WO2020091035A1 - Matériau pour la régénération osseuse présentant une surface comprenant des fibres biodégradables hydrophilisées et procédé de production associé - Google Patents

Matériau pour la régénération osseuse présentant une surface comprenant des fibres biodégradables hydrophilisées et procédé de production associé Download PDF

Info

Publication number
WO2020091035A1
WO2020091035A1 PCT/JP2019/043010 JP2019043010W WO2020091035A1 WO 2020091035 A1 WO2020091035 A1 WO 2020091035A1 JP 2019043010 W JP2019043010 W JP 2019043010W WO 2020091035 A1 WO2020091035 A1 WO 2020091035A1
Authority
WO
WIPO (PCT)
Prior art keywords
biodegradable fiber
biodegradable
fiber
calcium phosphate
bone
Prior art date
Application number
PCT/JP2019/043010
Other languages
English (en)
Japanese (ja)
Inventor
敏宏 春日
松原　孝至
将央 渡部
直也 大坂
Original Assignee
国立大学法人 名古屋工業大学
Ｏｒｔｈｏｒｅｂｉｒｔｈ株式会社
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by 国立大学法人 名古屋工業大学, Ｏｒｔｈｏｒｅｂｉｒｔｈ株式会社 filed Critical 国立大学法人 名古屋工業大学
Priority to JP2020554976A priority Critical patent/JP7429391B2/ja
Publication of WO2020091035A1 publication Critical patent/WO2020091035A1/fr

Links

Images

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/02—Inorganic materials
    • A61L27/12—Phosphorus-containing materials, e.g. apatite
    • 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
    • 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
    • A61L27/18—Macromolecular materials obtained otherwise than 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
    • A61L27/28—Materials for coating prostheses
    • A61L27/30—Inorganic materials
    • A61L27/32—Phosphorus-containing materials, e.g. apatite
    • 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
    • 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
    • D—TEXTILES; PAPER
    • D06—TREATMENT OF TEXTILES OR THE LIKE; LAUNDERING; FLEXIBLE MATERIALS NOT OTHERWISE PROVIDED FOR
    • D06M—TREATMENT, NOT PROVIDED FOR ELSEWHERE IN CLASS D06, OF FIBRES, THREADS, YARNS, FABRICS, FEATHERS OR FIBROUS GOODS MADE FROM SUCH MATERIALS
    • D06M11/00—Treating fibres, threads, yarns, fabrics or fibrous goods made from such materials, with inorganic substances or complexes thereof; Such treatment combined with mechanical treatment, e.g. mercerising
    • D06M11/68—Treating fibres, threads, yarns, fabrics or fibrous goods made from such materials, with inorganic substances or complexes thereof; Such treatment combined with mechanical treatment, e.g. mercerising with phosphorus or compounds thereof, e.g. with chlorophosphonic acid or salts thereof
    • D06M11/70—Treating fibres, threads, yarns, fabrics or fibrous goods made from such materials, with inorganic substances or complexes thereof; Such treatment combined with mechanical treatment, e.g. mercerising with phosphorus or compounds thereof, e.g. with chlorophosphonic acid or salts thereof with oxides of phosphorus; with hypophosphorous, phosphorous or phosphoric acids or their salts
    • D06M11/71—Salts of phosphoric acids

Definitions

  • the present invention relates to a bone regenerating material comprising biodegradable fibers having hydrophilicity imparted by coating the surface of biodegradable fibers with an amorphous calcium phosphate layer, and a method for producing the same.
  • an artificial bone of a type in which a bone regenerating material composed of a biodegradable fiber is implanted in a living body to form a bone is used.
  • the biodegradable fiber contains bone-forming particles such as calcium phosphate, and when the bone regeneration material is implanted in the body and comes into contact with body fluid and the biodegradable resin is hydrolyzed, the surface of the fiber causes bone formation. The effective ions are gradually released to promote bone formation in the affected area.
  • the biodegradable fiber can be spun using the melt spinning method or the electrospinning method.
  • the electrospinning method it is possible to spin by adding inorganic particles to a spinning solution in which a resin is dissolved in a solvent, so that a bone regenerating material composed of biodegradable fibers containing osteogenic particles such as calcium phosphate can be used. It can be easily manufactured.
  • biodegradable fiber Since the polymer surface of biodegradable fiber has water repellency and hydrophobicity, when implanting the material in the affected area, it will be repelled at the earliest stage of mixing with blood, which will increase the time required for shaping during surgery. The problem has been pointed out. In order to deal with this problem, an attempt to make the surface of the biodegradable fiber hydrophilic has been proposed.
  • a hydrophilic treatment method plasma treatment, hydrophilic polymer coating, inorganic film coating, calcium phosphate (CaP) coating are known. CaP coating is performed by immersing biodegradable fibers in an aqueous solution containing phosphate ions. Since the surface can be easily coated with calcium hydroxyphosphate (HA), it is a suitable method for hydrophilizing a fibrous bone regenerating material having a complicated three-dimensional structure.
  • alternate dipping method and biomimetic method are known.
  • the material is alternately immersed in a solution containing calcium ions and a solution containing phosphate ions to form HA on the surface. It can be performed at room temperature and normal pressure, and can be applied to a material having a three-dimensional structure (Non-Patent Document 1).
  • the biomimetic method forms HA on the surface of a material by immersing the material in a simulated body fluid (SBF) whose inorganic ion concentration is adjusted to be equal to that of human plasma. This method can form HA more easily than the alternate dipping method, and can also coat HA on a three-dimensional structure as in the alternate dipping method (Non-Patent Document 2).
  • SBF simulated body fluid
  • the phase produced by the conventional alternate dipping method or biomimetic method has a high content of HA crystals, so it is difficult to dissolve in contact with water. Further, in order to coat the surface of the material fiber with HA using these methods, the material must be immersed in the solution for several hours to several days, so that the bone-forming particles contained in the biodegradable fiber during There is a problem that the ions are eluted into the solution, and the stimulating effect on the tissue due to the elution of the bone-forming particles is weakened.
  • the inventors of the present invention have studied the means for solving the above-mentioned problems, and have concluded that it is best to coat the surface of the biodegradable fiber with the amorphous calcium phosphate layer in a short time. Since amorphous calcium phosphate has a high solubility in water, it does not dissolve and disappear in a short time in contact with body fluid or blood, and does not prevent the elution of osteogenic particles. Further, since this amorphous phase has high ionicity, the hydrophilicity of the surface of the polymer fiber coated with it is significantly improved.
  • the inventors of the present invention have conducted extensive experiments and studies, and as a result, heat the biodegradable resin in a state where the vaterite phase calcium carbonate particles are attached to the surface of the biodegradable fiber by using the triboelectric charging method.
  • the triboelectric charging method By adhering and fixing the particles to the fiber surface by doing so, when the material is immersed in an aqueous solution containing phosphate ions of a predetermined concentration in that state, the amorphous calcium phosphate layer starts from the vaterite phase calcium carbonate particles in a short time. It was envisioned that they could grow and coat the entire surface of the fiber.
  • the inventors of the present invention are a method for producing a bone regeneration material comprising a biodegradable fiber whose surface is hydrophilized, Vaterite phase calcium carbonate particles are electrostatically attached to the surface of the biodegradable fiber containing the bone-forming particles by using a triboelectric charging method, The vaterite phase calcium carbonate particles adhered to the biodegradable fiber, by heating at a temperature above the glass transition point of the biodegradable resin of the biodegradable fiber, the vaterite phase on the surface of the biodegradable fiber Adhere and fix calcium carbonate particles, The vaterite phase calcium carbonate particles are adhered and fixed The biodegradable fiber is immersed in an aqueous solution containing a phosphate ion of a predetermined concentration for a predetermined time, whereby the vaterite phase is fixedly adhered to the surface of the biodegradable fiber.
  • amorphous calcium phosphate is grown along the surface of the biodegradable fiber to cover the entire surface of the biodegradable fiber, A bone regenerating material consisting of the biodegradable fiber whose surface is coated with the amorphous calcium phosphate is taken out from the aqueous solution containing the phosphate ion and dried,
  • the present invention has arrived at a method for producing a bone regeneration material comprising a biodegradable fiber having a hydrophilic surface.
  • the inventors of the present invention further provide a bone regeneration material comprising a biodegradable fiber whose surface is hydrophilized,
  • the biodegradable fiber constituting the bone regeneration material contains bone-forming particles,
  • the entire surface of the biodegradable fiber is covered with an amorphous calcium phosphate layer containing substantially no crystalline phase,
  • the present invention has reached the invention of a bone regeneration material comprising a biodegradable fiber having a hydrophilic surface.
  • the amorphous calcium phosphate layer contains carbonate apatite containing carbonic acid in a part of calcium phosphate.
  • the inventors of the present invention further provide a method for producing a biodegradable fiber whose surface is hydrophilized, Vaterite phase calcium carbonate particles are electrostatically attached to the surface of the biodegradable fiber containing the bone-forming particles by using a triboelectric charging method, The vaterite phase calcium carbonate particles adhered to the biodegradable fiber, by heating at a temperature above the glass transition point of the biodegradable resin of the biodegradable fiber, the vaterite phase on the surface of the biodegradable fiber Adhere and fix calcium carbonate particles, The vaterite phase calcium carbonate particles are adhered and fixed The biodegradable fiber is immersed in an aqueous solution containing a phosphate ion of a predetermined concentration for a predetermined time, whereby the vaterite phase is fixedly adhered to the surface of the biodegradable fiber.
  • amorphous calcium phosphate is grown along the surface of the biodegradable fiber to cover the entire surface of the biodegradable fiber, A bone regenerating material consisting of the biodegradable fiber whose surface is coated with the amorphous calcium phosphate is taken out from the aqueous solution containing the phosphate ion and dried,
  • the invention has arrived at a method for producing a biodegradable fiber having a hydrophilic surface.
  • the inventors of the present invention further have a biodegradable fiber whose surface is hydrophilized,
  • the biodegradable fiber contains bone-forming particles,
  • the entire surface of the biodegradable fiber is covered with an amorphous calcium phosphate layer containing substantially no crystalline phase,
  • the invention has been reached, which is a biodegradable fiber whose surface is hydrophilized.
  • the amorphous calcium phosphate layer contains carbonate apatite containing carbonic acid in a crystal structure in a part of calcium phosphate.
  • the vaterite phase calcium carbonate particles include siloxane
  • the amorphous calcium phosphate layer grown from the vaterite phase calcium carbonate particles including siloxane includes silicon.
  • the aqueous solution containing phosphate ions is a disodium hydrogen phosphate solution
  • the bone regenerating material comprising biodegradable fibers having the vaterite phase calcium carbonate particles adhered and fixed to the surface thereof is used as the disodium hydrogen phosphate.
  • the amorphous calcium phosphate layer is formed containing sodium and coats the surface of the fiber.
  • the diameter of the biodegradable fiber is 10 to 100 ⁇ m, and the diameter of the vaterite phase calcium carbonate particles is 0.5 to 4 ⁇ m. More preferably, the diameter of the biodegradable fiber is 20 to 60 ⁇ m, and the diameter of the vaterite phase calcium carbonate particles is 0.7 to 2.0 ⁇ m.
  • the biodegradable resin of the biodegradable fiber is PLGA.
  • the osteogenic particles are ⁇ -phase tricalcium phosphate particles.
  • the bone-forming particles are silicon-eluting type vaterite phase calcium carbonate particles.
  • the inventors of the present invention further provide a method for hydrophilizing the surface of the biodegradable fiber constituting the cell culture substrate, Vaterite phase calcium carbonate particles on the surface of the biodegradable fiber constituting the cell culture substrate, electrostatically attached using a triboelectric charging method, By heating the biodegradable fiber to which the vaterite phase calcium carbonate particles are attached at a temperature equal to or higher than the glass transition point of the biodegradable resin of the biodegradable fiber, the vaterite phase carbonate is formed on the surface of the biodegradable fiber.
  • the vaterite phase calcium carbonate particles are adhered and fixed
  • the biodegradable fiber is immersed in an aqueous solution containing a phosphate ion of a predetermined concentration for a predetermined time, whereby the vaterite phase is fixedly adhered to the surface of the biodegradable fiber.
  • amorphous calcium phosphate is grown along the surface of the biodegradable fiber to cover the entire surface of the biodegradable fiber, Removing the biodegradable fiber whose surface is coated with the amorphous calcium phosphate from an aqueous solution containing the phosphate ion and drying the biodegradable fiber;
  • the invention has been reached, which is a method of hydrophilizing the surface of the biodegradable fiber constituting the cell culture substrate.
  • the inventors of the present invention further provide a cell culture substrate comprising a biodegradable fiber whose surface is hydrophilized,
  • the biodegradable fiber constituting the cell culture substrate contains an inorganic filler, Substantially the entire surface of the biodegradable fiber is covered with an amorphous calcium phosphate layer,
  • the amorphous calcium phosphate layer may include carbonate apatite containing carbonic acid in a crystal structure in a part of calcium phosphate,
  • the invention has been reached, which is a cell culture substrate comprising a biodegradable fiber whose surface is hydrophilized.
  • the inventors of the present invention further have a biodegradable fiber whose surface is hydrophilized,
  • the biodegradable fiber consists essentially of biodegradable resin, Substantially the entire surface of the biodegradable fiber is covered with an amorphous calcium phosphate layer, The invention has been reached, which is a biodegradable fiber whose surface is hydrophilized.
  • the amorphous calcium phosphate layer contains carbonate apatite containing carbonic acid in a crystal structure in a part of calcium phosphate.
  • the vaterite phase calcium carbonate particles include siloxane
  • the amorphous calcium phosphate layer grown from the vaterite phase calcium carbonate particles including siloxane includes silicon.
  • the aqueous solution containing phosphate ions is a disodium hydrogen phosphate solution
  • the bone regeneration material comprising biodegradable fibers having the vaterite phase calcium carbonate particles adhered and fixed on the surface thereof is used as the disodium hydrogen phosphate.
  • the amorphous calcium phosphate layer is formed containing sodium and coats the surface of the fiber.
  • the diameter of the biodegradable fiber is 10 to 100 ⁇ m, and the diameter of the vaterite phase calcium carbonate particles is 0.5 to 4 ⁇ m. More preferably, the diameter of the biodegradable fiber is 20 to 60 ⁇ m, and the diameter of the vaterite phase calcium carbonate particles is 0.7 to 2.0 ⁇ m.
  • the biodegradable resin of the biodegradable fiber is PLGA.
  • the vaterite phase calcium carbonate particles fixed on the surface of the biodegradable fiber are immersed in an aqueous solution containing phosphate ions and rapidly dissolved to supply carbonate ions to form a calcium phosphate layer. It can be formed on the fiber surface in a short time.
  • the calcium phosphate layer thus formed is substantially entirely occupied by the amorphous phase and has no crystal structure (see FIG. 6).
  • a large amount of carbonate ions are supplied from the calcium carbonate particles fixed on the surface of the fiber to form a highly soluble amorphous phase calcium phosphate.
  • soluble carbonate-containing apatite may be formed.
  • the amorphous calcium phosphate coating the surface of the biodegradable fiber is bioabsorbable and the layer is so thin that it is absorbed shortly after implanting the material in vivo. It decomposes and disappears from the fiber surface. As a result, the elution of ions from the bone-forming particles contained in the biodegradable fiber is not impeded by the presence of the amorphous calcium phosphate layer.
  • the calcium phosphate phase coated on the surface of the biodegradable fiber has a large specific surface area and high ionicity, so that many proteins can be adsorbed. As a result, high initial adhesiveness of cells is obtained when the bone regeneration material is implanted in the body.
  • silicon is supported on the vaterite phase calcium carbonate particles fixed on the surface of the fiber, and the silicon is eluted in the course of dissolution of the vaterite phase calcium carbonate by the phosphoric acid treatment. It is incorporated in the amorphous calcium phosphate layer formed on the surface.
  • silicon contained in the amorphous calcium phosphate layer is eluted and stimulates osteoblasts to promote bone formation.
  • a disodium hydrogen phosphate solution is used for the phosphoric acid treatment, and sodium is contained in the amorphous calcium phosphate layer during the process of forming the amorphous calcium phosphate layer on the surface of the fiber by the phosphoric acid treatment. Is taken into.
  • One embodiment of the present invention is a cell culture substrate comprising a biodegradable fiber whose surface is hydrophilized.
  • the biodegradable fiber coated with amorphous calcium phosphate has high protein adsorption performance and good initial cell adhesion performance.
  • One embodiment of the present invention is a biodegradable fiber whose surface is hydrophilized, said biodegradable fiber consisting essentially of a biodegradable resin and containing no inorganic particles.
  • FIG. 1 illustrates a method of depositing calcium carbonate particles on the surface of polymer fibers using the triboelectric charging method of the present invention.
  • FIG. 2 shows a method of embedding calcium carbonate particles attached to the surface of a polymer fiber by using the triboelectric charging method of the present invention by utilizing the residual stress of the fiber.
  • FIG. 3 shows a method of immersing a cotton-like bone regeneration material in an aqueous solution containing phosphate ions to perform phosphoric acid treatment.
  • FIG. 1 illustrates a method of depositing calcium carbonate particles on the surface of polymer fibers using the triboelectric charging method of the present invention.
  • FIG. 2 shows a method of embedding calcium carbonate particles attached to the surface of a polymer fiber by using the triboelectric charging method of the present invention by utilizing the residual stress of the fiber.
  • FIG. 3 shows a method of immersing a cotton-like bone regeneration material in an aqueous solution containing phosphate
  • FIG. 4 (A) is a phosphoric acid treatment in which a biodegradable fiber having calcium carbonate particles adhered to the surface of a polymer fiber is immersed in a 0.02 M disodium hydrogen phosphate solution using the triboelectric charging method of the present invention and treated with phosphoric acid The state of the surface of the fiber after applying is shown.
  • FIG. 4B is a phosphoric acid treatment in which a biodegradable fiber having calcium carbonate particles adhered to the surface of a polymer fiber by the triboelectric charging method of the present invention is immersed in a 0.2 M disodium hydrogen phosphate solution. The state of the surface of the fiber after applying is shown.
  • FIG. 4 (A) is a phosphoric acid treatment in which a biodegradable fiber having calcium carbonate particles adhered to the surface of a polymer fiber is immersed in a 0.02 M disodium hydrogen phosphate solution using the triboelectric charging method of the present invention and treated with phosphoric acid The state of the
  • FIG. 4C is a phosphoric acid treatment in which a biodegradable fiber having calcium carbonate particles adhered to the surface of a polymer fiber by the triboelectric charging method of the present invention is immersed in a 2.0 M concentration disodium hydrogen phosphate solution. The state of the surface of the fiber after applying is shown.
  • FIG. 5 shows the hydrophilicity of the phosphorous-treated cotton-shaped bone regeneration material.
  • FIG. 6 shows the results of STEM observation of a cotton-shaped bone regeneration material that has been treated with phosphoric acid. From the vague image in the lower left part a, it can be seen that the calcium phosphate layer is substantially entirely occupied by the amorphous phase.
  • FIG. 5 shows the hydrophilicity of the phosphorous-treated cotton-shaped bone regeneration material.
  • FIG. 6 shows the results of STEM observation of a cotton-shaped bone regeneration material that has been treated with phosphoric acid. From the vague image in the lower left part a, it can be seen that the calcium phosphate layer is substantially entirely
  • FIG. 7 (A) shows the surface of a biodegradable fiber obtained by immersing a sample of the amorphous calcium phosphate coating experiment in a disodium hydrogen phosphate solution having a concentration of 19.2 mM for 5 minutes to grow an amorphous calcium phosphate layer.
  • FIG. 7 (B) shows the surface of a biodegradable fiber obtained by immersing the sample of the amorphous calcium phosphate coating experiment in a disodium hydrogen phosphate solution having a concentration of 19.2 mM for 10 minutes to grow an amorphous calcium phosphate layer. .. FIG.
  • FIG. 8 (A) shows the surface of a biodegradable fiber in which an amorphous calcium phosphate coating experiment sample was immersed in a disodium hydrogen phosphate solution having a concentration of 192 mM for 5 minutes to grow an amorphous calcium phosphate layer.
  • FIG. 8 (B) shows the surface of a biodegradable fiber obtained by immersing a sample of the amorphous calcium phosphate coating experiment in a disodium hydrogen phosphate solution having a concentration of 192 mM for 10 minutes to grow an amorphous calcium phosphate layer.
  • FIG. 8 (B) shows the surface of a biodegradable fiber obtained by immersing a sample of the amorphous calcium phosphate coating experiment in a disodium hydrogen phosphate solution having a concentration of 192 mM for 10 minutes to grow an amorphous calcium phosphate layer.
  • FIG. 9 (A) shows the surface of a biodegradable fiber in which an amorphous calcium phosphate coating experiment sample was immersed in a disodium hydrogen phosphate solution having a concentration of 1.92 M for 5 minutes to grow an amorphous calcium phosphate layer. ..
  • FIG. 9 (B) shows the surface of a biodegradable fiber obtained by immersing a sample of the amorphous calcium phosphate coating experiment in a disodium hydrogen phosphate solution having a concentration of 1.92 M for 10 minutes to grow an amorphous calcium phosphate layer. .. In Fig.
  • Fig. 11 20 mg of the sample (192 mM) was set aside and placed in a test tube, and a weight (56.5 mg) was placed and pressed for 2 min. Then, the weight was removed, and 5 ⁇ l of a red solution prepared by dissolving Rhodamine B in distilled water was dropped on the sample, and after 5 minutes, the red solution permeated the sample.
  • Fig. 11 20 mg of the sample (192 mM) was set aside and placed in a test tube, and a weight (56.5 mg) was placed and pressed for 2 min. Then, the weight was removed, and 5 ⁇ l of a red solution prepared by dissolving Rhodamine B in distilled water was dropped on the sample, and after 5 minutes, the red solution permeated the sample.
  • Fig. 11 20 mg of the sample (192 mM) was set aside and placed in a test tube, and a weight (56.5 mg) was placed and pressed for 2 min. Then, the weight was removed, and 5 ⁇ l of a red
  • FIG. 12 20 mg of the sample (1.92 M) was set aside and placed in a test tube, and a weight (56.5 mg) was placed and pressed for 2 min. Then, the weight was removed, and 5 ⁇ l of a red solution prepared by dissolving Rhodamine B in distilled water was dropped on the sample, and after 10 minutes, the red solution permeated the sample.
  • FIG. 12 20 mg of the sample (1.92 M) was set aside and placed in a test tube, and a weight (56.5 mg) was placed and pressed for 2 min. Then, the weight was removed, and 5 ⁇ l of a red solution prepared by dissolving Rhodamine B in distilled water was dropped on the sample, and after 10 minutes, the red solution permeated the sample.
  • FIG. 13 shows the results of an adsorption test of bovine serum albumin (BSA) on the surface of the biodegradable fiber coated with the amorphous calcium phosphate of the present invention.
  • FIG. 14 shows an evaluation method for evaluating the cell adhesion / proliferation property of the bone regeneration material comprising the biodegradable fiber coated with the amorphous calcium phosphate of the present invention.
  • FIG. 15 shows a diagram of metabolic activity values of cells by Alamar Blue.
  • FIG. 16 shows the morphology of the fiber surface by FE-SEM and the state after the cells have grown. The part surrounded by the line is the part where cells exist.
  • FIG. 17 shows that the CaP phase is confirmed by morphology of the fiber surface by FE-SEM and strong expansion of the fiber surface after 24 hours of culture.
  • FIG. 18 shows the elution behavior of ions from the sample measured by ICP-AES.
  • FIG. 19 shows an example of usage of the cotton-like bone filling material of the present invention.
  • the bone regeneration material of the present invention is composed of biodegradable fibers spun by electrospinning or melt spinning.
  • the biodegradable fiber used in the present invention contains bone-forming particles such as calcium phosphate, and when the material is implanted in the body and the biodegradable resin is decomposed, the bone-forming particles contained in the fiber are changed in the process. Ions are eluted and promote bone formation in the affected area. By collecting the spun biodegradable fibers in the form of cotton or nonwoven fabric, a fibrous bone regeneration material is formed.
  • the thickness of the biodegradable fiber used in the present invention is preferably 10 to 100 ⁇ m in diameter, more preferably 20 to 60 ⁇ m in diameter.
  • the diameter of the biodegradable fiber should be 10 ⁇ m or more. preferable. When the fiber diameter is 10 ⁇ m or less, it becomes difficult to uniformly distribute and fix the particles on the surface of the fiber.
  • Biodegradable fibers having a diameter of 10 ⁇ m or more can be easily spun by using the melt spinning method.
  • the melt spinning method since the amount of inorganic particles that can be mixed with the molten resin is limited, it is difficult to include the bone-forming particles in an amount exceeding 5% by weight.
  • the electrospinning method since the spinning solution is prepared by dissolving the resin or the composite of the resin and the inorganic particles with the solvent, it is possible to include a larger amount of the bone-forming particles.
  • the inventors of the present invention succeeded in spinning a biodegradable fiber containing 70% by weight of inorganic particles by using a thermal kneading method together (Japanese Patent No. 6251462).
  • the solvent is volatilized during the winding process due to the unstable flight trajectory (bending instability) of the spinning solution discharged from the nozzle of the device before reaching the collector.
  • the diameter of the fiber spun by it is usually several tens nm to several ⁇ m, and it is difficult to make the diameter 10 ⁇ m or more.
  • the inventors of the present invention installed a nozzle having a large diameter downward, sent the spinning solution filled in a syringe to the exit of the nozzle at a high speed, and ejected the spinning solution vertically downward to obtain a diameter of 10 ⁇ m or more. Have succeeded in stably spinning the biodegradable fiber (Japanese Patent Application No. 2019-73453).
  • biodegradable resin such as PLA, PLGA, PCL
  • PLA PLGA
  • PCL poly(ethylene glycol)
  • PLGA which is an amorphous resin and has a glass transition point of about 40 ° C.
  • the biodegradable fiber used in the present invention contains a considerable amount of inorganic particles that are bone-forming particles such as calcium phosphate and silicon-eluting calcium carbonate, and the bone regeneration material implanted in the living body is in contact with body fluid. When the biodegradable resin is decomposed, the bone-forming particles contained in the process are eluted to promote bone formation.
  • the bone regeneration material includes an implant material in which biodegradable fibers are collected in a non-woven fabric or cotton form.
  • the operator fills the bone regenerating material with the bone regenerating material as it is, or mixes the material with blood in advance and embeds it in the bone defective portion (see FIG. 19). ).
  • the bone-forming particles refer to inorganic particles that are contained in a bone regenerating material and are implanted into the human body together with the material to elute ions that promote bone formation by coming into contact with body fluid or blood.
  • Bone-forming particles include, but are not limited to, tricalcium phosphate, hydroxyapatite, calcium carbonate, and the like.
  • vaterite phase calcium carbonate particles are preferably used as the inorganic particles to be adhered and fixed on the surface of the biodegradable fiber by using the triboelectric charging method.
  • Calcium carbonate has the most stable calcite phase at room temperature and normal pressure, the metastable aragonite phase, and the metastable vaterite phase.
  • the calcium phosphate phase starts from the particles when immersed in an aqueous solution containing phosphate ions.
  • the vaterite phase which has a high solubility in water, is most suitable for the production of
  • the vaterite phase calcium carbonate particles used in the present invention preferably have a diameter of 0.5 ⁇ m to 4.0 ⁇ m. It is important that the range of the particle size of the vaterite phase calcium carbonate recovered in the form of particles is smaller than the thickness of the biodegradable fiber to which the particles are attached. Furthermore, it is preferable that the particle size is smaller than the larger particle size because the reaction with the aqueous solution containing phosphate ions becomes faster.
  • Siloxane can be supported on the surface of the vaterite phase calcium carbonate particles by the carbonation process (Japanese Patent No. 5131724).
  • the amount of siloxane supported on the surface of the vaterite phase calcium carbonate particles by the carbonation process is increased, the particle diameter increases accordingly, and the maximum particle diameter increases to 4 ⁇ m. If the amount of siloxane is further increased from the level, it becomes difficult to recover the vaterite phase calcium carbonate as particles.
  • the triboelectric charging method refers to a method for adhering and fixing inorganic particles on the surface of polymer fibers. Specifically, when the powder of inorganic particles and the fiber material are housed in a container such as a plastic bag and the container is vibrated up and down or front and back and left and right at high speed with the container in that state, the inorganic particles and the fibers are The resin surface is rubbed to generate static electricity, and the generated electrostatic attraction uniformly attaches the inorganic particles to the entire surface of the fiber material (see FIG. 1). Alternatively, the material can be put into a ball mill and rotated without adding a ball (only by the pot) to uniformly attach the material.
  • the inorganic particles attached to the surface of the fibers by electrostatic attraction can be fixed.
  • the biodegradable fiber is pulled during spinning flight in electrospinning, and in melt spinning, the fiber is pulled when it is wound on a rotating drum and residual stress is generated inside the fiber.
  • heat is applied to the fiber, the fiber shrinks in the longitudinal direction, and during the shrinking process, the inorganic particles adhering to the polymer surface are embedded inside at the contact part with the fiber and firmly fixed at that position (Fig. 2).
  • ⁇ Phosphoric acid treatment> A fibrous material having inorganic particles adhered and fixed on the surface is immersed in an aqueous solution containing phosphate ions, and allowed to stand for a fixed time under a predetermined temperature condition. Then, the fiber material is taken out from the aqueous solution containing phosphate ions and dried (see FIG. 3).
  • an aqueous solution containing a phosphate ion used for the phosphoric acid treatment of the present invention
  • an aqueous solution containing a phosphate ion such as a disodium hydrogen phosphate aqueous solution or a diammonium hydrogen phosphate aqueous solution is preferable.
  • a phosphate ion such as a disodium hydrogen phosphate aqueous solution or a diammonium hydrogen phosphate aqueous solution.
  • amorphous calcium phosphate includes carbonate apatite (Ca 10-x (HPO 4 , CO 3 ) x (PO 4 ) 6-x (OH, CO 3 ) 2-x ⁇ nH 2 O) and / or calcium.
  • Deficient calcium phosphate (Ca 10-x (HPO 4 ) x (PO 4 ) 6-x (OH) 2-x ⁇ nH 2 O may be included.
  • Amorphous calcium phosphate is soluble at around neutral pH Is high and dissolves in a short time on contact with water.
  • an amorphous calcium phosphate layer is formed along the surface of the fiber from the vaterite phase calcium carbonate particles fixed on the surface of the fiber by the triboelectric charging method. And cover the entire surface of the fiber.
  • FIG. 6 shows that, using the method of the present invention, SiV (silicon-eluting calcium carbonate) particles are used as the vaterite phase calcium carbonate to be adhered and fixed on the fiber surface, and disodium hydrogen phosphate (concentration 0) as an aqueous solution containing phosphate ions.
  • the cell culture substrate is composed of a non-woven fabric made of biodegradable fibers spun by the electrospinning method.
  • amorphous calcium phosphate By coating the surface of the biodegradable fiber constituting the cell culture substrate with amorphous calcium phosphate, protein adsorption on the fiber surface is improved, and thus high cell initial adhesion can be obtained.
  • ReBOSSIS registered trademark
  • PLGA 30 wt% / ⁇ -TCP 40 wt % / SiV30 wt%
  • PLLA PGA
  • Evonik LG855S not including D body
  • ⁇ -TCP ⁇ -TCP-100 manufactured by Taihei Chemical Industry Co., Ltd.
  • a ⁇ -TCP crushed product obtained by crushing 1.7 mm in particle size to about 4 ⁇ m was used.
  • ⁇ SiV sicon elution type vaterite phase calcium carbonate: calcium hydroxide (reagent special grade purity of 96% or more Wako Pure Chemical Industries, Ltd.), methanol (reagent special grade purity of 99.8% or more Wako Pure Chemical Industries, Ltd.), ⁇ -Aminopropyltriethoxysilane (SILQUEST A-1100 with a purity of 98.5% or more Momentive Performance Materials Japan LLC), carbon dioxide (high purity liquefied carbon dioxide purity 99.9% Taiyo Chemical Co., Ltd.) was used to prepare a vaterite phase calcium carbonate having a Si content of 2.9% by weight.
  • SiV particle size 1 to 1.5 ⁇ m. Details of the method for producing SiV are disclosed in Japanese Patent Laid-Open No. 2008-100878 (silicon-eluting calcium carbonate and method for producing the same).
  • Step of Adhering SiV Particles to Biodegradable Fibers Constituting ReBOSSIS® 1.
  • Adhesion of SiV particles to the fiber surface 1.0 g of SiV powder and 0.1 g of ReBOSSIS (registered trademark) were put in a glass bottle (5 cm ⁇ ⁇ 10 cm), rotated (264 rpm, 2 min.) For temporary adhesion and heat treatment (60 ° C., 10 min.). Then, electrostatically non-adhering SiV particles existing on the sample surface were removed by a compressor to prepare a sample. Step 2.
  • Step 3 The dried sample was removed from the aqueous disodium hydrogen phosphate solution and then dried in a drier (50 ° C., 2 h). SEM images of the surface of the fiber of the sample obtained as a sample (19.2 mM), a sample (192 mM), and a sample (1.92 M) for each concentration of the disodium hydrogen phosphate aqueous solution are shown in FIG. 7 (A) ( B), FIGS. 8A and 8B, and FIGS. 9A and 9B.
  • the disodium hydrogen phosphate solution concentration was set to 100 mM / L, 150 mM / L for 5 minutes to form a calcium phosphate layer on the surface of the fiber, the disodium hydrogen phosphate solution At a concentration of 100 mM / L, the red solution was not soaked instantly, but at 150 mM / L water was soaked instantly. It is considered that 150 mM / L is more suitable than 100 mM / L as the optimum concentration of the disodium hydrogen phosphate aqueous solution for the water to soak into the water instantly.
  • a higher concentration of disodium hydrogen phosphate is better (eg 192 mM / L), but if the concentration is too high, PLGA will be attacked and SiV particles will fall off. there's a possibility that.
  • the upper limit of the temperature may be limited to 110 ° C, but if it is 60 ° C or higher, the precipitated calcium phosphate phase will be buried again in PLGA, so 50 to 60 ° C is considered optimal.
  • a culture test was performed using osteoblast-like cells MC3T3-E1 strain. The outline is shown in FIG. 80 mg of bone regeneration material was added per well of a 24-well plate, 2 mL of ⁇ -MEM medium was injected, cells were seeded, and a culture test was performed at 37 ° C. in the presence of 5% CO 2 .
  • the cell seeding density was 90,000 cells / mL, and the culture time points were 3 hr, 24 hr, and 72 hr, respectively.
  • the metabolic activity was significantly higher in the bone regenerating material-injected sample than in the control glass plate sample alone, and was significantly higher in the hydrophilized complex than in the untreated complex.
  • the metabolic activity was higher in 24 hr than in 3 hr and in 72 hr than in 24 hr in each sample.
  • the result is shown in FIG. From the morphological observation of the sample surface by FE-SEM, it was found that the hydrophilized composite had a rough surface with larger irregularities than the untreated composite. The cells were covered more with the hydrophilized complex, and the fiber surface was entirely covered with the cells. The result is shown in FIG.
  • the CaP phase was produced on both the untreated complex and the hydrophilized complex on the fiber surface after 24 hours of culturing, and in the latter, the CaP phase was proceeding.
  • the CaP phase was partially formed in the untreated complex, which is probably due to the mechanism similar to that of the biomimetic method. The results are shown in FIG.
  • a biodegradable fiber containing a biodegradable fiber constituting a material for use, and further, a biodegradable fiber containing no inorganic particles and / or a cell culture substrate using such a biodegradable fiber is also coated with an amorphous calcium phosphate layer. As long as it is a material that has been removed, it is similarly applicable.

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Medicinal Chemistry (AREA)
  • Oral & Maxillofacial Surgery (AREA)
  • Transplantation (AREA)
  • Epidemiology (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • Veterinary Medicine (AREA)
  • Dermatology (AREA)
  • Inorganic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Composite Materials (AREA)
  • Materials Engineering (AREA)
  • Textile Engineering (AREA)
  • Materials For Medical Uses (AREA)
  • Chemical Or Physical Treatment Of Fibers (AREA)

Abstract

La surface polymère de fibres biodégradables incluses dans des matériaux pour la régénération osseuse présente une répulsion de l'eau et une hydrophobie et, par conséquent, lorsqu'un tel matériau est implanté dans une zone touchée, le matériau repousse le sang à un stade très précoce du mélange avec le sang et, par conséquent, l'incorporation du matériau pendant une intervention peut prendre une longue période de temps. Selon l'invention, le matériau pour la régénération osseuse présente une surface comprenant des fibres biodégradables hydrophilisées, les fibres biodégradables incluses dans le matériau pour la régénération osseuse contenant des particules ostéogènes et les surfaces des fibres biodégradables sont toutefois pratiquement entièrement recouvertes d'une couche de phosphate de calcium amorphe et une partie du phosphate de calcium de la couche de phosphate de calcium amorphe comprend de l'apatite de carbonate dans laquelle la structure cristalline de phosphate de calcium est remplacée par du carbonate de calcium.
PCT/JP2019/043010 2018-11-02 2019-11-01 Matériau pour la régénération osseuse présentant une surface comprenant des fibres biodégradables hydrophilisées et procédé de production associé WO2020091035A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
JP2020554976A JP7429391B2 (ja) 2018-11-02 2019-11-01 表面が親水化された生分解性繊維からなる骨再生用材料、及びその製造方法

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US201862755387P 2018-11-02 2018-11-02
US62/755,387 2018-11-02

Publications (1)

Publication Number Publication Date
WO2020091035A1 true WO2020091035A1 (fr) 2020-05-07

Family

ID=70461928

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/JP2019/043010 WO2020091035A1 (fr) 2018-11-02 2019-11-01 Matériau pour la régénération osseuse présentant une surface comprenant des fibres biodégradables hydrophilisées et procédé de production associé

Country Status (2)

Country Link
JP (1) JP7429391B2 (fr)
WO (1) WO2020091035A1 (fr)

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH06285151A (ja) * 1993-04-01 1994-10-11 Nippon Sherwood Kk 非晶質リン酸カルシウムをコートした医療用具
WO2017188435A1 (fr) * 2016-04-28 2017-11-02 国立大学法人名古屋工業大学 Procédé de fabrication de matériau de régénération osseuse comprenant des fibres biodégradables au moyen d'un procédé d'électrofilage
JP2019183322A (ja) * 2018-04-10 2019-10-24 株式会社バイオアパタイト 微小繊維

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2004112856A1 (fr) 2003-06-24 2004-12-29 Kyushu Tlo Company Limited Materiau prothetique osseux medical et processus de production de ce materiau

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH06285151A (ja) * 1993-04-01 1994-10-11 Nippon Sherwood Kk 非晶質リン酸カルシウムをコートした医療用具
WO2017188435A1 (fr) * 2016-04-28 2017-11-02 国立大学法人名古屋工業大学 Procédé de fabrication de matériau de régénération osseuse comprenant des fibres biodégradables au moyen d'un procédé d'électrofilage
JP2019183322A (ja) * 2018-04-10 2019-10-24 株式会社バイオアパタイト 微小繊維

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
MUCALO, M.R. ET AL.: "Growth of calcium phosphate on surface-modified cotton", JOURNAL OF MATERIALS SCIENCE :MATERIALS IN MEDICINE, vol. 6, 1995, pages 597 - 605, XP55703978 *

Also Published As

Publication number Publication date
JP7429391B2 (ja) 2024-02-08
JPWO2020091035A1 (ja) 2021-10-21

Similar Documents

Publication Publication Date Title
US9498561B2 (en) Fiber wadding for filling bone defects
Maeda et al. Preparation of poly (L-lactic acid)-polysiloxane-calcium carbonate hybrid membranes for guided bone regeneration
US6340648B1 (en) Calcium phosphate porous sintered body and production thereof
MX2013003089A (es) Aparato de control de presion.
Kasuga et al. Siloxane-poly (lactic acid)-vaterite composites with 3D cotton-like structure
Wutticharoenmongkol et al. Novel bone scaffolds of electrospun polycaprolactone fibers filled with nanoparticles
Sachot et al. Hybrid organic-inorganic scaffolding biomaterials for regenerative therapies
CA2978002A1 (fr) Echafaudages a enrobage cellulosique et methodes de fabrication
EP2682135A1 (fr) Tissu non-tissé contenant une matière prothétique osseuse
JP2006517451A (ja) ナノ結晶表面被膜の方法及び材料ならびにその上へのペプチド両親媒性物質ナノ繊維の付着物
JPS62221358A (ja) 骨欠損部及び空隙部充填材
WO2009156226A2 (fr) Membrane de soie pour matériau de greffe osseuse et procédé de fabrication de celle-ci
EP3691700B1 (fr) Procédé de fabrication d'un article fibreux ostéoconducteur et implant médical comprenant un tel article fibreux ostéoconducteur
US8512732B2 (en) Method for producing bioactive composites
Ramírez-Rodríguez et al. Biomineralized recombinant collagen-based scaffold mimicking native bone enhances mesenchymal stem cell interaction and differentiation
Koh et al. Modification of polyglutamic acid with silanol groups and calcium salts to induce calcification in a simulated body fluid
CN203235057U (zh) 用于填充骨缺损的纤维填料
US20110245922A1 (en) Material for filling bone defects and production method thereof
WO2020091035A1 (fr) Matériau pour la régénération osseuse présentant une surface comprenant des fibres biodégradables hydrophilisées et procédé de production associé
US20120219595A1 (en) Biodegradable fiber and fiber wadding for filling bone defects and method for producing the same
Yamada et al. Development of magnesium and siloxane-containing vaterite and its composite materials for bone regeneration
Pishva et al. Gallic acid-grafted hybrid strontium fluoride/polycaprolactone nanocomposite fibers for bone regeneration
US20110009522A1 (en) Material for filling bone defects and production method thereof
AU2018374827A1 (en) Water-soluble salt particle containing compositions and porous materials made therefrom
JP3718708B2 (ja) リン酸カルシウム系生体用セラミック焼結体およびその製造方法

Legal Events

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

Ref document number: 19880802

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

ENP Entry into the national phase

Ref document number: 2020554976

Country of ref document: JP

Kind code of ref document: A

122 Ep: pct application non-entry in european phase

Ref document number: 19880802

Country of ref document: EP

Kind code of ref document: A1