observed, and if the wound is infected, the fluorescence on the wound becomes bright; the dressing is continuously applied, when needed, the change of the fluorescence intensity is observed by using an ultraviolet lamp, the change trend of the pH value of the wound can be determined through the change of the fluorescence intensity, the wound healing or infection state is further determined, and the real-time monitoring of the pH value of the wound is realized.
Example 2
Weighing 2g of citric acid and 0.4g of ciprofloxacin, dissolving in 5mL of water, transferring into a polytetrafluoroethylene reaction kettle, reacting for 3h at 180 ℃, and dialyzing for 24h by using a 3000KDa dialysis bag for later use.
1g of chitosan was dissolved in 20ml of aqueous solution, 1g of gelatin was dissolved in 10ml of aqueous solution, and 3ml of glutaraldehyde with a mass concentration of 0.25% and 0.5ml of glycerol were added. Reacting at 60 ℃ for 20min to obtain the chitosan hydrogel. And (3) freezing the chitosan hydrogel in a refrigerator at the temperature of-22 ℃ for 24h, and then freeze-drying the chitosan hydrogel in a freeze dryer for 72h to obtain the chitosan xerogel.
And soaking the dried gel in a carbon quantum dot solution, and fully reacting to enable the carbon quantum dots to enter a gel network to obtain the fluorescent hydrogel dressing. In the dressing, carbon quantum dots are present in the gel network by ionic interaction.
The prepared fluorescent pH-sensitive antibacterial hydrogel and each intermediate product were tested separately, and the results were similar to those in example 1.
Example 3
Weighing 2g of citric acid and 0.4g of ciprofloxacin, dissolving in 5mL of water, transferring into a polytetrafluoroethylene reaction kettle, reacting for 3h at 170 ℃, and dialyzing for 24h by using a 3000KDa dialysis bag for later use.
0.5g of chitosan was dissolved in 20ml of 1% aqueous acetic acid solution, 0.8g of gelatin was dissolved in 10ml of aqueous solution, and 2ml of glutaraldehyde with a mass concentration of 0.25% and 1ml of glycerol were added. Reacting at 70 ℃ for 20min to obtain the chitosan hydrogel. And (3) freezing the chitosan hydrogel in a refrigerator at the temperature of-22 ℃ for 24h, and then freeze-drying the chitosan hydrogel in a freeze dryer for 72h to obtain the chitosan xerogel.
And soaking the dried gel in a carbon quantum dot solution, and fully reacting to make the carbon quantum dots enter a gel network to obtain the fluorescent hydrogel dressing. In the dressing, carbon quantum dots are present in the gel network by ionic action.
The prepared fluorescent pH-sensitive antibacterial hydrogel and each intermediate product were tested separately, and the results were similar to those in example 1.
The preferred embodiments of the present invention have been described above in detail, but the present invention is not limited thereto. Within the scope of the technical idea of the invention, many simple modifications can be made to the technical solution of the invention, including various technical features being combined in any other suitable way, and these simple modifications and combinations should also be regarded as the disclosure of the invention, and all fall within the scope of the invention.
Claims (8)
1. A preparation method of a fluorescent pH-sensitive antibacterial hydrogel dressing is characterized by comprising the following steps:
s1, dissolving citric acid and ciprofloxacin in ultrapure water, placing the ultrapure water in a reaction kettle, heating to 160-180 ℃, reacting for 2-3 hours, taking out, cooling, and dialyzing by using a dialysis bag to obtain a transparent carbon quantum dot solution; the molar ratio of the citric acid to the ciprofloxacin is 3-7: 1;
s2, dissolving chitosan in 1% acetic acid aqueous solution to prepare chitosan solution, adding gelatin aqueous solution, cross-linking agent glutaraldehyde aqueous solution and plasticizer glycerol aqueous solution to obtain mixed solution, reacting for 10min at 50-80 ℃ to obtain chitosan hydrogel, and freeze-drying the gel to obtain xerogel; the mass ratio of the chitosan to the gelatin is 1: 1-3, and the volume ratio of the chitosan solution to the cross-linking agent glutaraldehyde aqueous solution is 10: 1-2;
s3, putting the xerogel prepared in the S2 into a carbon quantum dot solution to obtain the fluorescent pH-sensitive antibacterial hydrogel dressing containing the carbon quantum dots.
2. The method for preparing a fluorescent pH-sensitive antibacterial hydrogel dressing according to claim 1, wherein in step S2, the mass percent concentration of chitosan is 1-3%, the mass percent concentration of gelatin is 5-15%, and the mass percent concentration of glutaraldehyde is 0.1-1% in the mixed solution.
3. The method for preparing a fluorescent pH-sensitive antibacterial hydrogel dressing according to claim 2, wherein in step S2, the concentration of chitosan is 2% by mass, the concentration of gelatin is 10% by mass, and the concentration of glutaraldehyde is 0.5% by mass in the mixed solution.
4. The method for preparing a fluorescent pH-sensitive antibacterial hydrogel dressing according to claim 1, wherein in step S2, the mass ratio of chitosan to gelatin is 1: 2.
5. The method for preparing a fluorescent pH-sensitive antibacterial hydrogel dressing according to claim 1, wherein in step S2, the volume ratio of glycerol to water in the glycerol aqueous solution is 0.01-0.02: 1.
6. The fluorescent pH-sensitive antibacterial hydrogel dressing prepared by the method of any one of claims 1 to 5.
7. The fluorescent pH-sensitive antimicrobial hydrogel dressing according to claim 6, wherein the pH of the environment in which the hydrogel dressing is placed is positively correlated with the fluorescence intensity.
8. Use of the fluorescent pH-sensitive antimicrobial hydrogel dressing of claim 6 for promoting skin wound repair and monitoring changes in wound pH.
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