WO2020204230A1 - Composite de nanofibre et d'hydrogel, et échafaudage pour régénération tissulaire le comprenant - Google Patents

Composite de nanofibre et d'hydrogel, et échafaudage pour régénération tissulaire le comprenant Download PDF

Info

Publication number
WO2020204230A1
WO2020204230A1 PCT/KR2019/003978 KR2019003978W WO2020204230A1 WO 2020204230 A1 WO2020204230 A1 WO 2020204230A1 KR 2019003978 W KR2019003978 W KR 2019003978W WO 2020204230 A1 WO2020204230 A1 WO 2020204230A1
Authority
WO
WIPO (PCT)
Prior art keywords
hydrogel
nanofibers
cells
composite
tissue regeneration
Prior art date
Application number
PCT/KR2019/003978
Other languages
English (en)
Korean (ko)
Inventor
이강원
양충모
고원건
국윤민
Original Assignee
서울대학교산학협력단
연세대학교 산학협력단
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by 서울대학교산학협력단, 연세대학교 산학협력단 filed Critical 서울대학교산학협력단
Priority to PCT/KR2019/003978 priority Critical patent/WO2020204230A1/fr
Publication of WO2020204230A1 publication Critical patent/WO2020204230A1/fr

Links

Images

Classifications

    • 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
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/40—Composite materials, i.e. containing one material dispersed in a matrix of the same or different material
    • A61L27/44—Composite materials, i.e. containing one material dispersed in a matrix of the same or different material having a macromolecular matrix
    • A61L27/48—Composite materials, i.e. containing one material dispersed in a matrix of the same or different material having a macromolecular matrix with macromolecular fillers
    • 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
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
    • 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
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
    • A61L27/52—Hydrogels or hydrocolloids
    • 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
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
    • A61L27/54—Biologically active materials, e.g. therapeutic substances
    • 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
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
    • A61L27/56—Porous materials, e.g. foams or sponges

Definitions

  • the present invention relates to a composite of nanofibers and hydrogel and a scaffold for tissue regeneration including the same, and more specifically, a composite of nanofibers and hydrogel that can be effectively used for regeneration of biological tissues, particularly myocardial tissues, and It relates to a scaffold for tissue regeneration, and a method of manufacturing the same.
  • Cardiovascular diseases such as coronary artery disease, arrhythmia, and heart failure have rapidly increased due to westernized diet, lack of exercise, and aging. Death from working with heart disease now has the second highest mortality rate among major causes of death. Cardiovascular for the fundamental treatment of cardiovascular disease. Tissue regeneration is very important.
  • heart transplantation or cell transplantation therapy has been used for treatment.
  • the number of donors is significantly less than the number of patients required, and the cost of surgery is also a heavy burden on patients.
  • the cell transplantation treatment method is a method of allografting cardiomyocytes into damaged tissues, but it is still impossible to proliferate as many cardiomyocytes as necessary at an appropriate time, and thus, there is a need to develop a technology to replace them.
  • Patent Document 1 discloses a method of encapsulating human pancreatic cells and PDX1 (duodenal homeobox gene 1) positive human pancreatic progenitor cells using a device based on a polyethylene glycol hydrogel, a biocompatible polymer. According to this, there is an advantage that the encapsulated progenitor cells are well differentiated, but there is a disadvantage in that physical properties such as biodegradability of the hydrogel cannot be easily adjusted because differentiation is controlled using a differentiation medium.
  • Patent Document 2 discloses a method of encapsulating PDX1-positive human pancreatic progenitor cells by fabricating a three-dimensional cell encapsulation device using PDMS (polydimethylsiloxane). This method is effective for cell differentiation, but due to the nature of PDMS, a material constituting the device, cell adhesion is low and modification is difficult, so it is difficult to control physical properties.
  • PDMS polydimethylsiloxane
  • Patent Document 3 devised a hydrogel composite of nanofibers and alginate by electrospinning.
  • the nanofibers were used as the cell adhesion layer, and the nanofibers were encapsulated with hydrogel to prepare a scaffold for tissue regeneration.
  • alginate hydrogel due to the nature of alginate hydrogel, there is a problem in that it does not proliferate smoothly with respect to adherent and proliferating cells, and thus, it is difficult to generate very important blood vessels during tissue regeneration.
  • An object of the present invention is to provide a complex that can promote blood vessel formation, which is important for tissue regeneration, and can be used as an effective scaffold for tissue regeneration, which can have properties suitable for tissue transplantation.
  • Another object of the present invention is to provide a scaffold for tissue regeneration comprising the complex.
  • Another object of the present invention is to provide a method for preparing the composite.
  • a first hydrogel layer encapsulating the nanofibers in the first hydrogel
  • It provides a composite of nanofibers and a hydrogel, including a second hydrogel layer, encapsulating the first hydrogel layer in a second hydrogel.
  • Another aspect of the present invention provides a scaffold for tissue regeneration comprising a composite of nanofibers and hydrogel according to the above aspect.
  • Forming a first hydrogel layer encapsulating the nanofibers by putting the nanofibers into a frame, adding a first hydrogel precursor solution, and then crosslinking the nanofibers;
  • a composite of nanofibers and hydrogel according to the above aspect comprising the step of forming a second hydrogel layer encapsulating the first hydrogel layer by adding a second hydrogel precursor solution to the first hydrogel layer and then crosslinking reaction It provides a method of manufacturing.
  • the composite of nanofibers and hydrogel according to an aspect of the present invention introduces a first hydrogel layer capable of attaching and cultivating cells to the nanofibers, thereby culturing cells for tissue regeneration on the nanofibers, and the first hydrogel layer It is possible to cultivate vascular cells essential for tissue regeneration.
  • the complex includes a second hydrogel layer at the outermost surface of which cells are difficult to adhere to, so that when the complex is transplanted into a living body for tissue regeneration as a scaffold for tissue regeneration, fibroblasts in the living body and It can block the fibrosis process by preventing the adhesion of collagen, etc., enabling smooth transplantation.
  • the complex according to an aspect of the present invention can sufficiently form blood vessels, which are essential for tissue regeneration, and avoid the rejection reaction of the living body such as fibrosis that may occur during the scaffold transplantation process, so that more effective tissue regeneration It can be used as a scaffold for use, especially as a scaffold for cardiovascular tissue regeneration.
  • FIG. 1 is a schematic diagram of a scaffold for tissue regeneration according to an embodiment of the present invention, comprising electrospun nanofibers, a fibrin hydrogel layer encapsulating the nanofibers, and an alginate hydrogel layer encapsulating the fibrin hydrogel layer.
  • biomaterials may be released and exchanged, and protein and cell invasion from the outside may be prevented or controlled, and co-culture of cells may be performed on the electrospun nanofibers, and the fibrin hydrogel
  • the creation of vascular tissue is possible.
  • Figure 2 is the present, including the process of manufacturing a nanofiber scaffold by electrospinning (step 1), disinfecting the scaffold (step 2), encapsulating fibrin hydrogel (step 3), and encapsulating alginate hydrogel (step 4). It is a schematic diagram showing the manufacturing process of the composite according to one embodiment of the invention.
  • FIG. 3 shows the manufacturing process (a,b) of nanofibers by electrospinning, and the shape (c,d) of the electrospun nanofiber scaffold as viewed through a scanning electron microscope.
  • each fibrin hydrogel (a) prepared in Examples 1 to 3, a composite of electrospun nanofibers and fibrin hydrogel (b), an electrospun nanofiber, fibrin hydrogel, and a composite of alginate hydrogel ( This is a photograph of the result of observing c) with the naked eye.
  • Fibrin-PCL-alginate is a composite of electrospun nanofibers (Fibrin), electrospun nanofibers and fibrin hydrogel (Fibrin-PCL) prepared in Examples 1, 2 and 3, and electrospun nanofibers, fibrin hydrogel, and alginate hydro
  • a graph showing the results of measuring the mechanical strength of the gel composite (Fibrin-PCL-alginate) is shown.
  • Example 6 is a photograph taken by observing and taking a cross section of the nanocage prepared in Example 3 using a cryo-scanning electron microscopy (Cryo-SEM).
  • Figure 7 is to confirm the viability of HUVECs inside the fibrin hydrogel, without electrospun nanofibers, HUVECs were dispersed in the fibrin hydrogel at a ratio of 1 ⁇ 10 5 cells/ml, cultured for a week, and This is a picture of the results of confirming the viability by staining with the Live/Dead cell viability assay.
  • Figure 8 is, in order to confirm the appearance of the composite of the first hydrogel and the second hydrogel, after coating the HUVEC-containing fibrin hydrogel with alginate hydrogel without electrospun nanofibers, 1, 4, in EGM-2 Bullekit mixed medium. And after incubation for 7 days, the cross section of the two hydrogels was taken through an optical microscope.
  • FIG. 10 is a graph showing the results of measuring the activity of cells by culturing HUVEC and ADSC in fibrin hydrogel and electrospun nanofibers for 1 week, respectively, in order to observe the activity of cells inside the nanocage according to an embodiment of the present invention to be.
  • the present inventors have carefully studied not only the cultivation of cells for tissue regeneration, but also the scaffold that is advantageous for culturing vascular cells that can form vascular tissue essential for tissue regeneration, and as a result, nanofibers, the primary encapsulating the nanofibers
  • a first hydrogel layer encapsulating the nanofibers in the first hydrogel
  • It provides a composite of nanofibers and a hydrogel, including a second hydrogel layer, encapsulating the first hydrogel layer in a second hydrogel.
  • nano cage the composite of nanofibers and hydrogel according to an aspect of the present invention is also referred to as a “nano cage”.
  • the nanofibers are located inside the complex, and when the complex is used as a scaffold for tissue regeneration, it is used for culturing cells for tissue regeneration, for example, co-culture of stem cells and cardiomyocytes. It is a part.
  • the nanofiber is not particularly limited as long as it is a nanofiber in which cells for tissue regeneration can be cultured, but in one embodiment, the nanofiber is an electrospun nanofiber manufactured by electrospinning.
  • the electrospun nanofibers may be any electrospun nanofibers conventionally known to be suitable for culturing living cells.
  • the electrospun nanofibers are an aggregate of fibers having a diameter of several tens of nanometers to several micrometers, and the structure includes a plurality of pores of various sizes, and forms a three-dimensional structure capable of attaching cells. I can. In addition, these voids have a predetermined shape, size and volume.
  • the diameter of the electrospun nanofibers can be changed by adjusting conditions such as the concentration of the polymer solution used as the material of the electrospun nanofibers, the flow rate of the electrospinning, and the voltage, and can also be changed by adding a salt to the polymer solution.
  • the size of the pores of the electrospun nanofibers may be adjusted to 20 to 100 ⁇ m. Since most of the cells are fixed on the scaffold and exist at 10 to 15 ⁇ m before transformation occurs, cells for tissue regeneration enter the pores within the above size range and are fixed inside the scaffold, making it suitable for growth and differentiation. have.
  • the electrospun nanofibers may be nanofibers manufactured by a known electrospinning method.
  • the material used to produce the nanofibers may be any material suitable for culturing cells for tissue regeneration and capable of producing nanofibers.
  • the scaffold formed of nanofibers may be subjected to additional modifications to ensure that the biomaterial is well fixed to the nanofibers. For example, for the additional modification, an oxygen plasma treatment, a radiation grafting method, or a self-assembly monolayer (SAM) method may be used. In the case of oxygen plasma treatment, nanofibers with high hydrophobicity are used to increase hydrophilicity.
  • SAM self-assembly monolayer
  • a benzophenone and azide material for example, by UV irradiation on a nanofiber scaffold through a radiation grafting method, reactivity can be given to transform the surface of the nanofiber into a desired material.
  • the SAM method uses a spontaneous reaction between a surface where a hydroxy functional group exists and silane (Silane) to fix a portion bonded to the silane to the surface.
  • Silane silane
  • N-hydroxysuccinimide (NHS) N-hydroxysuccinimide
  • the additional modification to the scaffold formed of such nanofibers allows the biomaterial to be fixed well regardless of the type of the nanofiber-generating material, there is no limitation on the type of the nanofiber-generating material.
  • the type of polymer forming the electrospun nanofibers is not particularly limited as long as it does not inhibit the culture of cells for tissue regeneration, and various natural polymers and synthetic polymers may be used.
  • a polymer material that does not have biodegradable properties in terms of cell adhesion may be used, but preferably a polymer material having biodegradation properties in terms of drug delivery, cell transplantation and cell therapy may be used.
  • biodegradable polymers are, for example, chitosan, elastin, hyaluronic acid, alginate, gelatin, collagen, cellulose, polyethylene glycol (PEG), polyethylene oxide (PEO), polycaprolactone (PCL), polylactic acid (PLA), and polyglycol.
  • PGA Poly[(lactic-co-(glycolic acid))(PLGA), poly[(3-hydroxybutyrate)-co-(3-hydroxyvalerate) (PHBV), polydioxanone (PDO) ), poly[(L-lactide)-co-(caprolactone)], poly(ester urethane)(PEUU), poly[(L-lactide)-co-(D-lactide)], poly[ethylene -co-(vinyl alcohol)](PVOH), polyacrylic acid (PAA), polyvinyl alcohol (PVA), polyvinylpyrrolidone (PVP), polystyrene (PS), polyaniline (PAN), and any combination thereof It can be selected from the group consisting of.
  • the biodegradable polymer used in the manufacture of electrospun nanofibers is an electrospun nanofiber of a mixture of PCL and gelatin.
  • the electrospun nanofibers may also be suitable for attachment, proliferation and differentiation of stem cells as well as cells for tissue regeneration.
  • the nanofibers may be coated with fibronectin for smoother adhesion of cells.
  • growth factors used to help proliferation and differentiation of cells can be introduced into electrospun nanofibers.
  • the polymer solution may be mixed and injected into the fiber, or may be physically/chemically attached to the outside of the fiber to be introduced.
  • the electrospun nanofibers are prepared by electrospinning a polymer solution, and may be prepared using a method commonly used in the art.
  • a polymer solution for electrospinning may be prepared by selecting a suitable solvent capable of dissolving the polymers as listed above for the production of electrospun nanofibers.
  • a suitable solvent capable of dissolving the polymers as listed above for the production of electrospun nanofibers.
  • a solvent is not particularly limited as long as the above conditions are satisfied, but for example, chloroform, trifluoroethanol (2,2,2-trifluoroethanol, TFE), tetrahydrofuran (THF), dimethylform
  • TFE trifluoroethanol
  • THF tetrahydrofuran
  • dimethylform It may be selected from aldehydes (Dimethylformaldehyde, DMF), and any combination thereof, and may be appropriately selected by a person skilled in the art in consideration of factors such as viscosity and dielectric constant of the solution.
  • the electrospinning method for producing the electrospun nanofibers when a high voltage is applied to the polymer solution, one charge of + or-is accumulated in the polymer solution, and the surface tension of the polymer solution is exceeded by the mutual repulsion between the same charges. As the solvent evaporates, the fibers are charged with the opposite charge transferred to the solution or collected into a grounded substrate.
  • the diameter of the fiber can be adjusted by controlling the concentration, flow rate, voltage, etc. of the solution in which the polymer is dissolved using the solvent, and the density of the fiber can be controlled by controlling the electrospinning time and height of the nozzle and the substrate.
  • the solvent used in the polymer solution for preparing the electrospinning nanofibers may have toxicity that is not suitable for culturing cells for tissue regeneration, and is not completely evaporated during the electrospinning process and remains in the obtained nanofibers, causing cytotoxicity. There is concern. Therefore, in order to use the complex as a scaffold for tissue engineering, it is necessary to completely remove the residual solvent. In order to remove the residual solvent, there is a method of storing the fibers at a temperature of several tens of C for evaporation of the solvent, or storing the fibers in a vacuum state, and the above methods may be used in combination for more effective treatment. .
  • the nanofibers are encapsulated with the first hydrogel and then again encapsulated with the second hydrogel, thereby forming the nanofiber and hydrogel composite.
  • the first hydrogel encapsulating the nanofibers may be formed by inserting the nanofibers into the first hydrogel to encapsulate the nanofibers.
  • the nanofibers may be encapsulated with the first hydrogel by, for example, putting nanofibers in a hydrogel precursor solution and crosslinking the hydrogel precursor solution to form a hydrogel.
  • the nanofibers encapsulated with the first hydrogel may be encapsulated with the second hydrogel in the same manner.
  • the first hydrogel is a hydrogel suitable for culturing vascular cells required for blood vessel formation due to its high cell adhesion, and the second hydrogel has low cell adhesion and prevents adhesion proliferation when the complex is transplanted into a living body as a scaffold. It is a hydrogel that can prevent external invasion of cells. That is, the first hydrogel has excellent adhesion of vascular endothelial cells and plays a role that can greatly aid in proliferation and angiogenesis, and the second hydrogel has a significantly smaller pore size than the first hydrogel, and adheres to cells.
  • the ability to encapsulate cells is poor, but it is not helpful for the proliferation of adherent-proliferating cells, but has the advantage of preventing external invasion of proteins and cells during the proliferation of non-adherent proliferating cells and transplantation into the body.
  • the first hydrogel may be a porous hydrogel having pores of 1 ⁇ m to 100 ⁇ m
  • the second hydrogel may be a porous hydrogel having pores of 1 nm to 10 nm.
  • hydrogels have a three-dimensional structure like electrospun nanofibers, they help in three-dimensional cultivation of cells, but can show a large difference in the adhesion of cells depending on the properties of the polymers that make up the hydrogel.
  • macromolecules such as collagen exhibit good properties for cell adhesion in the form of a hydrogel, but polymers such as PEG and alginate have great restrictions on cell adhesion when produced as a hydrogel. Suitable. Therefore, the first hydrogel and the second hydrogel can be prepared by appropriately selecting a polymer.
  • the cell adhesion may vary depending on the conditions for producing the hydrogel. Therefore, the cell adhesion capability of the hydrogel may be adjusted by controlling the porosity according to the production conditions, and according to methods known in the art. Accordingly, a person skilled in the art can properly manufacture.
  • the first hydrogel may be formed of a biocompatible polymer suitable for culturing vascular cells required for angiogenesis, for example, selected from the group consisting of collagen, chitosan, fibrin, copolymers thereof, and any combination thereof. It may be a hydrogel of a biocompatible polymer, but is not limited thereto.
  • the first hydrogel is a fibrin hydrogel.
  • the composite of the nanofibers and the hydrogel includes a porous first hydrogel layer, it is possible to adhere and cultivate vascular cells, thereby enabling the formation of blood vessels essential for tissue regeneration.
  • the second hydrogel may be formed of a biocompatible polymer while being able to block adhesion and proliferation of cells.
  • a biocompatible polymer for example, polyethylene glycol (PEG), polyethylene oxide (PEO), polyhydroxyethyl methacrylate (PHEMA), Polyacrylic acid (PAA), polyvinyl alcohol (PVA), poly(N-isopropylacrylamide) (PNIPAM), polyvinylpyrrolidone (PVP), polylactic acid (PLA), polyglycolic acid (PGA), polycapro Lactone (PCL), gelatin, alginic acid, carrageenan, chitosan, hydroxyalkylcellulose, alkylcellulose, silicone, rubber, agar, carboxyvinyl copolymer, polydioxolane, polyacrylic acetate, polyvinyl chloride, maleic anhydride/vinyl ether, And it may be a hydrogel of a biocompatible polymer selected from the group consisting of any combination thereof, but is not limited
  • the second hydrogel is alginic acid hydrogel.
  • the composite of the nanofibers and the hydrogel further includes a secondary hydrogel layer
  • a secondary hydrogel layer when the composite is implanted as a scaffold, there is an advantage that it is possible to block rejection reactions in the body such as fibrosis.
  • fibrosis occurs and the effect of transplantation of external tissues is very poor.
  • passivation by encapsulating the hydrogel excellent effects can be obtained for in vivo transplantation.
  • the first hydrogel layer and the second hydrogel layer may form a hydrogel layer by adding a hydrogel precursor solution and performing a crosslinking reaction.
  • Polymers forming hydrogels have various crosslinking methods according to their respective characteristics. For example, photo-crosslinking by ultraviolet (UV) irradiation, crosslinking using a reactive crosslinker, etc. are mentioned.
  • the crosslinked hydrogel using this method is not easily decomposed and crosslinked.
  • a patterning technique can be applied by crosslinking the hydrogel only in a specific area using a mask.
  • the thermal-crosslinking method which crosslinks by temperature, is crosslinked at a specific temperature or higher to form a hydrogel, and can be applied to cell release and drug delivery through temperature change.
  • crosslinking methods using specific protein reactions and reactions between ions and polymers have the advantage of being able to crosslink hydrogels using well-known reactions between proteins, and to easily produce hydrogels.
  • fibrin hydrogel when fibrin hydrogel is included as the first hydrogel, fibrin hydrogel may be formed by using fiprinogen as a hydrogel precursor solution and adding thrombin as a crosslinking agent.
  • the alginate hydrogel when the alginate hydrogel is included as the second hydrogel, the alginate hydrogel may be formed by using the alginate salt solution as a precursor solution such as calcium chloride.
  • the hydrogel Since the hydrogel has a three-dimensional porous structure like the electrospun nanofibers, it can function as a drug delivery system through the release of drugs from the inside.
  • the hydrogel may be capable of sustained drug release.
  • the drug In order to introduce a drug into the hydrogel, if the drug is added before the crosslinking reaction for the hydrogel form and then crosslinked, the drug can be incorporated into the hydrogel, and the drug incorporated from the hydrogel can be continuously delivered.
  • the drug may be incorporated into the hydrogel from the medium by adding the drug to an external medium in the cell culture process using the hydrogel.
  • VEGF Vascular endothelial growth factor
  • S1P Sphingosin-1-phosphate
  • the composite of the nanofibers and the hydrogel may have sufficient strength such that it is not easily damaged during implantation in vivo for tissue regeneration.
  • the composite may have better strength due to the fact that it includes a first hydrogel layer and a second hydrogel layer encapsulating nanofibers, the strength may vary depending on the degree of crosslinking of the hydrogel, the thickness of the nanofibers, the thickness of the hydrogel, etc. It can be adjusted according to a known method by a person skilled in the art. Accordingly, if the complex has a sufficient strength, a scaffold for tissue regeneration can be produced similar to the physical strength of the extracellular matrix of each tissue to be regenerated. If the physical strength is similar, the tissue of the cells in the scaffold It has the advantage that specific specificity can be easily assigned.
  • the composite may have a strength of 0.1 to 0.5 MPa.
  • the intensity is higher than the above intensity, blood vessel regeneration may not be performed smoothly, and when it is low, it may be difficult to control the differentiation of stem cells.
  • the complex may include cells for tissue regeneration adhered and cultured on the nanofibers to be used as a scaffold for tissue regeneration.
  • Cells for tissue regeneration may be used for any desired tissue regeneration, for example, cardiomyocytes, smooth muscle cells, cartilage cells, bone cells, skin cells, fibroblasts, vascular endothelial cells, neurons, Schwann cells. , Stem cells, and any combination thereof, but is not limited thereto.
  • cardiomyocytes and stem cells may be co-cultured and introduced on the nanofibers.
  • the first hydrogel layer may contain cells, biomaterials, or any combination thereof.
  • the cells and/or biomaterials may be any cells or biomaterials that can contribute to tissue regeneration.
  • fibroblasts, vascular endothelial cells, neurons, Schwann cells, stem cells, and stromal cells selected from any combination thereof may be used in combination.
  • the biological material may be a biological material that may contribute to tissue regeneration, and for example, may be a growth factor such as VEGF or S1P mentioned above that promotes proliferation of vascular cells.
  • the first hydrogel layer may contain human umbilical cord blood vein endothelial cells (HUVEC), and because of containing HUVEC, angiogenesis may be promoted, and thus tissue regeneration may be remarkably advantageous. Additionally, the first hydrogel layer may contain a growth factor, and due to the inclusion of the growth factor, the proliferation of cells adhered to the nanofibers and proliferation of vascular cells in the first hydrogel layer may be promoted. In one embodiment, the growth factor is vascular endothelial growth factor (VEGF).
  • VEGF vascular endothelial growth factor
  • the complex contains cardiomyocytes and stem cells adhered and cultured on nanofibers, and the first hydrogel layer contains HUVEC.
  • the complex contains cardiomyocytes and stem cells adhered and cultured on nanofibers, and the first hydrogel layer is a complex containing HUVEC and VEGF.
  • the present invention provides a scaffold for tissue regeneration comprising a composite of nanofibers and a hydrogel according to the above aspect.
  • scaffold for tissue regeneration may be the description of the composite of nanofibers and hydrogels according to an aspect of the present invention.
  • the term "scaffold” refers to a structure that serves to provide an environment suitable for attachment, differentiation of cells, and proliferation and differentiation of cells moving from around tissues as one of important basic elements in the field of tissue regeneration engineering.
  • the tissue regeneration scaffold may determine a tissue capable of regeneration according to the type of cells adhered and cultured on the nanofibers.
  • a tissue capable of regeneration may be used.
  • cardiomyocytes for cardiovascular tissue regeneration cartilage cells for cartilage tissue regeneration, bone cells for bone tissue regeneration, skin cells for skin tissue regeneration, fibroblasts for wound tissue regeneration, and the like may be used.
  • the tissue regeneration scaffold is a cardiovascular tissue regeneration scaffold, and contains adherent-cultured cardiomyocytes or additional stem cells on the nanofibers.
  • the tissue regeneration scaffold is a wound treatment scaffold
  • the nanofibers contain fibroblasts, or additionally stem cells.
  • the scaffold for tissue regeneration according to the present invention can block the fibrosis process by blocking the adhesion of fibroblasts and collagen in a living body to the scaffold due to passivation due to the introduction of a secondary hydrogel layer, so that smooth transplantation is possible. .
  • the scaffold for tissue regeneration can form vascular tissue by culturing vascular cells in the first hydrogel layer, so that when the scaffold is implanted in vivo, it can stably and effectively regenerate living tissue.
  • the complex according to an aspect of the present invention is capable of sufficiently forming blood vessels essential for tissue regeneration and avoiding the rejection reaction of the living body such as fibrosis that may occur during the scaffold transplantation process. It can be used as a scaffold, in particular a scaffold for cardiovascular tissue regeneration.
  • the scaffold for tissue regeneration can enable continuous drug release by incorporating desired drugs into electrospun nanofibers and hydrogels, and scaffolds by incorporating drugs effective in constructing an extracellular matrix (ECM) It can promote the smooth formation of extracellular matrix in surrounding tissues to be transplanted. Due to the inclusion of these drugs, more effective applications are possible in fields such as wound healing.
  • ECM extracellular matrix
  • FIG. 1 is a schematic diagram of a scaffold for tissue regeneration according to an embodiment of the present invention.
  • the scaffold for tissue regeneration according to FIG. 1 has electrospun nanofibers, a fibrin hydrogel layer encapsulating the nanofibers, and an alginate hydrogel layer encapsulating the fibrin hydrogel layer.
  • biomaterials can be released and exchanged, protein and cell invasion from the outside can be prevented or controlled, and co-culture of cells can be performed on the electrospun nanofibers, and blood vessels in the fibrin hydrogel Organizational creation is possible.
  • Forming a first hydrogel layer encapsulating the nanofibers by putting the nanofibers into a frame, adding a first hydrogel precursor solution, and then crosslinking the nanofibers;
  • a nanofiber and hydrogel according to an aspect of the present invention comprising the step of forming a second hydrogel layer encapsulating the first hydrogel layer by adding a second hydrogel precursor solution to the first hydrogel layer and then crosslinking. It provides a method for preparing a gel complex.
  • the first hydrogel precursor is fibrinogen, and the crosslinking reaction may be performed by using thrombin as a crosslinking agent.
  • the second hydrogel precursor is an aqueous alginate solution, and the crosslinking reaction may be performed by using calcium chloride as a crosslinking agent.
  • the aqueous alginate solution is, for example, sodium daily acid.
  • the nanofibers may be used after seeding the cell dispersion of cells for tissue regeneration as described above on the nanofibers and then culturing them.
  • the cell dispersion is a mixed dispersion of cardiomyocytes and stem cells
  • the first hydrogel precursor solution may contain HUVEC, and may further contain VEGF.
  • Fig. 2 is a schematic diagram showing a step-by-step process of a method for preparing a composite of the nanofiber and hydrogel according to an embodiment.
  • preparation of nanofibers by electrospinning step 1), disinfection of nanofibers (step 2), formation of a fibrin hydrogel layer encapsulating nanofibers (step 3), and encapsulating the fibrin hydrogel layer It includes the process of forming an alginate hydrogel layer (step 4).
  • the solution is placed in a syringe and mounted in the electrospinning system, and then the solution is pushed at a constant flow rate of 0.7 ml/hr using a syringe pump, and a cylindrical needle made of metal (Needle). tip).
  • a voltage of 8kV was applied to a cylindrical needle made of metal using a high voltage device (power supply).
  • aluminum foil is laid on it to make a fixed size electrospun nanofiber scaffold with a cover glass of 1.8 cm x 1.8 cm ( A fibrous sheet was obtained through electrospinning for 30 minutes by placing a frame wrapped with aluminum foil on the dust collecting plate.
  • the distance between the cylindrical needle to which the voltage was applied and the dust collecting plate was about 10 cm.
  • the solvent of the polymer solution evaporated due to the voltage difference, the solution was pulled by the grounded dust collecting plate to form a fiber layer.
  • the resulting electrospun nanofibers were placed in a vacuum at 60° C. for 24 hours to remove residual solvent.
  • 3A and 3B are photographs of the electrospun fiber from which the residual solvent was removed in a vacuum state in the above method using a mobile phone camera, and c and d are the electrospun fibers coated with platinum of 15 mA for 1 minute and then scanning electrons This is a picture taken through a microscope.
  • Example 2 Fabrication of a composite of electrospun nanofibers and fibrin hydrogel
  • the electrospun nanofibers prepared in Example 1 were washed twice with a PBS (Phosphate-buffered saline) solution in order to completely remove the residual solvent, and then cut into 5 mm x 5 mm size and placed in a 96 well plate.
  • PBS Phosphate-buffered saline
  • a fibrin hydrogel precursor solution i.e., a fibrinogen solution
  • sodium chloride NaCl
  • fibrinogen from Human plasma, Sigma 100 mg was added and dissolved at room temperature for 10 minutes.
  • thrombin was used as a crosslinking agent to crosslink fibrinogen with fibrin.
  • a thrombin solution 0.2 mmol of calcium chloride (CaCl 2 ) was added to 5 ml of distilled water and dissolved by vortexing for 30 minutes at room temperature.
  • a fibrin solution was prepared by adding 100 U (Enzyme unit) of thrombin to the solution and dissolving it at room temperature for 10 minutes.
  • Example 3 Electrospun nanofiber, fibrin Hydrogel , And Alginate Hydrogel Composite fabrication
  • an alginate hydrogel precursor solution 0.1 g of sodium alginate (Wako) was added to 5 ml of distilled water, sealed so that distilled water did not evaporate, and completely dissolved at 37° C. for 2 hours.
  • 0.1 g of calcium chloride (CaCl 2 ) as a crosslinking agent was added to 10 ml of distilled water and completely dissolved at room temperature for 30 minutes.
  • Fibrin hydrogel prepared in Examples 1, 2 and 3 (a), composite of electrospun nanofiber and fibrin hydrogel (b), composite of electrospun nanofiber, fibrin hydrogel, and alginate hydrogel (c)
  • Figure 4 shows a picture of.
  • the composite of the electrospun fiber and the fibrin hydrogel, and the nanocage with a PBS solution to remove the residual hydrogel precursor, compressive stress and compressive strain were applied. It was placed on an instron (Instron corporation) equipment substrate for measurement.
  • compressive stress and compressive strain were measured by compressing until 80% of the existing height of each scaffold.
  • the compressive stress and compressive strain obtained by the above method were measured using an equation to measure the actual compressive stress and the actual compressive strain, draw a graph, and then measure the slope in a certain section of the graph to calculate the compressive modulus.
  • the formula for calculating the actual compressive stress and the actual compressive strain is as follows.
  • Electrospun nanofibers (Fibrin), electrospun nanofibers and fibrin hydrogel prepared in Examples 1, 2 and 3 (Fibrin-PCL), and electrospun nanofibers, fibrin hydrogel, and A graph of the mechanical strength of a composite of alginate hydrogel (Fibrin-PCL-alginate) is shown.
  • the cross section of the prepared nano cage was observed.
  • the nanocage was quenched using liquid nitrogen, and then vertically cut, and the vertical section of the nanocage was observed using a cryo-scanning electron microscopy (Cryo-SEM). The results are shown in FIG. 6.
  • Example 6 is a photograph taken by observing and taking a cross section of the nanocage prepared in Example 3 using a cryo-scanning electron microscopy (Cryo-SEM).
  • Example 4 Fabrication of a nanocage containing cells
  • the medium for culturing adipose-derived mesenchymal stem cells (ADSC) and cardiomyocytes is DMEM (Dulbecco's Modified Eagle's Medium) with 10% Fetal bovine serum (FBS) and 1% Penicillin.
  • a DMEM mixed medium mixed with /streptomycin was used.
  • a solution of 2% FBS and 1% Penicillin/streptomycin in EGM-2 Bullekit Endothelial Cell Growth Medium
  • a cell dispersion was prepared by dispersing in a DMEM mixed medium at a ratio of 1 to 10 5 cells/ml of total cells at 1:1.
  • the electrospun nanofibers prepared in Example 1 were put into a 96 well plate, and the cell dispersion prepared above was seeded thereon.
  • the electrospun nanofibers seeded with cells were cultured at 37° C. for 2 hours.
  • HUVECs are dispersed in an EGM-2 Bullekit mixed medium at a ratio of 1 ⁇ 10 5 cells/ml to prepare a HUVEC dispersion, and the cells are adhered and cultured.
  • the spun nanofibers and fibrinogen hydrogel precursor solution 40 ⁇ l, cell dispersion 10 ⁇ l, and thrombin solution 40 ⁇ l were added to a 96 well plate, mixed well, and crosslinked at 37° C. for 30 minutes.
  • the electrospun nanofiber and fibrin hydrogel complex in which the prepared cells are cultured with alginate hydrogel, 100 ⁇ l of a 2 wt% alginate hydrogel precursor solution is put into a 96 well plate, and the prepared cells are cultured. After the spinning nanofibers and fibrin hydrogel complex were added, 50 ⁇ l of a calcium chloride solution was added. By crosslinking at 37° C. for 5 minutes, a nanocage containing cells, a complex of electrospun nanofibers, fibrin hydrogel, and alginate hydrogel, in which three cells were co-cultured, was prepared.
  • HUVEC was dispersed in an EGM-2 Bullekit mixed medium at a ratio of 1 ⁇ 10 5 cells/ml to prepare a HUVEC dispersion, and without the presence of nanofibers, fibrinogen 40 ⁇ l of the hydrogel precursor solution, 10 ⁇ l of the cell dispersion, and 40 ⁇ l of the thrombin solution were added to a 96 well plate, mixed well, crosslinked at 37°C for 30 minutes, and incubated for a week (incubated in EGM-2 Bullekit mixed medium). Cell viability was confirmed. After incubation for a week, cells were stained using a Live/Dead cell viability assay, and then confirmed through a fluorescence microscope. The results are shown in FIG. 7
  • Figure 7 is to confirm the viability of HUVECs inside the fibrin hydrogel, without electrospun nanofibers, HUVECs were dispersed in the fibrin hydrogel at a ratio of 1 ⁇ 10 5 cells/ml, cultured for a week, and This is a picture of the results of confirming the viability by staining with the Live/Dead cell viability assay. Live cells are green and dead cells are red.
  • HUVEC exhibits a high survival rate inside the fibrin hydrogel.
  • the fibrin hydrogel was coated with alginate hydrogel in the same manner as in Example 4 except that there was no electrospun nanofiber, and then EGM-2 Bullekit mixed medium After incubation for 1, 4, and 7 days at, the cross sections of the two hydrogels were confirmed through an optical microscope. The results are shown in FIG. 8.
  • FIG. 8 is a first hydrogel and a secondary hydrogel after coating the HUVEC-containing fibrin hydrogel with alginate hydrogel without electrospun nanofibers, 1 and 4 in EGM-2 Bullekit mixed medium. , And after incubation for 7 days, the cross section of the two hydrogels was taken through an optical microscope. Based on the boundary, the inner side represents the fibrin hydrogel with cells, and the outer side represents the alginate hydrogel without cells.
  • HUVEC was dispersed in EGM-2 Bullekit mixed medium at a ratio of 1 ⁇ 10 5 cells/ml to prepare a HUVEC dispersion, fibrinogen hydrogel precursor solution 40 ⁇ l, cell dispersion 10 ⁇ l and 40 ⁇ l of thrombin solution were added to a 96 well plate, mixed well, and crosslinked at 37°C for 30 minutes.
  • Fibrin hydrogel containing cells prepared by the above method was cultured in EGM-2 Bullekit mixed medium for 1 week, and then angiogenesis of HUVEC was confirmed through an optical microscope. The results are shown in FIG. 9.
  • FIG. 9 shows a fibrin hydrogel containing HUVEC without electrospun nanofibers in order to confirm the suitability of angiogenesis of fibrin hydrogel, and then the HUVEC through optical microscopy at the time points of days 1, 4, and 7 The angiogenesis process was observed.
  • a cell dispersion was prepared by dispersing ADSC in a DMEM mixed medium at a ratio of 1 ⁇ 10 5 cells/ml in a ratio of 1:1.
  • the electrospun fibers prepared in Example 1 were placed in a 96 well plate, and the cell dispersion prepared above was seeded thereon.
  • the electrospun nanofibers seeded with cells were cultured at 37° C. for 2 hours.
  • a fibrin hydrogel composite was prepared in the same manner as in Example 2, and a nanocage was prepared in the same manner as in Example 3.
  • the prepared nano-cage containing ADSC was cultured in EGM-2 Bullekit mixed medium for 1 week, and the activity of ADSC was measured using CCK-8 reagent.
  • HUVEC was dispersed in an EGM-2 Bullekit mixed medium at a ratio of 1 ⁇ 10 5 cells/ml to prepare a HUVEC dispersion, and the cells prepared in Example 1 40 ⁇ l of the electrospun nanofibers and fibrinogen hydrogel precursor solution, 10 ⁇ l of cell dispersion, and 40 ⁇ l of thrombin solution were added to a 96 well plate, mixed well, and crosslinked for 30 minutes at 37°C.
  • a nanocage was manufactured according to the method in Example 3.
  • the nano-cage containing HUVEC thus prepared was cultured in EGM-2 Bullekit mixed medium for 1 week, and the activity of ADSC was measured using CCK-8 reagent. The results are shown in FIG. 10.

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Dermatology (AREA)
  • Animal Behavior & Ethology (AREA)
  • Veterinary Medicine (AREA)
  • Oral & Maxillofacial Surgery (AREA)
  • Transplantation (AREA)
  • Epidemiology (AREA)
  • Public Health (AREA)
  • General Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Dispersion Chemistry (AREA)
  • Materials Engineering (AREA)
  • Composite Materials (AREA)
  • Biomedical Technology (AREA)
  • Molecular Biology (AREA)
  • Materials For Medical Uses (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

Un aspect de la présente invention concerne un composite d'une nanofibre et d'un hydrogel, qui comprend une nanofibre, une première couche d'hydrogel encapsulant la nanofibre dans un premier hydrogel, et une seconde couche d'hydrogel encapsulant la première couche d'hydrogel dans un second hydrogel; un échafaudage pour la régénération tissulaire comprenant le composite; et son procédé de production.
PCT/KR2019/003978 2019-04-04 2019-04-04 Composite de nanofibre et d'hydrogel, et échafaudage pour régénération tissulaire le comprenant WO2020204230A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
PCT/KR2019/003978 WO2020204230A1 (fr) 2019-04-04 2019-04-04 Composite de nanofibre et d'hydrogel, et échafaudage pour régénération tissulaire le comprenant

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
PCT/KR2019/003978 WO2020204230A1 (fr) 2019-04-04 2019-04-04 Composite de nanofibre et d'hydrogel, et échafaudage pour régénération tissulaire le comprenant

Publications (1)

Publication Number Publication Date
WO2020204230A1 true WO2020204230A1 (fr) 2020-10-08

Family

ID=72666520

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/KR2019/003978 WO2020204230A1 (fr) 2019-04-04 2019-04-04 Composite de nanofibre et d'hydrogel, et échafaudage pour régénération tissulaire le comprenant

Country Status (1)

Country Link
WO (1) WO2020204230A1 (fr)

Cited By (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113559318A (zh) * 2021-07-05 2021-10-29 四川大学 一种促神经功能恢复的手性导电修复支架及其制备方法
CN114246987A (zh) * 2022-01-21 2022-03-29 吉林大学 一种支架类骨膜材料及其制备方法
CN114533231A (zh) * 2022-04-27 2022-05-27 杭州锐健马斯汀医疗器材有限公司 球囊体及其制备方法和应用
CN114557956A (zh) * 2021-12-30 2022-05-31 江苏拓弘生物科技有限公司 一种负载脐带间充质干细胞的温敏型水凝胶及其制备方法
CN114702695A (zh) * 2022-04-20 2022-07-05 珠海麦得发生物科技股份有限公司 一种pha水凝胶及其制备方法及其应用
CN115726060A (zh) * 2022-11-23 2023-03-03 武汉纺织大学 具有褶皱表面结构的凝胶纤维及其制备方法与应用
CN115748249A (zh) * 2022-11-23 2023-03-07 浙江诸暨聚源生物技术有限公司 重组胶原蛋白水凝胶纤维及其制备方法
CN116059441A (zh) * 2022-08-29 2023-05-05 广东省科学院生物与医学工程研究所 一种各向异性纳米纤维复合天然多糖水凝胶及其制备方法与应用
WO2023191272A1 (fr) * 2022-03-29 2023-10-05 주식회사 에이알씨코리아 Échafaudage polymère pour prothèse et procédé de fabrication associé

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20140271843A1 (en) * 2013-03-14 2014-09-18 Massachusetts Institute Of Technology Multi-Layer Hydrogel Capsules for Encapsulation of Cells and Cell Aggregates
KR20150084519A (ko) * 2014-01-14 2015-07-22 연세대학교 산학협력단 다층의 전기방사 섬유가 복합된 하이드로젤
KR20160035316A (ko) * 2014-09-23 2016-03-31 한국산업기술대학교산학협력단 나노섬유매트와 하이드로젤을 이용한 혈관내피세포 단일층 형성방법
KR20190002253A (ko) * 2017-06-29 2019-01-08 서울대학교산학협력단 나노섬유 및 하이드로젤의 복합체 및 이를 포함하는 조직 재생용 스캐폴드

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20140271843A1 (en) * 2013-03-14 2014-09-18 Massachusetts Institute Of Technology Multi-Layer Hydrogel Capsules for Encapsulation of Cells and Cell Aggregates
KR20150084519A (ko) * 2014-01-14 2015-07-22 연세대학교 산학협력단 다층의 전기방사 섬유가 복합된 하이드로젤
KR20160035316A (ko) * 2014-09-23 2016-03-31 한국산업기술대학교산학협력단 나노섬유매트와 하이드로젤을 이용한 혈관내피세포 단일층 형성방법
KR20190002253A (ko) * 2017-06-29 2019-01-08 서울대학교산학협력단 나노섬유 및 하이드로젤의 복합체 및 이를 포함하는 조직 재생용 스캐폴드

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
LI, Y. ET AL.: "Fibrin gel as an injectable biodegradable scaffold and cell carrier for tissue engineering", THE SCIENTIFIC WORLD JOURNAL, vol. 2015, 2015, pages 1 - 10, XP055601578, DOI: 10.1155/2015/685690 *

Cited By (12)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113559318A (zh) * 2021-07-05 2021-10-29 四川大学 一种促神经功能恢复的手性导电修复支架及其制备方法
CN113559318B (zh) * 2021-07-05 2022-09-13 四川大学 一种促神经功能恢复的手性导电修复支架及其制备方法
CN114557956A (zh) * 2021-12-30 2022-05-31 江苏拓弘生物科技有限公司 一种负载脐带间充质干细胞的温敏型水凝胶及其制备方法
CN114246987A (zh) * 2022-01-21 2022-03-29 吉林大学 一种支架类骨膜材料及其制备方法
CN114246987B (zh) * 2022-01-21 2022-05-17 吉林大学 一种支架类骨膜材料及其制备方法
WO2023191272A1 (fr) * 2022-03-29 2023-10-05 주식회사 에이알씨코리아 Échafaudage polymère pour prothèse et procédé de fabrication associé
CN114702695A (zh) * 2022-04-20 2022-07-05 珠海麦得发生物科技股份有限公司 一种pha水凝胶及其制备方法及其应用
CN114533231A (zh) * 2022-04-27 2022-05-27 杭州锐健马斯汀医疗器材有限公司 球囊体及其制备方法和应用
CN114533231B (zh) * 2022-04-27 2022-11-29 杭州锐健马斯汀医疗器材有限公司 球囊体及其制备方法和应用
CN116059441A (zh) * 2022-08-29 2023-05-05 广东省科学院生物与医学工程研究所 一种各向异性纳米纤维复合天然多糖水凝胶及其制备方法与应用
CN115726060A (zh) * 2022-11-23 2023-03-03 武汉纺织大学 具有褶皱表面结构的凝胶纤维及其制备方法与应用
CN115748249A (zh) * 2022-11-23 2023-03-07 浙江诸暨聚源生物技术有限公司 重组胶原蛋白水凝胶纤维及其制备方法

Similar Documents

Publication Publication Date Title
WO2020204230A1 (fr) Composite de nanofibre et d'hydrogel, et échafaudage pour régénération tissulaire le comprenant
KR102001120B1 (ko) 나노섬유 및 하이드로젤의 복합체 및 이를 포함하는 조직 재생용 스캐폴드
JP5855151B2 (ja) 絹糸生体材料およびその使用方法
Zhang et al. Electrospun PDLLA/PLGA composite membranes for potential application in guided tissue regeneration
Jin et al. Human bone marrow stromal cell responses on electrospun silk fibroin mats
Hwang et al. Poly (ɛ‐caprolactone)/gelatin composite electrospun scaffolds with porous crater‐like structures for tissue engineering
Ortega et al. Combined microfabrication and electrospinning to produce 3-D architectures for corneal repair
Stankus et al. Microintegrating smooth muscle cells into a biodegradable, elastomeric fiber matrix
US20090202616A1 (en) Composite, Method of Producing the Composite and Uses of the Same
Tan et al. Electrospun vein grafts with high cell infiltration for vascular tissue engineering
Skotak et al. Improved cellular infiltration into nanofibrous electrospun cross-linked gelatin scaffolds templated with micrometer-sized polyethylene glycol fibers
US20060153815A1 (en) Tissue engineering devices for the repair and regeneration of tissue
Chen et al. A three-dimensional dual-layer nano/microfibrous structure of electrospun chitosan/poly (d, l-lactide) membrane for the improvement of cytocompatibility
WO2011105724A2 (fr) Echafaudage pour la régénération du cartilage articulaire et procédé de fabrication associé
EP1835949A2 (fr) Dispositifs d'ingenierie tissulaire destines a reparer et a regenerer des tissus
Rajasekaran et al. Role of nanofibers on MSCs fate: Influence of fiber morphologies, compositions and external stimuli
US9956711B2 (en) Facile methods for fabricating a uniformly patterned and porous nanofibrous scaffold
Jang et al. Small diameter vascular graft with fibroblast cells and electrospun poly (L-lactide-co-ε-caprolactone) scaffolds: Cell Matrix Engineering
Zhang et al. Development of FGF-2-loaded electrospun waterborne polyurethane fibrous membranes for bone regeneration
WO2015008877A1 (fr) Procédé de préparation d'un échafaudage bicouche par un unique processus et procédé de régénération de tissu au moyen d'un échafaudage bicouche obtenu par le procédé de préparation
Ezhilarasu et al. Functionalized core/shell nanofibers for the differentiation of mesenchymal stem cells for vascular tissue engineering
Li et al. Applying electrospun gelatin/poly (lactic acid-co-glycolic acid) bilayered nanofibers to fabrication of meniscal tissue engineering scaffold
Spadaccio et al. A G-CSF functionalized PLLA scaffold for wound repair: an in vitro preliminary study
Zeybek et al. Electrospinning of nanofibrous polycaprolactone (PCL) and collagen-blended polycaprolactone for wound dressing and tissue engineering
KR20150021093A (ko) 스피룰리나를 포함하는 생분해성 세포지지체

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 19922258

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 19922258

Country of ref document: EP

Kind code of ref document: A1