CN112326743A - C-SF-FA flexible conductive film based on silk fibroin, wearable wound monitoring sensor and preparation method of wearable wound monitoring sensor - Google Patents
C-SF-FA flexible conductive film based on silk fibroin, wearable wound monitoring sensor and preparation method of wearable wound monitoring sensor Download PDFInfo
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- G—PHYSICS
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/02—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating impedance
- G01N27/04—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating impedance by investigating resistance
- G01N27/12—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating impedance by investigating resistance of a solid body in dependence upon absorption of a fluid; of a solid body in dependence upon reaction with a fluid, for detecting components in the fluid
- G01N27/121—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating impedance by investigating resistance of a solid body in dependence upon absorption of a fluid; of a solid body in dependence upon reaction with a fluid, for detecting components in the fluid for determining moisture content, e.g. humidity, of the fluid
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/02—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating impedance
- G01N27/04—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating impedance by investigating resistance
- G01N27/12—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating impedance by investigating resistance of a solid body in dependence upon absorption of a fluid; of a solid body in dependence upon reaction with a fluid, for detecting components in the fluid
- G01N27/125—Composition of the body, e.g. the composition of its sensitive layer
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Abstract
The invention discloses a C-SF-FA flexible conductive film based on silk fibroin and a monitoring sensor made of the same, wherein the C-SF-FA flexible conductive film comprises the following components in parts by weight: 1) degumming silk, dissolving in a salt-formic acid system to obtain a silk fibroin solution, drying to obtain silk fibroin, soaking the silk fibroin in deionized water, and dissolving the dried silk fibroin in formic acid again to obtain an SF-FA electrospinning solution; 2) dissolving dopamine in water, and adding a carbon nano tube to obtain a DA-CNT solution; 3) preparing the SF-FA electrospinning solution into a flexible film through electrostatic spinning, and spraying the DA-CNT solution on the film for multiple times in the electrostatic spinning process; drying the film to obtain a C-SF-FA conductive film; 4) adding DA and CNT into an alkaline solution to obtain a PDA-CNT solution, soaking the C-SF-FA conductive film in the PDA-CNT, cleaning and drying, soaking in an ethanol or methanol solution, and drying to obtain the C-SF-FA flexible conductive film.
Description
Technical Field
The invention relates to the technical field of biological materials, in particular to a C-SF-FA flexible conductive film based on silk fibroin, a wearable wound monitoring sensor and a preparation method thereof.
Background
The skin is the first barrier of human immunity and plays a crucial role in protecting human health. When the skin is irreparably injured, the rest-free system of the human body is greatly influenced. Chronic wounds are the more common skin injuries, and the skin can be orderly repaired for slight burns, but when the wound area is too large and too deep, the healing capacity of the patient is insufficient, and the life of the patient is threatened. Thus, the management and monitoring of chronic wounds has received widespread worldwide attention.
Wound healing is a continuous and overlapping dynamic process, the whole process including three phases: inflammatory, proliferative and remodeling phases. The inflammatory phase is mainly responsible for initiating the immune response to signal various signs of wound repair; the proliferation stage is to perform migration of various key cells according to signals so as to close wounds and rebuild blood vessels; the remodeling phase is the period during which myofibroblasts and wound scarring gradually form. Because the entire healing process is extremely complex, improper treatment can cause repetition of the healing process and infection of the wound, thereby exacerbating the inflammatory response. Therefore, it is important to monitor the wound.
In order to prevent wound infection, various indexes have been used as observation targets for monitoring wound healing, such as protease, humidity, temperature, and blood oxygen level. The moisture balance of the wound is a recognized critical factor. Any wound healing is associated with moisture, a dry environment is not conducive to wound healing, and excessive moisture also increases the risk of wound infection. Therefore, medical staff needs to monitor the wound humidity in real time. Some humidity sensors have been developed, but these sensors have some disadvantages, such as poor biocompatibility, poor air permeability, and poor real-time data. In recent years, due to the rapid development of various biomaterials, it has become possible to design a wound real-time monitoring sensor with excellent biocompatibility and high conductivity.
Silk is a common purely natural bio-organic material. Silk has gained increasing interest in recent years due to its good biocompatibility and mechanical properties. Silk Fibroin (SF) accounts for 70% of the weight of silk and is the most predominant protein. It has been shown by research that silk fibroin does not persistently cause inflammatory reactions, so it plays a great role in chondrocyte culture, bone tissue scaffolds, and antibacterial dressings. Meanwhile, the excellent physical properties of the material enable the material to be used as an additive to be added into the material to enhance the mechanical properties of the material, and the material can be made into films, hydrogels, fibers and other scaffolds.
Among the nanofillers, Carbon Nanotubes (CNTs) have the advantage of significantly improving the properties of nanocomposites and are an ideal material for the preparation of high quality nanocomposites. Carbon nanotubes contain graphitic layers with lydi diameters (1-100nm) and large lengths (1-10 μm), resulting in large specific surface areas. In addition, the tensile modulus of the carbon nanotube is 1000GPa, and the strength is 10-50 GPa. Furthermore, the superconducting CNTs (105-106S/m) introduce the required conductivity in the insulating matrix. Therefore, the CNT has excellent physical, mechanical and electrical conductivity and is a promising filling material.
Disclosure of Invention
The invention aims to solve the problems and provides a C-SF-FA flexible conductive film based on silk fibroin and a preparation method thereof, wherein the C-SF-FA flexible conductive film has excellent electrical and mechanical properties and biocompatibility.
In order to achieve the purpose, the invention adopts the technical scheme that:
a preparation method of a C-SF-FA flexible conductive film based on silk fibroin comprises the following steps:
1) degumming silkworm cocoon, drying the degummed silk, dissolving the degummed silk in a salt-formic acid dissolving system to obtain a silk fibroin solution, drying the obtained silk fibroin solution to obtain silk fibroin, soaking the silk fibroin in deionized water to remove salt and formic acid, and dissolving the dried silk fibroin in formic acid again to obtain an SF-FA electrospinning solution;
2) dissolving dopamine in water, adding carbon nanotubes, and uniformly dispersing to obtain a DA-CNT solution;
3) preparing the SF-FA electrospinning solution into a flexible film through electrostatic spinning, and spraying the DA-CNT solution on the film for multiple times in the electrostatic spinning process; after the electrospinning is finished, drying the film in the air to obtain a C-SF-FA conductive film;
4) adding DA and CNT into an alkaline solution to obtain a PDA-CNT solution, placing the C-SF-FA conductive film into the PDA-CNT solution, soaking for 2-10 hours, taking out, cleaning with ultrapure water, drying, soaking the film into an ethanol or methanol solution for 45-60 minutes after drying, taking out, and drying to obtain the C-SF-FA conductive film.
The salt in the salt-formic acid dissolving system is selected from one or more of calcium chloride, lithium bromide and magnesium chloride; preferably, the concentration of the salt in the salt-formic acid dissolving system is 2-10 wv%, and the concentration of the formic acid is 90-98 wv%;
the pH value of the alkaline solution in the step 4) is 8-11, and the alkaline solution is preferably a sodium hydroxide aqueous solution or an ammonia aqueous solution.
In the DA-CNT solution or the PDA-CNT solution, the concentration of DA is 5-20 mg/ml, and the concentration of CNT is 1-10 mg/ml.
The mass fraction of silk fibroin in the SF-FA electrospinning solution in the step 1) is 15-25%;
the dosage of the DA-CNT solution in the step 3) is that 1ml of the DA-CNT solution is used for every 5-10 ml of SF-FA electrospinning solution.
The silk fibroin solution obtained by drying the silk fibroin solution in the step 1) is dried at room temperature for 12-24 hours;
soaking the silk fibroin in the deionized water in the step 1) for 10-24 hours to remove calcium chloride and formic acid.
Degumming the silkworm cocoon silk in the step 1) by adopting a sodium carbonate solution with the concentration of 0.5-2.0 mol/ml;
the ethanol or methanol solution in the step 4) is 75 percent by volume of ethanol or methanol water solution.
The electrostatic spinning working conditions in the step 3) are as follows: the voltage is 10-24KV, the electrospinning speed is 0.002-0.015ml/min, the receiving distance is 10-15cm, the temperature is controlled at 20-25 ℃, and the humidity is controlled at 40-60%.
A C-SF-FA flexible conductive film based on silk fibroin is prepared by adopting the method.
The invention also aims to provide a flexible wearable wound monitoring sensor based on silk fibroin, which is made of the C-SF-FA flexible conductive film based on silk fibroin.
The preparation method of the silk fibroin-based flexible wearable wound monitoring sensor comprises the following steps: and assembling the C-SF-FA flexible conductive film based on silk fibroin and a lead to obtain the flexible wearable wound monitoring sensor.
The salt-formic acid system can induce the silk fibroin to form beta-folding, which can enhance the mechanical property of the silk fibroin film and enhance the conductivity of the silk fibroin film after the CNT is added. The change of the wound healing degree is judged by applying a flexible wearable sensor on a wound and monitoring the change of the conductivity of the sensor after the sensor is soaked by tissue fluid. Because the film is in a three-dimensional porous net structure, when the wound sensor is contacted with the surface of a wound, the silk fibroin film expands after absorbing wound tissue fluid due to the high swelling property of the silk fibroin film, and then the resistivity changes. And finally, the balance degree of the water environment of the wound can be judged according to the change of the resistivity, so that the healing of the wound is monitored in real time.
The invention has the beneficial effects that: the C-SF-FA conductive film is prepared by using a simple electrostatic spinning technology, the reaction condition is mild, and the operation is simple and easy; the C-SF-FA flexible conductive film has a highly dense mesh support structure, so that the C-SF-FA flexible conductive film has excellent mechanical properties; and the CNT conductive interlayer in the flexible film enables the flexible film to have excellent conductivity. The flexible sensor can detect the moisture change of the wound by leading the overall conductivity change after absorbing the wound tissue fluid; the used material has excellent electrical and mechanical properties and biocompatibility, and the detection method has high sensitivity and can be expected to be applied to portable, remote and real-time health monitoring systems.
In addition, the invention has the following remarkable advantages: 1) high manufacturing costs have always been the biggest challenge that plagues the potential for flexible sensor applications. The invention does not utilize any micro-manufacturing technology in the whole process of manufacturing the flexible sensor, thereby further reducing the manufacturing cost of the flexible sensor. 2) The uniform mixing of the conductive material has always been a great problem, and the conductive material CNT and the spraying method used in the invention well solve the problem. 3) The excellent flexibility and biocompatibility of the material can greatly improve the problem of mismatching between tissues and devices. 4) The prepared flexible sensor has high sensitivity, good air permeability and excellent mechanical properties. The sensor can detect the humidity balance of various wounds.
Drawings
FIG. 1 is an electron micrograph of a C-SF-FA conductive film according to the present invention.
FIG. 2 is a schematic diagram of a sensor assembled by the C-SF-FA conductive film and the copper tape of the present invention.
FIG. 3 is a graph showing the results of measuring the response of the sensor of the present invention to the same simulated body fluid amount using example 1, wherein (a) is a graph showing the change in resistance, (b) is a graph showing the change in sensitivity, and (C) is a fitted straight line graph showing the C-SF-FA conductive film and the simulated body fluid absorption amount.
FIG. 4 is a graph showing the measurement reproducibility of the sensor of the present invention measured by using example 2, wherein (a) is a graph showing a change in resistance, (b) is a graph showing a change in sensitivity, and (C) is a fitted straight line graph of a C-SF-FA conductive film and a simulated body fluid absorption amount.
Detailed Description
The invention is further illustrated by the following examples, which are not intended to be limiting.
The experimental procedures in the following examples are conventional unless otherwise specified.
The main reagent sources are as follows:
dopamine hydrochloride (DA): CAS number 62-31-7, available from Biotechnology engineering (Shanghai) Inc.;
carbon Nanotube (CNT): the production company is Nanjing Jicang nanometer technology Co., Ltd, the product number: JCNT-95-50-15-W;
simulating the body fluid: the production company is Beijing Ku Laibobu technology Limited, the goods number: SL6710-500 ml;
the remaining reagents, if not indicated, were not conventional and were all commercially available.
Example 1:
preparation of a silk fibroin-based flexible wearable wound monitoring sensor, which is operated according to the following steps:
1) boiling and degumming domestic silkworm cocoon silk by using 2mol/L sodium carbonate solution, cleaning and naturally drying after degumming, dissolving the domestic silkworm cocoon silk in a calcium chloride/Formic Acid (FA) dissolving system (wherein the concentration of calcium chloride is 2 wv% and the concentration of formic acid is 98 wv%) for more than 3 hours after air drying to form a silk fibroin solution (SF solution), and drying the obtained SF solution at room temperature for 24 hours to obtain the silk fibroin. And then placing the obtained dry silk fibroin into deionized water to be soaked for 10h to remove redundant calcium chloride and formic acid, and airing. And drying the obtained silk fibroin at 35 ℃ and dissolving the silk fibroin in formic acid again to obtain SF-FA electrospinning solution with the Silk Fibroin (SF) content of 20 wv%.
2) 5mg of Dopamine (DA) is dissolved in 1ml of water, 5mg of Carbon Nano Tube (CNT) is added, and ultrasonic dispersion is carried out to obtain a DA-CNT solution.
3) The SF-FA electrospinning solution is prepared into a flexible film through electrostatic spinning, and 1ml of DA-CNT solution is sprayed on the film for multiple times in the electrostatic spinning process. The working conditions of electrostatic spinning are as follows: the voltage is 18KV, the electrospinning speed is 0.008ml/min, the receiving distance is 12cm, the temperature is controlled at 20-25 ℃, the humidity is controlled at 40-60%, the volume of the electrospinning solution is 8ml, and after the electrospinning is finished, the film is air-dried at room temperature to obtain the C-SF-FA conductive film.
4) 50mg DA was dissolved in 10ml sodium hydroxide solution (pH 8), 10mg CNT was added and dispersed by ultrasound to give PDA-CNT solution. The C-SF-FA conductive film is soaked in the PDA-CNT solution for 2 h. After the removal, excess CNT on the surface was washed off with ultrapure water and dried at room temperature. And after the film is dried, soaking the film in 75% ethanol solution for 45min, taking out the film, and after the film is taken out, air-drying the film at room temperature to finally obtain the C-SF-FA conductive film, wherein an electron microscope scanning image of the C-SF-FA conductive film is shown in figure 1.
5) The C-SF-FA conductive film and the copper tape are assembled into the flexible wearable sensor, and the schematic structural diagram is shown in fig. 2.
Example 2:
preparation of a silk fibroin-based flexible wearable wound monitoring sensor, which is operated according to the following steps:
1) boiling and degumming domestic silkworm cocoon silk by using 2mol/L sodium carbonate solution, cleaning and naturally drying after degumming, dissolving the silkworm cocoon silk in a lithium bromide/Formic Acid (FA) dissolving system (wherein the concentration of lithium bromide is 10 wv% and the concentration of formic acid is 98 wv%) after air drying for more than 3 hours to form a silk fibroin solution (SF solution), and drying the obtained SF solution at room temperature for 12 hours to obtain the silk fibroin. And then placing the obtained dry silk fibroin in deionized water for soaking for 10h to remove redundant lithium bromide and formic acid, and airing. And drying the obtained silk fibroin at 50 ℃ and dissolving the silk fibroin in formic acid again to obtain SF-FA electrospinning solution with the Silk Fibroin (SF) content of 20 wv%.
2) 5mg of Dopamine (DA) is dissolved in 1ml of water, 1mg of Carbon Nano Tube (CNT) is added, and ultrasonic dispersion is carried out to obtain a DA-CNT solution.
3) The SF-FA electrospinning solution is prepared into a flexible film through electrostatic spinning, and 1ml of DA-CNT solution is sprayed on the film for multiple times in the electrostatic spinning process. The voltage is 24KV, the electrospinning speed is 0.015ml/min, the receiving distance is 15cm, the temperature is controlled to be 20-25 ℃, the humidity is controlled to be 40-60%, the volume of the electrospinning solution is 10ml, and after the electrospinning is finished, the film is air-dried at room temperature to obtain the C-SF-FA conductive film.
4) 200mgDA was dissolved in 10ml of an aqueous ammonia solution (pH 11), and 100mgCNT was added thereto and then ultrasonically dispersed to obtain a PDA-CNT solution. The C-SF-FA conductive film was soaked in the PDA-CNT solution for 4 h. After the removal, excess CNT on the surface was washed off with ultrapure water and dried at room temperature. And after the film is dried, soaking the film in 75% methanol solution for 45min, taking out the film, and after the film is taken out, air-drying the film at room temperature to finally obtain the C-SF-FA conductive film.
5) And assembling the C-SF-FA conductive film and the copper strip into the flexible wearable sensor.
Example 3:
preparation of a silk fibroin-based flexible wearable wound monitoring sensor, which is operated according to the following steps:
1) boiling and degumming domestic silkworm cocoon silk by using 2mol/L sodium carbonate solution, cleaning and naturally drying after degumming, dissolving the domestic silkworm cocoon silk in a calcium chloride/Formic Acid (FA) dissolving system (wherein the concentration of calcium chloride is 5 wv% and the concentration of formic acid is 98 wv%) for more than 3 hours after air drying to form a silk fibroin solution (SF solution), and drying the obtained SF solution at room temperature for 24 hours to obtain the silk fibroin. And then placing the obtained dry silk fibroin into deionized water to be soaked for 10h to remove redundant calcium chloride and formic acid, and airing. And drying the obtained silk fibroin at 45 ℃ and dissolving the silk fibroin in formic acid again to obtain SF-FA electrospinning solution with the Silk Fibroin (SF) content of 20 wv%.
2) 20mg of Dopamine (DA) is dissolved in 1ml of water, 10mg of Carbon Nano Tube (CNT) is added, and then the DA-CNT solution is obtained through ultrasonic dispersion.
3) The SF-FA electrospinning solution is prepared into a flexible film through electrostatic spinning, and 1ml of DA-CNT solution is sprayed on the film for multiple times in the electrostatic spinning process. The voltage is 18KV, the electrospinning speed is 0.008ml/min, the receiving distance is 12cm, the temperature is controlled at 20-25 ℃, the humidity is controlled at 40-60%, the volume of the electrospinning solution is 8ml, and after the electrospinning is finished, the film is air-dried at room temperature to obtain the C-SF-FA conductive film.
4) 50mgDA was dissolved in 10ml of sodium hydroxide solution (pH 10), and 10mgCNT was added thereto and ultrasonically dispersed to obtain PDA-CNT solution. The C-SF-FA conductive film is soaked in the PDA-CNT solution for 2 h. After the removal, excess CNT on the surface was washed off with ultrapure water and dried at room temperature. And after the film is dried, soaking the film in 75% ethanol solution for 45min, taking out the film, and after the film is taken out, air-drying the film at room temperature to finally obtain the C-SF-FA conductive film.
5) And assembling the C-SF-FA conductive film and the copper strip into the flexible wearable sensor.
Example 4 application example
Application example 1. flexible wearable wound monitoring sensors were used to measure the response of the sensor to the same simulated body fluid volume:
the both ends of the C-SF-FA conductive film prepared in example 1 were wrapped with copper tape and then connected to an electrochemical workstation to test the resistance change of the C-SF-FA conductive film. Simulated body fluid is used for replacing human wound exudate, and 5, 10, 15, 20 and 25 mul of simulated body fluid is respectively dripped into the C-SF-FA conductive film. As shown in FIG. 3, the conductive paths in the C-SF-FA conductive film increased and the resistance decreased with the dropping of different amounts of the simulant body fluid. And because the dropping amount of the simulated body fluid is different, the resistance change of the C-SF-FA conductive film is linearly changed, which shows that the C-SF-FA conductive film can test the humidity signal of the wound.
Application example 2. flexible wearable wound monitoring sensor was used to measure the repeatability of the sensor:
the both ends of the C-SF-FA conductive film prepared in example 1 were wrapped with copper tape and then connected to an electrochemical workstation to test the resistance change of the C-SF-FA conductive film. The simulated body fluid is used to replace the human wound exudate and is dripped into the C-SF-FA conductive film by 10 mul, and the process is repeated for 5 times. The results of the test are shown in FIG. 4, and show that C-SF-FA shows good reproducibility with the same amount of the simulated body fluid dropped.
Claims (10)
1. A preparation method of a C-SF-FA flexible conductive film based on silk fibroin is characterized by comprising the following steps:
1) degumming silkworm cocoon, drying the degummed silk, dissolving the degummed silk in a salt-formic acid dissolving system to obtain a silk fibroin solution, drying the obtained silk fibroin solution to obtain silk fibroin, soaking the silk fibroin in deionized water to remove salt and formic acid, and dissolving the dried silk fibroin in formic acid again to obtain an SF-FA electrospinning solution;
2) dissolving dopamine in water, adding carbon nanotubes, and uniformly dispersing to obtain a DA-CNT solution;
3) preparing the SF-FA electrospinning solution into a flexible film through electrostatic spinning, and spraying the DA-CNT solution on the film for multiple times in the electrostatic spinning process; after the electrospinning is finished, drying the film in the air to obtain a C-SF-FA conductive film;
4) adding DA and CNT into an alkaline solution to obtain a PDA-CNT solution, placing the C-SF-FA conductive film into the PDA-CNT solution, soaking for 2-10 hours, taking out, cleaning with ultrapure water, drying, soaking the film into an ethanol or methanol solution for 45-60 minutes after drying, taking out, and drying to obtain the C-SF-FA conductive film.
2. The method of claim 1, wherein: the salt in the salt-formic acid dissolving system is selected from one or more of calcium chloride, lithium bromide and magnesium chloride; preferably, the concentration of the salt in the salt-formic acid dissolving system is 2-10 wv%, and the concentration of the formic acid is 90-98 wv%;
the pH value of the alkaline solution in the step 4) is 8-11, and the alkaline solution is preferably a sodium hydroxide aqueous solution or an ammonia aqueous solution.
3. The method of claim 1, wherein: in the DA-CNT solution or the PDA-CNT solution, the concentration of DA is 5-20 mg/ml, and the concentration of CNT is 1-10 mg/ml.
4. The method of claim 1, wherein: the mass fraction of silk fibroin in the SF-FA electrospinning solution in the step 1) is 15-25%;
the dosage of the DA-CNT solution in the step 3) is that 1ml of the DA-CNT solution is used for every 5-10 ml of SF-FA electrospinning solution.
5. The method of claim 1, wherein: the silk fibroin solution obtained by drying the silk fibroin solution in the step 1) is dried at room temperature for 12-24 hours;
soaking the silk fibroin in the deionized water in the step 1) for 10-24 hours to remove calcium chloride and formic acid.
6. The method of claim 1, wherein: degumming the silkworm cocoon silk in the step 1) by adopting a sodium carbonate solution with the concentration of 0.5-2.0 mol/ml;
the ethanol or methanol solution in the step 4) is 75 percent by volume of ethanol or methanol water solution.
7. The method of claim 1, wherein: the electrostatic spinning working conditions in the step 3) are as follows: the voltage is 10-24KV, the electrospinning speed is 0.002-0.015ml/min, the receiving distance is 10-15cm, the temperature is controlled at 20-25 ℃, and the humidity is controlled at 40-60%.
8. A C-SF-FA flexible conductive film based on silk fibroin is characterized in that: is prepared by the method of any one of claims 1 to 7.
9. The utility model provides a flexible wearable wound monitoring sensor based on silk fibroin which characterized in that: is made of the C-SF-FA flexible conductive film based on silk fibroin according to claim 8.
10. The method of making a silk fibroin-based flexible wearable wound monitoring sensor of claim 9, characterized in that: assembling the silk fibroin-based C-SF-FA flexible conductive film of claim 8 and a wire to obtain the flexible wearable wound monitoring sensor.
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