CN113183594B - Preparation method of electrostatic spinning functional nanofiber film for epidermis repair - Google Patents

Preparation method of electrostatic spinning functional nanofiber film for epidermis repair Download PDF

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CN113183594B
CN113183594B CN202110372670.6A CN202110372670A CN113183594B CN 113183594 B CN113183594 B CN 113183594B CN 202110372670 A CN202110372670 A CN 202110372670A CN 113183594 B CN113183594 B CN 113183594B
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layer
solution
membrane
polylactic acid
fiber membrane
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CN113183594A (en
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熊俊杰
陈桪
王晗
陈梓瀚
吕泽思
毛睿欣
麦仁标
何景帆
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Guangdong University of Technology
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    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B32—LAYERED PRODUCTS
    • B32B—LAYERED PRODUCTS, i.e. PRODUCTS BUILT-UP OF STRATA OF FLAT OR NON-FLAT, e.g. CELLULAR OR HONEYCOMB, FORM
    • B32B38/00—Ancillary operations in connection with laminating processes
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    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
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    • A61L15/22—Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons containing macromolecular materials
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    • A61L15/00—Chemical aspects of, or use of materials for, bandages, dressings or absorbent pads
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    • A61L15/42—Use of materials characterised by their function or physical properties
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00—Preparations for care of the skin
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    • B32B38/00—Ancillary operations in connection with laminating processes
    • B32B38/0012—Mechanical treatment, e.g. roughening, deforming, stretching
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B32—LAYERED PRODUCTS
    • B32B—LAYERED PRODUCTS, i.e. PRODUCTS BUILT-UP OF STRATA OF FLAT OR NON-FLAT, e.g. CELLULAR OR HONEYCOMB, FORM
    • B32B38/00—Ancillary operations in connection with laminating processes
    • B32B38/0036—Heat treatment
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    • B32—LAYERED PRODUCTS
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    • B32B38/00—Ancillary operations in connection with laminating processes
    • B32B38/08—Impregnating
    • D—TEXTILES; PAPER
    • D01—NATURAL OR MAN-MADE THREADS OR FIBRES; SPINNING
    • D01F—CHEMICAL FEATURES IN THE MANUFACTURE OF ARTIFICIAL FILAMENTS, THREADS, FIBRES, BRISTLES OR RIBBONS; APPARATUS SPECIALLY ADAPTED FOR THE MANUFACTURE OF CARBON FILAMENTS
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    • D01F8/14—Conjugated, i.e. bi- or multicomponent, artificial filaments or the like; Manufacture thereof from synthetic polymers with at least one polyester as constituent
    • D—TEXTILES; PAPER
    • D01—NATURAL OR MAN-MADE THREADS OR FIBRES; SPINNING
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    • D01F8/00—Conjugated, i.e. bi- or multicomponent, artificial filaments or the like; Manufacture thereof
    • D01F8/04—Conjugated, i.e. bi- or multicomponent, artificial filaments or the like; Manufacture thereof from synthetic polymers
    • D01F8/16—Conjugated, i.e. bi- or multicomponent, artificial filaments or the like; Manufacture thereof from synthetic polymers with at least one other macromolecular compound obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds as constituent
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    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
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    • B32B38/00—Ancillary operations in connection with laminating processes
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Abstract

The invention discloses a preparation method of an electrostatic spinning functional nanofiber film for epidermis repair, belonging to the field of medical cosmetology and comprising the following steps: preparing a nanofiber membrane with polylactic acid caprolactone serving as a shell layer and polyethylene glycol serving as a core layer through coaxial electrostatic spinning; applying stress to two ends of the nanofiber membrane, unloading, uniformly spraying heated absolute ethyl alcohol on the surface of a sample, and rapidly cooling the fiber membrane by using low-temperature airflow to form a meandering fiber membrane; and (2) mixing the recombinant human epidermal growth factor into a sodium polystyrene sulfonate solution to form a negative electricity layer, immersing the meandering fiber membrane into a polyallylamine hydrochloride solution to form a positive electricity layer, and assembling layer by layer to form the multilayer composite fiber membrane. The polylactic acid and polycaprolactone copolymer polylactic acid caprolactone is used as a shell layer of the coaxial electrostatic spinning nanofiber, so that the mechanical property of the fiber film is improved, and the polyethylene glycol is used as a core layer of the fiber, so that the hydrophilicity and the moisture permeability of the fiber film are improved.

Description

Preparation method of electrostatic spinning functional nanofiber film for epidermis repair
Technical Field
The invention relates to the field of medical cosmetology, in particular to a preparation method of an electrostatic spinning functional nanofiber film for epidermis repair.
Background
Solution electrostatic spinning is a method for preparing micro-nano fibers from polymer solution or melt by using electrostatic force, and compared with the conventional spinning process, the prepared fibers have smaller diameters (from nano to micron) and larger specific surface areas. The electrospinning process is carried out by applying a voltage in the range of tens of kilovolts, which is mainly based on the principle that strong repulsive forces overcome weak surface tensions in charged polymer liquids. Currently, there are two standard electrospinning modes of operation: vertical and horizontal. As this technology has evolved, more complex nanofiber structures can be fabricated in a more controllable and efficient manner. The electrospinning system consists of three main components: a high voltage power supply, a spinneret and a grounded collector plate (usually a metal screen, plate or rotating mandrel) are used to inject charge of one polarity into the polymer solution or melt, which is then accelerated towards a collector of the opposite polarity. Most polymers are dissolved in certain solvents prior to electrospinning, and when it is completely dissolved, a polymer solution is formed.
In the electrospinning process, the polymer solution held at the end of the capillary by its surface tension is subjected to an electric field and a charge is induced on the surface of the liquid due to the electric field. When the applied electric field reaches a critical value, the repulsive force overcomes the surface tension. An electrically charged jet of solution is ejected from the tip of the taylor cone and unstable and rapid jet agitation occurs in the space between the capillary tip and the collector, forming polymer fibers as the solvent evaporates. While the coaxial electrospinning process can synthesize nanofibers with more forms and uses from two or more types of polymer solutions, the application range of coaxial electrospinning has been expanded to biomedicine, electrochemical development and environmental protection with the diversification of material selection and structure.
Most animal cells grow in a fibrous extracellular matrix, which is essentially a network of collagen fibers. Collagen generally exists in the form of serpentine-like parallel fibers and exhibits specific nonlinear mechanical properties. The serpentine structure may provide excellent elasticity and flexibility to the tissue. The nonlinear elasticity exhibited by collagen effectively slows the oscillation of mechanical stress, prevents tissue overstretching, and maintains a steady state of the blood flow environment, thereby preserving the normal activities of the biological individual. Therefore, the preparation of the fibrous scaffold with nonlinear characteristics by utilizing the bionic collagen fibers has important significance for repairing epidermal tissues. However, most synthetic polymeric materials are linear elastic and therefore it is necessary to obtain complex non-linear mechanical properties through structural adjustment.
The prior art has constructed different fiber space structures by weaving, bicomponent electrospinning, micro-forming and other methods, which make synthetic polymer materials have mechanical nonlinearity, but the deformation of the whole structure causes larger strain under low stress, and the recoverability of the deformation of the material is poor. Micro-forming forces straight parallel fibers into a serpentine shape, but the internal stresses applied during forming greatly increase the instability of mechanical properties. Researchers have electrospun two polymers with different shrinkage rates into a single fiber through side-by-side spinnerets, using residual stress to form a serpentine structure. There have also been researchers releasing electrospun fibers from collection devices above the glass transition temperature of the polymer to induce them to form a serpentine structure, but uncontrolled fiber meandering and structural instability severely limit their application in different fields of tissue engineering.
Layer-by-layer assembly is a powerful surface coating technique that has made substantial progress in loading and delivering drugs and growth factors to study basic biological processes, and the use of layer-by-layer assembly in tissue engineering and biomedical applications has expanded dramatically. Microfluidics, which allow selective deposition of multilayer films on micron-scale surfaces, with microchannel widths or diameters ranging from 50 to 800 μm, has been combined appropriately with layer-by-layer assembly techniques. By flowing a polyelectrolyte solution of polystyrene sulfonate (PSS) and polyallylamine hydrochloride (PAH) through a network of microchannels, a negatively charged film layer and a positively charged film layer are created, which can then be assembled together to form a composite film.
The Chinese patent document with the publication number of CN109172073A and the publication date of 2019, 1 month and 11 days discloses a coaxial electrostatic spinning nanofiber membrane for controlling growth factors and a preparation method thereof, but the structure of the fiber is not designed, functional materials are less, and the living environment of more bionic cells is not existed.
The Chinese patent document with the publication number of CN107397973B and the publication date of 2020, 7 and 24 discloses a four-layer coaxial fiber wound dressing and a preparation method thereof, wherein although functional materials are many, the thickness of a film is fixed, and a more bionic skin deformation mechanism and a cell living environment are not provided.
Most animal cells grow in a fibrous extracellular matrix, which is essentially a network of collagen fibers, usually in the form of serpentine parallel fibers, exhibiting special non-linear mechanical properties, while the deformation of certain parts of the individual organism upon activity is anisotropic, with the serpentine structure providing excellent elasticity and flexibility to the tissue. To date, most manufacturing techniques for making these scaffolds include lyophilization, solvent casting, and simple braiding, but they do not reconstitute the delicate fibrous structure of natural skin collagen fibers. Solution electrospinning is the most common different fiber forming technique used to make fiber scaffolds, and the basic working principle is to use electrostatic forces to generate continuous fibers from a polymer solution, which are then collected on a stationary or dynamic collector table. Solution electrospinning can produce fibers with topographical features that mimic collagen fiber networks, but has reproducibility problems and limited pattern control capabilities. Some studies have improved the manufacturing process by using parallel electrodes, rotating mandrels or sacrificial materials, but control of the fiber geometry is still limited, especially in three-dimensional structures. Fused electrographic printing is an effective method to achieve controlled deposition of highly ordered 3D scaffolds of non-linear fibers, but the influence of residual charge on fiber printing is detrimental to control of fiber stacking-the process is limited by charge dynamics. Meanwhile, the efficiency of preparing a film with a certain thickness only through electrostatic spinning is low, and the process of loading the functional material, such as the process of preparing the microsphere controlled release, is complex and time-consuming.
Disclosure of Invention
The invention aims to provide a preparation method of an electrostatic spinning functional nanofiber film for skin repair, so as to solve one or more technical problems in the prior art.
In order to achieve the purpose, the invention adopts the following technical scheme:
the preparation method of the electrostatic spinning functional nanofiber membrane for repairing the epidermis comprises the following steps:
step S1: preparing a polylactic acid caprolactone spinning solution and a polyethylene glycol spinning solution;
step S2: preparing a nanofiber membrane with polylactic acid caprolactone serving as a shell layer and polyethylene glycol serving as a core layer through coaxial electrostatic spinning;
step S3: applying stress to two ends of the nanofiber membrane through a stretching device, unloading, repeating the operation for multiple times, fixing the two ends of the nanofiber membrane, uniformly spraying the heated absolute ethyl alcohol on the surface of a sample until the absolute ethyl alcohol is completely soaked, and finally quickly cooling the fiber membrane by using low-temperature airflow to form a stable meandering fiber membrane;
step S4: and (4) mixing the recombinant human epidermal growth factor into a sodium polystyrene sulfonate solution to form a negative electric layer, immersing the meandering fiber membrane prepared in the step S3 into a polyallylamine hydrochloride solution to form a positive electric layer, and assembling the negative electric layer and the positive electric layer by layer to form the multilayer composite fiber membrane.
Preferably, in step S4, after the multilayer composite fiber film is formed, the multilayer composite fiber film is continuously immersed in the sodium polystyrene sulfonate solution and the polyallylamine hydrochloride solution for neutralization.
Preferably, in step S2, the collection distance of the electrostatic spinning is set to 13-15cm, the voltage is set to 11-14kV, the feeding speed of the solution is set to 1.8-2mL/h, and the rotation speed of the roller collector is 1800-2000 rpm.
Preferably, in step S2, after the nanofiber membrane is prepared, the nanofiber membrane is dried in a vacuum oven.
Preferably, the preparation process of the polycaprolactone polylactic acid spinning solution comprises the following steps: dichloromethane and dimethylformamide were mixed at a ratio of 7: 3, and then putting the polylactic acid caprolactone into the mixed solution to form the polylactic acid caprolactone spinning solution.
Preferably, the preparation process of the polyethylene glycol spinning solution comprises the following steps: dissolving polyethylene glycol in dimethylformamide to form polyethylene glycol spinning solution.
The beneficial effects of the invention are as follows: according to the invention, polylactic acid and polycaprolactone copolymer polylactic acid caprolactone (PLCL) is used as a shell layer of the coaxial electrostatic spinning nanofiber, so that the mechanical property of the fiber film is improved, and polyethylene glycol is used as a core layer of the fiber, so that the hydrophilicity and the moisture permeability of the fiber film are improved; the composite film prepared by the preparation method can be used as a wound dressing to repair damaged skin, and can also be used as a mask to maintain the skin.
Drawings
The drawings are further illustrative of the invention and the content of the drawings does not constitute any limitation of the invention.
FIG. 1 is a flow chart of the preparation method of the electrospun functional nanofiber membrane for repairing epidermis according to the present invention.
Detailed Description
The technical scheme of the invention is further explained by the specific implementation mode in combination with the attached drawings.
The preparation method of the electrospun functional nanofiber membrane for repairing the epidermis of the embodiment refers to fig. 1, and the method comprises the following steps:
step S1: preparing a polylactic acid caprolactone spinning solution and a polyethylene glycol spinning solution: dichloromethane and dimethylformamide were mixed at a ratio of 7: 3, then putting the polylactic acid caprolactone into the mixed solution to obtain a polylactic acid caprolactone spinning solution with the concentration of 10% w/V; dissolving 50% by mass of polyethylene glycol in dimethylformamide to prepare polyethylene glycol spinning solution;
step S2: respectively injecting the polylactic acid caprolactone spinning solution and the polyethylene glycol spinning solution into a coaxial nozzle of an electrostatic spinning machine, connecting the coaxial nozzle with the positive electrode of a high-voltage power supply, setting the fixed collection distance from a needle point to a roller collector to be 13cm, setting the voltage to be 11-14kV, setting the solution feeding speed to be 2mL/h, and setting the rotating speed of the roller collector to be 1800rpm, forming a core layer jet flow conical tip on the core layer needle point of the coaxial nozzle, forming a shell layer jet flow conical tip on the shell layer, further forming nano fibers, obtaining a nano fiber film with the polylactic acid caprolactone as the shell layer and the polyethylene glycol as the core layer after electrostatic spinning is finished, and drying the prepared nano fiber film in a vacuum oven set to be 45 ℃ to remove a solvent;
step S3: cutting the nanofiber membrane prepared in the step S2 into rectangular strips, applying stress to the samples through a clamp of a universal tensile testing machine, unloading, repeating for 3 times under the same low stress level, fixing two ends of each rectangular strip sample, presetting a certain shrinkage space, uniformly spraying heated absolute ethyl alcohol on the surfaces of the samples through a pipette until the samples are completely soaked, and preferably, rapidly cooling the fiber membrane by using low-temperature air flow to form a stable winding fiber membrane;
step S4: mixing Recombinant Human Epidermal Growth Factor (RHEGF) into a sodium polystyrene sulfonate solution (PSS, 1mg/ml, pH = 4.7) to form a negatively charged negative electric layer, immersing the serpentine fiber membrane into a polyallylamine hydrochloride (PAH, 1mg/ml, pH = 4.7) solution for 1 hour to form a positively charged positive electric layer, assembling the negative electric layer and the positive electric layer by layer to form a multilayer composite fiber membrane, and finally, continuously immersing the composite fiber membrane into the sodium polystyrene sulfonate solution and the polyallylamine hydrochloride solution for neutralization to prepare the multilayer composite membrane with controllable thickness.
The preparation method of the embodiment is used for preparing the functional nanofiber film capable of repairing damaged epidermis, the application of the electrostatic spinning nanofiber structure is innovated by the method, and the human tissue skin is anisotropic when deformed, so that the embodiment prepares the film with the fiber structure and nonlinear mechanical properties to more simulate the deformation mechanism of human skin, and meanwhile, the living environment of fibroblasts is similar to the network formed by winding fibers, so that the film prepared by the embodiment has better guiding significance for the growth of the fibroblasts on the damaged skin surface. The polylactic acid caprolactone is used as the shell layer of the nano fiber to improve the mechanical property of the fiber film, the polyethylene glycol is used as the core layer of the fiber to improve the hydrophilic property of the fiber film, the film has better moisture permeability when being applied to the skin, and in hot weather, the film can play a certain heat regulation function on the surface of the skin, and the skin feels very comfortable. Through the combination of a coaxial electrostatic spinning process and a layer-by-layer assembly technology, the recombinant human epidermal growth factor is loaded in the nanofiber membrane for repairing damaged skin tissues, and meanwhile, the functional nanofiber membrane can be prepared into a functional composite membrane with adjustable thickness through assembly manufacturability. The skin care product can be used as a wound dressing to repair damaged skin, and can also be used as a mask to treat the skin.
Compared with the nano meandering fiber printed by the existing solution electrostatic spinning technology, the preparation method of the embodiment can obtain a better oriented meandering nanofiber film, is more suitable for a skin deformation mechanism and a film for improving the comfortableness of damaged skin, and compared with other growth factor loading technologies, the layer-by-layer assembly process of the preparation method is simpler and has higher efficiency, the preparation of a functional film is realized, the overall mechanical property of the film can be improved, a three-dimensional growth environment for cell growth is provided, the preparation efficiency is high, and the thickness of the composite film is controllable.
The technical principle of the present invention is described above in connection with specific embodiments. The description is made for the purpose of illustrating the principles of the invention and should not be construed in any way as limiting the scope of the invention. Based on the explanations herein, those skilled in the art will be able to conceive of other embodiments of the present invention without inventive effort, which would fall within the scope of the present invention.

Claims (6)

1. The preparation method of the electrostatic spinning functional nanofiber film for repairing the epidermis is characterized by comprising the following steps of:
step S1: preparing a polylactic acid caprolactone spinning solution and a polyethylene glycol spinning solution;
step S2: preparing a nanofiber membrane with polylactic acid caprolactone serving as a shell layer and polyethylene glycol serving as a core layer through coaxial electrostatic spinning;
step S3: cutting the nanofiber membrane prepared in the step S2 into rectangular strips, applying stress to the samples through a clamp of a universal tensile testing machine, unloading, repeating for 3 times under the same low stress level, fixing two ends of each rectangular strip sample, presetting a certain shrinkage space, uniformly spraying heated absolute ethyl alcohol on the surfaces of the samples through a pipette until the absolute ethyl alcohol is completely soaked, and finally quickly cooling the fiber membrane by using low-temperature airflow to form a stable winding fiber membrane;
step S4: and (4) mixing the recombinant human epidermal growth factor into a sodium polystyrene sulfonate solution to form a negative electric layer, immersing the meandering fiber membrane prepared in the step S3 into a polyallylamine hydrochloride solution to form a positive electric layer, and assembling the negative electric layer and the positive electric layer by layer to form the multilayer composite fiber membrane.
2. The method of preparing an electrospun functional nanofiber membrane for skin repair according to claim 1, wherein in step S4, after the multilayer composite fiber membrane is formed, the multilayer composite fiber membrane is continuously immersed in a sodium polystyrene sulfonate solution and a polyallylamine hydrochloride solution for neutralization.
3. The method as claimed in claim 1, wherein the step S2 is performed by setting the collection distance of the electrostatic spinning to 13-15cm, the voltage to 11-14kV, the feeding speed of the solution to 1.8-2mL/h, and the rotation speed of the roller collector to 1800-2000 rpm.
4. The method of preparing an electrospun functional nanofiber membrane for skin repair according to claim 1 or 3, wherein in step S2, after the nanofiber membrane is prepared, the prepared nanofiber membrane is dried in a vacuum oven.
5. The method for preparing the electrospun functional nanofiber membrane for epidermis repair according to claim 1, wherein the preparation process of the polylactic acid caprolactone spinning solution is as follows: dichloromethane and dimethylformamide were mixed at a ratio of 7: 3, and then putting the polylactic acid caprolactone into the mixed solution to form the polylactic acid caprolactone spinning solution.
6. The method for preparing the electrospun functional nanofiber membrane for epidermis repair according to claim 1, wherein the polyethylene glycol spinning solution is prepared by the following steps: polyethylene glycol was dissolved in dimethylformamide.
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