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
  • A61L27/58—Materials at least partially resorbable by the body
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
  • A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
  • A61K35/37—Digestive system
  • A61K35/39—Pancreas; Islets of Langerhans
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
  • A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
  • A61K35/37—Digestive system
  • A61K35/407—Liver; Hepatocytes
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K9/00—Medicinal preparations characterised by special physical form
  • A61K9/0012—Galenical forms characterised by the site of application
  • A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
  • A61K9/0024—Solid, semi-solid or solidifying implants, which are implanted or injected in body tissue
• • 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
  • A61L27/14—Macromolecular materials
  • A61L27/16—Macromolecular materials obtained by reactions only involving carbon-to-carbon unsaturated bonds
• • 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
  • A61L27/14—Macromolecular materials
  • A61L27/22—Polypeptides or derivatives thereof, e.g. degradation products
  • A61L27/222—Gelatin
• • 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
  • A61L27/38—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 containing added animal 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
  • A61L27/36—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
  • A61L27/38—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 containing added animal cells
  • A61L27/3804—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 containing added animal cells characterised by specific cells or progenitors thereof, e.g. fibroblasts, connective tissue cells, kidney 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
  • A61L27/40—Composite materials, i.e. containing one material dispersed in a matrix of the same or different material
  • A61L27/44—Composite materials, i.e. containing one material dispersed in a matrix of the same or different material having a macromolecular matrix
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
  • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
  • A61L27/40—Composite materials, i.e. containing one material dispersed in a matrix of the same or different material
  • A61L27/44—Composite materials, i.e. containing one material dispersed in a matrix of the same or different material having a macromolecular matrix
  • A61L27/48—Composite materials, i.e. containing one material dispersed in a matrix of the same or different material having a macromolecular matrix with macromolecular fillers
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
  • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
  • A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
  • A61L27/52—Hydrogels or hydrocolloids
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
  • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
  • A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
  • A61L27/54—Biologically active materials, e.g. therapeutic substances
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
  • A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
  • A61L2300/60—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a special physical form
  • A61L2300/62—Encapsulated active agents, e.g. emulsified droplets
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
  • A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
  • A61L2300/60—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a special physical form
  • A61L2300/64—Animal cells
• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
  • C12N2513/00—3D culture
• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
  • C12N2537/00—Supports and/or coatings for cell culture characterised by physical or chemical treatment
  • C12N2537/10—Cross-linking
• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
  • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
  • C12N5/06—Animal cells or tissues; Human cells or tissues
  • C12N5/0602—Vertebrate cells
  • C12N5/0676—Pancreatic cells
  • C12N5/0677—Three-dimensional culture, tissue culture or organ culture; Encapsulated cells

Definitions

• the present invention relates to a novel implantation device and the like.
• a cell-containing device for example, contains living cells, biological tissues, etc., and supplies physiologically active substances such as hormones and proteins related to metabolic functions to patients instead of organs of diseased humans and animals, or may be harmful. It can be described as a device for preventing and/or treating disease in a patient by detoxifying substances.
• Such devices include (devices composed of) cells and tissues themselves, as well as devices with an immunoisolation layer (immunoisolation function) containing (embedded) cells and tissues (cells and tissues are device that can be implanted via a device) and the like.
• devices with such an immunoisolation layer can protect living cells and living tissues from the body's defense mechanisms, so compared to direct cell and tissue transplantation (e.g., living organ transplantation).
• immunosuppressants there is no need to administer immunosuppressants, so there is no need to worry about side effects due to immunosuppressants, and the treatment is minimally invasive, so it has the ability to solve the shortage of donors.
• Devices with an immunoisolation function come in various shapes, but one example is a microcapsule-type or macrocapsule-type preparation (e.g., cell preparation) in which living cells or living tissues are wrapped in a polymer. things are mentioned. These protect the cells and tissues contained therein from the body's defense mechanism by the strong crosslinked structure possessed by the high molecular weight polymer. It is characterized by supplying hormones and the like secreted from the body to the living body.
• a microcapsule-type or macrocapsule-type preparation e.g., cell preparation
• These protect the cells and tissues contained therein from the body's defense mechanism by the strong crosslinked structure possessed by the high molecular weight polymer. It is characterized by supplying hormones and the like secreted from the body to the living body.
• Patent Document 1 a cell or tissue capsule having an aqueous gel containing a modified polyvinyl alcohol resin (A) having an active carbonyl group and a cross-linking agent (B) as components as an immunoisolation layer is disclosed.
• An implanted device is disclosed.
• Patent Document 2 discloses a cell or tissue-embedded device having, as an immunoisolation layer, an aqueous gel containing a polyvinyl alcohol resin (A) having a syndiotacticity of 32 to 40% in terms of triad. ing.
• Patent Document 3 discloses a method using a gelatin hydrogel containing basic fibroblast growth factor.
• An object of the present invention is to provide a novel device or the like.
• the contact area between the cell-containing device and the newly formed blood vessels is small, so oxygen and nutrients are efficiently supplied to living cells, biological tissues, and the like. may not be supplied.
• angiogenesis when angiogenesis is performed using cell growth factors, there may be a problem that blood and exudates accumulate at the site of angiogenesis due to bleeding and inflammation. These blood and exudates form a liquid film between new blood vessels and cells or cell-containing devices, inhibiting the diffusion of oxygen and nutrients from the new blood vessels to the cell-containing devices. can be significantly reduced.
• Patent Document 3 based on a completely different idea from exogenous administration of growth factors to induce angiogenesis, as a result of intensive studies, they found that specific physical properties Implantation (dwelling) of a specific device (material, member) containing a material having a value can activate the implantation site [e.g. It is possible to suppress or prevent the problem and efficiently express the function of the cell-containing device], etc., and after repeated studies, the present invention was completed.
• An implant device comprising a material (A) having a density of 12 mg/cm 3 or more and an elution rate of 80% by mass or less after being immersed in physiological saline at 37° C. for 24 hours.
• A material having a density of 12 mg/cm 3 or more and an elution rate of 80% by mass or less after being immersed in physiological saline at 37° C. for 24 hours.
• Y fold, i.e., double the mass
• the ratio (X/Y) value of 0.22 or more and an elution rate of 80% or less after being immersed in physiological saline at 37°C for 24 hours.
• the ratio of these ( X /Y) value is 0.22 or more, and the elution rate after immersion in physiological saline at 37° C. for 24 hours is 80% by mass or less.
• the degree of swelling after being immersed in physiological saline at 37° C. for 60 minutes is 70 times (ie, 70 times by mass) or less in terms of mass.
• the device has a density of 14 mg/cm 3 or more, a density of X (mg/cm 3 ), and a degree of swelling of Y (fold, that is, weight-fold) after being immersed in physiological saline at 37° C. for 60 minutes.
• Material (A) [in particular, material (A) containing bioabsorbable material (A1)] and non-bioabsorbable material (member) (B), wherein the ratio of non-bioabsorbable material (B) is The device according to any one of [1] to [12], which is 1 part by volume or more per 100 parts by volume of material (A).
• [16] The device according to any one of [1] to [15], which is (substantially) free of growth factors.
• [17] A device according to any one of [1] to [16] for forming a film.
• the cell- or tissue-containing device is a modified polyvinyl alcohol-based resin (A1) having an active carbonyl group, a polyvinyl alcohol-based resin (A2) having a syndiotacticity of 32 to 40% in terms of triad, and a saponification degree of 97 mol%.
• pancreatic islets pancreatic islet cells
• hepatocytes stem cells thereof
• progenitor cells thereof a method of implanting (implanting, indwelling for a predetermined period of time) the device according to any one of [1] to [22].
• a method for preventing and/or treating a disease or condition comprising transplanting a cell- or tissue-containing device into the transplantation site (removal site) of the device according to any one of [1] to [22].
• [31] [29] comprising the step of implanting the device according to any one of [1] to [22] [further the step of removing at least the non-bioabsorbable material (member) (further forming an implantation spot)] Or the method described in [30].
• the present invention can provide a novel device.
• the implantation (indwelling) site can be activated.
• transplantation (indwelling) prior to transplantation of a cell-containing device can suppress or prevent the above-described problems, and efficiently exert the function of the cell-containing device.
• a coating can be formed on the implantation (indwelling) site (or its vicinity).
• the device of the present invention can activate the implantation site efficiently in this way, by setting specific physical properties (e.g., density, elution rate) in the device to a predetermined range (value), for example
• specific physical properties e.g., density, elution rate
• the cell-containing device can activate the transplanted site, such as by forming a strong membrane with an appropriate thickness at the transplanted site.
• the bioabsorbable material (A) is used as a part of the coating formed. , it is assumed that a film having a sufficient thickness is likely to be formed.
• effector cells that secrete growth factors are easily drawn in, and the effector cells that are drawn in tend to remain in the material (A), the effect of endogenously increasing (promoting secretion) of extracellular matrix (ECM) and growth factors can be obtained. considered to be easy.
• ECM extracellular matrix
• the device by constructing the device from a non-bioabsorbable material, it is also assumed that a foreign body reaction may be induced and the film formation and the secretion of growth factors may be promoted.
• such a device can be suitably used as an implantation device (device for implantation) and can also be used as a device for forming a capsule.
• the use of the device of the present invention is not particularly limited, and depending on the type of device, the degree and mode of activation [whether or not encapsulation is involved, etc.], the desired use, for example, wound repair (for example, after surgery tissue repair, etc.), scaffolds for injected cells, etc.
• the device of the present invention can be suitably used as a device for transplantation (implantation and indwelling for a predetermined period of time) (prior to transplantation of the cell-containing device) at the transplantation site of the cell-containing device.
• the function of the cell-containing device can be efficiently expressed (improved).
• the reproducibility (accuracy) of such function expression is high.
• the transplanted site can be activated by forming a strong capsule with an appropriate thickness at the transplanted site (furthermore, the cell-containing device can be easily transplanted to function). It is presumed that there is a function to prepare the environment). Further, if a coating can be formed by the device of the present invention, the coating improves the adhesion between the transplanted site and the cell-containing device. It is considered that the function of can be effectively expressed (improved). The coating is also presumed to play a role in protecting the implantation site.
• the coating easily prevents the transplanted cell-containing device from being displaced from the transplanted site, and facilitates the efficient functioning of the cell-containing device at the target site.
• the implantation site can be activated without using growth factors or the like. Therefore, it is completely different from exogenous, so to speak, activating the implantation site from the outside using a device containing a growth factor, as in Patent Document 3 above.
• exogenous administration In exogenous administration, it is difficult to control such as sustained release or local administration, and it spreads throughout the body from the transplant site, requiring large doses and causing unexpected side effects (e.g., bleeding, swelling, tumorigenesis, etc.). ). In addition, in exogenous administration, it is difficult to target and locally increase (exist) a growth factor or the like at the transplant site. Since the device of the present invention does not require such exogenous administration, it can efficiently activate the transplanted site (furthermore, the cell-containing device can be used) while suppressing or preventing such problems. When implanted, the function of the cell-containing device can be improved).
• the device of the present invention is useful because it can be implanted (and activated) even in a site, such as subcutaneous tissue, where there are few blood vessels and where cell-containing devices and the like are thought to be difficult to function.
• the site to which the cell-containing device is implanted can be activated, and central necrosis can be efficiently suppressed or prevented regardless of the degree of blood vessel formation (new formation), thereby significantly improving the function of the cell-containing device. sell.
• Such a device of the present invention does not require the administration of an exogenous growth factor or the like (and angiogenesis accompanying the administration) as described above, and prevents retention of blood and exudates due to bleeding or inflammation at the transplant site. can be effectively prevented. Therefore, according to such a device, functions such as a cell-containing device can be fully exhibited.
• the device of the present invention may also induce neovascularization at the site of implantation, the extent of such neovascularization, if at all, is less (or less extreme) than that associated with exogenous administration. In addition to this, it is due to endogenous factors, and bleeding and the like can usually be suppressed or prevented at a high level.
• Devices in other aspects of the invention include non-bioabsorbable materials. With such a device, even after implantation (dwelling), at least the non-bioabsorbable material can remain without being absorbed. ) can be efficiently formed. Such implantation pockets facilitate implantation (positioning) of cell-containing devices and the like.
• devices containing such non-bioabsorbable materials can usually be removed efficiently.
• such a device usually adheres less to the site of implantation (as well as to the formed capsule) and surrounding sites, and less damages to such sites upon removal. Therefore, it is possible to prevent or suppress bleeding, inflammation (occurrence of inflammation), and exudate at the transplant site or the like.
• devices containing non-bioabsorbable materials are extremely useful because they are both activatable and easy to remove.
• the device of the present invention (sometimes referred to as device 1, material, member, etc.) includes at least material (A).
• the material (member) (A) usually has specific physical property values.
• the density of the material (A) may be selected from a range of about 1 mg/cm 3 or higher (eg, 5 mg/cm 3 or higher), preferably 10 mg/cm 3 or higher (eg, 12 mg/cm 3 or higher). may be about 14 mg/cm 3 or more (e.g., 15 mg/cm 3 or more), 20 mg/cm 3 or more (e.g., 25 mg/cm 3 or more, 30 mg/cm 3 or more, 35 mg/cm 3 or more, 40 mg/cm 3 or more). cm 3 or more, 45 mg/cm 3 or more, 50 mg/cm 3 or more, 55 mg/cm 3 or more).
• the upper limit of the density of material (A) is not particularly limited, but is, for example, 5000 mg/cm 3 , 3000 mg/cm 3 , 2000 mg/cm 3 , 1500 mg/cm 3 , 1200 mg/cm 3 , 1000 mg/cm 3 , 800 mg/cm 3 .
• cm3 700 mg/ cm3 , 600 mg/ cm3 , 500 mg/ cm3 , 400 mg/ cm3 , 350 mg/ cm3 , 300 mg/ cm3 , 250 mg/ cm3 , 200 mg/ cm3 , 180 mg/ cm3 , 150 mg/cm3 cm 3 , 120 mg/cm 3 , 100 mg/cm 3 , 90 mg/cm 3 , 80 mg/cm 3 , 70 mg/cm 3 , 60 mg/cm 3 and the like.
• the density is not too large, particularly a value that is relatively small (e.g., 500 mg/cm 3 or less, 300 mg/cm 3 or less, 250 mg/cm 3 or less, 200 mg/cm 3 or less, cm 3 or less, 150 mg/cm 3 or less, 100 mg/cm 3 or less, 1-500 mg/cm 3 , 5-300 mg/cm 3 , 10-200 mg/cm 3 , 12-150 mg/cm 3 , 14-100 mg/cm 3 etc.).
• a value that is relatively small e.g., 500 mg/cm 3 or less, 300 mg/cm 3 or less, 250 mg/cm 3 or less, 200 mg/cm 3 or less, cm 3 or less, 150 mg/cm 3 or less, 100 mg/cm 3 or less, 1-500 mg/cm 3 , 5-300 mg/cm 3 , 10-200 mg/cm 3 , 12-150 mg/cm 3 , 14-100 mg/cm 3 etc.
• a specific density range can be set by appropriately combining the above upper limit and lower limit (for example, 12 to 2000 mg/cm 3 , 15 to 1000 mg/cm 3 , hereinafter the same in the description of the range). .
• Density can be determined (measured) based on volume and mass (weight).
• they may be determined by an indirect method.
• the density of a water-absorbing (water-absorbing) material is determined by measuring the water content (moisture content) by a conventional method, correcting (subtracting) the measured water content, and determining the density. good too.
• Having the above density facilitates activation of the transplant site.
• a coating when a coating is formed, it is easy to prevent the coating (e.g., fibroblasts, etc.) from entering the device (e.g., holes, voids, etc.). It is easy to obtain a film efficiently.
• the material (A) when the material (A) is composed of the bioabsorbable material (A1), if the density is as described above, the bioabsorbable material (A1) is used as part of the coating. Therefore, it is easy to form a film with sufficient thickness or a strong film.
• effector cells that secrete growth factors are easily drawn in, and the effector cells that are drawn in tend to remain in the material (A), the effect of endogenous increase (secretion promotion) of the extracellular matrix (ECM) and growth factors can be obtained.
• the material (A) is composed of a non-bioabsorbable material, with the density as described above, adhesion with surrounding tissue is less likely to occur, making it easier to remove.
• the material (A) has an elution rate (dissolution rate, residual rate) after immersion in physiological saline at 37° C. for 24 hours of 90% by mass or less (e.g., 80% by mass or less), preferably 70% by mass or less.
• the lower limit of the dissolution rate can be selected according to the type of material (A), and is, for example, 0% (substantially insoluble), 0.5% by mass, 1% by mass, 2% by mass, 3% by mass. %, 5% by mass, 8% by mass, 10% by mass, 12% by mass, 14% by mass, 15% by mass, 16% by mass, 17% by mass, 18% by mass, or the like.
• the elution rate should be a value that is not too small, particularly a value that is relatively large (e.g., 14% by mass or more, 15% by mass or more, 16% by mass or more, 17% by mass above, 18% by mass or more, 14 to 70% by mass, 15 to 50% by mass, 16 to 40% by mass, etc.).
• the lower limit of the elution rate is often a finite value, and when composed of a non-bioabsorbable material, The elution rate is often 0% by mass or a low value (eg, 5% by mass or less, 3% by mass or less, 1% by mass or less, etc.).
• the material (A) is composed of the bioabsorbable material (A1)
• the elution rate is as described above, the rate of absorption (decomposition) into the body is moderated, and a thick coating with a sufficient thickness is easy to form.
• effector cells are likely to be retained for a long time, the effect of endogenous increase (secretion promotion) of extracellular matrix (ECM) and growth factors is likely to be obtained.
• the material (A) when the material (A) is composed of a non-bioabsorbable material, the elution rate as described above makes it easy to suppress shape change during indwelling, making it easy to form a coating of the desired shape. In addition, it is easy to prevent a part of the non-bioabsorbable material from remaining in the body and being unable to be removed when necessary. In addition, it is preferable that the material (A) is satisfied by combining both such an elution rate and the density as described above. Such combination and sufficiency facilitates efficient activation of the transplanted site.
• the degree of swelling of material (A) after immersion in physiological saline at 37° C. for 60 minutes can be selected according to the type of material (A), and is, for example, 200 times or less (e.g., 150 times or less in terms of mass). , 120 times or less, 100 times or less, 80 times or less), 70 times or less (e.g., 65 times or less), preferably 60 times or less (e.g., 55 times or less), more preferably 50 times or less may be about 45 times or less (e.g., 40 times or less, 35 times or less, 32 times or less, 30 times or less, 28 times or less, 25 times or less, 22 times or less, 20 times or less, 18 times or less, 15 times or less, 12 times or less, 10 times or less).
• 200 times or less e.g., 150 times or less in terms of mass
• 120 times or less 100 times or less, 80 times or less
• 70 times or less e.g., 65 times or less
• 60 times or less e.g.,
• the lower limit of the swelling degree is, in terms of mass, for example, 1-fold (substantially no swelling), 1.5-fold, 2-fold, 3-fold, 5-fold, 6-fold, 7-fold, 8-fold, 8.5-fold. It may be double, nine times, or the like.
• the degree of swelling should be a value that is not too small, particularly a value that is somewhat large (for example, 6 times or more, 7 times or more, 8 times or more, 8.5 times or more, 9 times or more, 6 to 200 times, 7 to 100 times, 8 to 60 times, etc.).
• the lower limit of the degree of swelling is often a finite value, and when composed of a non-bioabsorbable material, In many cases, the degree of swelling is one-fold or a lower value.
• the density is X (mg/cm 3 ) and the degree of swelling (in terms of mass) after immersion in physiological saline at 37° C. for 60 minutes is Y (fold, i.e., mass-fold)
• Y fold, i.e., mass-fold
• the value of the ratio (X/Y) is, for example, 0.01 or more (e.g., 0.02 or more, 0.03 or more, 0.05 or more, 0.08 or more, 0.1 or more, 0.12 or more, 0 0.15 or more, 0.18 or more, 0.2 or more), and 0.21 or more (for example, 0.22 or more, 0.23 or more, 0.24 or more, 0.25 or more, 0.21 or more).
• 0.4 or more e.g., 0.5 or more, 0.6 or more, 0.7 or more, 0.8 or more, 0.9 or more
• more preferably 1 or more may be 2 or more (e.g., 2.5 or more, 3 or more, 3.5 or more, 4 or more, 4.5 or more, 5 or more, 5.5 or more, 6 or more, etc.) ).
• the upper limit of the X/Y value is, for example, 3000, 2500, 2000, 1500, 1200, 1000, 500, 400, 300, 200, 150, 120, 100, 90, although it depends on the material of material (A). , 80, 70, 60, 50, 40, 35, 30, 20, 18, 15, 12, 10, etc.
• the X/Y value should be a value that is not too large, particularly a value that is somewhat small (for example, 150 or less, 100 or less, 80 or less, 50 or less, 30 or less, 20 0.1 to 150, 0.2 to 120, 0.21 to 100, 0.25 to 50, etc.).
• the material (A) is not limited, but may be a polymer (polymer, resin).
• the material (A) may be a bioabsorbable material (bioabsorbable material) or a non-bioabsorbable material (non-bioabsorbable material).
• bioabsorbable materials include proteins (e.g., gelatin, collagen, collagen peptide), polysaccharides or derivatives thereof [e.g., mucopolysaccharides (e.g., hyaluronic acid, heparin), alginic acid, chitin , chitosan, starch, dextran, etc.], aliphatic polyesters (eg, polyglycolic acid, polylactic acid, glycolic acid/lactic acid copolymer, poly ⁇ -hydroxybutyrate), and the like.
• proteins e.g., gelatin, collagen, collagen peptide
• polysaccharides or derivatives thereof e.g., mucopolysaccharides (e.g., hyaluronic acid, heparin), alginic acid, chitin , chitosan, starch, dextran, etc.
• aliphatic polyesters eg, polyglycolic acid, polylactic acid, glycolic acid
• bioabsorbable material only needs to be bioabsorbable, and it does not matter whether the material is actually bioabsorbed when the device is used.
• non-bioabsorbable materials include the materials described later (silicone-based resins, polyvinyl alcohol-based resins, etc.).
• Material (A) may preferably comprise a bioabsorbable material (A1).
• a bioabsorbable material (A1) When the bioabsorbable material (A1) is included, when a coating is formed, the bioabsorbable material (A) is used as part of the coating, so a coating having a sufficient thickness is formed.
• Cheap In addition, since effector cells that secrete growth factors are easily drawn in, and the effector cells that are drawn in tend to remain in the material (A), the effect of endogenously increasing (promoting secretion) of extracellular matrix (ECM) and growth factors can be obtained. Cheap.
• the proportion of the bioabsorbable material (A1) is 10% by mass or more (e.g., 20% by mass or more), preferably 30% by mass or more (e.g., , 40% by mass or more), more preferably 50% by mass or more (e.g., 60% by mass or more), and 70% by mass or more (e.g., 80% by mass or more, 90% by mass or more, 95% by mass or more) or 100% by mass (material (A) may be composed substantially of bioabsorbable material (A1) only).
• material (A) may contain at least gelatin.
• Gelatin when gelatin is included as the bioabsorbable material (A1)) will be described in detail below.
• the gelatin is not particularly limited, and may be derived from any animal such as fish (fish), cattle, or pigs, and may be recombinant [recombinant gelatin (human recombinant gelatin, etc.)].
• the manufacturing method of gelatin is not particularly limited, and it may be, for example, alkali-processed gelatin, acid-processed gelatin, or the like.
• the isoelectric point, molecular weight, molecular weight distribution, viscosity, jelly strength, etc. of gelatin are not particularly limited and can be selected as appropriate.
• Gelatin is modified or derivatized ⁇ e.g., introduction of hydrophobic groups [e.g., hydrocarbon groups (e.g., alkyl groups), etc.] [e.g., acylation of amino groups constituting gelatin (e.g., hexanoylation, dodecanoylation alkanoylation, etc.)] may or may not be performed.
• hydrophobic groups e.g., hydrocarbon groups (e.g., alkyl groups), etc.
• acylation of amino groups constituting gelatin e.g., hexanoylation, dodecanoylation alkanoylation, etc.
• the site of modification or derivatization is not particularly limited, but may be a molecular terminal, a side chain, or the like. Further, even when modification or derivatization is performed, it is often performed before and after the modification or derivatization to the extent that the bioabsorbability is not impaired.
• gelatin is not modified (chemically modified) or derivatized (for example, introduction of a hydrophobic group), or modified (chemically modified) or derivative (for example, the introduction of a hydrophobic group, etc.), but the extent is small [for example, the amino group constituting gelatin is 20 mol% or less (e.g. modified or derivatized gelatin etc.] may be suitably used.
• the material (A) containing gelatin may generally be biocompatible.
• Material (A) containing gelatin may contain other bioabsorbable materials as necessary. Such other materials may also generally be biocompatible.
• materials other than gelatin such as proteins or peptides (e.g., collagen, collagen peptide), polysaccharides or derivatives thereof [e.g., mucopolysaccharides ( hyaluronic acid, heparin), alginic acid, chitin, chitosan, starch, dextran, etc.], aliphatic polyesters (e.g., polyglycolic acid, polylactic acid, glycolic acid/lactic acid copolymer, poly ⁇ -hydroxybutyrate), etc. mentioned.
• proteins or peptides e.g., collagen, collagen peptide
• polysaccharides or derivatives thereof e.g., mucopolysaccharides ( hyaluronic acid, heparin), alginic acid, chitin, chitosan, starch, dextran, etc.
• aliphatic polyesters e.g., polyglycolic acid, polylactic acid, glycolic acid/lactic acid cop
• the proportion of gelatin in the bioabsorbable materials is, for example, 30 mass, although it depends on the type of other materials. % or more, 50 mass % or more, 70 mass % or more, 80 mass % or more, 90 mass % or more, 95 mass % or more, and the like.
• the bioabsorbable material (A1) [especially gelatin, the bioabsorbable material (A1) containing gelatin, at least the gelatin contained in the bioabsorbable material (A1)] is preferably crosslinked.
• the material (A) (bioabsorbable material) is easily bioabsorbed over a predetermined period of time (for example, 1 week to 3 months). It can be advantageous in terms of [in particular, the formation of a coating (uniform coating), etc.].
• the cross-linking method is not particularly limited, and cross-linking using a chemical reagent [for example, cross-linking using a cross-linking agent such as aldehyde (e.g., formaldehyde, glutaraldehyde, etc.) or carbodiimide], cross-linking by heat (heat dehydration cross-linking, heat cross-linking, etc. ), enzyme cross-linking, cross-linking by energy rays (photo-crosslinking, ultraviolet rays, electron beams, radiation, etc.), physical cross-linking (eg, hydrogen bonding, hydrophobic interaction, ionic interaction, etc.), and the like can be used.
• a chemical reagent for example, cross-linking using a cross-linking agent such as aldehyde (e.g., formaldehyde, glutaraldehyde, etc.) or carbodiimide
• cross-linking by heat heat
• heat cross-linking heat
• heat heat
• the material (A) may contain other components.
• Other components include, but are not particularly limited to, cell culture components (eg, alkali metals, alkaline earth metals, halogens, glucose, physiologically active substances, etc.) and the like, which will be described later.
• the other component may be an angiogenic component (a component that contributes to angiogenesis).
• Such angiogenic components may be components that contribute to (promote) angiogenesis.
• factor VEGF
• FGF fibroblast growth factor
• HGF hepatocyte growth factor
• EGF epidermal growth factor
• IGF insulin-like growth factor
• PDGF platelet-derived cell growth factor
• PlGF placental growth factor
• insulin etc.
• cells eg, cells secreting a growth factor (cell growth factor)
• VEGF vascular endothelial growth factor
• FGF fibroblast growth factor
• HGF hepatocyte growth factor
• EGF epidermal growth factor
• IGF insulin-like growth factor
• PDGF platelet-derived cell growth factor
• PlGF placental growth factor
• insulin eg, cells secreting a growth factor (cell growth factor)), and the like.
• stem cells examples include mesenchymal stem cells (MSC) [e.g., adipose-derived stem cells (ADSC), bone marrow-derived stem cells (BMSC), placenta-derived stem cells, umbilical cord-derived stem cells, etc.], etc.), and the like.
• MSC mesenchymal stem cells
• ADSC adipose-derived stem cells
• BMSC bone marrow-derived stem cells
• placenta-derived stem cells e.g., umbilical cord-derived stem cells, etc.
• Material (A) (or device 1, bioabsorbable material (A1)) may be substantially free of such other components (e.g., angiogenic components), and, inter alia, substantially free of growth factors. may not be included in
• the device of the present invention can activate the implantation site as described above. Therefore, the implantation site can be activated without growth factor-induced angiogenesis (or in a manner different from growth factor-induced angiogenesis).
• ADSCs Unlike the use of cells (ADSCs), the use of growth factors (exogenous administration) causes problems such as bleeding, inflammation, and retention of blood and exudate associated with angiogenesis (excessive or extreme angiogenesis).
• angiogenesis excessive or extreme angiogenesis
• the material (A) may be gel (gel-like) or may have a porous structure (porosity).
• the material (A) may generally have an appropriate shape (or be molded into an appropriate shape).
• the form (shape) of the material (member) (A) is not particularly limited, but may be selected according to the desired physical properties (density, etc.). ) shape, columnar shape (for example, columnar shape, prismatic shape), rod shape, tube shape, and the like.
• the size (size) of the material (A) can be appropriately selected according to the transplant site, its size, etc., and is not particularly limited. 0.05 mm or more (for example, 0.1 to 50 mm), more preferably 0.15 mm or more (for example, 0.2 to 10 mm), and 0.3 mm or more (for example, 0.5 to 5 mm, 0.3 mm or more). 7-2 mm, 0.5-2 mm, 0.7-1.5 mm, etc.).
• the device only needs to contain the material (A), may be composed of the material (A) alone, or may contain other materials in addition to the material (A).
• the device includes at least a non-bioabsorbable material (member) (hereinafter sometimes referred to as non-bioabsorbable material (B), material (B), etc.) among such other materials (members). good too.
• a non-bioabsorbable material member
• the material (A) is composed of a non-bioabsorbable material, adhesion with surrounding tissue is unlikely to occur, and effects such as facilitating removal and the like can be obtained.
• the device may not contain other materials (non-bioabsorbable materials) (or contain non-bioabsorbable materials as the material (A)). many.
• a device is configured (formed) by combining a bioabsorbable material (A1) (or a material (A) containing a bioabsorbable material (A1)) and a non-bioabsorbable material (B), thereby implanting (indwelling) ), even if the bioabsorbable material (A1) is absorbed (dissolved), the non-bioabsorbable material can remain.
• a non-bioabsorbable material may function as a spacer (spacer-wise).
• Such residual non-bioabsorbable material may be the device [at least the non-bioabsorbable material, or, if (part or all of) the bioabsorbable material, the non-bioabsorbable portion of the device] (eg, non-bioabsorbable material)], a pocket (void) is formed at the implantation site.
• Such a pocket is likely to serve as a marker for the site of implantation (site where a capsule is formed or the like is activated), and can also serve as a pocket (implantation pocket) when a cell-containing device or the like described later is implanted. It is particularly useful when performing further transplants.
• non-bioabsorbable materials non-bioabsorbable materials
• the non-bioabsorbable material (B) usually contains a non-bioabsorbable polymer (non-bioabsorbable polymer) [is composed of (formed from) a non-bioabsorbable polymer].
• Non-bioabsorbable polymers include, for example, silicone-based resins, polyvinyl alcohol-based resins, polyvinyl acetal-based resins, polyurethane-based resins, and fluorine resins (e.g., polytetrafluoroethylene, perfluoroalkoxyalkane, etc.).
• olefin-based resins e.g., polyethylene-based resins, polypropylene-based resins
• polyacrylamide-based resins polyester-based resins (e.g., polyethylene terephthalate, etc.), polyacrylonitrile-based resins, polystyrene-butadiene copolymer resins, polysulfone-based resins, cellulose -based resin [eg, cellulose ether (eg, carboxymethylcellulose, hydroxypropylcellulose, etc.)], polyoxyalkylene-based resin (eg, resin containing ethylene oxide or propylene oxide as a polymer component, etc.), and the like.
• cellulose -based resin eg, cellulose ether (eg, carboxymethylcellulose, hydroxypropylcellulose, etc.)]
• polyoxyalkylene-based resin eg, resin containing ethylene oxide or propylene oxide as a polymer component, etc.
• non-bioabsorbable materials are non-bioabsorbable, they may generally be biocompatible (biocompatible polymers).
• the non-bioabsorbable material may be a hydrophilic polymer (or a water-soluble polymer).
• Such hydrophilic (water-soluble) polymers often become water-insoluble (for example, form a gel) in devices (non-bioabsorbable materials) due to cross-linking, chemical modification, or the like. .
• the non-bioabsorbable material does not easily adhere to the transplanted site (surrounding tissue), that is, has anti-adhesion ability.
• the anti-adhesion ability means, for example, the ability to prevent or suppress adhesion between the device and the surrounding tissue or the formed coating when the device is left in the implantation site.
• silicone-based resins, polyvinyl alcohol-based resins, and the like are preferable.
• a resin described later for example, a modified polyvinyl alcohol-based resin (A1) having an active carbonyl group, with a syndiotacticity of 32 to 40% in triad display
• preferred aspects and the like may be the same as those described later.
• the non-bioabsorbable material is not particularly limited, but the elution rate (dissolution rate, residual rate) after immersion in physiological saline at 37 ° C. for 24 hours is, for example, 5% by mass or less, preferably 3% by mass or less, more preferably 3% by mass or less. may be 1% by mass or less, or may be 0% by mass (substantially insoluble).
• the degree of swelling after immersion in physiological saline at 37 ° C. for 60 minutes in terms of mass, may be, for example, 3 times or less, 2 times or less, 1.5 times or less, or 1 time. (substantially does not swell) or the like.
• the volume change rate when the non-bioabsorbable material is immersed in physiological saline at 37° C. for 60 minutes is not particularly limited, but is, for example, 300% or less (e.g., 200% or less, 150% or less, 120% or less).
• the non-bioabsorbable material may generally have a suitable shape (or be molded into a suitable shape).
• the form (shape) of the non-bioabsorbable material (member) is not particularly limited, and examples thereof include particulate (granular), film (sheet), columnar (e.g., columnar, prismatic), rod, and tube. , etc.
• the non-bioabsorbable material may be fibrous, woven (non-woven fabric, etc.), or [forming a woven fabric (non-woven fabric, etc.)].
• the non-bioabsorbable material may be gel-like, rubber-like, or the like, and may have a porous structure (sponge) or network structure.
• the polyvinyl alcohol-based resin used as the non-bioabsorbable material may be in the form of gel or sponge.
• preferred aspects are the same as those described later.
• the size (size) of the non-bioabsorbable material can be appropriately selected according to the implantation site, its size, etc., and is not particularly limited.
• the non-bioabsorbable material has a thickness of 0.01 mm or more (eg, 0.03 mm or more), preferably 0.05 mm or more (eg, 0.1 to 50 mm). , more preferably 0.15 mm or more (eg, 0.2 to 10 mm), particularly 0.3 mm or more (eg, 0.5 to 5 mm, 0.7 to 2 mm, etc.).
• the non-bioabsorbable material is not particularly limited, and commercial products may be used depending on the type, form, presence form in the device, etc. of the non-bioabsorbable material (polymer), or a conventional method may be used. It may be manufactured (synthesized).
• a commercially available or synthesized silicone rubber sheet may be used as the silicone resin.
• a polyvinyl alcohol resin as a non-bioabsorbable material
• a commercially available or synthesized polyvinyl alcohol resin is used as a gel [especially aqueous gel (hydrogel)], sponge, etc. by a conventional method.
• the polyvinyl alcohol-based resin may be gelled by a method described later (method using a cross-linking agent, etc.) depending on the type and the like. In such a case, preferred aspects and the like are the same as those described later.
• non-bioabsorbable material regardless of whether it is a commercially available product or a synthetic product, may be appropriately molded (for example, punched into a predetermined size and shape) according to the shape of the implantation site. .
• the non-bioabsorbable material may contain other components.
• Other components are not particularly limited, and include components exemplified in the section of material (A).
• Non-bioabsorbable materials may also be substantially free of such other components (e.g., angiogenic components), among others, for similar reasons without growth factors. It does not have to be substantially contained.
• the device may contain at least the material (A), and may contain other materials (members) as described above.
• the device material (A) and other materials (non-bioabsorbable material (B))] may be substantially free of growth factors.
• the material (A) may generally constitute at least part of the surface of the device (may be exposed on the surface).
• the material (A) accounts for 10% or more (e.g., 20% or more), preferably 30% or more (e.g., 40% or more), more preferably 50% or more (e.g., 40% or more) of the surface (surface area) 60% or more), or 70% or more [eg, 80% or more, 90% or more, 95% or more, (substantially) 100%].
• the positional relationship (form of existence) between the material (A) and the other material (in particular, a non-bioabsorbable material) is also It is preferable that the positional relationship is such that
• the material (A) and the other material may be in contact or separated, but they are usually in contact in many cases.
• non-bioabsorbable material examples include devices in which at least a part of the surface of another material (non-bioabsorbable material) is coated with the material (A) [for example, at least one non-bioabsorbable material such as a sheet device having (laminated) material (A) such as a sheet on its surface (especially both surfaces or upper and lower surfaces)].
• the ratio of other materials is, for example, 1 part by volume or more (eg, 5 to 5000 parts by volume) per 100 parts by volume of material (A), preferably is 10 parts by volume or more (e.g., 15 to 3000 parts by volume), more preferably 20 parts by volume or more (e.g., 25 to 1000 parts by volume), 30 parts by volume or more (e.g., 35 parts by volume or more, 40 volume parts or more, 45 volume parts or more), etc., and 800 volume parts or less (e.g., 500 volume parts or less, 400 volume parts or less, 300 volume parts or less, 200 volume parts or less, 150 volume parts or less, etc.) There may be.
• Such a ratio facilitates well-balanced activation of the transplanted site (formation of a capsule, etc.) and formation of a transplanted pocket.
• material (A) and other materials may generally be fixed (integrated).
• material (A) and another material may be fixed (fixed to each other).
• fixing the material (A) and the non-bioabsorbable material it is easy to efficiently match (correspond, position) the site of activation (film formation, etc.) and the position of the pocket.
• the fixing method is not particularly limited, but examples include suture (suture with suture thread, etc.), adhesion [for example, adhesion (bonding) using a medical adhesive], and the like. Among these, suturing is preferably used.
• the sutures and adhesives may be biocompatible and/or bioabsorbable (degradable).
• the material (A) and other materials may be swollen with water or a predetermined liquid medium depending on the type. In this case, they may be combined after being swollen, or may be combined and then swollen.
• the device (material, member) of the present invention can be implanted (implanted in vivo) and used (used for implantation or for implantation).
• transplantation can activate the implantation site.
• transplantation may result in (1) encapsulation, (2) extracellular matrix increase (secretion enhancement), (3) growth factor increase (secretion enhancement), and the like.
• the device (material, member) of the present invention can be suitably used as a device for implantation (implantation device, implantation material, implantation member), and if it can be activated in this way, it can be used for at least one of these uses. It can also be used as a device for use.
• the device of the present invention particularly, the device containing the bioabsorbable material (A1) as the material (A)
• at least a coating can be formed. It can often be used as a device for film formation.
• the formation of the coating can be visually confirmed from the state before and after transplantation.
• the coating is usually formed at the implantation site of the device (bioabsorbable material) or in the vicinity (periphery) thereof, and may be composed of at least the bioabsorbable material derived from the device (bioabsorbable material) (bioabsorbable material).
• examples of the extracellular matrix include collagen (eg, collagen III, collagen IV, etc.), laminin, and the like.
• the extracellular matrix may contain these alone or in combination of two or more.
• the ECM can be increased (secreted) by the device of the present invention, the ECM can be supplemented, which is more advantageous in terms of activation (which leads to suppression or prevention of functional deterioration of the cell-containing device described later). )it is conceivable that.
• the increase in extracellular matrix (promotion of secretion) can be confirmed, for example, by immunohistochemical analysis.
• the surrounding tissue is harvested, fixed with 4% paraformaldehyde, and then embedded in paraffin.
• immunohistochemical staining for example, anti-collagen III (ab7778; Abcam), anti-collagen IV (ab6586; Abcam) and laminin (ab11575; Abcam) antibodies are preferably used.
• EnVision+System-HRP-labeled polymer rabbit antibody (4003; DAKO) is preferably used as a secondary antibody.
• growth factors include epidermal growth factor (EGF), insulin-like growth factor (IGF) (eg, IGF-1, IGF-2, etc.), fibroblast growth factor (FGF) (e.g., FGF12, etc.), hepatocyte growth factor (HGF), transforming growth factor (TGF) (e.g., TGF- ⁇ 1, etc.), platelet-derived growth factor (PDGF) (e.g., PDGF-A, PDGF- B etc.).
• EGF epidermal growth factor
• IGF insulin-like growth factor
• FGF fibroblast growth factor
• HGF hepatocyte growth factor
• TGF transforming growth factor
• PDGF platelet-derived growth factor
• the growth factor may contain these alone or in combination of two or more.
• IGF-2 may be increased at a high level (promoted secretion).
• the device of the present invention increases growth factors (eg, IGF-2, etc.) (promotes secretion). If it is possible to inhibit apoptosis, it is considered that it will be more advantageous in terms of activation (which in turn leads to suppression or prevention of functional deterioration of cell-containing devices).
• growth factors eg, IGF-2, etc.
• growth factors may contribute to the induction of angiogenesis to some extent. efficient expression).
• the device contains (4) components other than extracellular matrix and growth factors, such as cell adhesion molecules (N-cadherin, ICAM-1, VCAM-1, etc.), inducers [e.g., epithelial-mesenchymal transition inducer (c -MET), hypoxia-inducible factor 1 ⁇ subunit (HIF-1 ⁇ ), etc.], anti-angiogenic factors (eg, vasohibin 1), CD31, versican, thrombospondin (TSP) 2, etc. .
• inducers e.g., epithelial-mesenchymal transition inducer (c -MET), hypoxia-inducible factor 1 ⁇ subunit (HIF-1 ⁇ ), etc.
• anti-angiogenic factors eg, vasohibin 1
• CD31 e.g., CD31, versican, thrombospondin (TSP) 2, etc.
• the increase (secretion promotion) of growth factors, etc. can be confirmed, for example, by real-time PCR.
• RNA is extracted from the surrounding tissue.
• Relative gene expression is determined using, for example, TaqMan Array 96-well FAST plates (4413257; Applied Biosystems).
• the TaqMan Array contains 46 target genes and 2 endogenous control gene candidates. Samples were run at 50°C for 2 min, 95°C for 20 s, followed by 40 Analyze under cycling amplification conditions. The result is the Expression Suite Software ver. Analyze with 1.3 (Applied Biosystems).
• the comparative CT method can be used to assess growth factor enhancement (secretion enhancement) by comparative quantification (RQ). 18S is used as a housekeeping gene.
• Extracellular matrix and growth factors are usually formed at the implantation site of the device (bioabsorbable material) and in the vicinity (periphery) thereof, and are formed in the capsule (inside the capsule). may be
• the device of the present invention may realize the above (1) to (3) (furthermore, (4)) independently, but in particular, it may be realized by combining a plurality of them.
• a plurality of these factors furthermore, by combining angiogenesis, which will be described later
• various factors can work together to exert an effect, and the transplantation site can be activated more efficiently.
• the device can usually be used for implantation as described above.
• the device of the present invention can be used by being implanted (dwelled) in the body of animals including humans.
• the transplantation (dwelling) site can be activated.
• a device containing a bioabsorbable material (A1) as the material (A) such activation is performed by (1) coating formation, (2) extracellular matrix increase ( (3) an increase in growth factors (secretion enhancement) [further, an increase in components other than extracellular matrix and growth factors (secretion enhancement)].
• the device of the present invention may undergo angiogenesis (generate new blood vessels) through implantation (indwelling).
• angiogenesis gene derived new blood vessels
• implantation indwelling
• activation as described above can be achieved without exogenously administering growth factors, but activation [e.g., endogenous increase in growth factors ( Angiogenesis (mild angiogenesis) may occur along with secretion promotion), and such angiogenesis and capsule formation may be combined to activate the transplantation site.
• the degree of angiogenesis can be confirmed by, for example, immunohistochemical analysis, computed tomography (CT) angiography, and the like.
• vWF anti-von Willebrand factor
• ab7356 MerckMillipore
• EnVision+System-HRP-labeled polymer rabbit antibody 4003; DAKO
• Angiogenesis can be assessed by counting the cells themselves or vWF-positive cells in the interstitial area.
• CT computed tomography
• ExiTron nano 12000 an alkaline earth metal-based nanoparticle contrast agent specially formulated for animal CT, is preferably used.
• the nanoparticle contrast agent is intravenously injected from the tail vein or the like, and angiography is performed using an X-ray CT scanner for experimental animals (Latheta LCT-200; Hitachi).
• Angiogenesis can be assessed by calculating vascular volume for the tissue surrounding the cells or cell-containing device implanted.
• it is preferable to set the CT value of the blood vessel region to 100 or more.
• the device of the present invention may be used, and if necessary, another device [for example, a device containing a non-bioabsorbable material such as a polyvinyl alcohol-based resin (such as a resin described later), or the non-biological It may be implanted together with a device or the like containing growth factor-secreting cells (such as ADSC) in an absorbable material].
• a non-bioabsorbable material such as a polyvinyl alcohol-based resin (such as a resin described later), or the non-biological It may be implanted together with a device or the like containing growth factor-secreting cells (such as ADSC) in an absorbable material.
• ADSC growth factor-secreting cells
• the transplantation site is not particularly limited, and examples thereof include subcutaneous, subfascial, visceral surfaces (liver surface, splenic surface, etc.), intraperitoneal (eg, omentum), intramesenteric, intraperitoneal, and groin areas.
• the transplant site may be muscle tissue, adipose tissue (adipocytes), or the like.
• the implantation site is preferably subcutaneous (subcutaneous tissue).
• the device of the present invention can efficiently activate even a site with few blood vessels (regardless of angiogenesis).
• the method of implanting (dwelling) the device is not particularly limited, and may follow a commonly used or conventionally known method.
• implantable devices may also be known.
• the subject (transplantation subject) of the device of the present invention may be either humans or non-human animals (eg, mammals such as dogs and cats).
• the non-human animal may be a pet animal.
• a preferred subject is a human.
• the duration of placement of the device of the present invention depends on the site of implantation, etc., but for example, it is preferably left in place at the site of implantation for 1 week or more, more preferably 10 days to 3 months, and 2 weeks to 2 months. is particularly preferred.
• the implantation (dwelling) of the device of the present invention usually causes the coating containing the bioabsorbable material (A1) derived from the device (the material (A) containing the bioabsorbable material (A1)) to form. It is formed.
• a bioabsorbable material (A1) is considered to undergo bioabsorption over time after the film is formed, but at least the film containing the bioabsorbable material (A1) (the bioabsorbable material (A1) remains
• the indwelling period (or the period until removal, which will be described later) may be set within a range in which the membrane can be maintained.
• the implanted device may be removed after a predetermined retention period.
• a device containing a non-bioabsorbable material as the device of the present invention e.g., a device containing a non-bioabsorbable member as material (A), a device containing a non-bioabsorbable member (B) (particularly, When using a device containing a material (A) containing a bioabsorbable material (A1) and a non-bioabsorbable material (B)], after implantation, at least the non-bioabsorbable material (usually, only the non-bioabsorbable material) are often removed (removed from the implantation site).
• the present invention also includes such withdrawal methods.
• the bioabsorbable material (A1) When the device (material (A) containing the bioabsorbable material (A1)) is left in the body for a predetermined period of time (for example, 1 week to 3 months), the bioabsorbable material (A1) (one part or all) is absorbed by the body, and in this case, a film is often formed.
• the non-bioabsorbable material is not bioabsorbed and remains in the implanted site as it is, so it needs to be removed when the cell-containing device or the like is implanted later.
• Such removal is usually performed without causing bleeding, inflammation, blood vessels (newly formed blood vessels), breakage of the capsule (formed capsule), etc.
• bleeding, inflammation, and breakage of blood vessels and capsules cause blood and exudate to accumulate at the extraction site.
• non-bioabsorbable materials for example, silicone-based resins, polyvinyl alcohol-based resins, etc.
• indwelling period, etc. ensures that adhesion to the transplant site is unlikely to occur. Combined with this, in many cases, it can be removed efficiently without causing such bleeding, inflammation, or damage to the blood vessel.
• a graft pocket is formed at the graft site after removal.
• Such implant pockets may subsequently be implanted with cell-containing devices or the like, as described below.
• the implantation site of the device of the present invention is activated, another device or the like may be implanted in the implantation site.
• another device or the like may be implanted in the implantation site.
• implanting a device containing cells or tissues will be described in detail.
• the present invention also includes a method of implanting (indwelling) a device containing cells or tissues (a cell- or tissue-containing device, hereinafter sometimes referred to as a cell-containing device, device 2, etc.).
• a cell- or tissue-containing device hereinafter sometimes referred to as a cell-containing device, device 2, etc.
• cell-containing component may be performed in parallel (overlapping) with the implantation (dwelling) of the device (Device 1, gelatin-containing device), or may not be performed in parallel (overlapping). you don't have to)
• the device 1 may be implanted near (peripherally) or adjacent to the cell-containing device.
• the cell-containing device is implanted (removed from the device 1 (at least the non-bioabsorbable material)) at the transplanted (removed) site (implanted site, transplanted pocket) after the device 1 is transplanted [and the device 1 (at least the non-bioabsorbable material) is removed]. detention) may be used.
• the cell-containing device when a coating is formed by the device 1, the cell-containing device is implanted in a state (timing) in which the coating is formed (maintained) [in particular, gelatin is contained (remains) in the coating]. preferably.
• the device of the present invention is a device for use in combination with a cell-containing device, particularly when the cell-containing device is implanted at the implantation site (activated implantation site such as capsule formation) ( (implanted prior to implantation of a cell-containing device).
• the transplanted site (and its vicinity or periphery) can be activated, and the function of the cell-containing device can be efficiently exhibited.
• the function of the cell-containing device can be expressed. Cheap.
• the device 1 containing a non-bioabsorbable material by selecting the type of non-bioabsorbable material, adhesion is unlikely to occur, and the device 1 ( At least the non-bioabsorbable material) can be removed, and by transplanting to such a removed site, the cell-containing device is likely to function more efficiently.
• the cell-containing device can be efficiently transplanted to a transplant site where bleeding, inflammation, and blood vessel damage have not occurred.
• the cell-containing device can be transplanted (indwelled) without such a phenomenon, and the function of the cell-containing device can be efficiently exhibited in combination with the effect of activating the transplanted site as described above.
• Bleeding, inflammation, and rupture of blood vessels at the site of implantation can be visually confirmed, for example, before and after implantation [and removal (removal)] of the device 1 or before implantation of the cell-containing device.
• cell-containing devices include cells and tissues themselves, as well as devices in which cells and tissues are embedded (hereinafter sometimes referred to as cell-embedded devices).
• the cell-embedded device is not particularly limited, and for example, the cell-embedded devices described in Patent Documents 1 and 2 may be used. A specific description will be given below.
• the biological composition (cells or tissues) contained in the cell-containing device is not particularly limited, and can be appropriately selected according to the purpose of use.
• differentiated cells or stem cells derived from ectoderm, mesoderm, or endoderm can be used.
• Differentiated cells include, for example, epidermal cells, smooth muscle cells, osteocytes, bone marrow cells, chondrocytes, skeletal myoblasts, pancreatic real cells, pancreatic islet cells, pancreatic endocrine cells, pancreatic exocrine cells, pancreatic duct cells, liver cells (e.g. , hepatocytes), thyroid cells, parathyroid cells, adrenal cells, pituitary cells, splenocytes, pineal cells, kidney cells (renal cells), spleen cells, anterior lobe cells, growth hormone-secreting cells, dopamine-producing cells, blood Cells, cardiomyocytes, skeletal muscle cells, osteoblasts, nerve cells, pigment cells, adipocytes, etc. can be used.
• the above-mentioned cells may be cells isolated from living organisms as well as cells induced to differentiate from stem cells described later.
• the cells that can be induced to differentiate may be incorporated into a transplant or device, and then differentiated in vivo after transplant. may be used by incorporating it into the
• Stem cells include tissue stem cells (e.g., epidermal stem cells, hair follicle stem cells, pancreatic stem cells/pancreatic progenitor cells, hepatic stem cells, neural stem cells, retinal stem cells, hematopoietic stem cells, etc.), embryonic stem cells (ES cells), iPS cells ( Induced pluripotent stem cells, etc. can be used, but are not limited to these.
• tissue stem cells e.g., epidermal stem cells, hair follicle stem cells, pancreatic stem cells/pancreatic progenitor cells, hepatic stem cells, neural stem cells, retinal stem cells, hematopoietic stem cells, etc.
• ES cells embryonic stem cells
• iPS cells Induced pluripotent stem cells, etc.
• These cells are derived from mammals such as humans, monkeys, pigs, rats, mice, dogs, and cats, and produce and/or secrete physiologically active substances such as hormones and proteins that are useful for living bodies such as patients. , and the selection of cell type can be determined according to the type of disease in a living body such as a patient to be transplanted. Moreover, when these cells are other than human cells, they may be cells transfected with human genes for therapeutic purposes. Examples of hormones useful for living organisms include insulin, thyroid stimulating hormone, thyroid hormone, parathyroid hormone, growth hormone, thyroxine, glucocorticoid, glucagon, estradiol, testosterone, and the like.
• proteins useful for living organisms include blood coagulation factors, complement, albumin, globulin, and various enzymes (metabolic enzymes or digestive enzymes such as amylase, protease, or lipase).
• enzymes metabolic enzymes or digestive enzymes such as amylase, protease, or lipase
• neurotransmitters such as dopamine.
• pancreatic cells islet cells
• hepatocytes dopamine-producing cells
• pancreatic cells pancreatic cells
• pancreatic progenitor (stem) cells are more preferred.
• pancreatic islets pancreatic cells
• hepatocytes their stem/progenitor cells, and the like may be preferably used.
• the biological composition used in the cell-containing device may be either laboratory-established cells or biological tissue, cells isolated from biological tissue, or the like, but should be differentiated non-dividing cells. preferable.
• the separation method is not particularly limited, and a conventionally known method may be followed.
• pathogenic bacteria such as pathogenic viruses are removed from the cells separated from the living tissue.
• the content of the biological composition can be changed as appropriate according to the type of biological composition.
• the dosage is determined by the doctor in consideration of the patient's age, gender, symptoms, side effects, etc., so it is not possible to generalize, but usually about 1 to 10 devices per adult can be implanted in the body.
• IEQ international unit of pancreatic islet volume: the volume of an islet with a diameter of 150 ⁇ m is defined as 1 IEQ
• 5000 to 400000 IEQ the volume of an islet with a diameter of 150 ⁇ m is defined as 1 IEQ
• you can implant a device that
• a cell-embedded device forms a device in which a biological composition is embedded, as described above.
• the shape of such a cell-embedded device is not particularly limited, but it may be of the same system (especially the same shape) as the device 1 (especially non-bioabsorbable material). Examples of such a shape include the shapes exemplified above, including a disk shape, a spherical shape, a columnar shape, an ellipsoidal shape, etc., but a disk shape is preferable.
• the material of the cell-embedded device (the device part of the cell-embedded device) is not particularly limited, and polymers, metals, ceramics, etc. can be used.
• Polymers used in cell-embedded devices are not particularly limited, but examples include structural proteins (e.g., collagen, elastin, keratin, etc.), gelatin, glycosaminoglycans (e.g., hyaluronic acid, chondroitin sulfate, keratan sulfuric acid, dermatan sulfate, heparan sulfate, heparin, etc.), plyteoglycan, cell adhesion molecules (e.g., fibronectin, laminin, fibrinogen, hydronectin, etc.), fibrin, fibroin, sericin, alginic acid, chitosan, agarose, cellulose, cellulose derivatives (e.g., cellulose nanofibers, carboxymethylcellulose, hydroxypropylcellulose, etc.), synthetic polypeptides, polylactic acid, polyglycolic acid, polycaprolactone, polyethylene glycol, polypropylene glycol, 2-methacryl
• the polymer may be a hydrophilic polymer (or water-soluble polymer).
• hydrophilic (water-soluble) polymers often become water-insoluble (for example, form a gel) due to cross-linking or the like in cell-embedded devices.
• the polymer may be used alone or in combination of two or more.
• polyvinyl alcohol-based resins can be particularly preferably used. Therefore, the polymer may contain at least a polyvinyl alcohol-based resin.
• the polyvinyl alcohol-based resin will be described in detail below.
• the polyvinyl alcohol-based resin may generally be a saponified product of a polymer containing at least a vinyl ester-based monomer as a polymerization component.
• a polyvinyl alcohol-based resin may have a vinyl alcohol unit, and a vinyl ester unit (or a unit derived from a vinyl ester-based monomer, such as a vinyl acetate unit, a vinyl pivalate unit, etc., which will be described later).
• unit derived from a vinyl ester and other units (for example, a unit derived from an unsaturated monomer having an active carbonyl group described later, a unit derived from another unsaturated monomer).
• the 4% by mass aqueous solution viscosity (20° C.) of the polyvinyl alcohol resin is not particularly limited, but is, for example, 1 mPa s or more, 2 mPa s or more, 3 mPa s or more, 5 mPa s or more, 20 mPa s or more, 30 mPa s or more.
• s or more 40 mPa s or more, 50 mPa s or more, etc., 800 mPa s or less, 500 mPa s or less, 300 mPa s or less, 200 mPa s or less, 150 mPa s or less, 100 mPa s or less, 80 mPa s or less ⁇ It may be less than or equal to s.
• a polyvinyl alcohol-based resin having a 4% by mass aqueous solution viscosity of about 3 to 300 mPa ⁇ s may be suitably used.
• the viscosity of a 4% by mass aqueous solution of polyvinyl alcohol resin can be measured, for example, according to JIS K6726.
• the polyvinyl alcohol-based resin can be selected according to its type, composition, etc., and is not particularly limited. It may be a resin (for example, less than 97 mol % of saponification degree).
• the degree of saponification (average degree of saponification) can be measured, for example, according to JIS K6726.
• the polyvinyl alcohol-based resin (A) includes a modified polyvinyl alcohol-based resin (A1) having an active carbonyl group, a polyvinyl alcohol-based resin (A2) having a syndiotacticity of 32 to 40% in triad display, and a saponified At least one selected from polyvinyl alcohol-based resins (A3) having a molecular weight of 97 mol% or more may be preferably used.
• Modified polyvinyl alcohol resin having active carbonyl group A modified polyvinyl alcohol resin having an active carbonyl group (herein, also simply referred to as a "modified PVA-based resin") is obtained, for example, by copolymerizing a fatty acid vinyl ester and an unsaturated monomer having an active carbonyl group. obtained by directly contacting a compound having an active carbonyl group such as a liquid diketene or a diketene gas with a copolymer-modified PVA produced by saponifying the obtained copolymer, or a PVA or modified PVA-based resin produced by a known method. Although post-modified PVA can be used, copolymer-modified PVA is preferable from the viewpoint of the stability and safety of the PVA-based resin and workability in the gelation step.
• the fatty acid vinyl ester used in producing the copolymerized modified PVA is not particularly limited, but examples thereof include vinyl formate, vinyl acetate, vinyl propionate, vinyl pivalate and the like. preferred. These can be produced by conventionally known various polymerization methods such as bulk polymerization, solution polymerization, suspension polymerization, and emulsion polymerization. Among them, solution polymerization using an alcohol solvent such as methanol is industrially preferable.
• the unsaturated monomer having an active carbonyl group is not particularly limited, and examples thereof include diacetone acrylamide, diacetone methacrylamide, diacetone acrylate, diacetone methacrylate, acetoacetoxy acrylamide, acetoacetoxy methacrylamide, and the like. mentioned. These may be used alone or in combination of two or more. Among them, diacetone acrylamide is industrially preferred, and as the copolymer-modified PVA, diacetone acrylamide-modified PVA is preferred.
• a fatty acid vinyl ester and an unsaturated monomer having an active carbonyl group are used within a range that does not impair the effects of the present invention.
• Other copolymerizable unsaturated monomers may be used.
• unsaturated monomers include, for example, carboxyl group-containing unsaturated monomers [e.g., (meth)acrylic acid, maleic acid, maleic anhydride, fumaric acid, crotonic acid, itaconic acid, undecylenic acid, etc.], Unsaturated dibasic acid monoalkyl esters (e.g., monomethyl maleate, monomethyl itaconate, etc.), amide group-containing unsaturated monomers (e.g., acrylamide, dimethylacrylamide, dimethylaminoethylacrylamide, diethylacrylamide, dimethylaminopropylacrylamide , isopropylacrylamide, N-methylolacrylamide, N-vinylacetamide, etc.), vinyl halides (e.g., vinyl chloride, vinyl fluoride, etc.), unsaturated monomers having a glycidyl group (e.g., allyl glycidyl ether, glycidyl methacryl
• pentenes e.g., 4,5-dihydroxy-1-pentene, 4,5-diacyloxy-1-pentene, 4,5-dihydroxy-3-methyl-1-pentene, 4,5-diacyloxy-3-methyl - 1-pentene, etc.
• hexenes e.g., 5,6-dihydroxy-1-hexene, 5,6-diacyloxy-1-hexene, etc.
• amine unsaturated monomers e.g., N,N-dimethylallylamine, N-Allylpperazine, 3-piperidine acrylic acid ethyl ester, 2-vinylpyridine, 4-vinylpyridine, 2-methyl-6-vinylpyridine, 5-ethyl-2-vinylpyridine, 5-butenylpyridine, 4-pentenylpyridine, 2 -(4-pyridyl) allyl alcohol, etc.], an unsaturated monomer having a quaternary ammonium compound (
• the content of other unsaturated monomers is not particularly defined, it may be, for example, 10 mol or less per 100 mol of the vinyl ester monomer.
• the obtained copolymerized modified PVA may be subjected to acetalization, urethanization, etherification, grafting, phosphate esterification, acetoacetylation, cationization, etc. using a known method within a range that does not impair the effects of the present invention. It may be post-denatured by reaction.
• the polymerization catalyst used in producing the copolymerized modified PVA is not particularly limited, but an azo compound or a peroxide is usually used. Further, during polymerization, an organic acid such as tartaric acid, citric acid, or acetic acid may be added for the purpose of preventing hydrolysis of the fatty acid vinyl ester.
• a polymerization terminator can be used to terminate the polymerization, although it is not particularly limited.
• the polymerization terminator is not particularly limited, and examples thereof include m-dinitrobenzene and the like.
• the shape of the polymerization vessel when the fatty acid vinyl ester and the unsaturated monomer having an active carbonyl group are copolymerized, the shape of the polymerization vessel, the type of the polymerization stirrer, the polymerization temperature, the pressure in the polymerization vessel, etc. Any known method may be used.
• the method for saponifying the copolymer of the fatty acid vinyl ester and the unsaturated monomer having an active carbonyl group is not particularly limited, and conventionally known methods may be followed.
• An alcoholysis or hydrolysis reaction using a basic catalyst such as potassium hydroxide or sodium methoxide or an acid catalyst such as hydrochloric acid, sulfuric acid or p-toluenesulfonic acid can be applied.
• Solvents used in the saponification reaction include alcohols such as methanol and ethanol; esters such as methyl acetate; ketones such as acetone and methyl ethyl ketone; aromatic hydrocarbons such as benzene and toluene; can be used alone or in combination of two or more.
• the saponification temperature, time, etc. are not particularly limited.
• the method for drying, pulverizing, and washing the saponified product is not particularly limited, and known methods may be used.
• the unsaturated monomer (e.g., diacetoneacrylamide) unit content having an active carbonyl group is , an unsaturated monomer having an active carbonyl group (e.g., diacetone acrylamide)-modified PVA total amount (total amount of monomers), for example, 0.5 to 20 mol%, preferably 0.5 to 15 mol %, more preferably 1 to 12 mol %, still more preferably 2 to 10 mol % (eg, 3 to 8 mol %).
• the unsaturated monomer having an active carbonyl group for example, diacetone acrylamide
• the cross-linking agent there are many reaction sites with the cross-linking agent, and sufficient strength (stress ) can be obtained, and when it is 20 mol % or less, the solubility in water is improved.
• the saponification degree of the modified PVA-based resin is not particularly limited, but is preferably 80 mol% or more (eg, 80 to 99.9 mol%), more preferably 88 mol% or more (eg, 88 to 99.9 mol%). ), more preferably 95 mol % or more (for example, 95 to 99.9 mol %).
• the viscosity of the modified PVA-based resin can be varied, but the viscosity of a 4% by mass aqueous solution of the modified PVA-based resin (20° C.) is preferably 2 to 500 mPa ⁇ s, more preferably 3 to 300 mPa ⁇ s.
• the degree of saponification and the viscosity of the 4% by mass aqueous solution may be values measured according to JIS K-6726.
• polyvinyl alcohol resin whose syndiotacticity is 32 to 40% in terms of triad
• a polyvinyl alcohol resin having a syndiotacticity of 32 to 40% in terms of triad (herein, also simply referred to as "high syndio PVA resin”) can be preferably used.
• the syndiotacticity of the high syndio PVA-based resin is preferably 32 to 40%, more preferably 33 to 39%, and particularly preferably 34 to 38% in terms of triad.
• syndiotacticity is 32% or more, it is likely to form an aqueous gel, and if it is 40% or less, the production of an aqueous gel is facilitated.
• the syndiotacticity indicated by the triad can be obtained by dissolving the high syndio PVA-based resin in heavy DMSO (dimethylsulfoxide) and measuring the peak of the hydroxyl group by proton NMR measurement.
• the method for producing the high syndio PVA resin is not particularly limited as long as the syndiotacticity is 32 to 40% by triad display, but it can be easily obtained by saponifying a vinyl ester polymer obtained by a conventionally known method. be done. That is, the high syndio PVA-based resin is a saponified product of a vinyl ester polymer.
• the method for producing the vinyl ester polymer is not particularly limited as long as it is a method for polymerizing a vinyl ester monomer, and a conventionally known method may be used.
• any known method may be used for the shape of the polymerization vessel, the type of the polymerization stirrer, the polymerization temperature, the pressure inside the polymerization vessel, and the like.
• the polymerization method conventionally known various polymerization methods such as bulk polymerization, solution polymerization, suspension polymerization and emulsion polymerization are possible. Considering the control of the degree of polymerization and the saponification reaction performed after polymerization, solution polymerization using alcohol as a solvent, or suspension polymerization using water or water and alcohol as a dispersion medium is preferable, but is limited to these. is not.
• vinyl ester-based monomers examples include vinyl esters such as fatty acid vinyl esters and non-fatty acid vinyl esters (for example, vinyl formate, aromatic carboxylic acid vinyl esters, etc.), and PVA having high syndiotacticity.
• C 3-15 fatty acid vinyl esters for example, linear or branched C 3-15 fatty acid vinyl esters such as vinyl propionate, vinyl butyrate, and vinyl pivalate, preferably C 3- 10 fatty acid vinyl ester (e.g., linear or branched C 3-10 fatty acid vinyl ester, etc.)]
• C 3-15 fatty acid vinyl ester having a substituent (e.g., halogen group) e.g., trifluorovinyl acetate, vinyl trichloroacetate] vinyl formate and the like.
• These vinyl esters can be used individually by 1 type or in combination of 2 or more types.
• a vinyl ester having a bulky side chain such as vinyl propionate, vinyl butyrate, or vinyl pivalate is homopolymerized or copolymerized, and then saponified with an alkali catalyst.
• a method of homopolymerizing or copolymerizing a highly polar vinyl ester such as vinyl formate, vinyl trifluoroacetate and vinyl trichloroacetate, followed by saponification with an alkali catalyst.
• a method of polymerizing vinyl pivalate and then saponifying it with an alkali catalyst is preferably used.
• a production example in which the syndiotacticity by triad display is 37.1% is described in the following examples. Reducing the syndiotacticity by triad expression from 37.1% means, for example, that when vinyl pivalate is polymerized, a copolymer of vinyl pivalate and vinyl acetate can be obtained in the presence of vinyl acetate, and that the polymerization temperature is reduced. This can be achieved by raising Also, increasing the syndiotacticity by triad display from 37.1% can be achieved, for example, by lowering the polymerization temperature in the above production example. In either case, the obtained high syndio PVA resin can be dissolved in heavy DMSO and determined from the peak of hydroxyl group by proton NMR measurement. Can be used for inventions.
• the vinyl ester polymer may contain other unsaturated monomers copolymerizable with the vinyl ester as long as the effects of the present invention are not impaired.
• unsaturated monomers include, for example, carboxyl group-containing unsaturated monomers [e.g., (meth)acrylic acid, maleic acid, maleic anhydride, fumaric acid, crotonic acid, itaconic acid, undecylenic acid, etc.], Unsaturated dibasic acid monoalkyl esters (e.g., monomethyl maleate, monomethyl itaconate, etc.), amide group-containing unsaturated monomers (e.g., acrylamide, dimethylacrylamide, dimethylaminoethylacrylamide, diethylacrylamide, dimethylaminopropylacrylamide , isopropylacrylamide, N-methylolacrylamide, N-vinylacetamide, etc.), vinyl halides (e.g., vinyl chloride, vinyl fluoride, ethylene glycol glyco
• pentenes e.g., 4,5-dihydroxy-1-pentene, 4,5-diacyloxy-1-pentene, 4,5-dihydroxy-3-methyl-1-pentene, 4,5-diacyloxy-3-methyl - 1-pentene, etc.
• hexenes e.g., 5,6-dihydroxy-1-hexene, 5,6-diacyloxy-1-hexene, etc.
• amine unsaturated monomers e.g., N,N-dimethylallylamine, N-Allylpperazine, 3-piperidine acrylic acid ethyl ester, 2-vinylpyridine, 4-vinylpyridine, 2-methyl-6-vinylpyridine, 5-ethyl-2-vinylpyridine, 5-butenylpyridine, 4-pentenylpyridine, 2 -(4-pyridyl) allyl alcohol, etc.], an unsaturated monomer having a quaternary ammonium compound (
• a polymerization catalyst can be used in the polymerization.
• the polymerization catalyst is not particularly limited, azo compounds and peroxides are usually used.
• an organic acid such as tartaric acid, citric acid, or acetic acid may be added for the purpose of preventing hydrolysis of the fatty acid vinyl ester.
• a polymerization terminator can be used to terminate the polymerization.
• the polymerization terminator is not particularly limited, and examples thereof include m-dinitrobenzene and the like.
• the polymerization temperature is not particularly limited and may be any known polymerization temperature. More preferably, it is 0 to 120°C.
• a vinyl ester polymer is obtained as described above.
• the method for the saponification reaction of the obtained polymer is not particularly limited, and may be according to a conventionally known method. , p-toluenesulfonic acid, etc., can be used for alcoholysis or hydrolysis reaction. Before and after the saponification reaction, the syndiotacticity of the polymer usually hardly changes.
• Solvents used in the saponification reaction include alcohols such as methanol and ethanol; esters such as methyl acetate and ethyl acetate; ketones such as acetone and methyl ethyl ketone; aromatic hydrocarbons such as benzene and toluene; These can be used alone or in combination of two or more.
• the saponification temperature, time, etc. are not particularly limited.
• the method for drying, pulverizing, and washing the saponified product is not particularly limited, and known methods may be used.
• a saponified product of the vinyl ester polymer that is, the highly syndio PVA-based resin of the present invention is obtained.
• Reactions such as acetalization, urethanization, etherification, grafting, phosphate esterification, acetoacetylation, and cationization of the obtained high syndio PVA-based resin using known methods within a range that does not impair the effects of the present invention may be post-modified by
• the degree of saponification of the high syndio PVA-based resin is preferably 90 to 99.9 mol%, more preferably 98 to 99.9 mol%, still more preferably 99 to 99.9 mol%.
• the degree of saponification of the high syndio PVA-based resin can be obtained, for example, by measuring proton NMR in a heavy DMSO solution.
• the degree of polymerization of the high syndio PVA resin is preferably 100 to 10000, more preferably 500 to 8000, still more preferably 1000 to 5000, and particularly preferably 1000 from the viewpoint of relatively easy handling. ⁇ 3000. If the degree of polymerization is 100 or more, the resin strength (stress) is high and it is easy to prepare a shape-retaining aqueous gel. If the polymerization degree is 10000 or less, the aqueous solution is easy to handle.
• the degree of polymerization is the degree of polymerization in terms of polyvinyl acetate at 30°C in a benzene solution described in JIS K6725 in the resin before saponification.
• polyvinyl alcohol resin with saponification degree of 97 mol% or more As the polyvinyl alcohol resin, as described above, a polyvinyl alcohol resin having a degree of saponification of 97 mol % or more (in this specification, simply referred to as "completely saponified PVA resin") can also be used.
• the degree of saponification of the completely saponified PVA-based resin is preferably 97 mol% or more (eg, 97 to 99.9 mol%), more preferably 98 mol% or more (eg, 98 to 99.9 mol%), and 98.5. mol % or more (for example, 98.5 to 99.9 mol %) is particularly preferred.
• the degree of polymerization of the completely saponified PVA-based resin is, for example, 100 to 10,000, preferably 500 to 9,000, even more preferably 1,000 to 8,000, and particularly preferably 1,500 to 5,000.
• the cell-embedded device (the polymer that constitutes the cell-embedded device) may further contain a cross-linking agent.
• the cross-linking agent is not particularly limited, and can be selected according to the type of polymer and the like.
• a polymer having a functional group for example, a hydrazino group, etc.
• a polymer having a functional group for example, a hydrazino group, etc.
• cross-linking agents examples include hydrazide compounds and semicarbazide compounds.
• hydrazide compounds and semicarbazide compounds having two or more functional groups selected from the group represented by the following formulas (1) to (3) in the molecule are preferable. These can be used individually by 1 type or in combination of 2 or more types.
• -NH- NH2 (1) —CO—NH—NH 2 (2) —NH—CO—NH—NH 2 (3)
• hydrazide compounds include carbohydrazide, dicarboxylic acid hydrazide [fatty acid dicarboxylic acid hydrazide (e.g., oxalic acid dihydrazide, malonic acid dihydrazide, succinic acid dihydrazide, glutaric acid dihydrazide, adipic acid dihydrazide, pimelic acid dihydrazide, suberic acid dihydrazide, azelaic acid dihydrazide, sebacic acid dihydrazide, dodecanedioic acid dihydrazide, hexadecanedioic acid dihydrazide, etc.), aromatic dicarboxylic acid hydrazides (e.g., terephthalic acid dihydrazide, isophthalic acid dihydrazide, 2,6-naphthoic acid dihydrazide, 4,4' -bisbenzene dihydrazide, etc.),
• semicarbazide compound examples include N,N'-hexamethylenebissemicarbazide and biurettrietri (hexamethylenesemicarbazide). Derivatives obtained by reacting these hydrazide compounds and semicarbazide compounds with low boiling point ketones such as acetone and methyl ethyl ketone may also be used.
• dicarboxylic acid hydrazides, polyacrylic hydrazides and the like are preferred, adipic acid dihydrazides, polyacrylic acid hydrazides and the like are more preferred, and polyacrylic acid hydrazides are particularly preferred. It can be preferably used from the viewpoint of high efficiency.
• the cross-linking agent can be used alone or in combination of two or more of the above cross-linking agents.
• the amount of the cross-linking agent added is preferably 1 to 30 parts by mass, more preferably 2 to 25 parts by mass, and 3 to 20 parts by mass (eg, 4 to 15 parts by mass) is more preferable.
• the cross-linking density becomes high and sufficient strength (stress) as a cell-embedded device can be obtained. It is preferable from the viewpoint of being able to
• the molecular weight range is not particularly limited. More preferably, 800,000 (eg, about 10,000 to 300,000, about 1,000 to 200,000, about 10,000 to 100,000).
• the hydrazide conversion rate of polyacrylic acid hydrazide is not particularly limited, but is preferably 30% or more, more preferably 50% or more, further preferably 70% or more, and 80% or more. Especially preferred.
• the molecular weight and hydrazide conversion rate of polyacrylic acid hydrazide may be appropriately adjusted within a range in which the effects of the present invention are not hindered, for example, if the molecular weight is small, the hydrazide conversion rate is increased, and if the molecular weight is large, the hydrazide conversion rate is decreased.
• the polyvinyl alcohol-based resin may form a gel [especially an aqueous gel (hydrogel)] or a sponge.
• a gel depending on the type of polymer, for example, it may be crosslinked (gel crosslinked) with a crosslinking agent or the like described later. ) to form a gel.
• the polymer concentration may be, for example, 0.3-20%, preferably 0.5-10%, more preferably 1-8% (eg, 3-8%). Such a range is preferable from the viewpoint that the device shape can be maintained in the body for a long period of time after the device for forming a film is implanted in an animal.
• the content of the cross-linking agent can be appropriately selected according to the type of cross-linking agent and desired strength. It may be at least 1 part by mass, preferably at least 1 part by mass, more preferably at least 3 parts by mass, and may be at most 20 parts by mass, preferably at most 18 parts by mass, more preferably at most 15 parts by mass.
• the method for forming (forming) the gel includes, for example, a mixed solution containing a polymer [e.g., polyvinyl alcohol-based resin, and optionally a cross-linking agent, etc.]
• a method of pouring an aqueous solution which may be in a sol state
• a method of processing the obtained gel into the desired shape with a knife or the like is particularly advantageous.
• a mixed liquid (particularly an aqueous solution) containing a polymer goes through a sol state before reaching a gel state.
• a polymer eg, polyvinyl alcohol resin, and optionally a cross-linking agent, etc.
• sol states are also understood to be within the scope of the present invention as gel equivalents of the present invention.
• the solid content concentration of the mixed liquid is, for example, 0.3 to 20% by mass, preferably 0.5 to 10% by mass, more preferably 1 to 8% by mass. be. Such a range is preferable from the viewpoint of maintaining the shape of the device in the body for a long period of time and maintaining the anti-adhesion ability.
• the preparation method when preparing an aqueous solution containing a polyvinyl alcohol-based resin, the preparation method is not particularly limited. It can be prepared by a conventionally known method of dissolving PVA, which is followed by cooling.
• the cross-linking agent may be used as an aqueous solution.
• the method for preparing the aqueous solution of the cross-linking agent is not particularly limited. °C for 10 minutes, etc.), and then allowed to stand at room temperature.
• the aqueous solution of the polyvinyl alcohol resin and the aqueous solution of the cross-linking agent are desirably sterilized by a conventionally known method such as autoclave treatment, UV, ⁇ -ray or filter treatment.
• a conventionally known method such as autoclave treatment, UV, ⁇ -ray or filter treatment.
• a method of adding a pore-forming agent during the production of the gel and a method of forming pores by drying (eg, freeze-drying) the obtained gel are preferably used.
• a water-soluble polymer, a water-soluble inorganic substance, an organic solvent, or the like can be used as the pore-forming agent.
• starch is preferably used.
• a sponge is formed by adding a cross-linking agent (for example, formalin) to an aqueous solution containing polyvinyl alcohol-based resin and starch, gelling the polyvinyl alcohol-based resin, and then washing the starch with water.
• a cross-linking agent for example, formalin
• a drying treatment may be performed after forming the sponge.
• a cell-containing device may contain a cell culture component.
• cell culture components include, but are not limited to, alkali metals, alkaline earth metals, halogens and glucose. etc. are preferably used.
• the Na concentration may be adjusted to preferably 20-150 mEq/L, more preferably 80-140 mEq/L.
• the K concentration may be adjusted to preferably 2.5-130 mEq/L, more preferably 3.5-40 mEq/L.
• Cl the Cl concentration may be adjusted to preferably 15-170 mEq/L, more preferably 100-150 mEq/L.
• Ca the Ca concentration may be adjusted to preferably 0.5-5 mEq/L, more preferably 1-3 mEq/L.
• glucose the glucose concentration may be adjusted to preferably 1-11 mM, more preferably 3-7 mM.
• Cell culture components are not particularly limited. of Wisconsin solution), cytoprotective components (e.g., dimethyl sulfoxide (DMSO), serum albumin, etc.), components that prevent contamination by bacteria (e.g., antibiotics, etc.), components that maintain cell activity (e.g., nicotine vitamins such as amides), etc., preferably known cell culture media and the like.
• cytoprotective components e.g., dimethyl sulfoxide (DMSO), serum albumin, etc.
• DMSO dimethyl sulfoxide
• serum albumin e.g., etc.
• components that prevent contamination by bacteria e.g., antibiotics, etc.
• components that maintain cell activity e.g., nicotine vitamins such as amides
• nicotine vitamins such as amides
• cell culture components may be used in combination with other components (eg, sustained-release agents, tonicity agents, pH adjusters, etc.).
• the cell-containing device may further contain components other than these.
• cell-containing devices may contain growth factors (cell growth factors), cytokines, other physiologically active substances, blood flow promoting substances, neurotrophic factors, and the like.
• Growth factors include, for example, the above-exemplified growth factors such as epidermal growth factor (EGF), hepatocyte growth factor (HGF), and insulin.
• EGF epidermal growth factor
• HGF hepatocyte growth factor
• insulin insulin
• Cytokines include, for example, hematopoietic factors (eg, interleukins, chemokines, colony-stimulating factors, etc.), tumor necrosis factor, interferons, and the like.
• physiologically active substances include, for example, amino acids (eg, glycine, phenylalanine, lysine, aspartic acid, glutamic acid, etc.), vitamins (eg, biotin, pantothenic acid, vitamin D, etc.), serum albumin, antibiotics, and the like.
• amino acids eg, glycine, phenylalanine, lysine, aspartic acid, glutamic acid, etc.
• vitamins eg, biotin, pantothenic acid, vitamin D, etc.
• serum albumin e.g., antibiotics, and the like.
• Blood flow-promoting substances include, for example, citrulline or its salts, capsaicin, and capsaicinoids.
• neurotrophic factors examples include NGF (nerve growth factor), BDNF (brain-derived neurotrophic factor), NT-3 (neurotrophin-3), NT -4 (neurotrophin-4; neurotrophin-4), GDNF (Glial-Cell Derived Neurotrophic Factor; glial cell line-derived neurotrophic factor), neurturin, artemin, persephin, etc. .
• the amount of these components added is not particularly limited.
• a cell-containing device may form a gel.
• Such gels usually have a predetermined strength (stress). Such strength may be a stress that does not easily collapse during implantation.
• the strength of the gel depends on the type of polymer (for example, type of polyvinyl alcohol resin, viscosity of 4% aqueous solution, degree of saponification, degree of denaturation, syndiotacticity, etc.), type of cross-linking agent, amount added, solid content concentration of gel, etc. Although it cannot be generalized because it varies depending on the may be
• the gel stress can be measured, for example, using Shimadzu Corporation's small desktop tester EZTest EZ-SX according to its instruction manual.
• the shape of the gel is not particularly limited, but examples include sheet-like, plate-like, board-like, rod-like, tube-like, bead-like, and the like.
• the size of the gel (especially aqueous gel) can be appropriately selected according to the transplant site, its size, etc., and is not particularly limited. 0.5 to 2 mm is more preferred, and 0.7 to 1.5 mm is particularly preferred.
• a cell-containing device may contain a support substrate.
• gels eg, scaffolding gels
• a supporting substrate useful as a reinforcing material for their reinforcement and/or ease of manipulation.
• a gel especially aqueous gel
• a base material such as a resin mesh sheet for reinforcement and simplification of operability. good.
• the material of the supporting substrate is not limited, but examples thereof include polymers [e.g., PET (polyethylene terephthalate), PE (polyethylene), PP (polypropylene), Teflon (registered trademark), etc.], metals, and the like. It is preferable that the material does not decompose or degrade in vivo, but it may be decomposed in vivo after a certain period of time.
• polymers e.g., PET (polyethylene terephthalate), PE (polyethylene), PP (polypropylene), Teflon (registered trademark), etc.
• the mesh size of the mesh sheet is such that oxygen to be permeated, inorganic and organic nutrients, and molecules with a diameter of about 5 nm, which is assumed to be the largest among various hormones (for example, physiologically active substances including hormones such as insulin). It is usually 5 to 100 nm, preferably 10 nm, so as not to permeate molecules with a diameter of about 50 nm (eg, antibodies, complements, etc.), which are assumed to be the smallest among immune-related cells and immune-related substances to be permeated and blocked from permeation. ⁇ 50 nm, more preferably 20-30 nm.
• a cell-containing device may have, for example, an immunoisolation layer containing a polymer [eg, polyvinyl alcohol-based resin (A)].
• the polymer gel-forming polymer
• the immunoisolation layer or have an immunoisolation function).
• the immune isolation layer includes, for example, glucose; hormones such as insulin, thyroid stimulating hormone, thyroid hormone, parathyroid hormone, growth hormone, thyroxine, glucocorticoid, glucagon, estradiol, or testosterone; blood proteins such as coagulation factors, albumin, globulin, various enzymes (metabolic enzymes or digestive enzymes such as amylase, protease, or lipase); neurotransmitters such as dopamine; It means a layer (function) impermeable to proteins of the immune system such as.
• a cell-containing device may be, for example, a bioartificial organ.
• a cell-containing device (cell-embedded device) can be manufactured, for example, by coexisting (existing) a biological composition and a polymer (and other components).
• the method of forming (forming) the gel includes, for example, a polymer [e.g., a polyvinyl alcohol-based resin, and optionally a biocomposition, a cross-linking agent, a cell culture component, etc.].
• a polymer e.g., a polyvinyl alcohol-based resin, and optionally a biocomposition, a cross-linking agent, a cell culture component, etc.
• a method of pouring a mixed solution especially an aqueous solution (may be in a sol state)] containing into a mold of the desired shape before gelling, a method of processing the obtained gel into the desired shape with a knife etc. etc.
• a mixed solution (especially an aqueous solution) containing a polymer [e.g., polyvinyl alcohol-based resin, and optionally, a biological composition, a cross-linking agent, a cell culture component, etc.] is in a sol state before reaching a gel state.
• a polymer e.g., polyvinyl alcohol-based resin, and optionally, a biological composition, a cross-linking agent, a cell culture component, etc.
• sol states are also understood to be within the scope of the present invention as gel equivalents of the present invention.
• the solid content concentration of the mixed liquid is, for example, 0.3 to 20% by mass, preferably 0.5 to 10% by mass, more preferably 1 to 8% by mass. be. Such a range is preferable from the viewpoint of maintaining the shape of the cell-embedded device in the body for a long period of time after implantation and maintaining the anti-adhesion ability.
• each component may be mixed at the same time, and a mixed solution (for example, an aqueous solution of polyvinyl alcohol resin) containing a polymer (and optionally a cross-linking agent or cell culture component) is prepared in advance. , may be prepared by mixing biological compositions.
• a mixed solution for example, an aqueous solution of polyvinyl alcohol resin
• a polymer and optionally a cross-linking agent or cell culture component
• the preparation method when preparing an aqueous solution containing a polyvinyl alcohol-based resin, the preparation method is not particularly limited. It can be prepared by a conventionally known method of dissolving PVA, which is followed by cooling.
• the cross-linking agent may be used as an aqueous solution.
• the method for preparing the aqueous solution of the cross-linking agent is not particularly limited. °C for 10 minutes, etc.), and then allowed to stand at room temperature.
• the aqueous solution of the polyvinyl alcohol resin and the aqueous solution of the cross-linking agent are desirably sterilized by a conventionally known method such as autoclave treatment, UV, ⁇ -ray or filter treatment.
• a conventionally known method such as autoclave treatment, UV, ⁇ -ray or filter treatment.
• the mixture (or its aqueous solution) of the polyvinyl alcohol-based resin aqueous solution and the cross-linking agent (if necessary, the biocomposition and/or cell culture components are mixed) may be left alone.
• the leaving temperature may be a temperature suitable for storing the biological composition.
• the standing time (time for gelation, gelation time) when producing a gel can be appropriately selected depending on the concentration of the polymer (polyvinyl alcohol resin, etc.), the amount of the cross-linking agent, the standing temperature, etc. At normal temperature, it is usually about 1 hour to 1 week. A period of 1 hour or more is preferable from the viewpoint of not easily collapsing when the cell-embedded device is left in the body.
• a pH buffer solution or the like is added to a mixed solution of the modified PVA-based resin and a cross-linking agent (and, if necessary, MSCs and/or cell culture components are mixed).
• a cross-linking agent and, if necessary, MSCs and/or cell culture components are mixed.
• a polymer mixture containing cell culture components or its gel e.g., polyvinyl alcohol-based resin-containing aqueous solution or aqueous gel
• a substrate or substrate substrate (slide glass, etc.)
• a PET mesh e.g., Co., Ltd.
• a supporting substrate such as PET mesh sheet (trade name: TN120, etc., manufactured by Sunplatic Co., Ltd.) is covered, and the polymer mixture or its gel (for example, polyvinyl alcohol-based resin-containing aqueous solution or aqueous gel) is placed on the supporting substrate.
• a mixture obtained by dissolving or dispersing (suspending) a biological composition (dissolution or dispersion (suspension)) is placed on the substrate, and the mixture is spread on the support substrate using a gel loading tip or the like.
• a support substrate PET mesh, etc.
• a polymer mixture containing cell culture components or a gel thereof e.g., Separating (removing) the base material or substrate (slide glass, etc.) from a structure constructed by placing a polyvinyl alcohol-based resin-containing aqueous solution or aqueous gel) and covering it with a base material or substrate (slide glass, etc.)
• a cell-containing device of one embodiment is obtained.
• the present invention also includes a method of transplantation (a method of transplanting a cell-containing device).
• a method of transplantation a method of transplanting a cell-containing device.
• the cell-containing device is implanted at the implantation site (extraction site) after the device 1 has been implanted (and at least the non-bioabsorbable material has been removed).
• various diseases or symptoms can be prevented or ameliorated in response to biological compositions (cells or tissues). Therefore, the present invention also includes methods for preventing and/or treating (ameliorating) such diseases or conditions.
• the cell-containing device is implanted after implantation of the device 1 (and at least removal of non-bioabsorbable material), although of course the method involves implanting the device 1 prior to such implantation.
• a step of transplanting may be included.
• Symptoms or diseases include, for example, endocrine diseases (e.g., thyroid disease, parathyroid disease, adrenal disease, pituitary disease, pineal disease, etc.), metabolic diseases (e.g., ornithine transcarbamylase deficiency, hyperammonemia disease, hypercholesterolemia, homocystinuria, glycogen storage disease, Crigler-Nager syndrome, Wilson's disease), diabetes (e.g., type 1 diabetes, type 2 diabetes, pancreatic diabetes, etc.), neurodegenerative diseases (e.g., Parkinson's disease) , Alzheimer's disease, amyotrophic lateral sclerosis, spinocerebellar degeneration, etc.), hemophilia, bone diseases (eg, osteoporosis), cancer (eg, leukemia, etc.), and the like.
• endocrine diseases e.g., thyroid disease, parathyroid disease, adrenal disease, pituitary disease, pineal disease, etc.
• metabolic diseases e.g., or
• the implantation (dwelling) period of the cell-containing device is not particularly limited, but may be, for example, 10 days or longer, 1 month or longer, 2 months or longer, 3 months or longer, 6 months or longer, 1 year or longer, and the like.
• the device 1 of the present invention can be suitably used in combination with cell-containing devices. Therefore, the present invention also includes combination devices (kits, devices, combinations) comprising said device 1 and a cell-containing device.
• the material (A) was immersed in physiological saline at 37° C. to a concentration of 5% by mass, and after immersion for 24 hours, it was calculated from the mass of the solid content contained in the physiological saline.
• the moisture content was corrected in the same manner as the density.
• the I2 /MeOH solution was titrated with a 0.1M sodium thiosulfate standard solution. (It was 0.047M in this measurement.) 3. Each polymer sample was precisely weighed and dissolved in 20 mL of deionized water. 4. 2.0 mL of 0.047 M I 2 /MeOH solution was added to the solution of 3. 5. Iodine back titration was performed with 0.1 M sodium thiosulfate standard.
• the islet-containing device was implanted subcutaneously (adipose tissue) in rats, and the function of the islet-containing device was evaluated. Specifically, in streptozotocin-induced diabetic rats implanted with pancreatic islet-containing devices, blood glucose levels were measured over time to confirm the healing effect.
• Example 1 (Preparation of material (A)) A 5% by mass aqueous solution of commercially available gelatin (RM-50, derived from pigskin, acid-treated product manufactured by Zerais Co., Ltd.) was freeze-dried, punched into a cylindrical shape with a diameter of ⁇ 22 mm (thickness of 0.5 mm), and then at 145 ° C. The gelatin was cross-linked by vacuum drying for 24 hours to obtain material (A). The obtained material (A) had a density of 58 mg/cm 3 , an elution rate of 19.8%, a swelling degree of 9.1 times, and a value of density/swelling degree of 6.4.
• RM-50 commercially available gelatin
• a non-bioabsorbable material (B) was obtained by punching a commercially available silicone rubber (manufactured by AS ONE, model number 6-611-02, thickness 1 mm) into a cylindrical shape (thickness 0.5 mm) with a diameter of 26 mm.
• the non-bioabsorbable material (B) had a density of 1,200 mg/cm 3 , an elution rate of 0%, a swelling ratio of 1.0, and a density/swelling ratio of 1,200.
• Material (A) was allowed to stand in physiological saline at 22° C. for 1 minute to allow it to swell sufficiently.
• the swollen material (A) is attached to the upper and lower surfaces of the non-bioabsorbable material (B), and the material (A) and the non-bioabsorbable material (B) are immobilized by suturing to form a device (implantation device, Device 1) was obtained.
• the device (implantation device, device 1) was placed subcutaneously in 12- to 14-week-old male Wistar rats (Japan LSC) for 6 weeks. No hemorrhage or exudate was observed under the skin before the device was placed, and no capsule was observed. A coating was observed. In addition, the material (A) of the device was almost completely bioabsorbed as a coating (and into the surrounding tissue), and only the non-bioabsorbable material remained at the indwelling (implantation) site. Since adhesion between the surface of the non-bioabsorbable material and the subcutaneous tissue was sparse, peeling was extremely easy.
• transplantation for example, insertion of an islet-embedding device, which will be described later
• the transplantation/dwelling process was performed on two rats, and both showed similar results (sufficient capsule formation, no bleeding or exudate after removal of the device).
• pancreatic islet embedding device Male Lewis rats (Japan SLC) aged 11 to 14 weeks were used for islet isolation.
• Pancreatic islets were cultured overnight in RPMI1640 medium containing 5.5 mmol/L glucose and 10% Fetal Bovine Serum (FBS) at 37° C. under 5% CO 2 .
• FBS Fetal Bovine Serum
• Dinitrobenzene was added to stop the polymerization. The yield at the end of polymerization was 47.1%. The remaining vinyl acetate was distilled off while methanol vapor was added to the resulting reaction mixture to obtain a 35% methanol solution of diacetone acrylamide-vinyl acetate copolymer.
• To 500 parts of this solution were added 70 parts of methanol, 1 part of ion-exchanged water and 29.3 parts of a 4% methanol solution of sodium hydroxide, and the mixture was thoroughly mixed to carry out a saponification reaction at 45°C.
• the resulting gel-like material was pulverized, thoroughly washed with methanol and dried to obtain D-PVA1.
• the 4% aqueous solution viscosity was 53.4 mPa ⁇ s
• the saponification degree was 98.4 mol %
• the diacetone acrylamide unit content was 3.6 mol %.
• the concentration of D-PVA1 was 5.0%
• the concentration of APA1 was 0.5%
• the stress corresponding to the above composition was 5.2 kPa.
• the islet-embedded device constructed above was removed from the slide glass, and immersed in a 6-well plate at a rate of 5 mL/well in a preservation medium (RPMI1640 medium containing 10% FBS with a glucose concentration adjusted to 5.5 mM). It was stored at °C for about 16 hours.
• a preservation medium RPMI1640 medium containing 10% FBS with a glucose concentration adjusted to 5.5 mM. It was stored at °C for about 16 hours.
• Diabetes healing evaluation After the transplantation, the blood glucose level was measured over time to confirm the healing effect. Diabetes healing evaluation was performed on two rats.
• Example 2 Commercially available gelatin (Sponzel, manufactured by LTL Pharma Co., Ltd.) was punched into a cylindrical shape (thickness: 0.5 mm) with a diameter of 30 mm, and about 400 small holes were made with an injection needle to obtain material (A).
• the obtained material (A) had a density of 15 mg/cm 3 , an elution rate of 33.4%, a swelling degree of 48.5 times, and a value of density/swelling degree of 0.303.
• Example 1 The same operation as in Example 1 was performed, except that the material (A) in Example 1 was replaced with the material (A) obtained above, and diabetes healing evaluation was performed.
• the fabricated device showed the same tendency as Example 1 (sufficient film formation, no bleeding or exudate after device removal) from placement to removal.
• Example 3 In Example 1, except that a device without material (A) (only non-bioabsorbable material (B), only material (A) using non-bioabsorbable material as material (A)) was used as the device. , the same operation as in Example 1 was performed to evaluate diabetes cure. The fabricated device showed the same tendency as Example 1 (no bleeding or exudate after removal of the device) from indwelling to withdrawal. A coating was also formed, but compared to Examples 1 and 2, the thickness was smaller.
• Example 1 The same operation as in Example 1 was performed, except that the device 1 was not implanted or indwelled, and diabetes cure evaluation was performed.
• the results of diabetes healing evaluation obtained in Examples 1 to 3, Reference Example 1, and Comparative Example 1 are shown in the table below.
• the “pre-implantation blood glucose level” means the blood glucose level immediately before the device is removed and the pancreatic islet device is transplanted.
• the implanted site (subcutaneously) could be activated, and the pancreatic islet device implanted thereafter could function effectively.
• the implanted devices of Examples 1 to 3 (especially Examples 1 and 2) had a coating formed on the implanted site.
• it was surprising that such a pancreatic islet device could function effectively even though the device did not contain growth factors or the like. According to the study of the present inventor, this suggests an endogenous increase in growth factors, extracellular matrix, and the like.
• Such a device can be used as a device for activating the site of implantation, for example, as a device for recovering wounds or as a device for fully drawing out the inherent performance of a cell-containing device to be implanted separately.

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