CN114848887A - Nanofiber dressing and preparation method thereof - Google Patents
Nanofiber dressing and preparation method thereof Download PDFInfo
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- CN114848887A CN114848887A CN202210558507.3A CN202210558507A CN114848887A CN 114848887 A CN114848887 A CN 114848887A CN 202210558507 A CN202210558507 A CN 202210558507A CN 114848887 A CN114848887 A CN 114848887A
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- 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
- A61L15/00—Chemical aspects of, or use of materials for, bandages, dressings or absorbent pads
- A61L15/16—Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons
- A61L15/42—Use of materials characterised by their function or physical properties
- A61L15/44—Medicaments
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- 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
- A61L15/00—Chemical aspects of, or use of materials for, bandages, dressings or absorbent pads
- A61L15/16—Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons
- A61L15/22—Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons containing macromolecular materials
- A61L15/26—Macromolecular compounds obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds; Derivatives thereof
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- 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
- A61L15/00—Chemical aspects of, or use of materials for, bandages, dressings or absorbent pads
- A61L15/16—Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons
- A61L15/22—Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons containing macromolecular materials
- A61L15/32—Proteins, polypeptides; Degradation products or derivatives thereof, e.g. albumin, collagen, fibrin, gelatin
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- 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
- A61L15/00—Chemical aspects of, or use of materials for, bandages, dressings or absorbent pads
- A61L15/16—Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons
- A61L15/22—Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons containing macromolecular materials
- A61L15/32—Proteins, polypeptides; Degradation products or derivatives thereof, e.g. albumin, collagen, fibrin, gelatin
- A61L15/325—Collagen
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- 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
- A61L15/00—Chemical aspects of, or use of materials for, bandages, dressings or absorbent pads
- A61L15/16—Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons
- A61L15/42—Use of materials characterised by their function or physical properties
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- D—TEXTILES; PAPER
- D01—NATURAL OR MAN-MADE THREADS OR FIBRES; SPINNING
- D01D—MECHANICAL METHODS OR APPARATUS IN THE MANUFACTURE OF ARTIFICIAL FILAMENTS, THREADS, FIBRES, BRISTLES OR RIBBONS
- D01D5/00—Formation of filaments, threads, or the like
- D01D5/0007—Electro-spinning
- D01D5/0015—Electro-spinning characterised by the initial state of the material
- D01D5/003—Electro-spinning characterised by the initial state of the material the material being a polymer solution or dispersion
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- D—TEXTILES; PAPER
- D04—BRAIDING; LACE-MAKING; KNITTING; TRIMMINGS; NON-WOVEN FABRICS
- D04H—MAKING TEXTILE FABRICS, e.g. FROM FIBRES OR FILAMENTARY MATERIAL; FABRICS MADE BY SUCH PROCESSES OR APPARATUS, e.g. FELTS, NON-WOVEN FABRICS; COTTON-WOOL; WADDING ; NON-WOVEN FABRICS FROM STAPLE FIBRES, FILAMENTS OR YARNS, BONDED WITH AT LEAST ONE WEB-LIKE MATERIAL DURING THEIR CONSOLIDATION
- D04H1/00—Non-woven fabrics formed wholly or mainly of staple fibres or like relatively short fibres
- D04H1/70—Non-woven fabrics formed wholly or mainly of staple fibres or like relatively short fibres characterised by the method of forming fleeces or layers, e.g. reorientation of fibres
- D04H1/72—Non-woven fabrics formed wholly or mainly of staple fibres or like relatively short fibres characterised by the method of forming fleeces or layers, e.g. reorientation of fibres the fibres being randomly arranged
- D04H1/728—Non-woven fabrics formed wholly or mainly of staple fibres or like relatively short fibres characterised by the method of forming fleeces or layers, e.g. reorientation of fibres the fibres being randomly arranged by electro-spinning
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- 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
- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/40—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
- A61L2300/412—Tissue-regenerating or healing or proliferative agents
- A61L2300/414—Growth factors
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- A61L2400/00—Materials characterised by their function or physical properties
- A61L2400/12—Nanosized materials, e.g. nanofibres, nanoparticles, nanowires, nanotubes; Nanostructured surfaces
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
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Abstract
The invention provides a nanofiber dressing and a preparation method thereof, relating to the technical field of biomedical materials, wherein the preparation method comprises the following steps: synthesizing short peptides for recruitment of endogenous VEGF based on amino acid peptization reactions; dissolving the short peptide, the degradable high polymer material and the natural high polymer material in a solvent, and uniformly mixing to obtain an electrostatic spinning solution; carrying out electrostatic spinning on the electrostatic spinning solution by using an electrostatic spinning technology to obtain an electrostatic spinning nanofiber membrane; and drying and cutting the electrostatic spinning nanofiber membrane to obtain the nanofiber dressing. The preparation method of the nanofiber dressing provided by the invention can obviously promote the repair of tissues needing vascular regeneration by utilizing the VEGF recruitment performance of the short peptide, and has obvious healing effect on skin wounds and abnormal body diabetic wounds; meanwhile, the short peptide can be safely degraded in organisms, and the degradation product pair is amino acid and has no toxic action on organisms.
Description
Technical Field
The invention relates to the technical field of biomedical materials, in particular to a nanofiber dressing and a preparation method thereof.
Background
Skin trauma is a common tissue injury that is subjected to uncontrolled external forces, resulting in the detachment or loss of skin tissue. The body generally has spontaneous healing processes after skin trauma, including regeneration of various tissues and processes such as granulation tissue proliferation and scar tissue formation. However, when skin tissue is excessively damaged or skin damage occurs in an abnormal body (a patient with chronic healing such as diabetes) the spontaneous healing rate of the body cannot repair the wound in time, which may cause complications such as wound infection, aggravation of tissue inflammation, and the like. Based on severe trauma patients and chronic healing patients such as diabetes, a safe and feasible wound dressing support needs to be provided to accelerate wound healing.
At present, the medicines commonly used for accelerating the treatment of skin wounds comprise more components such as bioglass, phellodendron bark liquid and the like, but the medicines belong to mixtures, the action mechanism is not clear, the medicines directly act on wounds, and the defects of pharmacological toxicity, incapability of evaluating the safety and the like exist.
In view of the above, it is an urgent technical problem to provide a dressing that does not have toxic effect on the body to accelerate wound healing.
Disclosure of Invention
The invention aims to provide a dressing which has no toxic or harmful effect on organisms to accelerate wound healing.
In order to solve the above problems, the present invention provides a method for preparing a nanofiber dressing, comprising the following steps:
s1: synthesizing short peptides for recruitment of endogenous VEGF based on amino acid peptization reactions;
s2: dissolving the short peptide, the degradable high polymer material and the natural high polymer material in a solvent, and uniformly mixing to obtain an electrostatic spinning solution;
s3: carrying out electrostatic spinning on the electrostatic spinning solution by using an electrostatic spinning technology to obtain an electrostatic spinning nanofiber membrane;
s4: and drying and cutting the electrostatic spinning nanofiber membrane to obtain the nanofiber dressing.
Alternatively, the amino acid sequence of the short peptide is DRVQRQTTTVVA.
Optionally, at least one cysteine is introduced at the N-terminus of the short peptide.
Optionally, the degradable high molecular material is selected from at least one of polylactic acid-glycolic acid copolymer, polyglycolic acid, polylactic acid and polycaprolactone; the natural polymer material is selected from at least one of gelatin, silk fibroin and collagen.
Optionally, the solvent is hexafluoroisopropanol.
Optionally, the concentration of the short peptide in the electrospinning solution is 0.001% mg/mL to 1% mg/mL.
Optionally, the total concentration of the degradable polymer material and the natural polymer material in the electrospinning solution is 10% g/mL to 20% g/mL.
Optionally, the mass ratio of the degradable high polymer material to the natural high polymer material ranges from (3-7): 3.
optionally, the electrospinning technique is selected from one of a coaxial electrospinning technique, an oriented electrospinning technique, a water spinning electrospinning technique.
Another object of the present invention is to provide a nanofiber dressing prepared by the method for preparing a nanofiber dressing as described above.
Compared with the prior art, the preparation method of the nanofiber dressing provided by the invention has the following advantages:
the preparation method of the nanofiber dressing provided by the invention can obviously promote the repair of tissues needing vascular regeneration by utilizing the VEGF recruitment performance of the short peptide, and has obvious healing effect on skin wounds and abnormal body diabetic wounds; meanwhile, the short peptide can be safely degraded in a living body, and the degradation product pair is amino acid and has no toxic action on the body; further, the short peptide is co-spun in the nanofiber through an electrostatic spinning technology, on one hand, the structure of the short peptide is stabilized, on the other hand, by means of the high specific surface area and the extracellular matrix-like structure of the nanofiber, the short peptide can achieve the recruitment function at a wound part through a controllable slow release effect, the usage amount of the polypeptide is further reduced, the cost is saved, and meanwhile, the side effect caused by high dosage is avoided.
Drawings
FIG. 1 is an SEM image of a nanofiber dressing prepared in example 1 of the present invention;
FIG. 2 is an SEM image of a nanofiber dressing prepared in example 2 of the present invention;
FIG. 3 is an SEM image of a nanofiber dressing prepared in example 3 of the present invention;
FIG. 4 is a comparative graph of nanofiber dressings prepared according to various examples of the present invention and comparative examples for skin wound repair in SD rats;
fig. 5 is a comparative graph of the nanofiber dressing prepared in example 3 of the present invention and the comparative example for skin wound repair in diabetic mice.
Detailed Description
Reference will now be made in detail to embodiments of the present invention, examples of which are illustrated in the accompanying drawings, wherein like reference numerals refer to the same or similar elements or elements having the same or similar functions throughout. The embodiments described below with reference to the drawings are exemplary and intended to be illustrative of the present invention and should not be construed as limiting the present invention, and all other embodiments that can be obtained by one skilled in the art based on the embodiments of the present invention without inventive efforts shall fall within the scope of protection of the present invention.
In order to obtain a dressing which has no toxic and harmful effects on organisms to accelerate wound healing, the invention provides a preparation method of a nanofiber dressing, which comprises the following steps:
s1: synthesizing short peptides for recruitment of endogenous VEGF based on amino acid peptization reactions;
s2: dissolving the short peptide, the degradable high polymer material and the natural high polymer material in a solvent, and uniformly mixing to obtain an electrostatic spinning solution;
s3: carrying out electrostatic spinning on the electrostatic spinning solution by using an electrostatic spinning technology to obtain an electrostatic spinning nanofiber membrane;
s4: and drying and cutting the electrostatic spinning nanofiber membrane to obtain the nanofiber dressing.
VEGF (vascular endothelial growth factor) has the effects of promoting vascular permeability increase, vascular endothelial cell migration, proliferation and angiogenesis; the application introduces the short peptide for recruiting endogenous VEGF into the nanofiber dressing, so that when the nanofiber dressing is used for a wound site, the short peptide can help to increase the endogenous VEGF concentration of the wound site, so that the angiogenesis of the wound site can be accelerated, the tissue repair of the wound site can be promoted, and the wound healing can be accelerated; meanwhile, the short peptide can be safely degraded in organisms, the degradation product pair is amino acid, and has no toxic action on the organisms, and the amino acid generated by degradation can be consumed and removed along with the metabolism of the organisms in the organisms.
Furthermore, although the short peptide can accelerate wound healing by promoting angiogenesis at a wound site when acting on the wound site, the short peptide is easily subjected to disadvantages such as unstable structure, large dosage of the short peptide, and short action time if directly injected into the wound site; therefore, the short peptide is co-spun in the nanofiber through the electrostatic spinning technology, on one hand, the structure of the short peptide is stabilized, on the other hand, by means of the high specific surface area and the extracellular matrix-like structure of the nanofiber, the short peptide can achieve the recruitment function at a wound part through a controllable slow release effect, the usage amount of the polypeptide is further reduced, the cost is saved, and meanwhile, the side effect caused by high dosage is avoided.
The preparation method of the nanofiber dressing provided by the invention can obviously promote the repair of tissues needing vascular regeneration by utilizing the VEGF recruitment performance of the short peptide, and has obvious healing effect on skin wounds and abnormal body diabetic wounds; meanwhile, the short peptide can be safely degraded in a living body, and the degradation product pair is amino acid and has no toxic action on the body; further, the short peptide is co-spun in the nanofiber through an electrostatic spinning technology, on one hand, the structure of the short peptide is stabilized, on the other hand, by means of the high specific surface area and the extracellular matrix-like structure of the nanofiber, the short peptide can achieve the recruitment function at a wound part through a controllable slow release effect, the usage amount of the polypeptide is further reduced, the cost is saved, and meanwhile, the side effect caused by high dosage is avoided.
To ensure the effect of accelerating wound healing, the amino acid sequence of the short peptide for recruiting endogenous VEGF is DRVQRQTTTVVA, and the short peptide is marked as short peptide PR 1P.
Specifically, the short peptide PR1P for recruiting endogenous VEGF can be synthesized by chemical method, amino acid is gradually added, and synthesis is performed by dehydration condensation method.
In addition, the sequence of the short peptide PR1P is not limited to the 12 amino acid sequence, and the corresponding increase or decrease of the amino acid can be realized; for example, the function of the short peptide can be further increased by adding functional groups or other functional substances at two ends of the sequence, such as rhodamine grafted on the nitrogen terminal of the short peptide PR1P sequence, and the grafting condition of the short peptide PR1P in the prepared tissue engineering scaffold can be detected.
In order to promote skin wound repair, the application preferably introduces at least one cysteine (C) at the N-terminal of the short peptide; when a skin wound surface is generated, a large amount of peroxy groups are generated at the wound surface, blood vessels at the wound surface are damaged, and the generated peroxy groups cannot be metabolized normally, so that peroxide at the wound surface is accumulated, and wound surface repair is influenced; the application increases the antioxidant function in application by introducing at least one cysteine (C), thereby promoting the repair of skin wound.
The degradable polymer material is used for providing mechanical properties of the nanofiber dressing, and preferably, the degradable polymer material is selected from at least one of polylactic-co-glycolic acid (PLGA), polyglycolic acid (PGA), polylactic acid (PLA) and Polycaprolactone (PCL), and the degradable polymer material can be selected according to specific applications of the nanofiber dressing, for example, when the nanofiber dressing is used as a dressing for a wound site, PLGA and PGA which are easily degradable can be selected; when the compound is used for cartilage repair, a certain repair period is required, and PLA, PCL and the like can be selected.
The natural polymer material is used for providing the compatibility of the nanofiber dressing; the natural polymer material is preferably selected from at least one of gelatin (Gel), silk fibroin and collagen.
The solvent is used for dissolving all components so as to prepare the electrostatic spinning solution with uniform performance; the preferred solvent for this application is Hexafluoroisopropanol (HFIP).
In order to accelerate wound healing on the basis of no toxic action on organisms, the concentration of the short peptide in the electrostatic spinning solution is preferably 0.001-1% mg/mL; further, it is preferable that the concentration of the short peptide PR1P in the electrospinning solution is 0.1% mg/mL, 0.01% mg/mL or 0.001% mg/mL.
In order to give consideration to the mechanical property and compatibility of the nanofiber dressing, the total concentration of the degradable high polymer material and the natural high polymer material in the electrostatic spinning solution is preferably 10-20% g/mL, and the total concentration of the degradable high polymer material and the natural high polymer material is further preferably 15% g/mL; the mass ratio of the preferable degradable high polymer material to the natural high polymer material is (3-7): 3.
according to the invention, the short peptide PR1P is co-spun in the nano fiber through an electrostatic spinning technology to prepare the nano fiber dressing for treating skin wounds; the electrostatic spinning technology is selected from one of a coaxial electrostatic spinning technology, an oriented electrostatic spinning technology and a water spinning electrostatic spinning technology, different electrostatic spinning nanofiber scaffolds are prepared by different electrostatic spinning technologies, and the electrostatic spinning nanofiber scaffolds can be used for tissue engineering regeneration; the prepared dressing bracket is used for treating skin wounds and can be used for treating different wound types such as skin wounds caused by mechanical destructive force, diabetic skin wounds and the like; the nanofiber membrane support with uniform thickness is preferably prepared by an electrostatic spinning technology, the technological parameters of the electrostatic spinning technology are spinning voltage of 8-12 kv, spinning speed of 1.0-2.0 mL/h, a roller receiving device loads tin foil paper as a receiving device, the receiving distance is 5-10 cm, and the rotating speed of the roller is 40-150 rpm.
It is another object of the present invention to provide a nanofiber dressing prepared by the method for preparing a nanofiber dressing as described above.
The nanofiber dressing provided by the invention can remarkably promote the repair of tissues needing vascular regeneration by utilizing the VEGF recruitment performance of the short peptide, and has obvious healing effect on skin wounds and diabetes wounds of abnormal organisms; meanwhile, the short peptide can be safely degraded in a living body, and the degradation product pair is amino acid and has no toxic action on the body; further, the short peptide is co-spun in the nanofiber through an electrostatic spinning technology, on one hand, the structure of the short peptide is stabilized, on the other hand, by means of the high specific surface area and the extracellular matrix-like structure of the nanofiber, the short peptide can achieve the recruitment function at a wound part through a controllable slow release effect, the usage amount of the polypeptide is further reduced, the cost is saved, and meanwhile, the side effect caused by high dosage is avoided.
In the nanofiber dressing provided by the invention, the short peptide PR1P is synthesized relatively simply due to a short amino acid sequence, and meanwhile, the short peptide PR1P is synthesized from 12 amino acids, so that the dressing is safe and degradable in a living body, the degradation product amino acid has no toxic or harmful effect on the body, and finally the amino acid can be removed by random metabolism consumption in the living body; the short peptide PR1P can be further functionalized on the basis that the short peptide PR1P can currently recruit VEGF by grafting different groups or functional substances at the tail end, for example, a fluorescent substance rhodamine can be introduced to detect the binding condition of the short peptide PR1P and a tissue engineering scaffold; the short peptide PR1P is used as a small molecular short peptide, has simple carrier form and various application forms, and can be used for various tissue engineering repair applications; the VEGF recruitment performance of the short peptide PR1P can remarkably promote the repair of tissues needing revascularization, and has obvious healing effect on skin wounds and abnormal body diabetic wounds.
In order to make the aforementioned objects, features and advantages of the present invention comprehensible, embodiments accompanied with figures are described in detail below.
Example 1
The embodiment provides a preparation method of a nanofiber dressing, which comprises the following steps:
s1: preparation of short peptide PR 1P: according to the set amino acid sequence, synthesizing a short peptide PR1P amino acid sequence through an amino acid peptide forming reaction, and further obtaining short peptide PR1P powder through processes of purification, drying and the like;
s2: preparation of short peptide PR1P spinning solution: dissolving 1mg of short peptide PR1P powder in 10mL of HFIP (99%, Shanghai Dairy chemical Co., Ltd.) (the concentration of the short peptide PR1P is 0.01mg/mL), weighing 20 microliter of short peptide PR1P solution with the concentration of 0.01mg/mL, transferring the solution into 19.88mL of HFIP solution, further adding 2g of PLGA (the molar ratio is 75: 25, Jinan Dai bio-engineering Co., Ltd.)/Gel (the molecular weight is >8000, MP Biomedicals, LLC) (w: 7/3) mixture into the 20mL of HFIP solution, and treating the HFIP solution containing the mixture for 24h under a magnetic stirrer to obtain short peptide PR1P spinning solution with the mass concentration of 0.001% (mg/mL) to obtain electrostatic spinning solution;
s3: electrostatic spinning of short peptide PR1P spinning solution: preparing a nano-fiber membrane from the short peptide PR1P electrostatic spinning solution by an electrostatic spinning technology, wherein the spinning voltage is 10kv, the spinning speed is 1.0mL/h, a roller receiving device loads tin foil paper as a receiving device, the receiving distance is 8cm, and the roller rotating speed is 80 rpm;
s4: dressing support: and (3) drying the prepared nanofiber membrane in a vacuum drying oven at 25 ℃ for 2h, volatilizing residual HFIP in the nanofiber membrane, and cutting and customizing the nanofiber dressing in a personalized manner according to the size of a wound part and the shape of the wound.
The prepared nanofiber dressing is detected, and as shown in figure 1, the nanofiber dressing presents a nanofiber silk structure; the content of the short peptide PR1P in the spinning solution is 0.001% (mg/mL), and the nanofiber dressing is used for the skin wound repair of SD rats as shown in figure 4.
Example 2
The embodiment provides a preparation method of a nanofiber dressing, which comprises the following steps:
s1: preparation of short peptide PR 1P: according to the set amino acid sequence, synthesizing a short peptide PR1P amino acid sequence through an amino acid peptide forming reaction, and further obtaining short peptide PR1P powder through processes of purification, drying and the like;
s2: preparation of short peptide PR1P spinning solution: dissolving 1mg of short peptide PR1P powder in 10mL of HFIP (99%, Shanghai Dairy chemical Co., Ltd.) (the concentration of the short peptide PR1P is 0.01mg/mL), weighing 200 microliter of short peptide PR1P solution with the concentration of 0.01mg/mL, transferring the solution into 19.88mL of HFIP solution, further adding 2g of PLGA (the molar ratio is 75: 25, Jinan Dai bio-engineering Co., Ltd.)/Gel (the molecular weight is >8000, MP Biomedicals, LLC) (w: 7/3) mixture into 20mL of the HFIP solution, and treating the HFIP solution containing the mixture for 24h under a magnetic stirrer to obtain short peptide PR1P spinning solution with the mass concentration of 0.001% (mg/mL) to obtain electrostatic spinning solution;
s3: electrostatic spinning of short peptide PR1P spinning solution: preparing a nano-fiber membrane from the short peptide PR1P electrostatic spinning solution by an electrostatic spinning technology, wherein the spinning voltage is 10kv, the spinning speed is 1.0mL/h, a roller receiving device loads tin foil paper as a receiving device, the receiving distance is 8cm, and the roller rotating speed is 80 rpm;
s4: dressing support: and (3) drying the prepared nanofiber membrane in a vacuum drying oven at 25 ℃ for 2h, volatilizing residual HFIP in the nanofiber membrane, and cutting and customizing the nanofiber dressing in a personalized manner according to the size of a wound part and the shape of the wound.
The prepared nanofiber dressing is detected, and as shown in fig. 2, the nanofiber dressing presents a nanofiber silk structure; the content of the short peptide PR1P in the spinning solution is 0.01% (mg/mL), and the nanofiber dressing is used for the skin wound repair of SD rats as shown in figure 4.
Example 3
The embodiment provides a preparation method of a nanofiber dressing, which comprises the following steps:
s1: preparation of short peptide PR 1P: according to the set amino acid sequence, synthesizing a short peptide PR1P amino acid sequence through an amino acid peptide forming reaction, and further obtaining short peptide PR1P powder through processes of purification, drying and the like;
s2: preparation of short peptide PR1P spinning solution: dissolving 1mg of short peptide PR1P powder in 1mL of HFIP (99%, Shanghai Dairy chemical Co., Ltd.) (the concentration of the short peptide PR1P is 0.1mg/mL), weighing 200 microliter of short peptide PR1P solution with the concentration of 0.1mg/mL, transferring the solution into 19.8mL of HFIP solution, further adding 2g of PLGA (the molar ratio is 75: 25, Jinan Dai bio-engineering Co., Ltd.)/Gel (the molecular weight is >8000, MP Biomedicals, LLC) (w: 7/3) mixture into 20mL of HFIP solution, treating the HFIP solution containing the mixture for 24h under a magnetic stirrer to obtain short peptide PR1P spinning solution with the mass concentration of 0.1% (mg/mL) to obtain electrostatic spinning solution;
s3: electrostatic spinning of short peptide PR1P spinning solution: preparing a nano-fiber membrane from the short peptide PR1P electrostatic spinning solution by an electrostatic spinning technology, wherein the spinning voltage is 10kv, the spinning speed is 1.0mL/h, a roller receiving device loads tin foil paper as a receiving device, the receiving distance is 8cm, and the roller rotating speed is 80 rpm;
s4: dressing support: and (3) drying the prepared nanofiber membrane in a vacuum drying oven at 25 ℃ for 2h, volatilizing residual HFIP in the nanofiber membrane, and cutting and customizing the nanofiber dressing in a personalized manner according to the size of a wound part and the shape of the wound.
The prepared nanofiber dressing is tested, and as shown in fig. 3, the nanofiber dressing presents a nanofiber silk structure; the content of the short peptide PR1P in the spinning solution is 0.1% (mg/mL), and the nanofiber dressing is used for the skin wound repair of SD rats as shown in figure 4; in addition, the nanofiber dressing is used for wound repair of diabetic mice, and the condition is shown in figure 5.
Comparative example 1
The preparation method for preparing the pure PLGA/Gel nanofiber dressing comprises the following steps:
s1: preparing a spinning solution: adding 2g PLGA (molar ratio 75: 25, Jinan Dai handle bio engineering Co., Ltd.)/Gel (molecular weight >8000, MP Biomedicals, LLC) (w: w ═ 7/3) mixture into 20mL HFIP solution, treating the HFIP solution containing the mixture for 24h under a magnetic stirrer to obtain electrostatic spinning solution;
s2: preparing the electrostatic spinning solution into a nanofiber membrane by an electrostatic spinning technology, wherein the spinning voltage is 10kv, the spinning speed is 1.0mL/h, a roller receiving device loads tin foil paper as a receiving device, the receiving distance is 8cm, and the roller rotating speed is 80 rpm;
s3: dressing support: the prepared nanofiber membrane is placed in a vacuum drying oven at 25 ℃ for drying for 2h, residual HFIP in the nanofiber membrane is volatilized, and pure PLGA/Gel nanofiber dressing is customized according to the size of a wound part and the shape of the wound.
The pure PLGA/Gel nanofiber dressing is used for the skin wound repair of SD rats and is shown in figure 4 (control group); the conditions for wound repair in diabetic mice are shown in figure 5 (control).
As seen from fig. 4, when the nanofiber dressing prepared in examples 1 to 3 is used for the skin wound repair of SD rats, compared with the pure PLGA/Gel nanofiber dressing prepared in comparative example 1, the nanofiber dressing prepared in example 3 has a more obvious effect of promoting the wound healing of SD rats, and the nanofiber dressing prepared in example 3 has a better effect, so that the nanofiber dressing provided by the present invention can significantly promote the repair of tissues requiring blood vessel regeneration, and has an obvious effect of healing skin wounds; fig. 5 shows that, when the nanofiber dressing prepared in example 3 is used for repairing diabetic mouse wounds, compared with the pure PLGA/Gel nanofiber dressing prepared in comparative example 1, the nanofiber dressing prepared in example 3 has an effect of more obviously promoting the diabetic mouse wounds to heal, so that the nanofiber dressing provided by the invention can obviously promote the repair of tissues needing blood vessel regeneration, and has an obvious effect of healing the diabetic wounds of abnormal organisms.
Although the present disclosure has been described above, the scope of the present disclosure is not limited thereto. Those skilled in the art can make various changes and modifications without departing from the spirit and scope of the present disclosure, and such changes and modifications will fall within the scope of the present invention.
Claims (10)
1. The preparation method of the nanofiber dressing is characterized by comprising the following steps of:
s1: synthesizing a short peptide for recruiting endogenous VEGF based on an amino acid peptide formation reaction;
s2: dissolving the short peptide, the degradable high polymer material and the natural high polymer material in a solvent, and uniformly mixing to obtain an electrostatic spinning solution;
s3: carrying out electrostatic spinning on the electrostatic spinning solution by using an electrostatic spinning technology to obtain an electrostatic spinning nanofiber membrane;
s4: and drying and cutting the electrostatic spinning nanofiber membrane to obtain the nanofiber dressing.
2. The method of making a nanofiber dressing as claimed in claim 1, wherein the amino acid sequence of the short peptide is DRVQRQTTTVVA.
3. The method of preparing the nanofiber dressing of claim 2, wherein at least one cysteine is introduced at the N-terminus of the short peptide.
4. The preparation method of the nanofiber dressing as claimed in claim 3, wherein the degradable high molecular material is selected from at least one of polylactic acid-glycolic acid copolymer, polyglycolic acid, polylactic acid and polycaprolactone; the natural polymer material is selected from at least one of gelatin, silk fibroin and collagen.
5. The method of preparing a nanofiber dressing as claimed in claim 3, wherein the solvent is hexafluoroisopropanol.
6. The method of preparing the nanofiber dressing according to any one of claims 1 to 5, wherein the concentration of the short peptide in the electrospinning solution is 0.001% mg/mL to 1% mg/mL.
7. The preparation method of the nanofiber dressing of claim 6, wherein the total concentration of the degradable high polymer material and the natural high polymer material in the electrospinning solution is 10% g/mL-20% g/mL.
8. The preparation method of the nanofiber dressing as claimed in claim 7, wherein the mass ratio of the degradable high polymer material to the natural high polymer material is (3-7): 3.
9. the method of preparing the nanofiber dressing of claim 1, wherein the electrospinning technique is selected from one of a coaxial electrospinning technique, an oriented electrospinning technique, and a water-spinning electrospinning technique.
10. A nanofiber dressing prepared by the method for preparing a nanofiber dressing as claimed in any one of claims 1 to 9.
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