KR20170044250A - An artificial vessel comprising cell layer on the inside and outside surfaces and a process for preparing the same - Google Patents

An artificial vessel comprising cell layer on the inside and outside surfaces and a process for preparing the same Download PDF

Info

Publication number
KR20170044250A
KR20170044250A KR1020150143447A KR20150143447A KR20170044250A KR 20170044250 A KR20170044250 A KR 20170044250A KR 1020150143447 A KR1020150143447 A KR 1020150143447A KR 20150143447 A KR20150143447 A KR 20150143447A KR 20170044250 A KR20170044250 A KR 20170044250A
Authority
KR
South Korea
Prior art keywords
blood vessel
conduit
artificial blood
pattern
catheter
Prior art date
Application number
KR1020150143447A
Other languages
Korean (ko)
Other versions
KR101976238B1 (en
Inventor
양희석
이민석
Original Assignee
단국대학교 천안캠퍼스 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 단국대학교 천안캠퍼스 산학협력단 filed Critical 단국대학교 천안캠퍼스 산학협력단
Priority to KR1020150143447A priority Critical patent/KR101976238B1/en
Publication of KR20170044250A publication Critical patent/KR20170044250A/en
Application granted granted Critical
Publication of KR101976238B1 publication Critical patent/KR101976238B1/en

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
    • A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
    • A61L27/507—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials for artificial blood vessels
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61F—FILTERS IMPLANTABLE INTO BLOOD VESSELS; PROSTHESES; DEVICES PROVIDING PATENCY TO, OR PREVENTING COLLAPSING OF, TUBULAR STRUCTURES OF THE BODY, e.g. STENTS; ORTHOPAEDIC, NURSING OR CONTRACEPTIVE DEVICES; FOMENTATION; TREATMENT OR PROTECTION OF EYES OR EARS; BANDAGES, DRESSINGS OR ABSORBENT PADS; FIRST-AID KITS
    • A61F2/00—Filters implantable into blood vessels; Prostheses, i.e. artificial substitutes or replacements for parts of the body; Appliances for connecting them with the body; Devices providing patency to, or preventing collapsing of, tubular structures of the body, e.g. stents
    • A61F2/02—Prostheses implantable into the body
    • A61F2/04—Hollow or tubular parts of organs, e.g. bladders, tracheae, bronchi or bile ducts
    • A61F2/06—Blood vessels
    • A61F2/062—Apparatus for the production of blood vessels made from natural tissue or with layers of living cells
    • 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
    • A61L27/28—Materials for coating prostheses
    • 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
    • A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
    • A61L27/3604—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix characterised by the human or animal origin of the biological material, e.g. hair, fascia, fish scales, silk, shellac, pericardium, pleura, renal tissue, amniotic membrane, parenchymal tissue, fetal tissue, muscle tissue, fat tissue, enamel
    • A61L27/3625—Vascular tissue, e.g. heart valves
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61F—FILTERS IMPLANTABLE INTO BLOOD VESSELS; PROSTHESES; DEVICES PROVIDING PATENCY TO, OR PREVENTING COLLAPSING OF, TUBULAR STRUCTURES OF THE BODY, e.g. STENTS; ORTHOPAEDIC, NURSING OR CONTRACEPTIVE DEVICES; FOMENTATION; TREATMENT OR PROTECTION OF EYES OR EARS; BANDAGES, DRESSINGS OR ABSORBENT PADS; FIRST-AID KITS
    • A61F2230/00—Geometry of prostheses classified in groups A61F2/00 - A61F2/26 or A61F2/82 or A61F9/00 or A61F11/00 or subgroups thereof
    • A61F2230/0063—Three-dimensional shapes
    • A61F2230/0069—Three-dimensional shapes cylindrical
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61F—FILTERS IMPLANTABLE INTO BLOOD VESSELS; PROSTHESES; DEVICES PROVIDING PATENCY TO, OR PREVENTING COLLAPSING OF, TUBULAR STRUCTURES OF THE BODY, e.g. STENTS; ORTHOPAEDIC, NURSING OR CONTRACEPTIVE DEVICES; FOMENTATION; TREATMENT OR PROTECTION OF EYES OR EARS; BANDAGES, DRESSINGS OR ABSORBENT PADS; FIRST-AID KITS
    • A61F2240/00—Manufacturing or designing of prostheses classified in groups A61F2/00 - A61F2/26 or A61F2/82 or A61F9/00 or A61F11/00 or subgroups thereof
    • A61F2240/001—Designing or manufacturing processes
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61F—FILTERS IMPLANTABLE INTO BLOOD VESSELS; PROSTHESES; DEVICES PROVIDING PATENCY TO, OR PREVENTING COLLAPSING OF, TUBULAR STRUCTURES OF THE BODY, e.g. STENTS; ORTHOPAEDIC, NURSING OR CONTRACEPTIVE DEVICES; FOMENTATION; TREATMENT OR PROTECTION OF EYES OR EARS; BANDAGES, DRESSINGS OR ABSORBENT PADS; FIRST-AID KITS
    • A61F2240/00—Manufacturing or designing of prostheses classified in groups A61F2/00 - A61F2/26 or A61F2/82 or A61F9/00 or A61F11/00 or subgroups thereof
    • A61F2240/001—Designing or manufacturing processes
    • A61F2240/002—Designing or making customized prostheses
    • A61F2240/004—Using a positive or negative model, e.g. moulds

Abstract

The present invention relates to: a duct for preparing an artificial vessel, which is composed of a biocompatible polymer, and has a pattern providing adhesiveness and a direction property on an iron surface and an outer surface; a method for preparing the duct; the artificial vessel comprising the duct; and a method for preparing the artificial vessel. The artificial vessel provided in the present invention has cell layers composed of different cells formed on the iron surface and the outer surface of the duct, to exhibit forms similar to a blood vessel in a living body, thereby widely used for treating blood damages with a variety of forms.

Description

[0001] The present invention relates to an artificial blood vessel including a cell layer on both sides and a method for manufacturing the artificial vessel,

The present invention relates to a novel artificial blood vessel comprising a cell layer on both sides and a method for producing the same. More specifically, the present invention relates to a novel artificial blood vessel comprising a biocompatible polymer and a pattern imparting cell adhesiveness and directionality to the luminal surface and the outer surface A manufacturing method of the conduit, an artificial blood vessel including the conduit, and a method of manufacturing the artificial blood vessel.

Cardiovascular disease, a disease that occurs in the circulatory system, such as heart, heart valve, and blood vessels, is the number one cause of death in adults worldwide, and diseases related to blood vessels, including atherosclerosis, angina, myocardial infarction, and stroke, It occupies the most part. For example, according to statistics from the World Health Organization (WHO), the number of people who die from cardiovascular disease is estimated to be about 12 million a year, and in the United States about 57 million people are treated with one or more cardiovascular diseases And it is known that the cost is only about $ 260 billion a year.

The primary treatment for these cardiovascular diseases is autologous vascular grafting or an assistive device for vascular access such as stent. Autologous vascular grafting is mostly used for obstructive diseases caused by atherosclerosis, Aortic aneurysm, aortic dissection, arteritis, and external damage. Grafts that can be used as autologous grafts are extremely limited to the internal mammary artery, the superior mesenteric artery, the umbilical artery, the ulnar artery, etc., and the range of available venous grafts is limited. Therefore, A case may arise.

Accordingly, there is a demand for a device capable of replacing autologous blood vessels such as allografts or artificial blood vessels in order to improve the quality of life or quality of life of a patient, and researches for developing such devices have been actively conducted.

In the early days, artificial blood vessels made of polymer material having a simple conduit shape capable of providing safety and stability in vivo such as PET (poly (ethyleneterephalate) material and PTFE (poly tetra fluoro ethylene) material have been developed. Recently, as research results related to stem cells progressed, it has become clear that, after transplantation together with stem cells, they induce the differentiation of blood vessels from stem cells, and at the same time, they are gradually degraded in vivo, Research on the development of polymeric artificial blood vessels is actively under way. However, a method of completely differentiating blood vessels from stem cells has not yet been developed.

For example, Korean Patent Laid-Open Publication No. 2011-0112556 discloses a carbon nanomolecule pattern surface on which a carbon nanotube film (DLC) is deposited on a polymer surface such as polydimethylsiloxane (PDMS) to form a nanoembossing pattern on the polymer surface US Patent Publication No. 2014-566209 discloses a method for producing an artificial blood vessel using a polymer material having non-bloody ECM and PGS, biodegradable PLGA coated on the surface of the lumen and shell, There is disclosed a blood vessel for implantation in which cells are proliferated and a method for producing the same.

Meanwhile, artificial blood vessels developed to date correspond to large-diameter artificial blood vessels having an inner diameter of 6 mm or more, and small-diameter artificial blood vessels having an inner diameter smaller than that are not yet developed. This is because the small-diameter artificial blood vessel produced is not easy and is easily closed by factors such as thrombosis. Particularly, an artificial blood vessel made of a polymer material is subjected to a pretreatment such as coating a polymer material that enhances biocompatibility to the artificial blood vessel. However, since the treatment level of the coated polymer material is not uniform, thrombus formation is promoted, It is known that various problems such as causing an inflammatory reaction in the inserted transplantation site in vivo are caused.

Under these circumstances, the inventors of the present invention have made extensive efforts to develop an artificial blood vessel having a structure similar to that of blood vessels in a living body. As a result, they have found that an artificial blood vessel having a cell layer composed of different cells is formed on the luminal surface and the outer surface of a conduit composed of a biocompatible polymer Has a similar structure and function to blood vessels in vivo, and completed the present invention.

It is an object of the present invention to provide a catheter for artificial blood vessels, which is composed of a biocompatible polymer and has patterns for inducing cell adhesion and orientation on the luminal surface and the outer surface, respectively.

Another object of the present invention is to provide a method of manufacturing the artificial blood vessel manufacturing catheter.

It is still another object of the present invention to provide an artificial blood vessel in which a cell layer composed of cells constituting different blood vessels is formed on the luminal surface and the outer surface, respectively, of which the pattern of the artificial blood vessel manufacturing conduit is formed.

It is still another object of the present invention to provide a method for manufacturing the artificial blood vessel.

The inventors of the present invention paid attention to the structure of the blood vessel wall while carrying out various studies in order to develop an artificial blood vessel having a structure similar to a blood vessel in a living body. The cross-section of the blood vessel wall is composed of connective tissue composed of endothelium, tunica intima, elastin, collagen fiber, tunica media composed of hardwood roots, and tunica adventitia composed of collagen fiber layer. The asymmetric structure of the blood vessels is known to inhibit the production of blood clots, prevent blood vessel leakage, and impart blood vessel fluidity. The present inventors simulated the structures of blood vessels and tried to produce artificial blood vessels in which a cell layer composed of endothelial cells was formed on the luminal surface of the catheter and a cell layer composed of the smooth muscle cells were formed on the outer surface.

First, the catheter that is the base of the artificial blood vessel is manufactured in such a manner that a cell layer can be formed on the luminal surface and the outer surface, respectively, and a pattern capable of imparting cell adhesiveness and directionality is formed. After the cells adhere to the luminal surface and the outer surface of the catheter, they can proliferate to form a cell layer.

Up to now, artificial blood vessel forming catheters have been manufactured using common solvent casting methods and electrospinning methods. When such a method is used, a conduit having a pattern formed on the luminal surface and the outer surface is formed There is a problem that it can not be manufactured.

Accordingly, the present inventors selected the rolling method as a method of manufacturing a conduit in which a pattern is formed on the luminal surface and the outer surface, respectively. In the rolling method, a flat plate is wrapped around a central axis, a portion to which a plate is connected is bonded, and then a center axis is removed to manufacture a conduit. Specifically, a flat plate-type polymer scaffold is manufactured, a pattern is formed on both sides of the scaffold, and a rolling process is performed on a flat plate-like polymer scaffold on which the pattern is formed. And the like.

Next, an artificial blood vessel was prepared using a conduit in which a pattern was formed on the luminal surface and the outer surface, respectively. Specifically, a cell layer composed of vascular endothelial cells is formed by inoculating and culturing vascular endothelial cells on the luminal surface of the catheter, and the outer surface of the catheter is inoculated with the smooth muscle cells and cultured to form a cell layer composed of smooth muscle cells , An artificial blood vessel having a cell layer composed of cells constituting different blood vessels on the luminal surface and the outer surface was prepared.

Artificial blood vessels in which a cell layer composed of cells constituting different blood vessels are formed on the luminal surface and the outer surface provided by the present invention have been developed for the first time by the present inventors as a new type artificial blood vessel which has not been developed at all.

In order to accomplish the above object, the present invention provides, as one embodiment, a catheter for artificial blood vessel, which is made of a biocompatible polymer and has a pattern for imparting cell adhesiveness and directionality to the luminal surface and the outer surface.

The term "biocompatibility polymer" of the present invention means a polymeric substance which is harmless in vivo or in vitro and easily adapted to an in vivo environment and does not generate a rejection reaction. The biocompatible polymer is used in various fields ranging from implants, implants, substitute organs, etc., which can be inserted or implanted in a living body, to drug delivery carriers.

In the present invention, the biocompatible polymer may be used as a support material used in the artificial blood vessels provided in the present invention. The biocompatible polymer is not particularly limited and includes, for example, lactide, Caprolactone, glycolide, dioxanone, propylene, ethylene, vinylchloride, butadiene, methly methacrylate, methyl methacrylate, Acrylic acid, 2-hydroxyethlymethacrylate, carbonate, polyethylene terephthalate, or copolymers thereof may be used alone or in combination. As another example, chitosan / glycerol phosphate; Polyphosphazene; Polycaprolactone; Polycarbonate; Polycyanoacrylate; Polyorthoesters; Poly (N- (2-hydroxyethyl) methacrylamide-lactate)); Poly (propylene phosphate)); Poly (lactic-co-glycolic acid) (PLGA); Polyethylene glycol-polyester copolymers (poly (ethyleneglycol) / polyester); Poly (ethylene glycol) / poly (propylene glycol), PEG / PPG) copolymers; Polyethylene glycol-polycaprolactone copolymers; Methoxypolyethylene glycol-polycaprolactone copolymer; Polyethylene glycol- (polylactic-glycolic acid) -polyethylene glycol triple polymer; (Polylactic-glycolic acid) -polyethylene glycol- (polylactic-glycolic acid) triple polymer; Polyethylene glycol-polycaprolactone-polyethylene glycol triple polymer; Polyacrylonitrile-polycaprolactone-polyethylene glycol-polycaprolactone ternary polymer, and the like, alone or in combination. As another example, it may be a polylactic-glycolic acid copolymer (PLGA). For example, in the present invention, a catheter for artificial blood vessel production was prepared using a polylactic-glycolic acid copolymer (PLGA) as a biocompatible polymer.

The term "pattern" of the present invention means a three-dimensional structure formed on the surface of the biocompatible polymer in a relief or engraved form. The pattern may be formed in the form of a ridge (for example, a floor having a size of 800 nm) embossed on the surface of the polymer or a groove engraved on the surface of the polymer, And it plays a key role in the manufacture of artificial blood vessels.

First, the pattern improves cell adhesion to the polymer surface. That is, the cells can be efficiently attached to the surface of the polymer on which the pattern is formed, the proliferation of the attached cells can be promoted, and the cell migration rate can be improved. For example, in a conduit composed of a polymer of the same material, a conduit in which a pattern with a size of 800 nm is formed on the surface has increased cell attachment efficiency and proliferation efficiency as compared with a conduit in which no pattern is formed on the surface, The moving speed of the cells can be increased in a given state. The movement speed may be influenced by the presence or absence of the pattern and the directionality.

Next, the pattern provides directionality to the cells attached to the polymer surface. That is, when cells are attached to and propagate on the surface of the polymer on which the pattern is formed, the cells proliferated by the pattern are oriented. For example, when a cell is attached to a surface having a pattern formed in the longitudinal direction of the catheter, when the cell is proliferated, the shape of the cell is changed in a form that is elongated in the longitudinal direction of the catheter.

On the other hand, the pattern may be formed such that the lumen surface and the outer surface are the same shape, and the lumen surface and the outer surface may be formed to show different shapes. As described above, the cross-section of the blood vessel wall is composed of an inner blood vessel layer, a connective tissue, and an outer blood vessel, and each layer has a certain directionality. The endothelial cells are arranged in the longitudinal direction of the inner blood vessel, and the tissues and cells of the connective tissue and the vascular outer membrane are arranged in a direction perpendicular to the longitudinal direction. The present inventors simulated the structures of blood vessels and tried to produce artificial blood vessels in which a cell layer composed of endothelial cells was formed on the luminal surface of the catheter and a cell layer composed of the smooth muscle cells were formed on the outer surface. The luminal surface of the catheter may have a specific directionality by placing a longitudinal pattern in the catheter so as to exhibit a blood vessel-like shape and a shape similar to a blood vessel, and attaching and propagating the vascular endothelial cells. In addition, on the outer surface of the catheter, the adventitious muscle cells are adhered and proliferated so as to show a shape similar to that of the blood vessels, so that the adventitious muscle cells that give stability and elasticity of blood vessels are formed in a direction wrapping the conduit A pattern can be formed in a direction perpendicular to the longitudinal direction of the conduit.

The term "catheter for artificial blood vessel preparation" in the present invention means a tube-like structure composed of a biocompatible polymer material as a foundation for manufacturing the artificial blood vessel provided in the present invention. Since a cell layer must be formed on the luminal surface and the outer surface of the catheter, a pattern for supporting the formation and orientation of the cell layer is provided on the luminal surface and the outer surface. In this case, the biocompatible polymer material is the same as described above, and the pattern is not particularly limited as long as it can provide the formation and directionality of a cell layer.

In particular, since the conduit provided in the present invention has a pattern formed on both the luminal surface and the outer surface, a cell layer can be formed on both sides of the conduit. For example, when a cell layer formed on both sides of the catheter is a vascular endothelial cell layer formed on the luminal surface and a hard-tissue cell layer formed on the outer surface, the cell layer is formed The conduit can be used as an artificial blood vessel.

According to another aspect of the present invention, there is provided a method of manufacturing a catheter for artificial blood vessel production.

Specifically, the method for manufacturing a catheter for artificial blood vessel of the present invention comprises the steps of: (a) forming a pattern on both sides of a flat plate-like support composed of a biocompatible polymer; And (b) forming a conduit using a support having a pattern formed on both sides thereof.

In the step (a), the thickness of the support is not particularly limited, but may be, for example, 10 to 15 占 퐉, and the inner diameter of the conduit is also not particularly limited. For example, A catheter having an inner diameter of 5 mm or less can be used for manufacturing artificial blood vessels to replace small-diameter blood vessels, and an inner diameter of 5 to 50 mm for manufacturing artificial blood vessels that replace general blood vessels can be used Can be used. The present invention is advantageous in that it can be manufactured from small diameter to large diameter blood vessel.

In the step (a), a method of forming a pattern on both sides of the support is not particularly limited. For example, a capillary force lithography (CFL) method may be used. As another example, (Polydimethylsiloxane) mold and PUA (polyurethane acrylate) mold can be used. Further, the pattern may be a pattern similar to that of a blood vessel.

The term " capillary force lithography (CFL) method "of the present invention means a method of forming a pattern in a blanket manner by pulling up a polymer to an empty space of a mold using a natural force called capillary action, The capillary action can be used to form a finer and more elaborate pattern, compared with the conventional lithography method which is performed by applying the capillary action. At this time, the capillary action used may be caused by capillary depression (θ> 90 °). For example, a pattern can be formed on the surface of the plate-like support by placing the mold on a plate-like support of a biocompatible polymer material as shown in FIG. 1, applying pressure and applying gravity and capillary force together .

Meanwhile, when the pattern is formed using the CFL method, the mold used is not particularly limited. For example, a PUA (Young's modulus: 100-400 MPa) mold or a PDMS (Young's modulus: 2 MPa) mold is used .

In order to improve the adhesion of the cells to the patterned surface, it is preferable that the capillary or the capillary is coated on the patterned surface before performing the step (b) of forming the pattern by performing the step (a) And crosslinking these combinations to the surface on which the pattern is formed.

In the step (b), the method of forming the conduit is not particularly limited, but a rolling method may be used as an example.

The term "rolling method" of the present invention means a method of manufacturing a conduit by wrapping a flat plate around a central axis, bonding and fixing a portion where the flat plate is connected, and removing the central axis. Specifically, by performing a rolling method using a glass capillary as a central axis in a flat plate-like polymer scaffold in which the pattern is formed on both sides of the support after the plate-shaped polymer scaffold is manufactured, A conduit having a pattern on its surface and on its outer surface can be produced. Further, after the conduit is formed, it may further be left in a vacuum oven to remove the residual organic solvent. For example, in the present invention, a support made of a PLGA material having a double-sided pattern is obtained. The support is rolled so as to surround a glass capillary tube used as a central axis to form a conduit shape. Then, chloroform is applied to the joint portion of the support, This was dried in an oven for a certain period of time to remove the remaining chloroform, fix the shape of the conduit, and separate the glass capillary used as the central axis from the lumen of the conduit to prepare a conduit for artificial blood vessel preparation.

(A) a catheter for artificial blood vessel, which is composed of a biocompatible polymer and has a pattern for imparting cell adhesiveness and directionality to the luminal surface and the outer surface; (b) a first cell layer located on the luminal surface of the conduit, the first cell layer comprising vascular endothelial cells; And (c) a second cell layer located on the outer surface of the conduit, the second cell layer being composed of smooth muscle cells.

The term "artificial vessel " of the present invention means an artificial vessel designed to replace a damaged vessel tissue due to various causes such as an aneurysm, thrombosis, and trauma. As long as the artificial blood vessel can satisfy the characteristics such as superior anti-thrombogenic property, bending and elongation characteristic similar to that of natural blood vessel, biocompatibility, bonding with conventional blood vessels, excellent rod formation, The material and composition of the artificial blood vessel are not particularly limited.

If the artificial blood vessel is derived or separated from the patient to be transplanted or inserted, it can be a patient-made artificial blood vessel. In other words, the biocompatible polymer constituting the artificial blood vessel does not cause a specific rejection reaction in vivo, but in the case of a cell layer formed therefrom, a biodejection reaction can be induced. Therefore, a blood vessel derived from a patient to which the artificial blood vessel is to be implanted or inserted When vascular endothelial cells, vascular smooth muscle cells or fibroblasts differentiated using endothelial cells, vascular smooth muscle cells or fibroblasts or using stem cells derived from the patient are used, the occurrence of rejection can be suppressed from the patient, A patient-tailored artificial blood vessel can be provided.

According to another aspect of the present invention, there is provided a method of manufacturing the artificial blood vessel. Specifically, the artificial blood vessel manufacturing method of the present invention

(a) preparing a catheter for artificial blood vessel, which is composed of a biocompatible polymer and has a pattern imparting cell adhesiveness and directionality to the luminal surface and the outer surface; And (b) inoculating and culturing vascular endothelial cells on the luminal surface of the catheter, and intestinal muscle cells on the outer surface of the catheter, respectively. In this case, the inoculation and cultivation of vascular endothelial cells or hard-rooted endothelial cells may be performed simultaneously or separately.

The artificial blood vessel provided in the present invention has a cell layer composed of different cells on the luminal surface and the outer surface of the catheter and shows a shape similar to a blood vessel in a living body and thus can be widely used for treating various types of blood vessel injuries will be.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a schematic view showing a manufacturing process of a catheter for artificial blood vessel preparation according to an embodiment of the present invention. FIG.
FIG. 2A is a photograph showing a size of a conduit for artificial blood vessel preparation provided in an embodiment of the present invention. FIG.
2B is a scanning electron microscope photograph (X2000) showing a cross section of a catheter for artificial blood vessel preparation provided in an embodiment of the present invention.
2C is a scanning electron microscope photograph (X4000) showing a cross section of a catheter for artificial blood vessel preparation provided in an embodiment of the present invention.
FIG. 2D is a scanning electron microscope (SEM) image showing the upper surface of a support used for manufacturing a catheter for artificial blood vessel preparation provided in an embodiment of the present invention.
FIG. 2E is a scanning electron microscope photograph showing a lower surface of a support used for manufacturing a catheter for artificial blood vessel production provided in an embodiment of the present invention. FIG.
FIG. 2F is a scanning electron microscope photograph showing a curled shape of a conduit for artificial blood vessel preparation provided in an embodiment of the present invention. FIG.
FIG. 2G is a scanning electron microscope photograph showing the outer surface and the lumen surface of the artificial blood vessel-producing duct provided in one embodiment of the present invention.
FIG. 3 is a graph showing the results of evaluating the tensile force of a support used for manufacturing a catheter for artificial blood vessel production provided by the present invention. FIG.
4A is a photograph showing the HUVEC inoculated on the luminal surface of the artificial blood vessel-producing duct provided by the present invention.
4B is a photograph showing C2C12 inoculated on the outer surface of a catheter for artificial blood vessel production provided in an embodiment of the present invention.

Hereinafter, the present invention will be described in more detail with reference to examples. However, these examples are for illustrative purposes only, and the scope of the present invention is not limited to these examples.

Example  1: Fabrication of artificial blood vessel conduit

A biocompatible polymer matrix substrate was prepared, and a CFL pattern was formed on both sides of the substrate, and a catheter was prepared using the CFL method (FIG. 1). 1 is a schematic view showing a manufacturing process of a catheter for artificial blood vessel production provided by the present invention. The specific production process is as follows.

Example  1-1: For pattern making Mold  Ready

Molds were used to form patterns on both sides of a flat plate-like support of biocompatible polymeric material by the CFL method.

First, a PUA (polyurethane acrylate) solution was dispensed on a silicon wafer master and cured by light irradiation of a UV lamp to obtain a PUA mold.

Next, PDMS (Polydimethylsiloxane) solution and curing solution were mixed at a ratio of 10: 1 (W / W) and stirred to obtain a mixed solution. The mixed solution was poured into a 100 mm Petri dish fixed with a PUA mold, Curing for 3 hours or more, a PDMS mold was obtained. At this time, the size of the groove: ridge of each of the manufactured molds was set to 800 nm: 800 nm.

Example  1-2: Fabrication of artificial blood vessel conduit

PLGA solution was dissolved in chloroform to obtain a 15% (W / V) PLGA solution, and 80 PL of the PLGA solution was added to the PDMS mold obtained in Example 1-1, and a PDMS sheet without pattern was covered thereon, The PLGA solution was uniformly applied onto the PDMS mold. Then, a weight of 500 g was placed on the PDMS sheet and allowed to stand for 15 minutes to cure the PLGA solution. Then, the PDMS sheet and weight were removed, and the remaining chloroform was removed by heating at 120 DEG C for 10 minutes, To obtain a plate-shaped PLGA support having a pattern formed thereon.

Then, the PUA mold obtained in Example 1-1 was fixed on the PDMS sheet and the further removed surface, and the PDMS sheet was covered thereon. A weight of 1000 g was placed on the PDMS sheet, Min to obtain a plate-shaped PLGA support having a pattern formed on both sides thereof.

The obtained PLGA support was subjected to a rolling method using a glass capillary as a central axis to prepare a conduit. That is, a glass capillary (center axis) having a diameter of 2 mm was placed on a PLGA support, and a conduit was formed by axially rolling the same. A 3% chloroform solution was added to the bonding portion of the support, and the resultant was kept in a vacuum oven for 24 hours. After the chloroform was removed, the shape of the conduit was fixed, and the glass capillary was released from the lumen of the conduit to prepare a conduit for artificial blood vessel production.

Example  2: Analysis of the shape of conduit for artificial blood vessel preparation

The pattern of the double-sided pattern and the shape of the conduit of the plate-shaped PLGA support and the artificial blood vessel preparation catheter manufactured using the same were analyzed by a scanning electron microscope (FIGS. 2A to 2G).

2A is a photograph showing the size of a conduit for artificial blood vessel production provided by the present invention. As shown in FIG. 2A, it was confirmed that the conduit was manufactured to have a length of about 15 mm and a diameter of 2 mm.

FIG. 2B is a scanning electron microscope (X2000) sectional view showing a cross section of a catheter for artificial blood vessel preparation provided by the present invention, and FIG. 2C is a scanning electron microscope (X4000) showing a cross section of a catheter for artificial blood vessel production provided by the present invention. As shown in FIGS. 2B and 2C, it was confirmed that the manufactured conduit had nanopatterns formed on its surface.

FIG. 2 (d) is a scanning electron microscope (SEM) image showing the top surface of a support for use in the manufacture of a catheter for artificial blood vessel preparation according to the present invention, and FIG. 2 It is a scanning electron microscope photograph. As shown in FIGS. 2d and 2e, it was confirmed that the support used in the manufacture of the conduit had uniform nanopatterns formed on both sides thereof.

FIG. 2F is a scanning electron micrograph showing the curled shape of the artificial blood vessel manufacturing catheter provided in the present invention, FIG. 2G is a scanning electron microscope photograph showing the outer surface and the lumen surface of the artificial blood vessel- It is a photograph. As shown in FIGS. 2F and 2G, it was confirmed that the support could form the shape of the conduit, and the nanopatterns formed on the outer surface and the luminal surface of the conduit formed at this time had different directions.

Example  3: Tensile strength analysis of supports for manufacturing conduits

It was analyzed whether or not the tensile force of the plate-shaped PLGA support having the patterns formed on both sides of the Example 1 was affected by the nanopattern formed on the surface.

That is, in the method of Example 1, a support (flat) on which nano patterns were not formed on the surface, a support in which nano patterns in the same direction were formed on both side surfaces, vertical) were cut and cut into 1 cm x 3 cm. The tensile strength of the cut support was measured using a 5966 dual column testing system (INSTRON, USA) and the Young's modulus was calculated from the measured values (Fig. 3). At this time, the crosshead speed was set to 10 mm / min and the weight of the load cell was set to 10 KN maximum.

FIG. 3 is a graph showing the results of evaluating the tensile force of a support used for manufacturing a catheter for artificial blood vessel production provided by the present invention. FIG. As shown in FIG. 3, the support having nano patterns at right angles to each other has a relatively high tensile force, as compared with a support in which a support is not formed on the surface or a parallel nano pattern is formed in the same direction Respectively.

The supporters exhibiting a high level of tensile strength were analyzed to be low in strain due to the in vivo environment even after insertion into the living body.

Example  4: Cell culture using the surface of the catheter preparation support

Typically, the vessel wall exhibits asymmetry, and it is known that the cells forming the luminal surface and the outer surface of the blood vessel are different from each other. Thus, it was tried to confirm whether different cells could be inoculated on both sides of the support for preparing a catheter manufactured by the present invention and cultured and proliferated.

First, HUVEC (human umbilical vein endothelial cells), a kind of endothelial cells, and C2C12 cells (used for replacing the smooth muscle root), undifferentiated stem cells derived from mice were cultured, and the HUVECs were labeled with a dil fluorescent probe , C2C12 cells were labeled with a Dio fluorescent probe (green).

Each labeled cell was treated with trypsin to separate the cells. HUVEC was inoculated on the back surface of the support forming the lumen of the catheter, PDMS with grooves of supporter size was mounted on the culture container, A support was attached to the PDMS so that the bottom surface of the support did not contact the bottom, followed by culture. After the cultivation is completed, the above-mentioned tear body is turned upside down and mounted on the PDMS again, and C2C12 cells are inoculated on the front surface of the support forming the outer surface of the conduit, and then cultured again so that different cells can be proliferated on both sides of the support (Figs. 4A and 4B). At this time, the medium used was HUVEC culture medium (FBS 2%, VEGF 0.1%, Hydrocortisone 0.04%, hFGF-B 0.4%, R3-IGF-1 0.1%, Ascorbic acid 0.1%, hEGF 0.1% %, Heparin 0.1%) was used.

4A is a photograph showing HUVEC inoculated on the luminal surface of a catheter for artificial blood vessel preparation provided by the present invention, and FIG. 4B is a photograph showing C2C12 inoculated on the outer surface of a catheter for artificial blood vessel production provided in the present invention. As shown in FIGS. 4A and 4B, it was confirmed that the support provided in the present invention was able to proliferate by inoculating different cells on both sides.

As described above, the support provided in the present invention can be used for the production of artificial blood vessels since different cells are inoculated on both sides and proliferated to exhibit blood vessel asymmetry.

Claims (12)

A catheter for the manufacture of an artificial blood vessel, comprising a biocompatible polymer and having a pattern imparting cell adhesion and directionality to the luminal surface and the outer surface.
The method according to claim 1,
The biocompatible polymer may be selected from the group consisting of lactide, caprolactone, glycolide, dioxanone, propylene, ethylene, vinylchloride, butadiene, Methacrylic acid, 2-hydroxyethlymethacrylate, carbonate, polyethylene terephthalate, copolymers thereof, and combinations thereof. The term " polymer "≪ / RTI >
The method according to claim 1,
Wherein the biocompatible polymer is a poly (lactic-co-glycolic acid) (PLGA) copolymer which is a copolymer of lactide and glycolid.
(a) forming a pattern on both sides of a flat plate-like support composed of a biocompatible polymer; And
(b) forming a conduit using a support having a pattern formed on both sides thereof.
5. The method of claim 4,
Wherein the thickness of the support in step (a) is 10 to 15 占 퐉, and the diameter of the lumen of the conduit in step (b) is 1 to 50 mm.
5. The method of claim 4,
Wherein the pattern of step (a) is formed by a capillary force lithography (CFL) method.
The method according to claim 6,
Wherein the capillary force lithography method is performed using a PDMS (Polydimethylsiloxane) mold or a PUA (polyurethane acrylate) mold.
5. The method of claim 4,
Further comprising the step of cross-linking the heparin, catechol or combinations thereof to the patterned surface after step (a), before performing step (b).
5. The method of claim 4,
Wherein the conduit of step (b) is formed by rolling.
(a) a conduit for artificial blood vessel, which is composed of a biocompatible polymer and has a pattern for imparting cell adhesiveness and directionality to the luminal surface and the outer surface;
(b) a first cell layer located on the luminal surface of the conduit, the first cell layer comprising vascular endothelial cells; And
(c) a second cell layer located on the outer surface of the conduit, the second cell layer being composed of smooth muscle cells.
11. The method of claim 10,
Wherein the vascular endothelial cells or the smooth muscle cells are derived from an object to be transplanted with the artificial blood vessel.
(a) preparing a catheter for artificial blood vessel, which is composed of a biocompatible polymer and has a pattern imparting cell adhesiveness and directionality to the luminal surface and the outer surface; And
(b) inoculating and culturing vascular endothelial cells on the luminal surface of the catheter, and intestinal muscle cells on the outer surface of the catheter, respectively.
KR1020150143447A 2015-10-14 2015-10-14 An artificial vessel comprising cell layer on the inside and outside surfaces and a process for preparing the same KR101976238B1 (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
KR1020150143447A KR101976238B1 (en) 2015-10-14 2015-10-14 An artificial vessel comprising cell layer on the inside and outside surfaces and a process for preparing the same

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
KR1020150143447A KR101976238B1 (en) 2015-10-14 2015-10-14 An artificial vessel comprising cell layer on the inside and outside surfaces and a process for preparing the same

Publications (2)

Publication Number Publication Date
KR20170044250A true KR20170044250A (en) 2017-04-25
KR101976238B1 KR101976238B1 (en) 2019-05-08

Family

ID=58703464

Family Applications (1)

Application Number Title Priority Date Filing Date
KR1020150143447A KR101976238B1 (en) 2015-10-14 2015-10-14 An artificial vessel comprising cell layer on the inside and outside surfaces and a process for preparing the same

Country Status (1)

Country Link
KR (1) KR101976238B1 (en)

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20190088712A (en) * 2018-01-19 2019-07-29 제주대학교 산학협력단 Artificial biomembrane having cocultured endothelial cell and epithelial cell and methof for preparing the same
KR20200075936A (en) * 2018-12-12 2020-06-29 가천대학교 산학협력단 Method for preparing or tube type scaffold
WO2023200098A1 (en) * 2022-04-12 2023-10-19 재단법인 아산사회복지재단 Artificial structure for preventing or treating lymphedema and construction method therefor

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR100748038B1 (en) * 2006-10-09 2007-08-09 아주대학교산학협력단 Bioactive agent immobilized artificial vascular graft
KR20090117138A (en) * 2008-05-08 2009-11-12 한국기계연구원 A manufacturing method of a three dimensional scaffold

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR100748038B1 (en) * 2006-10-09 2007-08-09 아주대학교산학협력단 Bioactive agent immobilized artificial vascular graft
KR20090117138A (en) * 2008-05-08 2009-11-12 한국기계연구원 A manufacturing method of a three dimensional scaffold

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
KIM, H. N. et al., Annals of Biomedical Engineering (2012) Vol.40, No.6, pp.1339-1355. 1부.* *

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20190088712A (en) * 2018-01-19 2019-07-29 제주대학교 산학협력단 Artificial biomembrane having cocultured endothelial cell and epithelial cell and methof for preparing the same
KR20200075936A (en) * 2018-12-12 2020-06-29 가천대학교 산학협력단 Method for preparing or tube type scaffold
WO2023200098A1 (en) * 2022-04-12 2023-10-19 재단법인 아산사회복지재단 Artificial structure for preventing or treating lymphedema and construction method therefor

Also Published As

Publication number Publication date
KR101976238B1 (en) 2019-05-08

Similar Documents

Publication Publication Date Title
Lee et al. Development of a composite vascular scaffolding system that withstands physiological vascular conditions
Wu et al. Living nano-micro fibrous woven fabric/hydrogel composite scaffolds for heart valve engineering
de Valence et al. Plasma treatment for improving cell biocompatibility of a biodegradable polymer scaffold for vascular graft applications
Rayatpisheh et al. Combining cell sheet technology and electrospun scaffolding for engineered tubular, aligned, and contractile blood vessels
US11596512B2 (en) Decellularized biologically-engineered tubular grafts
Rai et al. Synthesis, properties and biomedical applications of poly (glycerol sebacate)(PGS): A review
Kim et al. Fabrication of poly (ethylene glycol): gelatin methacrylate composite nanostructures with tunable stiffness and degradation for vascular tissue engineering
Lv et al. Bacterial cellulose-based biomimetic nanofibrous scaffold with muscle cells for hollow organ tissue engineering
Wang et al. Micropatterning of proteins and mammalian cells on biomaterials
US20090248145A1 (en) Method of forming a three-dimensional structure of unidirectionally aligned cells
KR101976238B1 (en) An artificial vessel comprising cell layer on the inside and outside surfaces and a process for preparing the same
Xu et al. Biodegradable scaffolds for urethra tissue engineering based on 3D printing
EP3209340A1 (en) Microfabricated tissue scaffolds and methods of making and using the same
Wang et al. Stereo coverage and overall stiffness of biomaterial arrays underly parts of topography effects on cell adhesion
EP3952934A1 (en) Tissue equivalent tubular scaffold structure, and methods of production thereof
Liu et al. Fabrication of a dual-layer cell-laden tubular scaffold for nerve regeneration and bile duct reconstruction
Yang et al. Double-modified bacterial cellulose/soy protein isolate composites by laser hole forming and selective oxidation used for urethral repair
US10933163B2 (en) In situ tissue engineering
Daxini et al. Micropatterned polymer surfaces improve retention of endothelial cells exposed to flow-induced shear stress
KR102473438B1 (en) Membrane for forming biomaterial structure comprising artificial blood vessel, artificial biomaterial structure comprising the membrane and method for manufacturing the same
Heath et al. Biomaterials patterned with discontinuous microwalls for vascular smooth muscle cell culture: biodegradable small diameter vascular grafts and stable cell culture substrates
Shojaee et al. Design and characterization of a porous pouch to prevent peritoneal adhesions during in vivo vascular graft maturation
JP2022552097A (en) Novel porous scaffold and method of making same
CN117065096B (en) Rapid preparation method and application of biological film
Xue Micro-/Nanotechnologies to Engineer Microenvironmental Cues for Regenerative Heart Valve Therapies

Legal Events

Date Code Title Description
A201 Request for examination
E902 Notification of reason for refusal
AMND Amendment
E601 Decision to refuse application
AMND Amendment
X701 Decision to grant (after re-examination)
GRNT Written decision to grant