JP5822266B2 - Patterned porous material and method for producing the same - Google Patents

Description

The present invention relates to a patterned porous material and a method for producing the same.
The present invention repairs and regenerates tissues and organs such as skin, bone, cartilage, ligament, muscle, trachea, esophagus, nose, ear, blood vessel, pancreas, liver, etc. damaged or lost due to a disease or accident It relates to a porous material.
More specifically, the present invention relates to a porous material having a pattern structure and a manufacturing method in order to spatially control functions such as adhesion, proliferation, and differentiation of cells that differentiate and organize into these biological tissues and organs.

  To repair or treat skin, bone, cartilage, ligament, muscle, trachea, esophagus, nose, ear, blood vessel, pancreas, liver, and other biological tissues / organs damaged or lost due to an accident or illness. In the past, artificial organs and living organs have been transplanted. However, in the case of an artificial organ, the function is not sufficient, and there are problems such as wear, loosening and breakage due to the artificial object. In addition, in the case of living organ transplantation, in addition to the problem of lack of donors, when the donor is another person, there are many medical risks such as problems of rejection based on immune responses and complications associated with immunosuppression.

Due to the existence of such various problems, repairing and treating damage etc. by regenerating living tissue / organs by means of tissue engineering does not require a donor as compared with organ transplantation. So it is considered an ideal way. In this method, living cells are proliferated in vitro, seeded on a scaffold material that serves as a scaffold for living cells and tissues, cultured in vitro, and formed as a living tissue, and then transplanted in vivo. Alternatively, living cells are seeded on a scaffold material, embedded in the living body, and regeneration of living tissue is induced in the living body.

Therefore, scaffold materials that induce and promote the formation of biological tissue and maintain the morphology of the biological tissue play a very important role. The porous material used as the scaffold material is required to have biocompatibility as a property that does not affect the living body, bioabsorbability that is decomposed and absorbed while a new living tissue is formed.

In a living body, many types of cells are accumulated according to a certain spatial pattern to form tissues and organs. The direction of differentiation of each cell is determined at an appropriate location and at an appropriate timing. In this process, spatial signals that control pattern formation give positional information to the cell, and when the cell stores this information, it forms a complex and elaborate structure. For example, the blood vessels and nerve networks in the body are controlled by this pattern information, and a more precise pattern is formed. Therefore, it is very important to apply this spatial pattern information to the porous material and to spatially control the cell distribution in order to regenerate functional biological tissues / organs.

As a scaffold material for tissue regeneration, a porous material having bioabsorbability is often used.
Materials include polylactic acid, polyglycolic acid, copolymers of lactic acid and glycolic acid, bioabsorbable synthetic polymers such as poly-ε-caprolactone, collagen, gelatin, cellulose, polyalginic acid, chitin, chitosan, starch, hyaluron Examples include bioabsorbable natural polymers such as acid, chondroitin sulfate, fibronectin, and laminin.

Examples of a technique for producing a porous material of these bioabsorbable polymers include a porogen leaching method, a phase separation method, a freeze drying method, an emulsion freeze drying method, a fiber fusion method, a needle punching method, and an electrospinning method.

However, with these manufacturing techniques, it has been difficult to manufacture a porous material having a pattern structure. For example, in the freeze-drying method, a temperature distribution is generated when the sample is cooled, and therefore, the maximum pore size and the minimum pore size may vary several times. There is also an attempt (Non-Patent Document 1) to control the orientation of the pores by controlling the cooling direction, but it is extremely difficult to control a porous body having an arbitrary shape.

FIG. 19 is an electron micrograph of a tissue regeneration collagen sponge produced using a conventional freeze-drying method. Holes having different shapes and sizes are formed. Due to the large variation in the shape and diameter of the pores, the distribution of cells could not be sufficiently controlled with this collagen sponge. There was a tendency for cells to adhere to the large pores. As a result, the tissue becomes non-uniform, and it is difficult to regenerate complex organs.

Due to the heterogeneity of the structure, it has been extremely difficult to control the cell distribution. It was also difficult to give cells a spatially dependent chemical signal. It is indispensable to control these factors spatially in order to control cell functions and induce regeneration of living tissue.

Non-Patent Document 2 is a document relating to a collagen sponge in which a blood vessel growth factor is chemically immobilized. The blood vessel growth factor is distributed throughout the collagen sponge and is not controlled in a pattern.

Non-Patent Document 3 is a document relating to a collagen-based sponge in which a blood vessel growth factor gene is physically adsorbed. DNA is distributed throughout the entire sponge and is not controlled in a pattern.

Heike Schoof, Jorn Apel, Ingo Heschel, by Gunter Rau, J Biomed Mater Res (Appl Biometer) 58, 2001, p. 352-7 Y. H. Shen et al., Acta Biomaterialia, 2008, 4, p. 477-89 Z. Mao et al., Acta Biomaterialia 2009, 5, p. 2983-94

An object of the present invention is to provide a patterned porous material capable of controlling functions such as cell adhesion, proliferation and differentiation according to a three-dimensional pattern, and a method for producing the same.

In view of the above circumstances, the present inventor repeated trial and error and completed the present invention.
The present invention has the following configuration.

(1) A pattern characterized in that a bioabsorbable polymer is aggregated in a three-dimensional network, and a concave portion is provided on one surface of a plate-like porous material provided with a plurality of holes communicating with each other. Porous material.
(2) The patterned porous material according to (1), wherein a plurality of the concave portions are provided, and the shape and area in plan view are the same.
(3) The patterned porous material according to (1) or (2), wherein the bioabsorbable polymer is crosslinked.

(4) The patterned porous material according to any one of (1) to (3), wherein the distance between the nearest concave portions is the same.
(5) The patterned porous material according to any one of (1) to (4), wherein the concave portion has a semicircular shape in cross section.

(6) The patterned porous material according to any one of (1) to (5), wherein the concave portions are arranged in a lattice shape in a plan view or a hexagonal close-packed shape in a plan view.

(7) The patterned porous material according to any one of (1) to (5), wherein the concave portions are arranged adjacent to each other and have a planar view shape.
(8) The patterned porous material according to any one of (1) to (7), wherein another concave portion having a plan view shape and / or an area different from the concave portion is provided on the one surface.

(9) The patterned porous material according to any one of (1) to (8), wherein the bioabsorbable polymer is a bioabsorbable synthetic polymer or a bioabsorbable natural polymer.
(10) The bioabsorbable natural polymer is any one of collagen, gelatin, cellulose, polyalginic acid, chitin, chitosan, starch, hyaluronic acid, chondroitin sulfate, fibronectin, or laminin. The patterned porous material as described.

(11) The patterned porous material according to any one of (1) to (10), wherein the concave portion is filled with a bioabsorbable polymer.
(12) The patterned porous material according to (11), wherein a bioactive substance or a cell growth factor is added to the bioabsorbable polymer filled in the recess.
(13) The cell growth factor is epidermal growth factor (EGF), vascular cell growth factor (VEGF), nerve growth factor (NGF), basic fibroblast growth factor (bFGF), platelet growth derived factor (PDGF), Hepatocyte growth factor (HGF), insulin-like growth factor (IGF), transforming growth factor (TGF), osteogenesis-inducing protein (BMP), vascular cell growth factor or nerve cell growth factor The patterned porous material according to (12), characterized in that

(14) A step of freezing a plurality of droplets on one surface of the substrate to form a mold substrate, and a wall-like member is disposed on the outer peripheral portion of the substrate so as to cover the plurality of droplets that are frozen. A patterned porous material comprising: a step of freezing the bioabsorbable polymer aqueous solution after filling with the bioabsorbable polymer aqueous solution; and a step of taking out the frozen polymer aqueous solution and drying under reduced pressure. Material manufacturing method.
(15) The method for producing a patterned porous material according to (14), further comprising a step of crosslinking the bioabsorbable polymer dried under reduced pressure.

(16) The temperature of the bioabsorbable polymer aqueous solution is controlled in a range of not less than the melting temperature of the bioabsorbable polymer aqueous solution and less than the melting temperature of the droplet, and the bioabsorbable polymer aqueous solution is filled (14) or (15 ) For producing a patterned porous material.
(17) The method for producing a patterned porous material according to any one of (14) to (16), wherein a material having high thermal conductivity is used as the substrate.
(18) The method for producing a patterned porous material according to any one of (14) to (17), wherein water or t-BuOH is used as a main component of the droplet.

(19) The method for producing a patterned porous material according to (18), wherein a layer having an affinity opposite to that of the main component of the droplet is formed on one surface of the substrate.
(20) The method according to any one of (14) to (19), wherein the mold substrate is created by controlling the size and position of the droplets using an inkjet method, a manual production method, or a photolithographic method. A method for producing the patterned porous material according to 1.

In the patterned porous material of the present invention, the bioabsorbable polymer is aggregated in a three-dimensional network, and a recess is provided on one surface of a plate-like porous material provided with a plurality of holes communicating with each other. Therefore, functions such as cell adhesion, proliferation, and differentiation can be controlled according to a three-dimensional pattern. Specifically, a porous material with a patterned recess size, shape, and arrangement controls the distribution of signal molecules to cells in a top-down manner, and functions such as cell adhesion, proliferation, and differentiation are tertiary. Can be controlled according to the original pattern.

Since the patterned porous material of the present invention has a configuration in which a bioactive substance or a cell growth factor is added to the bioabsorbable polymer filled in the recess, the recess is made of a polymer together with the bioactive substance or the cell growth factor. By filling, a region defined by a special chemical composition is patterned, and functions such as cell adhesion, proliferation, and differentiation can be controlled according to a three-dimensional pattern. It can be a patterned porous material.

A method for producing a patterned porous material according to the present invention includes a step of freezing a plurality of droplets on one surface of a substrate to create a mold substrate, a wall-shaped member disposed on the outer periphery of the substrate, Filling the bioabsorbable polymer aqueous solution so as to cover the plurality of droplets formed, then freezing the bioabsorbable polymer aqueous solution, and taking out the frozen bioabsorbable polymer aqueous solution and drying under reduced pressure Therefore, it is possible to easily manufacture a porous material that can control functions such as cell adhesion, proliferation, and differentiation according to a three-dimensional pattern.

It is a schematic diagram which shows an example of the patterned porous material which is embodiment of this invention. It is process drawing which shows an example of the manufacturing method of the patterned porous material which is embodiment of this invention. It is process drawing which shows an example of the manufacturing method of the patterned porous material which is embodiment of this invention. It is a schematic diagram which shows another example of the patterned porous material which is embodiment of this invention. It is a schematic diagram which shows another example of the patterned porous material which is embodiment of this invention. It is process drawing which shows another example of the manufacturing method of the patterned porous material which is embodiment of this invention. It is process drawing which shows another example of the manufacturing method of the patterned porous material which is embodiment of this invention. (A): Stereomicroscopic images of equidistant arrangement patterns of ice microparticles obtained in Example 1, (b), (c): Electron microscopic images of patterned collagen sponge prepared using the ice microparticles as a template. is there. (A): Stereomicroscopic image of a vertical and horizontal equally spaced arrangement pattern of ice microparticles obtained in Example 2, (b): Stereomicroscopic image of a patterned collagen sponge prepared using the ice microparticles as a template, and (c) ), (D), (e): electron microscope images. (A): Stereomicroscopic image of a vertical and horizontal equidistant arrangement pattern of ice microparticles obtained in Example 3, (b): Stereomicroscopic image of a patterned collagen sponge prepared using the ice microparticles as a template, and (c) ), (D), (e): electron microscope images. Obtained from Example 4 (a): Stereomicroscopic image of a pattern in which large and small ice particles are arranged at equal intervals in the vertical and horizontal directions, (b): Stereomicroscopic image of a patterned collagen sponge prepared using the ice microparticles as a template. , And (c), (d): electron microscope images. Obtained by Example 5 (a): Stereomicroscopic image of a pattern in which large and small ice microparticles are arranged in a lattice pattern, (b): Stereomicroscopic image of a patterned collagen sponge prepared using the ice microparticles as a template, And (c), (d), (e): electron microscope images. Obtained by Example 6 (a): Stereomicroscopic image of a pattern in which ice particles are arranged along each side of the hexagon, (b), (c), (d), (e): It is an electron microscope image of the patterned collagen sponge produced using the casting_mold | template. Obtained by Example 7 (a): Stereomicroscopic image of a pattern in which ice fine particles are arranged along each side of the hexagon and has a hole in the center, (b): The ice fine particles are used as a template. 3 is a stereoscopic microscope image of the patterned collagen sponge prepared in the above, and (c) and (d): an electron microscopic image. (A): Stereomicroscopic image of an ice pattern having a lattice-like groove obtained in Example 8, (b): Stereomicroscopic image of a patterned collagen sponge produced using this ice pattern as a mold, and (c) ): Electron microscope image. Example 9 (a): Stereomicroscopic image of an ice pattern having a groove and a center hole along each side of the hexagon, (b): Patterned collagen produced using this ice pattern as a template Stereomicroscopic images of sponges, and (c) and (d): electron microscopic images. (A) obtained by Example 10: a frozen pattern structure of an angiogenic factor VEGF in a collagen aqueous solution, (b): a stereomicroscopic image of a collagen sponge in which VEGF is localized in a linear pattern, (c): Electron microscopic image, (d): immunostained image. (A): a frozen pattern structure of a collagen aqueous solution containing nerve growth factor NGF obtained in Example 11, (b): a stereoscopic microscope image of a collagen sponge localized in a linear pattern of NGF, (c) : Electron microscope image, (d): immunostained image. It is an electron micrograph of collagen sponge, which is a conventional porous material for biological tissue engineering.

(First embodiment of the present invention)
<Patterned porous material>
An example of the patterned porous material that is an embodiment of the present invention will be described.
FIG. 1 is a schematic view showing an example of a patterned porous material according to an embodiment of the present invention. 1A is a plan view, and FIG. 1B is a cross-sectional view taken along the line AA ′ in FIG.
As shown in FIG. 1, the patterned porous material 1 according to the first embodiment of the present invention is provided with a plurality of pores 11c in which bioabsorbable polymers are aggregated in a three-dimensional network and communicated with each other. A plurality of recesses 21 having the same shape and the same area in plan view are provided on the one surface 11a of the plate-like porous material 11 and are schematically configured.

In FIG. 1, the porous material 11 has a substantially rectangular shape in plan view, but may have a circular shape in plan view, a polygonal shape in plan view, or the like. The size of the porous material 11 is, for example, a vertical length of 10 cm, a horizontal length of 8 cm, and a plate thickness of 1 mm.

In FIG. 1, twelve recesses 21 are provided on one surface 11a of the substrate.
The shape of the recess 21 in plan view is not limited to a circle, and may be a square shape, a polygonal shape, a star shape, or the like. Even when only a droplet having a circular shape in a plan view can be ejected by the inkjet method, a star shape in a plan view can be formed by forming the outer frame with a linear pattern and then filling the inside with a circular pattern. it can. The interval between the nearest recesses 21 is the same length t. Thereby, a fixed pattern can be limited. The length t is, for example, 0.5 mm.

The recesses 21 are arranged in a lattice shape in plan view. Thereby, a fixed pattern can be limited.
The arrangement of the recesses 21 is not limited to this, and the recesses 21 may be arranged in a hexagonal close-packed shape in a plan view. Thus, a certain pattern can be limited.
The recess 21 is semicircular in sectional view. A cross-sectional semi-elliptical shape may be used. In any case, the cross-sectional view shape is determined by the shape of the mold that is prepared by freezing water, t-BuOH, or an aqueous polymer solution. The planar view shape is circular or elliptical.

The bioabsorbable polymer is a polymer that is easily absorbed in the living body and has high safety to the living body. There are bioabsorbable synthetic polymers or bioabsorbable natural polymers. Thereby, while being able to produce a bioabsorbable polymer aqueous solution easily, it can be frozen, dried, and bridge | crosslinked as needed, and the porous material 11 can be created.
The bioabsorbable synthetic polymer is polylactic acid, polyglycolic acid, a copolymer of lactic acid and glycolic acid, poly-ε-caprolactone, or the like.
The bioabsorbable natural polymer is collagen, gelatin, cellulose, polyalginic acid, chitin, chitosan, starch, hyaluronic acid, chondroitin sulfate, fibronectin, laminin, or the like.
One kind of these bioabsorbable polymers or a mixture of several kinds can be used.

A physiologically active substance such as a cell growth factor, nanoparticles, or the like can be mixed with the bioabsorbable polymer. Cell growth factors include epidermal growth factor (EGF), vascular cell growth factor (VEGF), nerve growth factor (NGF), basic fibroblast growth factor (bFGF), platelet growth factor (PDGF), hepatocyte growth Examples include, but are not limited to, factor (HGF), insulin-like growth factor (IGF), transforming growth factor (TGF), osteogenesis-inducing protein (BMP) and the like. Among cell growth factors, at least one kind or a combination of two or more kinds can be used. A gene DNA encoding a cell growth factor can also be used. In addition, a synthetic polymer having a physiological activity, a natural polymer such as DNA and RNA, a peptide, a low molecule, and the like can be used alone or in combination of two or more.
These bioabsorbable polymers can easily produce the plate-like porous material 11 and can be used as a substrate for biological tissue engineering.

<Method for producing patterned porous material>
An example of the manufacturing method of the patterned porous material which is embodiment of this invention is demonstrated.
2 and 3 are process diagrams showing an example of a method for producing a patterned porous material according to the first embodiment of the present invention. FIG. 2A is a plan view, and FIG. 2B is a cross-sectional view taken along line BB ′ in FIG.
The manufacturing method of the patterned porous material which is the 1st Embodiment of this invention has process S1 which produces a template board | substrate, and process S2 which freezes bioabsorbable polymer aqueous solution.

(Step S1 for creating a mold substrate)
First, in step S1 of creating a template substrate, an ice mold having a pattern structure is prepared in advance, and a void portion in the porous material is controlled.
Specifically, first, as shown in FIG. 2, a plurality of droplets 34 are frozen and arranged on one surface of the substrate 31 to create a template substrate 41. In addition, a wall member 33 is disposed on the outer peripheral portion of the substrate 31.
For the arrangement of the droplets 34, for example, the size and position of the droplets are precisely controlled using an ink jet method. The size and position of the droplets determine the pattern of the patterned porous material that is finally created.
In this way, the chemical composition of the polymer 51 serving as the support portion can be controlled by previously applying the first solution of the porous body to the template in a pattern in the same manner as described above and freezing. .

Specifically, there are (1) an ink jet method (a method for producing with a dispenser robot), (2) a manual production method, and (3) a photolithographic method as a method for arranging the droplets 34.
(1) Inkjet method (method of manufacturing with a dispenser robot)
The ink jet method is a method for producing a liquid dropping pattern using a device (dispenser robot) that discharges a small amount of a small amount of liquid. In the dispenser robot, the head portion provided with the liquid discharge nozzle can be moved to an arbitrary position in the xy direction by a computer program, and can discharge liquid toward the template.
Therefore, an arbitrary dropping pattern can be drawn.
First, a polymer film is laid on or wrapped around a metal plate, and the polymer film is placed on the metal plate. Next, a pattern of water or a polymer solution is drawn according to the computer program. Next, by freezing at a temperature below the freezing point of water, an ice mold having a pattern structure is obtained.
When drawing an ice pattern directly, the environment in contact with the ice is changed to a gas such as nitrogen or argon, and the drawn metal is immediately frozen while the metal plate on which the polymer film is placed is cooled to a temperature lower than the freezing point. A template pattern is prepared.
As the polymer film, a perfluorinated alkoxy film, a polyethylene film, a polypropylene film, polyphenylene sulfide, polybutylene terephthalate, Teflon film, or the like is used.

(2) Manual production method In the manual production method, first, a polymer film is laid on or wrapped around a metal plate, and the polymer film is placed on the metal plate.
Next, a water or ice pattern is drawn on the polymer film with a sharp tip.
Next, an ice mold having a pattern structure is obtained by freezing at a temperature below the freezing point.
When drawing an ice pattern directly, the environment in contact with the ice is changed to a gas such as nitrogen or argon, and the drawn metal is immediately frozen while the metal plate on which the polymer film is placed is cooled to a temperature lower than the freezing point. A template pattern is prepared.

(3) Photolithographic method In the photolithographic method, first, a photoreactive polymer is applied on the surface of a film made of the above-described polymer.
Next, a photomask is placed and irradiated with ultraviolet rays. As a result, the photoreactive polymer is grafted on the transparent portion of the photomask, and the polymer in the portion that does not allow UV light to pass through is removed by washing to form a photoreactive polymer pattern.
When pure water is distributed on the patterned surface, the water forms a pattern structure according to the pattern of the photoreactive polymer.
When this is placed below the freezing point, an ice mold having a pattern structure is obtained. Examples of the photoreactive polymer include hydrophilic photoreactive polymers such as photoreactive polyethylene glycol, polyvinyl alcohol, polyacrylic acid, and polyallylamine.

Water or t-BuOH is used as the main component of the droplet 34. Further, a reverse affinity layer 32 having an affinity opposite to that of the main component of the droplet 34 is formed on one surface of the substrate 31. That is, when water is used as the main component of the droplet 34, a water-repellent layer such as a Teflon sheet is formed as the reverse affinity layer 32, and t-BuOH is used as the main component of the droplet 34. In some cases, a hydrophilic layer is formed as the reverse affinity layer 32. Thereby, the droplet 34 can be formed so that the contact angle with respect to the reverse affinity layer 32 becomes large, and a droplet having a semicircular sectional shape or a semielliptical sectional shape can be formed. The planar view shape is a circular shape or an elliptical shape, but it can be a planar view elliptical shape by ejecting liquid droplets in a state where the substrate is inclined.

A material having high thermal conductivity is used as the substrate 31. An example of a material having high thermal conductivity is copper. Further, a cooling mechanism (not shown) is connected to the substrate 31. For example, the temperature of the substrate 31 is set to −10 ° C. or more and −5 ° C. or less by the cooling mechanism. As a result, the droplets 34 dropped on the substrate 31 can be immediately frozen and placed.

(Step S2 of freezing the bioabsorbable polymer aqueous solution)
Next, in step S2 of freezing the bioabsorbable polymer aqueous solution, the bioabsorbable polymer aqueous solution 35e is poured and frozen to freeze the ice mold and the bioabsorbable polymer together. The ice mold and solvent are removed by lyophilizing the frozen structure.
Specifically, first, a bioabsorbable polymer aqueous solution 35e is prepared and placed in a low temperature chamber or a refrigerator at 0 ° C. or lower for several hours to bring the temperature of the solution to 0 ° C. or lower.
The cooling temperature is not required to freeze the bioabsorbable polymer aqueous solution 35e, and a desirable temperature is 0 ° C. to −10 ° C.
The cooling time is sufficient if the bioabsorbable polymer aqueous solution 35e is sufficiently cooled, and a desirable cooling time is 1 hour to 12 hours.

Next, the bioabsorbable polymer aqueous solution 35e is filled so as to cover the plurality of droplets 34 that are frozen. Since the wall-shaped member 33 is disposed on the outer peripheral portion of the substrate 31, the droplet 34 can be completely covered with the bioabsorbable polymer aqueous solution 35e. The wall-shaped member 33 is a frame made of silicone rubber or the like, and can adjust the height of the solution surface.
At the time of filling, it is preferable to control the temperature in the range of not lower than the melting temperature of the bioabsorbable polymer aqueous solution and lower than the melting temperature of the droplets 34. Thereby, the bioabsorbable polymer aqueous solution can be filled in detail, and the shape of the droplet 34 can be transferred with high accuracy. When the temperature condition is not within the above range, the transfer accuracy is insufficient.
After filling, as shown in FIG. 3 (a), the top surface of the poured bioabsorbable polymer aqueous solution 35e is flattened, and glass, metal, polymer plates, etc. covered with the polymer film are placed thereon. Then, the polymer aqueous solution 35e is blocked from the outside.
Note that the polymer aqueous solution 35e as the second solution is added and frozen so that the droplets 61 as the first frozen material do not melt.

Next, the bioabsorbable polymer aqueous solution 35e is frozen by being placed in a freezer and set to a temperature lower than the temperature at which the bioabsorbable polymer solution freezes. If necessary, the structure can be frozen using dry ice or liquid nitrogen.
As shown in FIG. 3 (b), the frozen bioabsorbable polymer aqueous solution 35k is frozen with a portion of water 37k in which water contained in the bioabsorbable polymer aqueous solution is frozen in a needle shape, a columnar shape, or the like. And bioabsorbable polymer 11k.

Next, the frozen bioabsorbable polymer aqueous solution 35k is taken out.
FIG. 3C is a cross-sectional view of the frozen bioabsorbable polymer aqueous solution 35k that has been taken out. The liquid droplet 34 is frozen and disposed on the surface of the frozen bioabsorbable polymer aqueous solution 35k.
Next, the frozen bioabsorbable polymer aqueous solution (frozen structure) 35k is dried under reduced pressure.
Drying under reduced pressure is performed by holding at a predetermined temperature in a vacuum vessel connected to a vacuum pump.
By drying under reduced pressure, the frozen water 37k and the frozen droplet 34 are distilled off, leaving only the bioabsorbable polymer, and the hole 11c and the recess 21 are formed in the removed portion.
Through the above steps, the patterned porous material 1 according to the first embodiment of the present invention shown in FIG. 1 is produced.

Furthermore, the bioabsorbable polymer of the patterned porous material 1 according to the first embodiment of the present invention may be subjected to a crosslinking treatment.
Specifically, the bioabsorbable polymer dried under reduced pressure is subjected to a crosslinking treatment in a three-dimensional network. After the crosslinking, a porous material of a bioabsorbable polymer having a void pattern structure is obtained by removing or blocking the crosslinking agent.
As a method for crosslinking the extracellular matrix, conventionally known methods shown below can be used as long as the extracellular matrix is not broken. For example, (1) a crosslinking method by radiation, (2) a crosslinking (photocrosslinking) method by ultraviolet irradiation, (3) a crosslinking (thermal crosslinking) method by heating, and (4) a chemistry using a gaseous or solution crosslinking agent. There is a cross-linking method.

(1) Crosslinking method by radiation The crosslinking reaction by radiation performs a soluble reaction by irradiating a porous material with gamma rays or electron beams. When high-energy radiation such as gamma rays or electron beams is irradiated to a polymer compound, a part of the molecular chain is changed to a radical having high reactivity, and these molecular chains are cross-linked by reacting with each other. However, irradiation may cause molecular chain degradation. Therefore, it is desirable to irradiate under conditions that increase the efficiency of the crosslinking reaction while suppressing the decomposition reaction of the polymer chain as much as possible. In order to increase the crosslinking efficiency, it is preferable to irradiate in a wet state to such an extent that the porous material does not swell. Irradiation can be performed at 1.0 to 10.0 Mrad, but an irradiation dose of 1.0 to 2.0 Mrad is particularly preferable in order to suppress the decomposition reaction.

(2) Crosslinking method by ultraviolet irradiation In the crosslinking method by ultraviolet irradiation, a porous material is placed at a certain distance and crosslinked by irradiating with ultraviolet rays for a certain time.
Specifically, in the crosslinking by ultraviolet irradiation, the polymer aqueous solution is frozen, freeze-dried, and then crosslinked by ultraviolet irradiation.
Irradiation with ultraviolet light of 240 nm to 280 nm is performed for 10 minutes to 24 hours, but irradiation with ultraviolet light of 250 nm to 260 nm is desirable for 30 minutes to 10 hours.
The thermal crosslinking is performed by freezing the polymer aqueous solution, freeze-drying, and then heating the polymer aqueous solution at a high temperature from 100 ° C. to 140 ° C. for 48 hours to 96 hours under a reduced pressure condition of 0.01 Torr to 1 Torr.

(3) Crosslinking method by heating In the crosslinking method by heating, the porous material is heated and crosslinked under reduced pressure.
The degree of vacuum is 1 × 10 ⁻⁴ Torr to 50 Torr, preferably 1 × 10 ^{−3 to} 10 Torr. When the degree of vacuum is lowered, the crosslinking effect is worsened, and the porous material is easily denatured. The temperature is 5 × 10 to ² × 10 ² ° C., preferably 8 × 10 to 1.5 × 10 ² ° C. When the temperature is lowered, the crosslinking effect is deteriorated and the crosslinking time is also increased. On the other hand, if the temperature is too high, the porous material is denatured. However, the degree of vacuum and temperature are not limited as long as the porous material can be cross-linked without removing the moisture.

(4) Chemical cross-linking method using a gas-like or solution-like cross-linking agent Cross-linking treatment with a solution-like cross-linking agent involves immersing the aqueous polymer solution in the above-mentioned cross-linking agent solution for 30 minutes to 72 hours. To do. The crosslinking treatment temperature is from 4 ° C to 37 ° C. Desirably, crosslinking is performed at 4 ° C. for 1 hour to 48 hours and at room temperature for 30 minutes to 24 hours.
In the crosslinking treatment using the gaseous crosslinking agent, the above-mentioned crosslinking agent is used in a gaseous state.

Specifically, the cross-linking treatment using a gaseous cross-linking agent is performed at a constant temperature at a constant temperature when the collagen, the bioabsorbable natural polymer, the cell growth factor, the cell differentiation factor, or a derivative thereof is cross-linked. Crosslinking is carried out for a certain period of time in an atmosphere of a crosslinking agent vapor saturated with a crosslinking agent or an aqueous solution thereof.

Solution or gaseous crosslinking agents include aldehydes such as glutaraldehyde, formaldehyde, paraformaldehyde, ethylene propylene diglycidyl ether, glycerol polyglycidyl ether, diglycerol polyglycidyl ether, sorbitol polyglycidyl ether, ethylene glycol Glycidyl ethers such as diglycidyl ether, hexamethylene diisocyanate, α-tolidine isocyanate, tolylene diisocyanate, naphthylene 1,5-diisocyanate, 4,4-diphenylmethane diisocyanate, triphenylmethane-4, 4, 4, -tri Examples include isocyanates such as isocyanate, alcohols such as methanol and ethanol, and calcium gluconate. Preferably used crosslinking agents are aldehydes such as glutaraldehyde, formaldehyde and paraformaldehyde, in particular glutaraldehyde.

The crosslinking temperature may be selected within a range in which the porous material does not dissolve and the vapor of the crosslinking agent can be formed, and is usually set to 2 × 10 to 5 × 10 ° C. Desirably, it is 37 degreeC.
The cross-linking time depends on the type of cross-linking agent and the cross-linking temperature, but cross-linking and immobilization is performed so as not to inhibit the hydrophilicity and bioabsorbability of the porous material and to prevent the porous material from being dissolved at the time of living transplantation. It is desirable to set the range.
The preferred crosslinking time is about 1/6 to 12 hours. Although it depends on the kind and size of the porous body, it is more preferably 4 hours.

After the polymer aqueous solution is frozen and freeze-dried, crosslinking is performed for a certain period of time in an atmosphere of a crosslinking agent vapor saturated with a certain concentration of a crosslinking agent aqueous solution at a certain temperature. The crosslinking temperature is usually set to 20 ° C to 40 ° C. The crosslinking time is 1 hour to 12 hours.
In the chemical crosslinking method using a gaseous or solution-like crosslinking agent, it is necessary to deactivate the crosslinking agent with a glycine aqueous solution or the like at the final stage of the crosslinking treatment.

In order to inactivate an unreacted aldehyde group, an excess amount of amines such as glycine and ethanolamine may be reacted as a quenching agent.
Glycine is particularly preferable. The concentration is 0.05 to 1.0 mol / L, preferably 0.05 to. 0.1 mol / L.

In order to wash the cross-linked product and remove the unreacted cross-linking agent, the polymer produced by the cross-linking agent, and the unreacted quenching agent if quenched, with pure water at least 10 times the outer volume of the porous body. Wash.
Through the above steps, the patterned porous material 1 according to the first embodiment of the present invention shown in FIG. 1 is produced.

(Second embodiment of the present invention)
<Patterned porous material>
Another example of the patterned porous material according to the embodiment of the present invention will be described.
FIG. 4 is a schematic view showing another example of the patterned porous material according to the embodiment of the present invention. FIG. 4A is a plan view, and FIG. 4B is a cross-sectional view taken along the line CC ′ of FIG.

As can be seen by comparing FIG. 1 and FIG. 4, the patterned porous material 2 according to the second embodiment of the present invention has a concave portion 22 having a smaller planar view area than the concave portion 21 instead of the concave portion 21. Other than that provided in 11a, the configuration is the same as that of the patterned porous material 1 according to the first embodiment of the present invention.

In addition, as the recessed part 22, the recessed part from which the planar view shape differs from the recessed part 21 may be provided in the one surface 11a, and the recessed part from which both a planar view shape and an area differ may be provided in the one surface 11a.
Furthermore, three or more types of recesses having different planar shapes or / and areas may be provided on the one surface 11a.

(Third embodiment of the present invention)
<Patterned porous material>
Another example of the patterned porous material according to the embodiment of the present invention will be described.
FIG. 5 is a schematic view showing still another example of the patterned porous material according to the embodiment of the present invention. 5A is a plan view, and FIG. 5B is a cross-sectional view taken along the line DD ′ of FIG. 5A.

As can be seen from a comparison between FIG. 1 and FIG. 5, the patterned porous material 3 according to the third embodiment of the present invention has the concave portion 21 filled with the bioabsorbable polymer 51, and the bioabsorbable polymer 51. A hole 51c is formed in the tube, and a cell growth factor 55 is added, and the configuration is the same as that of the first embodiment of the present invention.

The bioabsorbable polymer 51 filled in the recess 21 may be the same bioabsorbable polymer as the bioabsorbable polymer used for the porous material 11 or a different bioabsorbable polymer.
When a different bioabsorbable polymer is used, the bioabsorbable polymer 51 itself is used as a selective growth region of cells as in the case of the recess.
Even when the same bioabsorbable polymer is used, a selective growth region of cells can be formed by adding a physiologically active substance or a cell growth factor.
Examples of the cell growth factor include vascular cell growth factor and nerve cell growth factor.

<Method for producing patterned porous material>
Another example of the manufacturing method of the patterned porous material which is embodiment of this invention is demonstrated.
6 and 7 are process diagrams showing still another example of a method for producing a patterned porous material according to an embodiment of the present invention. 6A is a plan view, and FIG. 6B is a cross-sectional view taken along the line EE ′ of FIG. 6A.
The method for producing a patterned porous material according to the third embodiment of the present invention, except that a droplet 61 made of a bioabsorbable polymer aqueous solution to which a cell growth factor 55 is added is used instead of the droplet 34. It is set as the structure similar to the manufacturing method of the patterned porous material which is the 1st Embodiment of this invention, Process S1 which produces a template board | substrate, Process S2 which freezes bioabsorbable polymer aqueous solution, Have.

(Step S1 for creating a mold substrate)
First, as shown in FIG. 6, a plurality of droplets 61 are frozen and arranged on one surface of the substrate 31 to create a template substrate 42. In the droplet 61, in addition to the frozen bioabsorbable polymer 51k, the water 38k is also frozen.

The main component of the droplet 61 is a bioabsorbable polymer. Further, a reverse affinity layer 32 having an affinity opposite to that of the main component of the droplet 61 is formed on one surface of the substrate 31. That is, since the bioabsorbable polymer is used as the main component of the droplet 61, a water-repellent layer such as a Teflon sheet is formed as the reverse affinity layer 32. Thereby, the droplet 61 can be formed so that the contact angle with respect to the reverse affinity layer 32 is increased, and a droplet having a semicircular shape in cross section or a semielliptical shape in cross section can be formed. The planar view shape is a circular shape or an elliptical shape, but it can be a planar view elliptical shape by ejecting liquid droplets in a state where the substrate is inclined.

(Step S2 of freezing the bioabsorbable polymer aqueous solution)
First, a bioabsorbable polymer aqueous solution 35e is prepared.
Next, the cooled bioabsorbable polymer aqueous solution 35e is filled so as to cover the plurality of droplets 61 that are frozen. A wall-shaped member 33 is disposed on the outer periphery of the substrate 31. After the filling, as shown in FIG. 7 (a), glass is placed on the upper part to block the bioabsorbable polymer aqueous solution 35e from the outside.

Next, it arrange | positions in a freezer and freezes the bioabsorbable polymer aqueous solution 35e.
As shown in FIG. 7 (b), the frozen bioabsorbable polymer aqueous solution 35k is frozen with a portion of water 37k in which water contained in the bioabsorbable polymer aqueous solution is frozen in a needle shape, a columnar shape, or the like. And bioabsorbable polymer 11k.
Next, the frozen bioabsorbable polymer aqueous solution 35k is taken out.
FIG. 7C is a cross-sectional view of the extracted frozen polymer aqueous solution 35k. The droplet 61 is frozen on the surface of the bioabsorbable polymer aqueous solution 35k.
Next, the frozen bioabsorbable polymer aqueous solution 35k is dried under reduced pressure.
Drying under reduced pressure is performed by holding at a predetermined temperature in a vacuum vessel connected to a vacuum pump.
By drying under reduced pressure, the frozen water 37k in the frozen bioabsorbable polymer aqueous solution 35k and the frozen water 38k in the frozen droplet 61 are removed, and the holes 11c and 51c are removed in the removed portions. Form. The concave portion 21 is filled with the aggregated bioabsorbable polymer 51. In the bioabsorbable polymer 51, a cell growth factor 55 is added.
Through the above steps, the patterned porous material 3 according to the third embodiment of the present invention shown in FIG. 5 is created.

Furthermore, the bioabsorbable polymer of the patterned porous material 3 which is the third embodiment of the present invention may be subjected to a crosslinking treatment.
Specifically, the bioabsorbable polymer dried under reduced pressure is subjected to a crosslinking treatment in a three-dimensional network. At this time, the bioabsorbable polymer 51 in the droplet 61 is also cross-linked.
Through the above steps, the patterned porous material 3 according to the third embodiment of the present invention shown in FIG. 5 is created.

The patterned porous materials 1 to 3 according to the embodiment of the present invention are plate-like porous materials 11 in which bioabsorbable polymers are aggregated in a three-dimensional network and provided with a plurality of holes 11c communicating with each other. Since the concave portion 21 is provided on the one surface 11a, functions such as cell adhesion, proliferation and differentiation can be controlled according to a three-dimensional pattern. Specifically, a porous material with a patterned recess size, shape, and arrangement controls the distribution of signal molecules to cells in a top-down manner, and functions such as cell adhesion, proliferation, and differentiation are tertiary. Can be controlled according to the original pattern.

Since the patterned porous materials 1 to 3 according to the embodiment of the present invention are provided with a plurality of concave portions 21 and have the same shape and area in plan view, functions such as cell adhesion, proliferation and differentiation are tertiary. Can be controlled according to the original pattern.

Patterned porous materials 1 to 3 according to an embodiment of the present invention have a configuration in which the bioabsorbable polymer is cross-linked, and therefore can be formed into a porous material in which the arrangement of the recesses is patterned. By controlling the distribution of cells in a top-down manner, functions such as cell adhesion, proliferation and differentiation can be controlled according to a three-dimensional pattern.

Since the patterned porous materials 1 to 3 according to the embodiment of the present invention have a configuration in which the intervals between the nearest recesses 21 are the same, the arrangement of the recesses can be made into a patterned porous material, and to the cells. By controlling the distribution of signal molecules in a top-down manner, functions such as cell adhesion, proliferation and differentiation can be controlled according to a three-dimensional pattern.

Since the patterned porous materials 1 to 3 according to the embodiment of the present invention have a configuration in which the concave portion 21 is semicircular in cross section, the concave portion can be formed into a porous material, and signal molecules to cells can be formed. By controlling the distribution in a top-down manner, functions such as cell adhesion, proliferation and differentiation can be controlled according to a three-dimensional pattern.

Since the patterned porous materials 1 to 3 according to the embodiment of the present invention have a configuration in which the concave portions 21 are arranged in a lattice shape in a plan view or a hexagonal close-packed shape in a plan view, the porous material in which the arrangement of the concave portions is patterned. It is possible to control the distribution of signal molecules to cells in a top-down manner and control functions such as cell adhesion, proliferation and differentiation according to a three-dimensional pattern.

Since the patterned porous material according to an embodiment of the present invention has a configuration in which the concave portions are arranged adjacent to each other and have a line shape in plan view, the arrangement of the concave portions can be made into a patterned porous material, By controlling the distribution of signal molecules in a top-down manner, functions such as cell adhesion, proliferation and differentiation can be controlled according to a three-dimensional pattern.

Since the patterned porous material 2 according to the embodiment of the present invention has a configuration in which another concave portion 22 having a planar shape or / and an area different from the concave portion 21 is provided on one surface 11a, the size, shape, and arrangement of the concave portions. Can be made into a patterned porous material, and the distribution of signal molecules to cells can be controlled in a top-down manner, and functions such as cell adhesion, proliferation and differentiation can be controlled according to a three-dimensional pattern.

In the patterned porous materials 1 to 3 according to the embodiment of the present invention, the bioabsorbable polymer is configured to be a bioabsorbable synthetic polymer or a bioabsorbable natural polymer. It can be used as a material, and the distribution of signal molecules to cells can be controlled in a top-down manner, and functions such as cell adhesion, proliferation and differentiation can be controlled according to a three-dimensional pattern.

In the patterned porous materials 1 to 3 according to the embodiment of the present invention, the bioabsorbable natural polymer is collagen, gelatin, cellulose, polyalginic acid, chitin, chitosan, starch, hyaluronic acid, chondroitin sulfate, fibronectin, or laminin. Therefore, it can be made into a porous material with concave parts patterned, and the distribution of signal molecules to cells can be controlled in a top-down manner to provide functions such as cell adhesion, proliferation, and differentiation in a three-dimensional pattern. Can be controlled according to.

The patterned porous material 3 according to the embodiment of the present invention has a configuration in which the concave portion 21 is filled with a bioabsorbable polymer, so that the bioabsorbable polymer in the concave portion can be a patterned porous material, By controlling the distribution of signal molecules to cells in a top-down manner, functions such as cell adhesion, proliferation and differentiation can be controlled according to a three-dimensional pattern.

Since the patterned porous material 3 according to the embodiment of the present invention has a configuration in which a bioactive substance or a cell growth factor 55 is added to the bioabsorbable polymer filled in the recesses 21, A porous material in which molecules are patterned with a special chemical composition can be formed, and a patterned porous material in which functions such as cell adhesion, proliferation, and differentiation can be controlled in accordance with a three-dimensional pattern.

In the patterned porous material 3 according to the embodiment of the present invention, cell growth factor 55 is epidermal cell growth factor (EGF), vascular cell growth factor (VEGF), nerve growth factor (NGF), basic fibroblast growth factor. (BFGF), platelet growth factor (PDGF), hepatocyte growth factor (HGF), insulin-like growth factor (IGF), transforming growth factor (TGF), osteogenesis-inducing protein (BMP), vascular cell growth factor or nerve Since it is composed of one or more cell growth factors, it can be made into a porous material in which the bioabsorbable polymer in the recess is patterned with a special chemical composition, and functions such as cell adhesion, proliferation and differentiation are tertiary. It can be a patterned porous material that can be controlled according to the original pattern.

The method for producing a patterned porous material according to an embodiment of the present invention includes a step of freezing a plurality of droplets 34 and 61 on one surface of a substrate 31 to form mold substrates 41 and 42, The wall-shaped member 33 is disposed on the outer peripheral portion, and the bioabsorbable polymer aqueous solution 35e is filled so as to cover the plurality of frozen droplets 34 and 61, and then the bioabsorbable polymer aqueous solution 35e is frozen. Since the structure includes a step, a step of taking out the frozen bioabsorbable polymer aqueous solution 35k and drying under reduced pressure, and a step of crosslinking the bioabsorbable polymer dried under reduced pressure, the size, shape, It is possible to easily manufacture a patterned porous material that can precisely control the arrangement and control functions such as cell adhesion, proliferation, and differentiation according to a three-dimensional pattern.

Since the method for producing a patterned porous material according to an embodiment of the present invention includes a step of crosslinking the bioabsorbable polymer dried under reduced pressure, the information on the droplets is transferred to the bioabsorbable polymer. Thus, a stabilized patterned porous material that can control functions such as cell adhesion, proliferation, and differentiation according to a three-dimensional pattern can be easily manufactured.

In the method for producing a patterned porous material according to an embodiment of the present invention, the temperature is controlled in the range of not lower than the melting temperature of the bioabsorbable polymer aqueous solution 35e and lower than the melting temperature of the droplets 34 and 61, and bioabsorbable. Since the structure is filled with the polymer aqueous solution 35e, the bioabsorbable polymer aqueous solution is introduced without impairing the size, shape, and arrangement of the droplet, and the droplet information is transferred to the bioabsorbable polymer. Patterned porous materials that can control functions such as cell adhesion, proliferation, and differentiation according to a three-dimensional pattern can be easily manufactured.

Since the method for producing a patterned porous material according to an embodiment of the present invention uses a material having high thermal conductivity as the substrate 31, functions such as cell adhesion, proliferation, differentiation, etc. can be immediately frozen and placed in a droplet. It is possible to easily manufacture a patterned porous material that can be controlled according to a three-dimensional pattern.

Since the manufacturing method of the patterned porous material according to the embodiment of the present invention uses water or t-BuOH as a main component of the droplets 34 and 61, the size, shape, and arrangement of the droplets can be precisely controlled. In addition, it is possible to easily produce a patterned porous material in which functions such as cell adhesion, proliferation, and differentiation can be controlled according to a three-dimensional pattern.

Since the method for producing a patterned porous material according to an embodiment of the present invention is configured to form a layer having an affinity opposite to the main components of the droplets 34 and 61 on one surface of the substrate 31, the size of the droplets, It is possible to easily manufacture a patterned porous material in which the shape and arrangement can be precisely controlled, and functions such as cell adhesion, proliferation, and differentiation can be controlled according to a three-dimensional pattern.

The method for producing a patterned porous material according to an embodiment of the present invention uses an ink-jet method, a manual production method, or a photolithographic method to control the size and position of the droplets 34 and 61, thereby forming a template substrate 41. 42, the size, shape, and arrangement of the droplets can be precisely controlled, and a patterned porous material that can control functions such as cell adhesion, proliferation, and differentiation according to a three-dimensional pattern can be easily manufactured.

The patterned porous material and the manufacturing method thereof according to the embodiment of the present invention are not limited to the above-described embodiment, and can be implemented with various modifications within the scope of the technical idea of the present invention. Specific examples of this embodiment are shown in the following examples. However, the present invention is not limited to these examples.

As a patterned porous material of Example 1, a collagen aqueous solution is placed on a Teflon mold plate on which ice fine particles are formed, and this is freeze-dried, whereby a patterned collagen sponge 1 having a constant interval between pores is obtained. Was made.

After placing the Teflon sheet on a copper template and cooling at −5 ° C., using a dispenser robot equipped with a 28-gauge syringe, pure water is dropped onto the surface of the Teflon sheet. An ice fine particle pattern was prepared. Here, the number of times of dropping was one for each dropping position, and the interval between the dropping positions was 1100 μm. The distance between the discharge tip of the nozzle and the surface of the Teflon sheet was 2.0 mm.
Next, a Teflon sheet on which a pattern of fine ice particles was formed was placed after 5 hours at -30 ° C. This was moved to a low temperature chamber of −1 ° C., and a mold of 1.0 mm thick silicone rubber plate cut out into a rectangle of 100 mm × 60 mm was piled up.
To this mold, 3 mL of 1.0 wt% porcine type I atelocollagen acidic aqueous solution (pH = 3.0) was added. This was frozen at −80 ° C. for 24 hours and then freeze-dried under reduced pressure (0.01 Torr) for 24 hours to form a porous structure.

The porous structure thus formed was subjected to a crosslinking reaction at 37 ° C. for 4 hours under a glutaraldehyde vapor saturated with a 25 wt% glutaraldehyde aqueous solution, and then washed five times with pure water.
Furthermore, after blocking the unreacted aldehyde group with 0.1 M glycine aqueous solution for 24 hours, it was washed 20 times with pure water.
This was frozen at −80 ° C. for 4 hours and freeze-dried for 24 hours to prepare patterned collagen sponge 1.

A stereomicrograph of the ice fine particle pattern is shown in FIG. 8A, and electron micrographs of the obtained patterned collagen sponge 1 are shown in FIGS. 8B and 8C. From FIG. 8 (a), it was found that a dot-like pattern of ice particles was formed on the surface of the Teflon sheet. Large vacancies reflecting the pattern of ice particles were confirmed on the surface of the patterned collagen sponge 1, and it was found that small vacancies exist between the large vacancies. It was confirmed that there were internal bulk vacancies under larger surface vacancies. The large pores on the outer surface of the sponge showed a funnel-like porous structure connected with the inner bulk pores.

As a patterned porous material of Example 2, a collagen aqueous solution is placed on a Teflon mold plate on which ice fine particles are formed, and this is freeze-dried, whereby a patterned collagen sponge 2 having a constant interval between pores is obtained. Was made.

After placing the Teflon sheet on the metal mold plate and cooling it at -5 ° C, by using a dispenser robot equipped with a 28 gauge syringe, pure water is dropped toward the Teflon surface, to the surface of the Teflon sheet. Ice fine particles were prepared. The number of times of dropping was 5 with respect to each dropping position, and the interval between the dropping positions was 1500 μm. The distance between the discharge tip of the nozzle and the surface of the Teflon sheet was 2.0 mm. Next, a Teflon sheet on which a pattern of fine ice particles was formed was placed after 5 hours at -30 ° C. This was moved to a low temperature chamber of −1 ° C., and a mold of 1.0 mm thick silicone rubber plate cut out into a rectangle of 100 mm × 60 mm was piled up. To this mold, 3 mL of 1.0 wt% porcine type I atelocollagen acidic aqueous solution (pH = 3.0) was added. This was frozen at −80 ° C. for 24 hours and then freeze-dried under reduced pressure (0.01 Torr) for 24 hours to form a porous structure.

The porous structure thus formed was subjected to a crosslinking treatment at 37 ° C. for 4 hours under a glutaraldehyde vapor saturated with a 25 wt% aqueous glutaraldehyde solution, and then washed five times with pure water. Furthermore, after blocking the unreacted aldehyde group with 0.1 M glycine aqueous solution for 24 hours, it was washed 20 times with pure water. This was frozen at −80 ° C. for 4 hours and freeze-dried for 24 hours to prepare a patterned collagen sponge 2.

9A is a stereomicrograph of the ice fine particle pattern, FIG. 9B is a stereomicrograph of the obtained patterned collagen sponge 2, and FIGS. 9C, 9D, and 9E are electron micrographs. ). Large pores reflecting the pattern of ice particles were confirmed on the surface of the patterned collagen sponge 2, and it was found that there were small pores between the large pores. It was confirmed that there were internal bulk vacancies under larger surface vacancies. The large pores on the outer surface of the sponge showed a funnel-like porous structure connected with the inner bulk pores.

As a patterned porous material of Example 3, a collagen aqueous solution is placed on a Teflon mold plate on which ice fine particles are formed, and freeze-dried to form a patterned collagen sponge 3 having a constant interval between pores. Was made.

After placing the Teflon sheet on the metal mold plate and cooling it at -5 ° C, by using a dispenser robot equipped with a 28 gauge syringe, pure water is dropped toward the Teflon surface, to the surface of the Teflon sheet. Ice fine particles were prepared. The number of times of dropping was 10 times for each dropping position, and the interval between the dropping positions was 1700 μm. The distance between the discharge tip of the nozzle and the surface of the Teflon sheet was 2.0 mm. Next, a Teflon sheet on which a pattern of fine ice particles was formed was placed after 5 hours at -30 ° C. This was moved to a low temperature chamber of −1 ° C., and a mold of 1.0 mm thick silicone rubber plate cut out into a rectangle of 100 mm × 60 mm was piled up. To this mold, 3 mL of 1.0 wt% porcine type I atelocollagen acidic aqueous solution (pH = 3.0) was added. This was frozen at −80 ° C. for 24 hours and then freeze-dried under reduced pressure (0.01 Torr) for 24 hours to form a porous structure.

The porous structure thus formed was subjected to a crosslinking treatment at 37 ° C. for 4 hours under a glutaraldehyde vapor saturated with a 25 wt% aqueous glutaraldehyde solution, and then washed five times with pure water. Furthermore, after blocking the unreacted aldehyde group with 0.1 M glycine aqueous solution for 24 hours, it was washed 20 times with pure water. This was frozen at −80 ° C. for 4 hours and freeze-dried for 24 hours to prepare a patterned collagen sponge 3. An electron micrograph of the resulting patterned collagen sponge is shown in FIG. It was found that the surface of the patterned collagen sponge had a pattern with a constant spacing between pores.

A stereomicrograph of the ice fine particle pattern is shown in FIG. 10A, a stereomicrograph of the obtained patterned collagen sponge 3 is shown in FIG. 10B, and electron micrographs are shown in FIGS. 10C, 10D, 10E. ). A hole pattern reflecting the shape of the ice fine particle pattern was confirmed on the surface of the patterned collagen sponge 3, and it was found that there were small holes between the large holes. It was confirmed that there were internal bulk vacancies under larger surface vacancies. The large pores on the outer surface of the sponge showed a funnel-like porous structure connected with the inner bulk pores.

As a patterned porous material of Example 4, an aqueous collagen solution is placed on a Teflon mold plate on which ice fine particles are formed, and this is freeze-dried to arrange large vacancies and small vacancies in an alternating pattern. A patterned collagen sponge 4 was prepared.

After placing the Teflon sheet on the metal mold plate and cooling it at -5 ° C, by using a dispenser robot equipped with a 28 gauge syringe, pure water is dropped toward the Teflon surface, to the surface of the Teflon sheet. Ice fine particles were prepared. Here, the number of drops for large droplets was 10 for each drop position, and the number of drops for small droplets was 1. The dropping interval was 1250 μm. The distance between the discharge tip of the nozzle and the surface of the Teflon sheet was 2.0 mm. Next, a Teflon sheet on which a pattern of fine ice particles was formed was placed after 5 hours at -30 ° C. This was moved to a low temperature chamber of −1 ° C., and a mold of 1.0 mm thick silicone rubber plate cut out into a rectangle of 100 mm × 60 mm was piled up. To this mold, 3 mL of 1.0 wt% porcine type I atelocollagen acidic aqueous solution (pH = 3.0) was added. This was frozen at −80 ° C. for 24 hours and then freeze-dried under reduced pressure (0.01 Torr) for 24 hours to form a porous structure.

The porous structure thus formed was subjected to a crosslinking treatment at 37 ° C. for 4 hours under a glutaraldehyde vapor saturated with a 25 wt% aqueous glutaraldehyde solution, and then washed five times with pure water. Furthermore, after blocking the unreacted aldehyde group with 0.1 M glycine aqueous solution for 24 hours, it was washed 20 times with pure water. This was frozen at −80 ° C. for 4 hours and freeze-dried for 24 hours to prepare a patterned collagen sponge 4.

A stereomicrograph of the ice fine particle pattern is shown in FIG. 11 (a), a stereomicrograph of the obtained patterned collagen sponge 4 is shown in FIG. 11 (b), and electron micrographs are shown in FIGS. 11 (c) and 11 (c). A hole pattern reflecting the shape of the ice fine particle pattern was confirmed on the surface of the patterned collagen sponge 4, and it was found that there were small holes between the large holes. It was confirmed that there were internal bulk vacancies under larger surface vacancies. The large pores on the outer surface of the sponge showed a funnel-like porous structure connected with the inner bulk pores.

As a patterned porous material of Example 5, a collagen aqueous solution is placed on a Teflon mold plate on which ice fine particles are formed, and this is freeze-dried to form patterned collagen in which pores are arranged in a lattice pattern. A sponge 5 was produced.

After placing the Teflon sheet on the metal mold plate and cooling it at -5 ° C, by using a dispenser robot equipped with a 28 gauge syringe, pure water is dropped toward the Teflon surface, to the surface of the Teflon sheet. A pattern of ice particles was prepared. Here, the number of drops for large droplets was 10 for each drop position, and the number of drops for small droplets was 1. The dropping interval was 1000 μm between the small droplets and 1500 μm between the large and small droplets. The distance between the discharge tip of the nozzle and the surface of the Teflon sheet was 2.0 mm. Next, a Teflon sheet on which a pattern of fine ice particles was formed was placed after 5 hours at -30 ° C. This was moved to a low temperature chamber of −1 ° C., and a mold of 1.0 mm thick silicone rubber plate cut out into a rectangle of 100 mm × 60 mm was piled up. To this mold, 3 mL of 1.0 wt% porcine type I atelocollagen acidic aqueous solution (pH = 3.0) was added. This was frozen at −80 ° C. for 24 hours and then freeze-dried under reduced pressure (0.01 Torr) for 24 hours to form a porous structure.

The porous structure thus formed was subjected to a crosslinking treatment at 37 ° C. for 4 hours under a glutaraldehyde vapor saturated with a 25 wt% aqueous glutaraldehyde solution, and then washed five times with pure water. Furthermore, after blocking the unreacted aldehyde group with 0.1 M glycine aqueous solution for 24 hours, it was washed 20 times with pure water. This was frozen at −80 ° C. for 4 hours and freeze-dried for 24 hours to prepare a patterned collagen sponge 5.

A stereomicrograph of the ice fine particle pattern is shown in FIG. 12 (a), a stereomicrograph of the obtained patterned collagen sponge 5 is shown in FIG. 12 (b), and electron micrographs are shown in FIGS. 12 (c), (d), and e). Shown in A hole pattern reflecting the shape of the ice fine particle pattern was confirmed on the surface of the patterned collagen sponge 5, and it was found that there were small holes between the large holes. It was confirmed that there were internal bulk vacancies under larger surface vacancies. The large pores on the outer surface of the sponge showed a funnel-like porous structure connected with the inner bulk pores.

A pattern in which pores are arranged in a hexagonal pattern by placing an aqueous collagen solution on a Teflon mold plate on which ice fine particles are formed as a patterned porous material of Example 6 and lyophilizing it. A collagen collagen sponge 6 was produced.

After placing the Teflon sheet on a metal mold plate and cooling it at -5 ° C, using a dispenser robot equipped with a 28-gauge syringe, 5 ° C pure water is dropped toward the Teflon surface to make the Teflon sheet. A pattern of ice fine particles was prepared on the surface of. The distance between the discharge tip of the nozzle and the surface of the Teflon sheet was 2.0 mm. Next, a Teflon sheet on which a pattern of fine ice particles was formed was placed after 5 hours at -30 ° C. This was moved to a low temperature chamber of −1 ° C., and a mold of 1.0 mm thick silicone rubber plate cut out into a rectangle of 100 mm × 60 mm was piled up. To this mold, 3 mL of 1.0 wt% porcine type I atelocollagen acidic aqueous solution (pH = 3.0) was added. This was frozen at −80 ° C. for 24 hours and then freeze-dried under reduced pressure (0.01 Torr) for 24 hours to form a porous structure.

The porous structure thus formed was subjected to a crosslinking treatment at 37 ° C. for 4 hours under a glutaraldehyde vapor saturated with a 25 wt% aqueous glutaraldehyde solution, and then washed five times with pure water. Furthermore, after blocking the unreacted aldehyde group with 0.1 M glycine aqueous solution for 24 hours, it was washed 20 times with pure water. This was frozen at −80 ° C. for 4 hours and freeze-dried for 24 hours to prepare a patterned collagen sponge 6.

A stereomicrograph of the ice fine particle pattern is shown in FIG. 13 (a), and electron micrographs of the obtained patterned collagen sponge 6 are shown in FIGS. 13 (b), (c), (d), and (e). A hole pattern reflecting the shape of the ice fine particle pattern was confirmed on the surface of the patterned collagen sponge 6, and it was found that there were small holes between the large holes. It was confirmed that there were internal bulk vacancies under larger surface vacancies. The large pores on the outer surface of the sponge showed a funnel-like porous structure connected with the inner bulk pores.

As a patterned porous material of Example 7, an aqueous collagen solution was placed on a Teflon mold plate on which ice fine particles were formed, and this was freeze-dried, whereby pores were placed on the sides and the center of the hexagon. A patterned collagen sponge 7 was prepared.
After placing the Teflon sheet on a metal mold plate and cooling at −5 ° C., by using a dispenser robot equipped with a 28-gauge syringe, 5 ° C. pure water was dropped toward the surface of the Freon to Ice particles were prepared on the surface. The distance between the discharge tip of the nozzle and the surface of the Teflon sheet was 2.0 mm. Next, a Teflon sheet on which a pattern of fine ice particles was formed was placed after 5 hours at -30 ° C. This was moved to a low temperature chamber of −1 ° C., and a mold of 1.0 mm thick silicone rubber plate cut out into a rectangle of 100 mm × 60 mm was piled up. To this mold, 3 mL of 1.0 wt% porcine type I atelocollagen acidic aqueous solution (pH = 3.0) was added. This was frozen at −80 ° C. for 24 hours and then freeze-dried under reduced pressure (0.01 Torr) for 24 hours to form a porous structure.

The porous structure thus formed was subjected to a crosslinking treatment at 37 ° C. for 4 hours under a glutaraldehyde vapor saturated with a 25 wt% aqueous glutaraldehyde solution, and then washed five times with pure water. Furthermore, after blocking the unreacted aldehyde group with 0.1 M glycine aqueous solution for 24 hours, it was washed 20 times with pure water. This was frozen at −80 ° C. for 4 hours and freeze-dried for 24 hours to prepare a patterned collagen sponge 7.

A stereomicrograph of the ice fine particle pattern is shown in FIG. 14 (a), a stereomicrograph of the obtained patterned collagen sponge 7 is shown in FIG. 14 (b), and electron micrographs are shown in FIGS. 14 (c) and 14 (d). A hole pattern reflecting the shape of the ice fine particle pattern was confirmed on the surface of the patterned collagen sponge 7, and it was found that there were small holes between the large holes. It was confirmed that there were internal bulk vacancies under larger surface vacancies. The large pores on the outer surface of the sponge showed a funnel-like porous structure connected with the inner bulk pores.

As a patterned porous material of Example 8, a collagen aqueous solution was placed on a Teflon mold plate on which ice fine particles were formed, and this was freeze-dried, whereby a patterned collagen sponge 8 having grooves in an orthogonal lattice pattern was obtained. Was made.

After placing the Teflon sheet on a metal mold plate and cooling at −5 ° C., pure water was dropped toward the surface of the dispenser robot Teflon to produce an ice pattern on the surface of the Teflon sheet. At this time, the distance between the discharge tip of the nozzle and the surface of the Teflon sheet was set to 2.0 mm. Next, a Teflon sheet on which an ice pattern was formed was placed at −30 ° C. for 5 hours. This was moved to a low temperature chamber of −1 ° C., and a mold of 1.0 mm thick silicone rubber plate cut out into a rectangle of 100 mm × 60 mm was piled up. To this mold, 3 mL of 1.0 wt% porcine type I atelocollagen acidic aqueous solution (pH = 3.0) was added. This was frozen at −80 ° C. for 24 hours and then freeze-dried under reduced pressure (0.01 Torr) for 24 hours to form a porous structure.

The porous structure thus formed was subjected to a crosslinking treatment at 37 ° C. for 4 hours under a glutaraldehyde vapor saturated with a 25 wt% aqueous glutaraldehyde solution, and then washed five times with pure water. Furthermore, after blocking the unreacted aldehyde group with 0.1 M glycine aqueous solution for 24 hours, it was washed 20 times with pure water. This was frozen at −80 ° C. for 4 hours and freeze-dried for 24 hours to prepare a patterned collagen sponge 8.

A stereomicrograph of the ice fine particle pattern is shown in FIG. 15 (a), a stereomicrograph of the resulting patterned collagen sponge 8 is shown in FIG. 15 (b), and an electron micrograph is shown in FIG. 15 (c). A hole pattern reflecting the shape of the ice pattern was confirmed on the surface of the patterned collagen sponge 8, and it was found that there were small holes between the large holes. It was confirmed that there were internal bulk vacancies under larger surface vacancies. The large pores on the outer surface of the sponge showed a funnel-like porous structure connected with the inner bulk pores.

As a patterned porous material of Example 9, a collagen aqueous solution was placed on a Teflon mold plate on which ice fine particles were formed, and this was freeze-dried, thereby forming a pattern having hexagonal grooves and pores in the center. Patterned collagen sponge 9 having

After placing the Teflon sheet on the metal mold plate and cooling at -5 ° C, pure water is dropped on the surface of the Teflon sheet using a dispenser robot equipped with a 28 gauge syringe. The pattern of was produced. At this time, the distance between the discharge tip of the nozzle and the surface of the Teflon sheet was set to 2.0 mm. Next, a Teflon sheet on which a pattern of fine ice particles was formed was placed after 5 hours at -30 ° C. This was moved to a low temperature chamber of −1 ° C., and a mold of 1.0 mm thick silicone rubber plate cut out into a rectangle of 100 mm × 60 mm was piled up. To this mold, 3 mL of 1.0 wt% porcine type I atelocollagen acidic aqueous solution (pH = 3.0) was added. This was frozen at −80 ° C. for 24 hours and then freeze-dried under reduced pressure (0.01 Torr) for 24 hours to form a porous structure.

The porous structure thus formed was subjected to a crosslinking treatment at 37 ° C. for 4 hours under a glutaraldehyde vapor saturated with a 25 wt% aqueous glutaraldehyde solution, and then washed five times with pure water. Furthermore, after blocking the unreacted aldehyde group with 0.1 M glycine aqueous solution for 24 hours, it was washed 20 times with pure water. This was frozen at −80 ° C. for 4 hours and freeze-dried for 24 hours to prepare a patterned collagen sponge 9.

A stereomicrograph of the ice fine particle pattern is shown in FIG. 16 (a), a stereomicrograph of the obtained patterned collagen sponge 9 is shown in FIG. 16 (b), and electron micrographs are shown in FIGS. 16 (c) and 16 (d). A hole pattern reflecting the shape of the ice fine particle pattern was confirmed on the surface of the patterned collagen sponge 9, and it was found that there were small holes between the large holes. It was confirmed that there were internal bulk vacancies under larger surface vacancies. The large pores on the outer surface of the sponge showed a funnel-like porous structure connected with the inner bulk pores.

As the patterned porous material of Example 10, a patterned collagen sponge 10 in which blood vessel growth factor (VEGF) was localized in a linear pattern was produced.

Human VEGF was added to a 1.0 wt% acidic aqueous solution of porcine type I atelocollagen (pH = 3.0) so as to be 5 μg / mL. Using a dispenser robot, this solution was applied to the surface of a copper template (100 mm × 80 mm × 5 mm thick) covered with a polyfluoroalkoxy film (thickness 25 μm) so that the line width was 1 mm and the line spacing was 2 mm. After leaving still in a -30 degreeC freezer for 1 hour, it moved to the low temperature chamber set to -5 degreeC, and left still for 1 hour. A silicone rubber frame with a thickness of 1.0 mm cut out to 80 mm × 65 mm was placed, and 7.5 mL of 1.0 wt% porcine type I atelocollagen aqueous solution (pH = 3.0) was added to the inside of the frame. A glass plate (100 mm × 80 mm × thickness 5 mm) was placed and sealed, and after removing an excessive amount of solution, it was cooled at −5 ° C. for 1 hour. After cooling, the silicone rubber frame, the glass plate, and the PFA film-covered copper mold plate were removed and cooled at −80 ° C. for 6 hours. Subsequently, a porous structure was formed by lyophilization at about 5 Pascals for 48 hours.

The porous structure thus formed was allowed to stand under glutaraldehyde vapor saturated with a 25 wt% aqueous glutaraldehyde solution to carry out a crosslinking reaction at 37 ° C. for 4 hours. The porous body after the reaction was cut into a square having a side of 12 mm. Furthermore, in order to quench the unreacted aldehyde group, the porous material was placed in a 0.1 M glycine aqueous solution and reacted at 4 ° C. for 12 hours. The plate was washed with 100 mL of pure water for 15 minutes, and this washing was repeated 10 times.

FIG. 17 (a) is a stereoscopic micrograph of a frozen aqueous collagen solution containing VEGF, (b) is a stereoscopic microscope image of the patterned collagen sponge 10 obtained, (d) is an electron micrograph, and (d) is immunostained. Show the image. From the electron microscope image, a porous structure was observed. From the immunostained image, VEGF was visualized according to the drawn pattern, indicating that VEGF was localized according to the pattern.

As Example 11, a collagen sponge 11 in which nerve growth factor (NGF) was localized in a linear pattern was prepared.

Human NGF was added to a 1.0 wt% acidic aqueous solution of porcine type I atelocollagen (pH = 3.0) so as to be 50 μg / mL. Using a dispenser robot, this solution was applied onto the surface of a copper template (100 mm × 80 mm × thickness 5 mm) covered with a polyfluoroalkoxy film (thickness 25 μm) so that the line width was 300 μm and the line spacing was 2 mm. After leaving still in a -30 degreeC freezer for 1 hour, it moved to the low temperature chamber set to -5 degreeC, and left still for 1 hour. A silicone rubber frame with a thickness of 1.0 mm cut out to 80 mm × 65 mm was placed, and 7.5 mL of 1.0 wt% porcine type I atelocollagen aqueous solution (pH = 3.0) was added to the inside of the frame. A glass plate (100 mm × 80 mm × thickness 5 mm) was placed and sealed, and after removing an excessive amount of solution, it was cooled at −5 ° C. for 1 hour. After cooling, the silicone rubber frame, the glass plate, and the PFA film-covered copper mold plate were removed and cooled at −80 ° C. for 6 hours. Subsequently, a porous structure was formed by lyophilization at about 5 Pascals for 48 hours.

The porous structure thus formed was allowed to stand under glutaraldehyde vapor saturated with a 25 wt% aqueous glutaraldehyde solution to carry out a crosslinking reaction at 37 ° C. for 4 hours. The porous body after the reaction was cut into a square having a side of 12 mm. Furthermore, in order to quench the unreacted aldehyde group, the porous material was placed in a 0.1 M glycine aqueous solution and reacted at 4 ° C. for 12 hours. The plate was washed with 100 mL of pure water for 15 minutes, and this washing was repeated 10 times.

FIG. 18 (a) is a stereoscopic microscope photograph of a frozen collagen aqueous solution containing NGF, (b) is a stereoscopic microscope image of the patterned collagen sponge 11 obtained, (c) is an electron microscope image, and (d) is immunostained. Show the image. From the electron microscope image, a porous structure was observed. From the immunostained image, NGF was visualized according to the drawn pattern, indicating that NGF was localized according to the pattern.

The present invention relates to a patterned porous material and a method for producing the same, and can spatially control functions such as cell adhesion, proliferation, and differentiation, and can be used in the regenerative organ industry, the regenerative medicine industry, and the like. There is.

1, 2, 3 ... patterned porous material, 11 ... porous material, 11a ... one side, 11c ... pore, 11k ... frozen bioabsorbable polymer, 21, 22 ... recess, 31 ... substrate, 32 ... Droplet forming layer, 33 ... wall-like member, 34 ... droplet, 35e ... bioabsorbable polymer aqueous solution, 35k ... frozen bioabsorbable polymer aqueous solution, 36 ... glass plate, 37k, 38k ... frozen water 41, 42 ... template substrate, 51 ... bioabsorbable polymer, 51c ... hole, 51k ... frozen bioabsorbable polymer, 55 ... cell growth factor, 61 ... droplet.

Claims (18)

Bioabsorbable polymer is aggregated in a three-dimensional net-like, Ri Contact plurality of concave portions are provided on one surface of the plurality of holes are provided plate-shaped porous material in communication with each other,
The plurality of recesses constitutes a predetermined pattern in plan view,
A patterned porous material characterized in that at least some of the plurality of recesses constituting the predetermined pattern are filled with a bioabsorbable polymer to which a physiologically active substance or a cell growth factor is added. . Before SL plurality of recesses patterned porous material according to claim 1, characterized in that in plan view the shape and area of the same.   The patterned porous material according to claim 1, wherein the bioabsorbable polymer is crosslinked.   The patterned porous material according to any one of claims 1 to 3, wherein the distance between the nearest concave portions is the same. Patterned porous material according to any one of claims 1 to 4, prior Symbol plurality of concave portions is characterized in that it is a cross section semicircle. Patterned porous material according to any one of claims 1 to 5, before Symbol plurality of concave portions is characterized in that it is arranged in plan view lattice or viewed hexagonal close-shaped. Before SL plurality of concave portions are arranged adjacent, patterned porous material according to any one of claims 1 to 5, characterized in that there is a flat sight line shape. Patterned porous material according to any one of claims 1 to 7 is a plan view shape and / or area of another different from the previous SL plurality of recessed portions recessed portion and being provided on the one surface .   The patterned porous material according to any one of claims 1 to 8, wherein the bioabsorbable polymer is a bioabsorbable synthetic polymer or a bioabsorbable natural polymer.   The pattern according to claim 9, wherein the bioabsorbable natural polymer is any one of collagen, gelatin, cellulose, polyalginic acid, chitin, chitosan, starch, hyaluronic acid, chondroitin sulfate, fibronectin, or laminin. Porous material. The cell growth factors include epidermal growth factor (EGF), vascular cell growth factor (VEGF), nerve growth factor (NGF), basic fibroblast growth factor (bFGF), platelet growth derived factor (PDGF), hepatocyte growth. It is any one or more of a factor (HGF), an insulin-like growth factor (IGF), a transforming growth factor (TGF), a bone formation-inducing protein (BMP), a vascular cell growth factor or a nerve cell growth factor The patterned porous material according to claim 1 . A plurality of droplets on a surface of a substrate and freeze arranged, there is more Engineering you prepare a template substrate,
Maintaining the substrate cooled to a temperature lower than the freezing point, dropping the plurality of droplets so as to form a predetermined pattern in plan view while controlling the size and position on one surface of the substrate, At least some of the plurality of dropped droplets are composed of a bioabsorbable polymer aqueous solution to which a physiologically active substance or a cell growth factor is added; and
Placing a wall-like member on the outer periphery of the substrate, filling the bioabsorbable polymer aqueous solution so as to cover the plurality of frozen droplets, and then freezing the bioabsorbable polymer aqueous solution;
A method of producing a patterned porous material, comprising: taking out a frozen bioabsorbable polymer aqueous solution and drying under reduced pressure. The method for producing a patterned porous material according to claim 12 , further comprising a step of crosslinking the bioabsorbable polymer dried under reduced pressure. The bioabsorbable polymer solution melting temperature or more, the solution was temperature controlled in the range below the melting temperature of the droplets, or claim 12, characterized in that filling the bioabsorbable polymer solution to Claim 13 A method for producing the patterned porous material as described. The method for producing a patterned porous material according to claim 12, wherein a material having high thermal conductivity is used as the substrate. The method for producing a patterned porous material according to any one of claims 12 to 15, wherein water or t-BuOH is used as a main component of the droplets. The method for producing a patterned porous material according to claim 16 , wherein a layer having an affinity opposite to that of the main component of the droplet is formed on one surface of the substrate. An ink-jet method, a manufacturing method of patterning a porous material according to any one of claims 12 to 17, characterized that you prepare a cast type substrate by the manufacturing method or photolithography method manually.