CN112326743B - C-SF-FA flexible conductive film based on silk fibroin, wearable wound monitoring sensor and preparation method thereof - Google Patents
C-SF-FA flexible conductive film based on silk fibroin, wearable wound monitoring sensor and preparation method thereof Download PDFInfo
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- 238000004090 dissolution Methods 0.000 claims description 4
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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
- G01N27/121—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 manufactured by the same, wherein the C-SF-FA flexible conductive film comprises a conductive film body and a conductive film body: 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 silk fibroin obtained by drying in formic acid again to obtain an SF-FA electrospinning solution; 2) Dissolving dopamine in water, and adding carbon nanotubes to obtain DA-CNT solution; 3) Manufacturing SF-FA electrospun solution into a flexible film through electrostatic spinning, and spraying DA-CNT solution on the film for multiple times in the electrostatic spinning process; airing the film to obtain a C-SF-FA conductive film; 4) Adding DA and CNT into alkaline solution to obtain PDA-CNT solution, soaking C-SF-FA conductive film in PDA-CNT, washing and drying, soaking in ethanol or methanol solution, and drying to obtain 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
Skin is the first barrier to human immunity and plays a vital role in guarding the health of people. When the skin is irreparably damaged, it will have a great influence on the human body's immune system. Chronic wounds are a relatively common skin lesion, and for relatively minor burns, the skin is orderly repaired, but when the wound area is too large and deep, the healing capacity of the patient himself will be insufficient for repair, which may be a threat to the life of the patient. Thus, treatment and monitoring of chronic wounds has received widespread attention worldwide.
Wound healing is a continuous and overlapping dynamic process, the whole process comprising three phases: inflammatory, proliferative and remodelling phases. The inflammatory phase is mainly responsible for initiating immune responses to signal wound repair; the proliferation period is to migrate various key cells according to the signals, so that the wound is closed and the blood vessel is rebuilt; the remodeling stage is the stage in which myofibroblasts and wound scar develop. Because the entire healing process is extremely complex, if improperly handled, it will cause duplication of the healing process and wound infection, thereby exacerbating the inflammatory response. Therefore, it is particularly important to monitor wounds.
In order to prevent wound infection, various indexes such as protease, humidity, temperature, blood oxygen level and the like have been used as observation targets for monitoring wound healing. The moisture balance of the wound is a well-recognized critical factor. Any wound healing is associated with moisture, a dry environment that is detrimental to wound healing and too moist may increase the risk of wound infection. Therefore, health care personnel need to monitor wound moisture in real time. Humidity sensors have been developed today, but these sensors suffer from disadvantages such as poor biocompatibility, poor air permeability, and poor data real-time. In recent years, due to the high-speed development of various biological materials, it becomes possible to design a wound real-time monitoring sensor with excellent biocompatibility and high conductivity.
Silk is a common purely natural bio-organic material. There has been increasing interest in silk 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 predominant protein. Studies have shown that silk fibroin does not continuously cause inflammatory reactions, so that it plays a great role in chondrocyte culture, bone tissue scaffolds, antibacterial dressings and the like. Meanwhile, the excellent physical properties of the composite material can be used as an additive to be added into materials to enhance the mechanical properties of the materials, and the composite material can be made into scaffolds such as films, hydrogels and fibers.
Among the nanofillers, carbon Nanotubes (CNT) have the advantage of significantly improving the properties of the nanocomposite, and are ideal materials for preparing high quality nanocomposites. The carbon nanotubes contain graphite layers of a Lidi diameter (1-100 nm) and a large length (1-10 μm), resulting in a large specific surface area. The carbon nanotubes have a tensile modulus of 1000GPa and a strength of 10 to 50GPa. In addition, superconducting CNTs (105-106S/m) introduce the desired conductivity into the insulating matrix. Therefore, CNT has excellent physical, mechanical and conductive properties, and is a promising filler.
Disclosure of Invention
The invention aims at providing a C-SF-FA flexible conductive film based on silk fibroin and having excellent electrical and mechanical properties and biocompatibility and a preparation method thereof.
In order to achieve the purpose, the invention adopts the following technical scheme:
a preparation method of a C-SF-FA flexible conductive film based on silk fibroin comprises the following steps:
1) Degumming silk of silkworm cocoon, drying degummed silk, dissolving in a salt-formic acid dissolving system, dissolving to obtain a silk fibroin solution, drying the silk fibroin solution to obtain silk fibroin, soaking the silk fibroin in deionized water to remove salt and formic acid, and dissolving the silk fibroin obtained after drying in formic acid again to obtain SF-FA electrospinning solution;
2) Dissolving dopamine in water, adding carbon nanotubes, and uniformly dispersing to obtain DA-CNT solution;
3) Manufacturing SF-FA electrospun solution into a flexible film through electrostatic spinning, and spraying DA-CNT solution on the film for multiple times in the electrostatic spinning process; airing the film after the electrospinning is finished to obtain a C-SF-FA conductive film;
4) Adding DA and CNT into alkaline solution to obtain PDA-CNT solution, soaking C-SF-FA conductive film in PDA-CNT solution for 2-10 hours, taking out, washing with ultrapure water, drying, soaking in ethanol or methanol solution for 45-60 minutes after the film is dried, taking out, and drying to obtain C-SF-FA conductive film.
The salt in the salt-formic acid dissolution system is one or more selected from calcium chloride, lithium bromide and magnesium chloride; preferably, the salt concentration in the salt-formic acid dissolution system is 2-10wv% and the formic acid concentration is 90-98wv%;
the pH of the alkaline solution in step 4) is 8 to 11, preferably an aqueous sodium hydroxide solution or an aqueous ammonia 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 DA-CNT solution is used in the step 3) in an amount of 1ml of DA-CNT solution per 5 to 10ml of SF-FA electrospinning solution.
The silk fibroin obtained by drying the silk fibroin solution in the step 1) means that the silk fibroin solution is dried for 12 to 24 hours at room temperature;
in the step 1), the soaking time for removing calcium chloride and formic acid by soaking silk fibroin in deionized water is 10-24 hours.
Degumming the silk of the silk cocoons in the step 1) by adopting sodium carbonate solution with the concentration of 0.5-2.0 mol/ml;
the ethanol or methanol solution in the step 4) is 75 volume percent 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, prepared by the method of any one of the above.
The invention also aims to provide a flexible wearable wound monitoring sensor based on the silk fibroin, which is made of the C-SF-FA flexible conductive film based on the silk fibroin.
The preparation method of the flexible wearable wound monitoring sensor based on silk fibroin comprises the following steps: and assembling the C-SF-FA flexible conductive film based on the silk fibroin and a lead to obtain the flexible wearable wound monitoring sensor.
The salt-formic acid system induces the formation of beta-sheet of silk fibroin, which can enhance the mechanical properties of silk fibroin film, and enhance its electrical conductivity after CNT addition. The change of the wound healing degree is judged by applying the flexible wearable sensor to the 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 reticular structure, when the wound sensor contacts the wound surface, 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 is changed. And finally, judging the balance degree of the water environment of the wound according to the change of the resistivity, thereby monitoring the healing of the wound in real time.
The beneficial effects of the invention are as follows: the invention uses simple electrostatic spinning technology to manufacture the C-SF-FA conductive film, and has mild reaction conditions and simple operation; the C-SF-FA flexible conductive film has a highly dense reticular bracket structure, so that the C-SF-FA flexible conductive film has excellent mechanical properties; and the CNT conductive interlayer inside the flexible film gives it excellent conductivity. The flexible sensor can detect the change of the humidity of the wound by the change of the overall conductivity caused by absorbing the tissue fluid of the wound; the material has excellent electrical and mechanical properties and biocompatibility, and the detection method has high sensitivity, and is 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 continue to be the greatest challenge plaguing the application potential of flexible sensors. The manufacturing cost of the flexible sensor is further reduced without utilizing any micro-manufacturing technology in the whole process of manufacturing the flexible sensor. 2) The uniform mixing of the conductive material has 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 mismatch 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 microscope scan of a conductive film of the present invention C-SF-FA.
FIG. 2 is a schematic structural diagram of a sensor assembled by a C-SF-FA conductive film and a copper strip according to the present invention.
FIG. 3 is a graph showing the response of the sensor of the present invention to the same simulated body fluid amount, wherein (a) is a resistance change chart, (b) is a sensitivity change chart, and (C) is a fitted straight line chart of the C-SF-FA conductive film and the simulated body fluid absorption amount, using example 1.
FIG. 4 shows the results of measurement of the reproducibility of the measurement of the sensor of the present invention using example 2, wherein (a) is a resistance change chart, (b) is a sensitivity change chart, and (C) is a fitted line chart of the C-SF-FA conductive film and the simulated body fluid absorption amount.
Detailed Description
The invention is further illustrated, but is not limited, by the following examples.
The experimental methods in the following examples are conventional methods unless otherwise specified.
The main reagent sources are as follows:
dopamine hydrochloride (DA): CAS number 62-31-7, available from biological engineering (Shanghai) Co., ltd;
carbon Nanotubes (CNT): the production company is Nanjing Jicang nanotechnology Co., ltd., product number: JCNT-95-50-15-W;
simulated body fluid: the production company is Beijing cool Lai Bo technology Co., ltd., product number: SL6710-500ml;
the remaining reagents, if not indicated, were not conventional and were all commercially available.
Example 1:
preparation of a flexible wearable wound monitoring sensor based on silk fibroin, which is operated according to the following steps:
1) Boiling silk cocoon silk with 2mol/L sodium carbonate solution, degumming, washing, naturally air drying, dissolving in calcium chloride/Formic Acid (FA) dissolving system (wherein the calcium chloride concentration is 2wv%, and the formic acid concentration is 98 wv%) for more than 3h to form silk fibroin solution (SF solution), and drying the obtained SF solution at room temperature for 24h to obtain silk fibroin. And then placing the obtained dry silk fibroin in deionized water for soaking for 10 hours to remove redundant calcium chloride and formic acid, and airing. And then drying the obtained silk fibroin at 35 ℃ and dissolving the silk fibroin in formic acid again to obtain SF-FA electrospun solution with 20wv% of Silk Fibroin (SF).
2) 5mg of Dopamine (DA) was dissolved in 1ml of water, and after adding 5mg of Carbon Nanotubes (CNT), the solution was dispersed by ultrasound to obtain DA-CNT solution.
3) The SF-FA electrospinning solution was fabricated into a flexible film by electrospinning, and 1ml of DA-CNT solution was sprayed onto the film several times during electrospinning. The electrostatic spinning working conditions 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 amount of electrospinning solution is 8ml, and the C-SF-FA conductive film is obtained after the film is air-dried at room temperature after the electrospinning is finished.
4) 50mg of DA was dissolved in 10ml of sodium hydroxide solution (pH 8), and after adding 10mg of CNT, the solution was subjected to ultrasonic dispersion to obtain a PDA-CNT solution. The C-SF-FA conductive film is soaked in PDA-CNT solution for 2h. After removal, excess CNT on the surface was rinsed with ultra pure water and dried at room temperature. And after the film is dried, soaking the film in 75% ethanol solution for 45min, taking out, and air-drying at room temperature after taking out to finally obtain the C-SF-FA conductive film, wherein the scanning image of the C-SF-FA conductive film is shown in figure 1.
5) The C-SF-FA conductive film and the copper strip are assembled into the flexible wearable sensor, and the structural schematic diagram of the flexible wearable sensor is shown in fig. 2.
Example 2:
preparation of a flexible wearable wound monitoring sensor based on silk fibroin, which is operated according to the following steps:
1) Boiling silk cocoon silk with 2mol/L sodium carbonate solution for degumming, washing, naturally air-drying, dissolving in lithium bromide/Formic Acid (FA) dissolving system (wherein the concentration of lithium bromide is 10wv%, and the concentration of formic acid is 98 wv%) for more than 3h to form silk fibroin solution (SF solution), and drying the obtained SF solution at room temperature for 12h to obtain silk fibroin. And then placing the obtained dry silk fibroin in deionized water for soaking for 10 hours to remove redundant lithium bromide and formic acid, and airing. And then drying the obtained silk fibroin at 50 ℃ and dissolving the silk fibroin in formic acid again to obtain SF-FA electrospun solution with 20wv% of Silk Fibroin (SF).
2) 5mg of Dopamine (DA) was dissolved in 1ml of water, and after adding 1mg of Carbon Nanotubes (CNT), the solution was dispersed by ultrasound to obtain DA-CNT solution.
3) The SF-FA electrospinning solution was fabricated into a flexible film by electrospinning, and 1ml of DA-CNT solution was sprayed onto the film several times during electrospinning. The voltage is 24KV, the electrospinning speed is 0.015ml/min, the receiving distance is 15cm, the temperature is controlled at 20-25 ℃, the humidity is controlled at 40-60%, the amount of electrospinning solution is 10ml, and the C-SF-FA conductive film is obtained after the film is air-dried at room temperature after the electrospinning is finished.
4) 200mgDA was dissolved in 10ml of an aqueous ammonia solution (pH 11), and 100mgCNT was added thereto and then subjected to ultrasonic dispersion to obtain a PDA-CNT solution. The C-SF-FA conductive film was immersed in the PDA-CNT solution for 4 hours. After removal, excess CNT on the surface was rinsed with ultra pure water and dried at room temperature. And (3) soaking the film in 75% methanol solution for 45min after the film is dried, taking out, and air-drying at room temperature after taking out 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 flexible wearable wound monitoring sensor based on silk fibroin, which is operated according to the following steps:
1) Boiling silk cocoon silk with 2mol/L sodium carbonate solution, degumming, washing, naturally air drying, dissolving in calcium chloride/Formic Acid (FA) dissolving system (wherein calcium chloride concentration is 5wv%, and formic acid concentration is 98 wv%) for more than 3h to form silk fibroin solution (SF solution), and drying the obtained SF solution at room temperature for 24h to obtain silk fibroin. And then placing the obtained dry silk fibroin in deionized water for soaking for 10 hours to remove redundant calcium chloride and formic acid, and airing. And then drying the obtained silk fibroin at 45 ℃ and dissolving the silk fibroin in formic acid again to obtain SF-FA electrospun solution with 20wv% of Silk Fibroin (SF).
2) 20mg of Dopamine (DA) was dissolved in 1ml of water, and 10mg of Carbon Nanotubes (CNT) were added thereto and then ultrasonically dispersed to obtain a DA-CNT solution.
3) The SF-FA electrospinning solution was fabricated into a flexible film by electrospinning, and 1ml of DA-CNT solution was sprayed onto the film several times during electrospinning. 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 amount of electrospinning solution is 8ml, and the C-SF-FA conductive film is obtained after the film is air-dried at room temperature after the electrospinning is finished.
4) 50mgDA was dissolved in 10ml of sodium hydroxide solution (pH 10), and after adding 10mgCNT, the solution was subjected to ultrasonic dispersion to obtain PDA-CNT solution. The C-SF-FA conductive film is soaked in PDA-CNT solution for 2h. After removal, excess CNT on the surface was rinsed with ultra pure water and dried at room temperature. And (3) soaking the film in 75% ethanol solution for 45min after the film is dried, taking out, and air-drying at room temperature after taking out 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 sensor was used to measure the response of the sensor to the same simulated body fluid volume:
the C-SF-FA conductive film prepared in example 1 was wrapped with copper tape at both ends, and then was 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 wound liquid seepage of the human body, and 5, 10, 15, 20 and 25 mu l of the simulated body fluid are respectively dripped into the C-SF-FA conductive film. As shown in FIG. 3, the conductive paths in the conductive film of C-SF-FA increased and the resistance decreased with the dropping of different amounts of simulated body fluid. And the resistance change of the C-SF-FA conductive film is linear change due to different dripping amounts of the simulated body fluid, which indicates 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 sensor repeatability:
the C-SF-FA conductive film prepared in example 1 was wrapped with copper tape at both ends, and then was connected to an electrochemical workstation to test the resistance change of the C-SF-FA conductive film. The simulated body fluid was used to replace the wound exudate of the human body and 10 μl was added dropwise to the C-SF-FA conductive film, and repeated 5 times. The test results are shown in FIG. 4, and the results show that the C-SF-FA exhibits good reproducibility with the same amount of simulated body fluid instillation.
Claims (3)
1. The preparation method of the C-SF-FA flexible conductive film based on the silk fibroin is characterized by comprising the following steps:
1) Degumming silk of silkworm cocoons, drying the degummed silk, dissolving the degummed silk in a salt-formic acid dissolving system, dissolving 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 electrospun solution with the mass fraction of the silk fibroin of 15-25%; the salt in the salt-formic acid dissolution system is one or more selected from calcium chloride, lithium bromide and magnesium chloride; the concentration of salt in the salt-formic acid dissolution system is 2-10wv%, and the concentration of formic acid is 90-98wv%;
2) Dissolving dopamine in water, adding carbon nanotubes, and uniformly dispersing to obtain DA-CNT solution;
3) Manufacturing SF-FA electrospun solution into a flexible film through electrostatic spinning, and spraying DA-CNT solution on the film for multiple times in the electrostatic spinning process; airing the film after the electrospinning is finished to obtain a C-SF-FA conductive film; the DA-CNT solution is used in an amount of 1ml of DA-CNT solution per 5-10 ml of SF-FA electrospinning solution; the electrostatic spinning working conditions 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%;
4) Adding DA and CNT into alkaline solution to obtain PDA-CNT solution, soaking C-SF-FA conductive film in PDA-CNT solution for 2-10 hours, taking out, washing with ultrapure water, drying, soaking in ethanol or methanol solution for 45-60 minutes after the film is dried, taking out, and drying to obtain C-SF-FA conductive film; the pH value of the alkaline solution is 8-11, and the alkaline solution is sodium hydroxide aqueous solution or ammonia water 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.
2. The method of manufacturing according to claim 1, wherein: the silk fibroin obtained by drying the silk fibroin solution in the step 1) means that the silk fibroin solution is dried for 12 to 24 hours at room temperature;
in the step 1), the soaking time for removing calcium chloride and formic acid by soaking silk fibroin in deionized water is 10-24 hours.
3. The method of manufacturing according to claim 1, wherein: degumming the silk of the silk cocoons in the step 1) by adopting sodium carbonate solution with the concentration of 0.5-2.0 mol/ml;
the ethanol or methanol solution in the step 4) is 75 volume percent ethanol or methanol water solution.
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