CN113278495A - Model and method for simulating infection wound for testing antibacterial effect of antibacterial dressing - Google Patents
Model and method for simulating infection wound for testing antibacterial effect of antibacterial dressing Download PDFInfo
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Abstract
The invention discloses a model and a method for simulating an infected wound for testing the antibacterial effect of an antibacterial dressing, wherein the model comprises the following steps: the wound infection simulation chamber is used for placing a substrate for simulating human tissues, the dressing placing chamber is clamped at the mouth of the wound infection simulation chamber and is positioned above the substrate, the through hole is arranged below the dressing placing chamber and is used for simulating an infected wound, and the sealing cover is arranged on the dressing placing chamber; according to the invention, infected wounds with different degrees can be accurately simulated through structural design, and the antibacterial performance of the antibacterial dressing for different infected wounds can be objectively and quantitatively evaluated by matching with a test method.
Description
Technical Field
The invention relates to the field of dressing production, in particular to a model and a method for simulating an infected wound to be used for testing the antibacterial effect of an antibacterial dressing.
Background
Wound infection refers to local tissue and systemic inflammatory response of aseptic wounds and lightly infected wounds caused by invasion of pathogens such as bacteria, viruses, fungi, parasites and the like, and is generally divided into acute wound infection and chronic wound infection. Acute wound infections are generally caused by a single microorganism, whereas chronic wound infections often involve multiple pathogens.
Infection complicates the wound situation and delays the healing process, and recent studies have shown that the presence of microbial biofilms plays a more significant role in the development and progression of chronic wounds than planktonic bacteria. More than 90% of chronic wounds are infected with biofilm, while 6% of acute wounds are also biofilm. The self-protective capacity and toxicity of biofilms appear to be extremely difficult to remove completely from chronically infected wound beds.
The dressing covers the wound surface, provides a mechanical protection barrier for the wound surface, is used for absorbing or draining the secretion of the wound, thereby controlling the microbial environment of the wound surface and directly or indirectly promoting the healing of the wound. The medical dressings are various in types, and the early days, the traditional dressings such as gauze, cotton pads and the like exist. With the acceptance of the theory of 'moist wound healing' and the rapid development of various novel high polymer materials in recent years, various functionalized and diversified novel dressings such as various synthetic dressings, biological dressings and the like are widely applied, typically representing various foam dressings and alginate dressings, and most of the existing antibacterial dressings are silver-containing dressings.
Currently, the in vitro detection method for the antibacterial performance of the antibacterial dressing generally comprises a diffusion method, a quinine method, a Maximum Inhibitory Concentration (MIC) method, a flask oscillation method, a dipping method and the like. The first three are qualitative methods, the last two are quantitative methods. The drawbacks of these methods are: (1) infected wounds of varying degrees cannot be accurately simulated, for example, acute infected wounds are generally infected with planktonic bacteria, while 80% of chronic wounds are infected with biofilms. (2) Can not be accurately and objectively evaluated, and is used for the antibacterial effect of wound dressings with different infections. The market needs an antibacterial dressing which can accurately simulate infected wounds with different degrees and objectively and quantitatively evaluate the antibacterial performance of the antibacterial dressing for the different infected wounds, and the invention solves the problems.
Disclosure of Invention
In order to overcome the defects of the prior art, the invention aims to provide a model and a method for simulating an infected wound for testing the antibacterial effect of an antibacterial dressing.
In order to achieve the above object, the present invention adopts the following technical solutions:
a model for simulating an infected wound for testing the antimicrobial effect of an antimicrobial dressing, comprising: the wound infection simulation chamber is used for placing a substrate for simulating human tissues, the dressing placing chamber is clamped at the mouth of the wound infection simulation chamber and positioned above the substrate, a through hole is arranged below the dressing placing chamber and used for simulating an infected wound, and a sealing cover is arranged on the dressing placing chamber.
The model for simulating the infection wound to be used for the antibacterial effect test of the antibacterial dressing is characterized in that the through hole is a circular hole, and the diameter of the circular hole simulates the size of the infection wound.
In the model for simulating the infected wound and testing the antibacterial effect of the antibacterial dressing, the sealing cover is a bandage, gauze or transparent application.
The model for simulating the infected wound for the antibacterial effect test of the antibacterial dressing further comprises: a movable barrier extending across the dressing placement chamber and engaged to the wound infection simulation chamber.
In the model for simulating an infected wound and testing the antibacterial effect of the antibacterial dressing, the movable barrier is a barrier plate.
A method for simulating an infected wound for testing the antibacterial effect of an antibacterial dressing comprises the following steps:
step one, sterilizing a model for later use;
the model comprises the following steps: the wound infection simulation chamber is used for placing a substrate for simulating human tissues, the dressing placing chamber is clamped at the mouth of the wound infection simulation chamber and is positioned above the substrate, the through hole is arranged below the dressing placing chamber and is used for simulating an infected wound, and the sealing cover is arranged on the dressing placing chamber;
step two, preparing a matrix solution;
step three, preparing a neutralizer;
pouring the prepared matrix into a wound infection simulation chamber, putting the wound infection simulation chamber into a dressing placing chamber after the surface of the matrix is smooth and flat, enabling the matrix to be in full contact with the through hole at the bottom of the dressing placing chamber, and standing for 30 min;
preparing a wound-feel dye, and culturing the wound-feel dye on a substrate below a through hole at the bottom of the dressing placing chamber;
sixthly, placing the dressing at the bottom of the dressing placing chamber to enable the dressing to be in full contact with the wound infection substances;
step seven, preparing the exudate, and adding the exudate into the dressing according to different gradients;
step eight, culturing the sealed model;
and step nine, taking out the model and analyzing the antibacterial effect.
According to the method for simulating the infected wound for testing the antibacterial effect of the antibacterial dressing, if the antibacterial effect of the antibacterial dressing in an acute infected wound or an acute wound with exudate needs to be tested, the wound infection is bacterial suspension.
According to the method for simulating the infected wound for testing the antibacterial effect of the antibacterial dressing, if the antibacterial effect of the antibacterial dressing in a chronic infected wound or a chronic wound with exudate needs to be tested, the wound infection substance is bacterial suspension.
In the method for simulating the infected wound for testing the antibacterial effect of the antibacterial dressing, the neutralizing agent is PBS phosphate buffer solution.
In the method for simulating the infected wound to test the antibacterial effect of the antibacterial dressing, the model sterilization adopts a pressure steam sterilization pot for sterilization at 132 ℃ for 30min.
The invention has the advantages that:
the through hole of the dressing placing chamber can accurately simulate infected wounds with different degrees, such as acute infected wounds and chronic infected wounds; therefore, the antibacterial effect of the antibacterial dressing on the acute infection wound can be objectively and quantitatively evaluated;
the method can accurately evaluate the damage and removal effect of the antibacterial dressing on the chronic infection wound biomembrane, can truly reflect the continuous antibacterial effect of the antibacterial dressing on the infected wound, and scientifically reflect the antibacterial effect of the antibacterial dressing on the wound infected with different exudates;
the wound infection simulation chamber and the dressing placing chamber are made of materials, so that constant temperature can be realized by combining with other equipment;
the wound infection simulation chamber and the dressing placing chamber are made of transparent materials, so that the influence of different exudates on the antibacterial effect of the dressing can be reflected visually;
the wound infection simulation room and the dressing placing room are made of high-temperature-resistant materials such as glass and can be sterilized by pressure steam, on one hand, the accuracy of a test result can be guaranteed, and on the other hand, the model can be used repeatedly.
Drawings
FIG. 1 is a schematic structural diagram of an embodiment of the present invention;
FIG. 2 is a schematic cross-sectional view of one embodiment of the present invention;
FIG. 3 is a schematic diagram of the configuration of one embodiment of the dressing placement chamber and the movable barrier of the present invention;
the meaning of the reference symbols in the figures:
1 sealing cover, 2 removable barrier, 3 dressing placing chamber, 301 through hole, 4 wound infection simulation chamber, 5 substrate.
Detailed Description
The invention is described in detail below with reference to the figures and the embodiments.
A model for simulating an infected wound for testing the antimicrobial effect of an antimicrobial dressing, comprising: the wound infection simulation chamber comprises a wound infection simulation chamber 4 for placing a substrate 5 for simulating human tissue, a dressing placing chamber 3 clamped at the mouth of the wound infection simulation chamber 4 and positioned above the substrate 5, a through hole 301 arranged below the dressing placing chamber 3 and used for simulating an infected wound, a sealing cover 1 arranged on the dressing placing chamber 3, and a movable blocking piece 2 extending to the dressing placing chamber 3 and clamped on the wound infection simulation chamber 4.
Preferably, the wound infection simulation chamber 4 and the dressing placing chamber 3 are both designed to be bottled made of glass materials; the transparent material can visually reflect the influence of different exudates on the antibacterial effect of the dressing; because the high-temperature resistant material is adopted, pressure steam sterilization can be carried out, on one hand, the accuracy of a test result can be ensured, and on the other hand, the model can be repeatedly used; the material is not limited, and the transparent high temperature resistant glass can be applied to the invention.
Preferably, the through holes 301 are circular holes, and the diameter of the circular holes simulates the size of an infected wound; the through hole 301 of the dressing placing chamber 3 can accurately simulate infected wounds with different degrees, such as acute infected wounds and chronic infected wounds; therefore, the antibacterial effect of the antibacterial dressing on the acute infection wound can be objectively and quantitatively evaluated.
The material of the sealing cover 1 is selected according to the simulated wound, such as bandage, gauze, transparent application and the like, and the material is not limited, so long as the sealing effect can be achieved, and the invention can be applied.
The shape and material of the movable blocking piece 2 are not limited, and the movable blocking piece can be a blocking plate as shown in fig. 1, a lifting handle, a handle or a clamping flange, and the movable blocking piece can be applied to the wound infection simulation chamber 4 as long as the movable blocking piece can extend to the dressing placing chamber 3 and is stably fixed on the wound infection simulation chamber. Such design is convenient to pick up the baffler and is taken out whole dressing placing chamber 3, realizes that the dressing easily gets rid of after using different days.
Example one, test method of antimicrobial dressing in an acute infected wound:
1. and (4) sterilizing the model, wherein the model is sterilized in a pressure steam sterilizing pot for 30min at 132 ℃ before test.
2. And after the sterilization is finished, taking out and placing the biological safety cabinet for later use.
3. Preparing a matrix 5 liquid, wherein the matrix 5 liquid is provided with nutrient components with different concentrations according to different test requirements, the main components of the matrix 5 liquid are bovine serum albumin, tryptone, sodium chloride and agar powder, and the preparation is carried out under an aseptic condition.
4. Preparing a neutralizing agent, wherein the neutralizing agent is PBS phosphate buffer solution.
5. And pouring the prepared matrix 5 into the wound infection simulation chamber 4, putting the matrix 5 into the dressing placing chamber 3 after the surface of the matrix 5 is smooth and flat, enabling the matrix 5 to be in full contact with the round hole at the bottom of the dressing placing chamber 3, and standing for 30min.
6. And (3) preparing a bacterial suspension, and selecting four bacteria of candida albicans, escherichia coli, pseudomonas aeruginosa and staphylococcus aureus for verification because 90% of acute infection wounds are infected by single bacteria. The bacterial suspension was prepared according to the sterilization specification 2002 edition 2.1.1.2.3 bacterial suspension preparation procedure.
7. 100 microliter of the prepared bacterial suspension is uniformly smeared in the matrix 5 in the round hole at the bottom of the dressing placing chamber 3 and stands for 10 min.
8. The antibacterial dressing is cut to a proper size and placed on the round hole at the bottom in the dressing placing chamber 3, and the dressing is lightly pressed to be in full contact with the infected wound.
9. The mold was sealed with a suitable material and incubated at a constant temperature of 36.5 ℃. The dressings are taken out on different days, then the dressings are placed in a chamber 3, a neutralizing agent is added into a wound infection simulation chamber 4 to be fully contacted with the infected wound surface, 1m of the dressings are taken out and placed in a sterile culture dish, and the sterile culture dish is placed at 37 ℃ for culture. .
10. And quantitatively evaluating the antibacterial effect of the antibacterial dressing according to the bacterial quantity in the culture dish.
Example two, test method of antimicrobial dressing in chronically infected wounds:
1. and (4) sterilizing the model, wherein the model is sterilized in a pressure steam sterilizing pot for 30min at 132 ℃ before test.
2. And after the sterilization is finished, taking out and placing the biological safety cabinet for later use.
3. Preparing a matrix 5 liquid, wherein the matrix 5 liquid is provided with nutrient components with different concentrations according to different test requirements, the main components of the matrix 5 liquid are bovine serum albumin, tryptone, sodium chloride and agar powder, and the preparation is carried out under an aseptic condition.
4. Preparing a neutralizing agent, wherein the neutralizing agent is PBS phosphate buffer solution.
5. And pouring the prepared matrix 5 into the wound infection simulation chamber 4, putting the matrix 5 into the dressing placing chamber 3 after the surface of the matrix 5 is smooth and flat, enabling the matrix 5 to be in full contact with the round hole at the bottom of the dressing placing chamber 3, and standing for 30min.
6. Preparation of biofilm, 80% of chronic wounds are infected by biofilm, so the biofilm needs to be prepared for testing.
7. And (3) culturing the biological membrane in a round hole at the bottom in the dressing placing chamber 3, and after the biological membrane is mature, cutting the dressing into a proper size and placing the dressing on the biological membrane to make the dressing fully contact with the biological membrane.
8. Sealing the model, placing in an incubator at 36.5 ℃, and culturing at constant temperature.
9. Different time gradients are set, the model is taken out, the dressing placing chamber 3 is taken out, then the biological membrane is treated, and the damage effect of the antibacterial dressing on the biological membrane is analyzed.
EXAMPLE III test method of antimicrobial dressing in acute wound with exudate
1. And (4) sterilizing the model, wherein the model is sterilized in a pressure steam sterilizing pot for 30min at 132 ℃ before test.
2. And after the sterilization is finished, taking out and placing the biological safety cabinet for later use.
3. Preparing a matrix 5 liquid, wherein the matrix 5 liquid is provided with nutrient components with different concentrations according to different test requirements, the main components of the matrix 5 liquid are bovine serum albumin, tryptone, sodium chloride and agar powder, and the preparation is carried out under an aseptic condition.
4. Preparing a neutralizing agent, wherein the neutralizing agent is PBS phosphate buffer solution.
5. And pouring the prepared matrix 5 into the wound infection simulation chamber 4, putting the matrix 5 into the dressing placing chamber 3 after the surface of the matrix 5 is smooth and flat, enabling the matrix 5 to be in full contact with the round hole at the bottom of the dressing placing chamber 3, and standing for 30min.
6. And (3) preparing a bacterial suspension, and selecting four bacteria of candida albicans, escherichia coli, pseudomonas aeruginosa and staphylococcus aureus for verification because 90% of acute infection wounds are infected by single bacteria. The bacterial suspension was prepared according to the sterilization specification 2002 edition 2.1.1.2.3 bacterial suspension preparation procedure.
7. 100 microliter of the prepared bacterial suspension is uniformly smeared in the matrix 5 in the round hole at the bottom of the dressing placing chamber 3 and stands for 10 min.
8. The antibacterial dressing is cut to a proper size and placed on the round hole at the bottom in the dressing placing chamber 3, and the dressing is lightly pressed to be in full contact with the infected wound.
9. And (3) preparing the exudate, wherein the exudate is configured according to 'comparison of water absorption, water locking and air permeability of various foam dressings'.
10. The exudate is added into the dressing according to different gradients.
11. The mold was sealed with a suitable material and incubated at a constant temperature of 36.5 ℃. The dressings are taken out on different days, then the dressings are placed in a chamber 3, a neutralizing agent is added into a wound infection simulation chamber 4 to be fully contacted with the infected wound surface, 1m of the dressings are taken out and placed in a sterile culture dish, and the sterile culture dish is placed at 37 ℃ for culture. .
12. And quantitatively evaluating the antibacterial effect of the antibacterial dressing according to the bacterial quantity in the culture dish.
Example four, test method of antimicrobial dressing in chronic wound with exudate.
1. And (4) sterilizing the model, wherein the model is sterilized in a pressure steam sterilizing pot for 30min at 132 ℃ before test.
2. And after the sterilization is finished, taking out and placing the biological safety cabinet for later use.
3. Preparing a matrix 5 liquid, wherein the matrix 5 liquid is provided with nutrient components with different concentrations according to different test requirements, the main components of the matrix 5 liquid are bovine serum albumin, tryptone, sodium chloride and agar powder, and the preparation is carried out under an aseptic condition.
4. Preparing a neutralizing agent, wherein the neutralizing agent is PBS phosphate buffer solution.
5. And pouring the prepared matrix 5 into the wound infection simulation chamber 4, putting the matrix 5 into the dressing placing chamber 3 after the surface of the matrix 5 is smooth and flat, enabling the matrix 5 to be in full contact with the round hole at the bottom of the dressing placing chamber 3, and standing for 30min.
6. Preparation of biofilm, 80% of chronic wounds are infected by biofilm, so the biofilm needs to be prepared for testing.
7. And (3) culturing the biological membrane in a round hole at the bottom in the dressing placing chamber 3, and after the biological membrane is mature, cutting the dressing into a proper size and placing the dressing on the biological membrane to make the dressing fully contact with the biological membrane.
8. And (3) preparing the exudate, wherein the exudate is configured according to 'comparison of water absorption, water locking and air permeability of various foam dressings'.
9. The exudate is added into the dressing according to different gradients.
10. Sealing the model, placing in an incubator at 36.5 ℃, and culturing at constant temperature.
11. Different time gradients are set, the model is taken out, the dressing placing chamber 3 is taken out, then the biological membrane is treated, and the damage effect of the antibacterial dressing on the biological membrane is analyzed.
According to the invention, through the cooperation of the structural design and the method, the infected wounds with different degrees can be simulated accurately, and the antibacterial performance of the antibacterial dressing for different infected wounds can be evaluated objectively and quantitatively.
The foregoing illustrates and describes the principles, general features, and advantages of the present invention. It should be understood by those skilled in the art that the above embodiments do not limit the present invention in any way, and all technical solutions obtained by using equivalent alternatives or equivalent variations fall within the scope of the present invention.
Claims (10)
1. A model for simulating an infected wound for testing the antibacterial effect of an antibacterial dressing, comprising: the wound infection simulation chamber is used for placing a substrate for simulating human tissues, the dressing placing chamber is clamped at the mouth of the wound infection simulation chamber and positioned above the substrate, a through hole is arranged below the dressing placing chamber and used for simulating an infected wound, and a sealing cover is arranged on the dressing placing chamber.
2. The model for simulating an infected wound for testing the antibacterial effect of the antibacterial dressing according to claim 1, wherein the through holes are circular holes, and the diameter of each circular hole simulates the size of the infected wound.
3. The model for simulating an infected wound for testing the antibacterial effect of an antibacterial dressing according to claim 1, wherein the sealing cover is a bandage, gauze or transparent application.
4. The model for simulating an infected wound for testing the antibacterial effect of the antibacterial dressing according to claim 1, further comprising: a movable barrier extending across the dressing placement chamber and engaged to the wound infection simulation chamber.
5. The model for simulating an infected wound for testing the antibacterial effect of an antibacterial dressing according to claim 4, wherein the movable barrier is a barrier plate.
6. A method for simulating an infected wound for testing the antibacterial effect of an antibacterial dressing is characterized by comprising the following steps:
step one, sterilizing a model for later use;
the model comprises: the wound infection simulation chamber is used for placing a substrate for simulating human tissues, the dressing placing chamber is clamped at the mouth of the wound infection simulation chamber and is positioned above the substrate, the through hole is arranged below the dressing placing chamber and is used for simulating an infected wound, and the sealing cover is arranged on the dressing placing chamber;
step two, preparing a matrix solution;
step three, preparing a neutralizer;
pouring the prepared matrix into a wound infection simulation chamber, putting the wound infection simulation chamber into a dressing placing chamber after the surface of the matrix is smooth and flat, enabling the matrix to be in full contact with the through hole at the bottom of the dressing placing chamber, and standing for 30 min;
preparing a wound-feel dye, and culturing the wound-feel dye on a substrate below a through hole at the bottom of the dressing placing chamber;
sixthly, placing the dressing at the bottom of the dressing placing chamber to enable the dressing to be in full contact with the wound infection substances;
step seven, preparing the exudate, and adding the exudate into the dressing according to different gradients;
step eight, culturing the sealed model;
and step nine, taking out the model and analyzing the antibacterial effect.
7. The method for simulating an infected wound for testing the antibacterial effect of the antibacterial dressing according to claim 6, wherein if the antibacterial dressing needs to be tested for the antibacterial effect in an acute infected wound or an acute wound with exudates, the wound-feeling infection is an bacterial suspension.
8. The method for simulating an infected wound for testing the antibacterial effect of the antibacterial dressing according to claim 6, wherein if the antibacterial dressing needs to be tested for the antibacterial effect in a chronic infected wound or a chronic wound with exudates, the wound-feeling pollutant is an antibacterial suspension.
9. The method for simulating an infected wound for testing the antibacterial effect of the antibacterial dressing according to claim 6, wherein the neutralizing agent is PBS phosphate buffer.
10. The method for simulating an infected wound for testing the antibacterial effect of the antibacterial dressing according to claim 6,
the model sterilization adopts a pressure steam sterilization pot for sterilization at 132 ℃ for 30min.
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