WO2018072679A1 - Matériau biomimétique artificiel biominéralisé de réparation osseuse et son procédé de préparation et son utilisation - Google Patents

Matériau biomimétique artificiel biominéralisé de réparation osseuse et son procédé de préparation et son utilisation Download PDF

Info

Publication number
WO2018072679A1
WO2018072679A1 PCT/CN2017/106513 CN2017106513W WO2018072679A1 WO 2018072679 A1 WO2018072679 A1 WO 2018072679A1 CN 2017106513 W CN2017106513 W CN 2017106513W WO 2018072679 A1 WO2018072679 A1 WO 2018072679A1
Authority
WO
WIPO (PCT)
Prior art keywords
collagen
biomineralized
mineralized
biomimetic
solution
Prior art date
Application number
PCT/CN2017/106513
Other languages
English (en)
Chinese (zh)
Inventor
张自强
张以河
陈飞旭
Original Assignee
张自强
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 张自强 filed Critical 张自强
Publication of WO2018072679A1 publication Critical patent/WO2018072679A1/fr
Priority to ZA2019/03065A priority Critical patent/ZA201903065B/en

Links

Images

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/02—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
    • A61L27/22—Polypeptides or derivatives thereof, e.g. degradation products
    • A61L27/24—Collagen
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
    • A61L27/56—Porous materials, e.g. foams or sponges
    • 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
    • A61L2400/00—Materials characterised by their function or physical properties
    • A61L2400/12—Nanosized materials, e.g. nanofibres, nanoparticles, nanowires, nanotubes; Nanostructured surfaces
    • 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
    • A61L2430/00—Materials or treatment for tissue regeneration
    • A61L2430/02—Materials or treatment for tissue regeneration for reconstruction of bones; weight-bearing implants

Definitions

  • the invention relates to a biomimetic biomineralized artificial bone and a preparation method and application thereof, and belongs to the field of biomedical materials.
  • Bone implant materials can be roughly divided into autologous bone, allogeneic bone, xenogenic bone, and synthetic materials.
  • Autologous bone grafts are almost limited by the limitations of their own donors; allogeneic bones have better osteogenic effects, but there are also risks of rejection and viral infection, and due to limited sources, it is difficult to achieve a large number of clinically Use; xenogeneic bone is rarely used due to large rejection. Therefore, artificially synthesized bone repair materials with good biological activity and similar to natural bone components have emerged.
  • This material can provide a microenvironment that contributes to the adhesion, proliferation and function of osteoblasts and is similar to natural bone. It can not only be used directly as a bone defect repair material, but also a good bone tissue engineering carrier material, which has opened up a broad prospect for the development of bone tissue engineering.
  • Synthetic bone materials are currently the hotspots of research, and can be divided into metal alloy materials, inorganic ceramic materials, polymer materials and composite materials containing metal/inorganic ceramics/polymer organic composites.
  • metal materials materials mainly used in the past are stainless steel, cobalt alloys, titanium alloys, niobium alloys, memory alloys and the like to replace gold. Since ordinary stainless steel is more susceptible to corrosion as an iron-based alloy, it is now mostly used to repair stainless steel containing molybdenum (Mo) bone, which is more resistant to biochemical corrosion than ordinary stainless steel. The excellent mechanical properties of metal materials can meet the demand, but it is too high to cause stress shielding.
  • ceramic materials mainly includes several types of inorganic ceramic materials such as calcium sulfate filler, inert bioceramic, bioactive ceramics and absorbable ceramics.
  • the composition of general inorganic ceramic materials is not toxic, but the strength and biological activity of the materials do not seem to have both.
  • materials with good bioactivity have poor mechanical properties, and materials with good mechanical properties are not biologically active.
  • polymer materials applied in orthopedics can be roughly classified into two types, namely, natural polymer materials and synthetic polymer materials.
  • Orthopaedic polymer materials are generally not toxic, but their biological activity is low, and the strength of the material is also far from the bone tissue. Therefore, people have developed many new jobs by using the advantages and disadvantages of these materials. Arts and methods have synthesized a variety of composite materials, and materials with different properties are combined in a clever way, in order to meet the different applications. New processes such as nanocomposite, gradient composite, 3D printing, etc. have gradually become the mainstream of orthopedic repair materials, such as L-polylactic acid/hydroxyapatite whisker/collagen liquid crystal scaffold (Wu Di, L-polylactic acid/hydroxyapatite crystal) Preparation and characterization of the whisker/collagen liquid crystal scaffold [D]. Jinan University, 2014.).
  • CN105457097A, CN101554493A are thoroughly mixed with anti-separation agents, dispersants and/or modifiers to obtain nano-hydroxyapatite/collagen scaffolds, but anti-seizure agents, dispersants and/or modifiers in scaffolds.
  • Components that belong to non-natural bone structures affect material structure and osteogenesis.
  • the inventors have realized that there is a need in the art for a bone repair scaffold material having a good biomimetic effect in a three-dimensional structure, but it has not been effectively satisfied for a long time. Therefore, it is urgent to develop a new bone repair scaffold material to meet the reality. need.
  • the present application discloses a biomimetic biomineralized artificial bone repair material, which is a three-dimensional porous scaffold material, which comprises a crosslinked collagen matrix and a biomineralized collagen/hydroxyapatite composite powder wrapped in the matrix.
  • the mass ratio of the collagen matrix to the biomineralized collagen/hydroxyapatite composite powder is from 9:1 to 1:9.
  • the present application further discloses a method for preparing the biomimetic artificial bone repairing material, the method comprising the following steps:
  • step (c) adjusting the pH of the mixed system obtained in the step (b) to 7 to 9, and then stirring to form a mineralized collagen composite sol;
  • the application further discloses the application of the biomimetic mineralized artificial bone repair material.
  • Figure 1 is a flow chart of Embodiments 1 to 7;
  • Example 2 is an electron micrograph of the product obtained in Example 1;
  • 3 and 4 are XRD comparison diagrams in the ninth embodiment.
  • the present application discloses a biomimetic biomineralized artificial bone repair material, which is a three-dimensional porous scaffold material, which comprises a crosslinked collagen matrix and a biomineralized collagen/hydroxyapatite composite powder wrapped in the matrix.
  • the mass ratio of the collagen matrix to the biomineralized collagen/hydroxyapatite composite powder is 9:1 to 1:9, for example, 1 to 5:5 to 9, and for example, 3 to 5:5 to 7. .
  • the mass ratio of the collagen matrix to the composite powder in the range of 3 to 5:5 to 7, the mechanical properties, flexibility, and porosity of the material of the present application are excellently balanced, and the practical application requirements are more satisfied.
  • the bionic biomineralized artificial bone repairing material described in the present application is a collagen-based biomimetic mineralized artificial bone, which solves the long-term desire to provide a three-dimensional bone repair scaffold material mainly composed of collagen and hydroxyapatite, but has not been
  • the technical problems achieved can meet the needs of artificial bone and tissue suture in GTR or GBR clinical operation, including hydroxyapatite and pure collagen, which are the main components of human natural tissue, non-toxic, and do not affect the material. Structure and osteogenic effect, with good biocompatibility. Due to the existence of cross-linked collagen matrix, the problems of material collapse, excessive degradation rate and poor mechanical properties are solved, and the biomimetic construction of nano-hydroxyapatite and collagen is realized.
  • the bionic biomineralized artificial bone of the present application has a good three-dimensional structure, the mature collagen fibers of the triple helix are arranged in an orderly manner, and the biomineralized collagen/hydroxyapatite composite powder is closely attached to the collagen fiber, and is fully filled with Between the collagen fibers, there is a collagen-hydroxyapatite-collagen tertiary composite structure, which is a combination of multiple dimensions, so that the biomineralized collagen/hydroxyapatite composite powder can be stabilized in the three-dimensional porous scaffold structure and is not easy to fall off. , the stability of the three-dimensional support structure is increased, thereby improving the mechanical properties of the material and being not easily damaged.
  • the biomineralized collagen/hydroxyapatite composite powder described in the present application is prepared by biomineralization process, and controls hydroxyapatite at the molecular level through the interaction of collagen organic macromolecules and inorganic ions at the interface. Crystallization and growth of the inorganic mineral phase, so that the collagen/hydroxyapatite composite powder has a special hierarchical structure and assembly method, which can be prepared according to the existing method, for example, by the step (1) in the specific embodiment of CN1106861C ⁇ (4) It is prepared, which is equivalent to the calcium phosphate salt dry powder in the case, and those skilled in the art know that the order of addition of phosphate and calcium ions has no substantial effect on biomineralization.
  • the collagen/biomineralized hydroxyapatite composite powder described in the present application is prepared by biomineralization of collagen, and hydroxyapatite is formed by orderly crystal growth of collagen on the molecular structure of collagen using collagen as a template. Collagen and hydroxyapatite are molecularly oriented. Through biomimetic biomineralization, the molecules re-order into an ordered structure with a more consistent and broader orientation of collagen fibers, resulting in high mechanical strength of the artificial bone.
  • the biomimetic mineralized artificial bone repair material has a pore diameter of 10 to 500 ⁇ m and a porosity of 50% to 97%.
  • the mass fraction of hydroxyapatite in the biomineralized collagen/hydroxyapatite composite powder is from 50 to 99% by weight. In some embodiments it is 80 to 99 wt%.
  • the mass fraction of hydroxyapatite in the composite powder is controlled within the range of 80 to 99 wt%, so that the particle size of the composite powder becomes smaller and the dispersion is more uniform in the collagen matrix.
  • the XRD pattern of the biomimetic mineralized artificial bone repair material is substantially as shown in any of col-HA in FIG. 4 or bionic bone 19, bionic bone 37 or bionic bone 55 in FIG.
  • the biomimetic mineralized artificial bone repair material has a high degree of similarity to natural bone, particularly bionic bone 37 or bionic bone 55.
  • the collagen matrix or the collagen in the biomineralized collagen/hydroxyapatite composite powder is a native type I collagen of an atelopeptide and/or a recombinant human type I collagen of an atelopeptide.
  • the natural type I collagen can extract skin and/or Achilles tendon of an animal (eg, pig, cow, sheep, horse, etc.), and is subjected to an enzymatic process (which may further include dialysis purification, etc.). The resulting depeptide peptide collagen.
  • the present application discloses a preparation method of the biomimetic mineralized artificial bone repairing material, wherein the method comprises the following steps:
  • step (c) adjusting the pH of the mixed system obtained in step (b) is 7 to 9, followed by stirring to form a mineralized collagen composite sol; in some embodiments, the pH of the resulting mixed system is adjusted to 7 to 9 using NaOH, KOH or ammonia;
  • the mineralized collagen composite sol obtained in the step (c) is pre-frozen to obtain a mineralized collagen jelly, and then the freeze-drying is performed;
  • the resulting biomimetic mineralized artificial bone repair material can be cut into a desired shape and packaged aseptically.
  • the crosslinking in step (e) comprises physical crosslinking and/or chemical crosslinking.
  • the physical crosslinking includes, but is not limited to, one or more of ultraviolet radiation, thermal dehydrogenation, and radiation sterilization crosslinking.
  • the chemical crosslinking includes, but is not limited to, the use of carbodiimide, diamine, epoxy compound, hydroxysuccinimide, diphenyl phosphate (DPPA), glutaraldehyde, formaldehyde, B.
  • DPPA diphenyl phosphate
  • glutaraldehyde glutaraldehyde
  • formaldehyde B.
  • One or more of aldehyde acid and genipin are crosslinked, and after using a chemical crosslinking agent, the residual crosslinking agent is removed by an elution procedure.
  • biomineralized collagen/hydroxyapatite composite powder described herein is prepared as follows:
  • step (iii) adding a phosphoric acid solution to the solution obtained in step (ii), stirring and mixing to form a mixed system; the phosphoric acid solution is added in an amount such that the molar ratio of calcium ions to phosphate ions in the system is, for example, 1.5 to 2;
  • the white suspension obtained in the step (iii) is allowed to stand, for example, at 15 to 25 ° C for 22 to 26 hours, filtered, and the filter cake is freeze-dried, for example, sufficiently frozen at -50 to -20 ° C. Drying for 24 to 48 hours, followed by grinding the prepared dry powder is biomineralized collagen / hydroxyapatite composite powder;
  • the acid in step (i) includes, but is not limited to, one or more of sulfuric acid, hydrochloric acid, nitric acid, acetic acid, propionic acid, and citric acid;
  • the calcium ion-containing aqueous solution described in step (ii) is formed using one or more of CaCl 2 , Ca(NO 3 ) 2 , and CaCO 3 .
  • the above method for preparing biomineralized collagen/hydroxyapatite in the present application can effectively control the precipitation effect of the powder, and is more advantageous for obtaining a composite powder having a smaller particle diameter than the method known to the inventors, thereby making the obtained composite powder Disperse more evenly in the collagen matrix.
  • the method for preparing the biomimetic mineralized artificial bone repairing material of the present application comprises the following steps:
  • (S1-1) dissolving collagen in an acid solution to prepare an acidic aqueous solution of collagen; in some embodiments, the mass fraction of collagen in the acidic aqueous solution is 0.01% to 2%, and the pH is 2.5 to 5.5. ;
  • step (S1-2) continuously stirring the solution obtained in the step (S1-1), slowly adding an aqueous solution containing calcium ions; in some embodiments, the amount of calcium ions added per gram of collagen in the acidic aqueous solution of collagen is 0.01 to 0.1 mol;
  • step (S1-3) continuously stirring the solution obtained in the step (S1-2), adding a phosphoric acid solution, and stirring and mixing to form a mixed system; in some embodiments, the amount of the phosphoric acid solution added depends on the calcium ion in the step (S1-2). The amount of addition, so that the molar ratio of calcium ions to phosphate ions reaches 1.5 to 2;
  • the mixed system obtained in the step (S1-4) is allowed to stand, and in some embodiments, it is allowed to stand at 15 to 25 ° C for 22 to 26 hours, filtered, and the filter cake is freeze-dried, in some embodiments. Fully freeze-drying at -50 to -20 ° C for 24 to 48 hours, followed by grinding to obtain a dry powder which is a biomineralized collagen/hydroxyapatite composite powder;
  • (S2) a composite of collagen and biomineralized collagen/hydroxyapatite composite powder, comprising:
  • (S2-1) dissolving collagen in purified water to prepare an aqueous solution of collagen; in some embodiments, the mass fraction of collagen in the aqueous collagen solution is 0.8% to 5%;
  • step (S2-2) continuously stirring the solution obtained in the step (S2-1), adding the biomineralized collagen/hydroxyapatite composite powder obtained in the step (S1-5), stirring and mixing to form a mixed system, biomineralized collagen/
  • the ratio of the amount of the hydroxyapatite composite powder added to the amount of collagen protein in the aqueous collagen solution obtained in the step (S2-1) is from 9:1 to 1:9, and in some embodiments, from 1 to 5:5 to 9, In some embodiments, it is 3 to 5: 5 to 7;
  • the mixed system obtained in the step (S2-3) is allowed to stand and stirred to form a mineralized collagen composite solution; in some embodiments, it is allowed to stand at 15 to 25 ° C for 10 to 48 hours, and stirred for 5 to 30. Minutes to make it fully mixed to form a mineralized collagen composite solution;
  • (S3-1) pre-freezing the mineralized collagen composite solution obtained in the step (S2-4), and in some embodiments, pre-freezing at 1 to 5 ° C for 3 to 24 hours to obtain a mineralized collagen jelly;
  • step (S3-2) freeze-drying the mineralized collagen of step (S3-1), and in some embodiments, performing sufficient freeze-drying at -50 to -20 ° C for 22 to 26 hours to obtain a mineralized collagen composite porous material;
  • the biomimetic collagen composite porous material is crosslinked by physical and/or chemical methods to obtain the biomimetic mineralized artificial bone repair material.
  • the method described in the present application has a simple production process, is easy to be mass-produced, and has low production cost, and is very beneficial for reducing the medical cost of the patient.
  • the present application discloses the use of the biomimetic mineralized artificial bone repair material or the biomimetic biomineralized artificial bone repair material prepared in the present application for preparing an implant at a location supporting a bone defect of a human or animal.
  • the bone repair material of the present application When the bone repair material of the present application is placed as an implant at a bone defect position, it is used as a three-dimensional stent for bone formation, bone regeneration, bone repair or bone replacement, or for repair and regeneration of bone defects such as bones and extraction sockets.
  • the biomimetic artificial bone has a collagen-hydroxyapatite-collagen tertiary composite structure, the triple-helix mature collagen fibers are arranged in an orderly manner, and the biomineralized collagen/hydroxyapatite composite powder is closely arranged. Attached to collagen fibers and fully filled between collagen fibers, it has a collagen-hydroxyapatite-collagen tertiary composite structure, which is a combination of multiple dimensions, enabling biomineralized collagen/hydroxyapatite composite powder to Stabilized in the three-dimensional porous scaffold structure, so that the hydroxyapatite crystal is not easy to fall off, and the stability of the hydroxyapatite in the three-dimensional scaffold structure is increased.
  • the biggest feature of the biomimetic artificial bone is the ordered three-dimensional interconnected structure of collagen and hydroxyapatite.
  • the interconnected pore structure facilitates the communication of the blood vessels deep into the material to ensure the growth of the material.
  • the nutrient supply of deep tissues; in addition to the combination of materials and implanted beds, the porous structure is the growth of the body's bone tissue, the formation of mechanical interlocks, and the combination of implant materials, creating conditions for retention; porous surfaces are beneficial for blood vessels And the soft tissue grows in, and the pores communicate with each other and are the prerequisites for the inward growth of the bone.
  • Interconnecting is beneficial to the flow of cells and body fluids in the body, tissue metabolism, bone growth into the micropores and a large number of external epiphyses to form a solid "biological fixation.” "Micropores constitute a large surface area for bone sedimentation Provide a good matrix.
  • the biomimetic artificial bone is composed of hydroxyapatite and pure collagen, and does not use polylactic acid, chitosan, chitin, anti-killing agent, templating agent, dispersing agent, modifier, etc., and does not affect Material structure and osteogenic effect, non-toxic, with good biocompatibility; and made with the use of the terminal peptide collagen, no immunogenicity; achieved simultaneous biomimetic composition and structure.
  • Biomineralized collagen/hydroxyapatite composite powder is mostly nano-scale; nano-sized hydroxyapatite particles are more common than ordinary hydroxy-phosphorus due to the unique properties of nano-materials such as surface effect, small size effect and quantum effect. Graystone has stronger biological activity.
  • the artificial bone made of nano-sized hydroxyapatite particles is white or yellowish-fibrous.
  • the surface is smooth and opaque. It has a porous three-dimensional network structure, which is beneficial to cell adhesion and growth.
  • FT/IR-6800 FT-IR spectrometer Jasco, Japan
  • D8 Focus X-ray diffractometer (Bruker, Germany)
  • HP-500 digital display force gauge (Yueqing Aidebao Instrument Co., Ltd.)
  • GLZ-2 vacuum freeze dryer (Shanghai Pudong Freeze Drying Equipment Co., Ltd.)
  • DZF-6050AF vacuum drying oven Teianjin Gongxing Laboratory Instrument Co., Ltd.
  • 2XZ-2 rotary vane vacuum pump Shanghai Double Goose Refrigeration
  • the preparation of the bionic biomineralized artificial bone is carried out by using an acid solution using hydrochloric acid, a calcium ion solution using a CaCl 2 aqueous solution, an alkaline solution using an aqueous NaOH solution, and an ultraviolet irradiation physical crosslinking method.
  • FIG. 1 is a flow chart showing a method for preparing a biomimetic biomineralized artificial bone of the present application.
  • the preparation method of the biomimetic biomineralized artificial bone is:
  • Step S1 preparation of biomineralized collagen/hydroxyapatite composite powder specifically includes:
  • Step S1-1 dissolved collagen: 2g of collagen is dissolved in 1000mL of a concentration of 0.001mol / L of hydrochloric acid solution, formulated into an acid solution of collagen, wherein the collagen concentration is 0.2%, the pH of the solution is 3;
  • Step S1-2 (adding calcium ions): continuously stirring the solution obtained in the step S1-1, slowly adding 200 mL of a 0.2 mol/L CaCl 2 aqueous solution;
  • Step S1-3 (adding phosphoric acid): continuously stirring the solution obtained in the step S1-2, adding 400 mL of a phosphoric acid solution having a concentration of 0.06 mol/L, stirring and mixing to form a mixed system, and the amount of the phosphoric acid solution added depends on the step S1- 2 the amount of calcium ions added, so that the molar ratio of calcium ions to phosphate ions reaches 1.67;
  • Step S1-5 filtering, freeze-drying, and grinding to obtain a composite powder:
  • the mixed system obtained in the step S1-4 is allowed to stand at 15 to 25 ° C for 24 hours, filtered, and the filter cake is sufficiently freeze-dried at -40 ° C. After hours, the dry powder obtained by the subsequent grinding is a biomineralized collagen/hydroxyapatite composite powder.
  • Step S2 (composite of collagen and biomineralized collagen/hydroxyapatite composite powder) specifically includes:
  • Step S2-1 dissolved collagen: 3 g of collagen is dissolved in 97 mL of purified water to prepare a solution of collagen, wherein the collagen concentration is 3%;
  • Step S2-2 (blending): continuously stirring the solution obtained in the step S2-1, adding 3 g of the biomineralized collagen/hydroxyapatite composite powder prepared in the step S1-5, stirring and mixing to form a mixed system;
  • Step S2-4 Standing, stirring: The mixed system obtained in the step S2-3 is allowed to stand at 25 ° C for 24 hours, poured into a blender and stirred for 15 minutes to be thoroughly mixed, and the mixed system becomes a white emulsion to form a ore. Collagen complex solution;
  • Step S3 preparation of mineralized collagen composite porous material specifically includes:
  • Step S3-1 filling, pre-freezing: taking the mineralized collagen composite solution obtained in step S2-4 into a rectangular mold and flattening, pre-freezing at 4 ° C for 3 hours to obtain mineralized collagen jelly;
  • Step S3-2 freeze-drying: the mineralized collagen gelatinized in step S3-1 is freeze-dried at -50 ° C for 24 hours to obtain a mineralized collagen composite porous material;
  • Step S3-3 crosslinking: crosslinking the mineralized collagen composite porous material by ultraviolet irradiation, and the ultraviolet irradiation condition is: ultraviolet light with a wavelength of 254 nm, and irradiation with 10 W for 24 hours;
  • Step S3-4 cutting, aseptic packaging, sterilization: The cross-linked material is cut into a desired shape, and then sterilized and used after being aseptically packaged.
  • the biomimetic artificial bone (numbered as bionic bone 55) has a three-layer composite structure of collagen-hydroxyapatite-collagen, and adopts a block-shaped material with regular shape, and measures the length, width, height and weight, respectively, and is calculated.
  • the apparent density is 0.13 g/cm 3 and the porosity is 65%.
  • the electron micrograph is shown in Fig. 2.
  • the biomimetic artificial bone is prepared by using hydrochloric acid, a calcium ion solution using a CaCl 2 aqueous solution, an alkaline solution using an aqueous NaOH solution, and glutaraldehyde for chemical crosslinking.
  • the mineralized collagen composite porous material is prepared according to steps S1-1 to S3-2 in Example 1, and then, the mineralized collagen composite porous material is chemically crosslinked using glutaraldehyde, specifically, the ore obtained in step S3-2.
  • the collagen composite porous material is immersed in 0.05 wt% of glutaraldehyde in anhydrous ethanol for 48 hours for crosslinking; then the crosslinked mineralized collagen composite porous material is taken out and placed in a chromatography column to flow pure
  • the water was washed for 48 hours to remove residual cross-linking agent; the cross-linked mineralized collagen composite porous material was vacuum dried at 110 ° C for 48 hours to obtain the bionic biomineralized artificial bone of the present example.
  • the bionic biomineralized artificial bone of the present embodiment was tested to have an apparent density of 0.24 g/cm 3 and a porosity of 85%.
  • the preparation of the biomimetic biomineralized artificial bone is carried out by using an aqueous solution of nitric acid in an acid solution, an aqueous solution of Ca(NO 3 ) 2 in a calcium ion solution, and an aqueous solution of KOH in an alkaline solution, and physically crosslinking using a thermal dehydrogenation method.
  • the biomimetic biomineralized artificial bone was prepared according to the substantially same procedure as in Example 1, except that the acid solution in the step S1-1 was replaced with 1000 mL of a 0.001 mol/L aqueous solution of nitric acid, and the calcium ion solution in the step S1-2 was 20 mL.
  • the Ca(NO 3 ) 2 aqueous solution having a concentration of 0.1 mol/L is substituted for the CaCl 2 aqueous solution, and the alkaline aqueous solution in the step S1-4 and the step S2-3 is replaced with the NaOH aqueous solution having a concentration of 0.01 mol/L, after the step S3-2.
  • the mineralized collagen composite porous material is physically crosslinked by a method of thermal dehydrogenation, and the parameters of the method for thermal dehydrogenation are a temperature of 130 ° C and a vacuum degree of -0.1 MPa, and the remaining steps are the same as in the first embodiment.
  • the biomimetic biomineralized artificial bone prepared in the present example was tested to have an apparent density of 0.15 g/cm 3 and a porosity of 70%.
  • the biomimetic biomineral artificial bone is prepared, and the calcium ion aqueous solution is CaCO 3 and physically crosslinked by a thermal dehydrogenation method.
  • the biomimetic biomineralized artificial bone is prepared according to the substantially same procedure as in the embodiment 1, except that the calcium ion solution in the step S1-2 is CaCO 3 particles, and after the step S3-2, the method for thermally dehydrogenating the mineralized collagen composite porous material is used.
  • the parameters of the method for thermal dehydrogenation were as follows: temperature 130 ° C, vacuum degree -0.1 MPa, and the remaining steps were the same as in Example 1.
  • the bionic biomineralized artificial bone of the present embodiment was tested to have an apparent density of 0.14 g/cm 3 and a porosity of 72%.
  • the biomimetic artificial bone is prepared by using an aqueous solution of acetic acid in an acid solution, an aqueous solution of CaCl 2 in an aqueous solution of calcium ions, and an aqueous solution of Na 2 CO 3 in an aqueous alkaline solution, which is chemically crosslinked using formaldehyde.
  • the preparation of the biomimetic biomineralized artificial bone is carried out by using an aqueous solution of acetic acid as an aqueous solution, an aqueous solution of calcium ions as CaCl 2 , an aqueous alkaline solution using KOH, and chemical crosslinking using formaldehyde.
  • the biomimetic biomineralized artificial bone according to the substantially same procedure as in Example 1, except that the acid solution in the step S1-1 is replaced with 1000 mL of 0.01 mol/L acetic acid instead of the hydrochloric acid solution, and the alkali in the step S1-4 and the step S2-3.
  • the solution was replaced by KOH with a concentration of 0.02 mol/L.
  • the mineralized collagen composite porous material was chemically crosslinked using a 0.025 wt% formaldehyde anhydrous ethanol solution, and the other steps were the same as in Example 1.
  • the bionic biomineralized artificial bone of the present embodiment was tested to have an apparent density of 0.31 g/cm 3 and a porosity of 66%.
  • the preparation of the biomimetic biomineralized artificial bone is carried out by using an aqueous solution of citric acid as an acid solution, an aqueous solution of CaSO 4 in an aqueous solution of calcium ions, and an aqueous solution of ammonia in an alkaline solution, and chemical crosslinking using genipin.
  • biomimetic biomineralized artificial bone according to substantially the same procedure as in Example 1, except that the acid solution in step S1-1 is replaced by 1000 mL of 0.01 mol/L citric acid instead of hydrochloric acid solution, in steps S1-4 and S2-3.
  • the alkaline solution was replaced with aqueous ammonia in the aqueous solution of NaOH.
  • the mineralized collagen composite porous material was chemically crosslinked using a 0.5 wt% aqueous solution of genipin, and the remaining steps were the same as in Example 1.
  • the bionic biomineralized artificial bone of the present embodiment was tested to have an apparent density of 0.25 g/cm 3 and a porosity of 74%.
  • step S2-1 and S2-2 are different from the first embodiment; the specific steps of the steps S2-1 and S2-2 are:
  • Step S2-1 Dissolving 1 g of collagen in 99 mL of purified water to prepare a solution of collagen, wherein the collagen concentration is 1%;
  • Step S2-2 (blending): continuously stirring the solution obtained in the step S2-1, adding 9 g of the biomineralized collagen/hydroxyapatite composite powder obtained in the step S1-5, stirring and mixing to form a mixed system;
  • biomimetic biomineralized artificial bone prepared in this example is numbered as bionic bone 19.
  • Step S2-1 and Step S2-2 are different from Embodiment 1;
  • the specific steps of S2-1 and S2-2 are:
  • Step S2-1 Dissolving 3 g of collagen in 97 mL of purified water to prepare a solution of collagen, wherein the collagen concentration is 3%;
  • Step S2-2 (blending): continuously stirring the solution obtained in the step S2-1, adding 7 g of the biomineralized collagen/hydroxyapatite composite powder prepared in the step S1-5, stirring and mixing to form a mixed system;
  • biomimetic biomineralized artificial bone prepared in this example is numbered as bionic bone 37.
  • Example 3 is an XRD pattern of natural bone (porcine bone), hydroxyapatite (HA), and a sample prepared in Example 1 (Col-HA). Comparing the two maps of HA and Col-HA in Fig. 3, it is found that the diffraction peak of the biomimetic artificial bone is consistent with the diffraction peak spectrum of the standard hydroxyapatite crystal, indicating that the inorganic phase in the biomimetic artificial bone is mainly hydroxyl. apatite. Comparing the two maps of Col-HA and pig bone in Figure 3, it is found that the positions and numbers of the diffraction peaks of the biomimetic artificial bone and the natural bone are consistent, indicating that the inorganic phase in the composite is similar to the natural bone inorganic phase.
  • Figure 4 is an XRD pattern of collagen, hydroxyapatite, bionic bone 19, bionic bone 37, bionic bone 55 and natural rabbit bone. From Figure 4, the ratio of biomineralized collagen/hydroxyapatite composite powder is At 3:7 and 5:5, the crystal phase structure and crystallinity of natural bone are more similar, and the bionic effect is more obvious.
  • the stability of the prepared biomimetic bone 19, bionic bone 37 and bionic bone 55 in the prepared biomimetic bone 19 was observed by immersion shaking in physiological saline to observe the stability of the hydroxyapatite.
  • the specific steps are as follows:

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Epidemiology (AREA)
  • Dermatology (AREA)
  • Medicinal Chemistry (AREA)
  • Oral & Maxillofacial Surgery (AREA)
  • Transplantation (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Biophysics (AREA)
  • Inorganic Chemistry (AREA)
  • Dispersion Chemistry (AREA)
  • Materials For Medical Uses (AREA)

Abstract

L'invention concerne un matériau biomimétique artificiel biominéralisé de réparation osseuse et son procédé de préparation et son utilisation. Le matériau comprend une matrice de collagène réticulé et une poudre composite biominéralisée à base de collagène/hydroxyapatite dispersée dans la matrice. Le rapport pondéral entre la matrice de collagène et la poudre composite biominéralisée à base de collagène/hydroxyapatite est égal à 9:1 à 1:9. Ce matériau résout les problèmes que représentent une trop grande tendance à la collapsibilité, une vitesse de dégradation rapide et des propriétés mécaniques médiocres des matériaux, et permet une construction biomimétique à base de nano-hydroxyapatite et de collagène.
PCT/CN2017/106513 2016-10-17 2017-10-17 Matériau biomimétique artificiel biominéralisé de réparation osseuse et son procédé de préparation et son utilisation WO2018072679A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
ZA2019/03065A ZA201903065B (en) 2016-10-17 2019-05-16 Biomimetic biomineralized artificial bone repair material and preparation method therefor and use thereof

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN201610902145.X 2016-10-17
CN201610902145.XA CN107952115B (zh) 2016-10-17 2016-10-17 一种仿生生物矿化人工骨修复材料及其制备方法与应用

Publications (1)

Publication Number Publication Date
WO2018072679A1 true WO2018072679A1 (fr) 2018-04-26

Family

ID=61953366

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2017/106513 WO2018072679A1 (fr) 2016-10-17 2017-10-17 Matériau biomimétique artificiel biominéralisé de réparation osseuse et son procédé de préparation et son utilisation

Country Status (3)

Country Link
CN (1) CN107952115B (fr)
WO (1) WO2018072679A1 (fr)
ZA (1) ZA201903065B (fr)

Cited By (13)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN112773938A (zh) * 2019-11-06 2021-05-11 湘潭大学 一种基于马钱子碱的3d打印人工骨组合物和制备基于马钱子碱的3d打印人工骨的方法
CN114404669A (zh) * 2022-02-08 2022-04-29 博纳格科技(天津)有限公司 一种合成矿化胶原骨塞及其制备方法
WO2022089287A1 (fr) * 2020-10-26 2022-05-05 杜明春 Procédé de préparation et procédé d'utilisation d'une solution de collagène biomimétique pour la surface de la peau
CN114504677A (zh) * 2022-01-11 2022-05-17 武汉亚洲生物材料有限公司 一种3d打印颅骨修复钛网及其制备方法
CN114525630A (zh) * 2022-02-28 2022-05-24 中国人民解放军东部战区总医院 一种复合纤维支架及其制备方法和应用
CN114621484A (zh) * 2022-04-01 2022-06-14 华中科技大学同济医学院附属协和医院 一种仿生矿化骨修复支架的制备方法
CN115105630A (zh) * 2022-08-08 2022-09-27 池州学院 一种内嵌有壳聚糖/明胶复合水凝胶的3d打印材料及其制备方法
CN115120774A (zh) * 2022-06-28 2022-09-30 奥精医疗科技股份有限公司 一种碳纳米管强化矿化胶原材料及其制备方法和应用
CN115607737A (zh) * 2022-09-08 2023-01-17 上海市第六人民医院 一种复合骨仿生支架及其制备方法
CN115721776A (zh) * 2022-11-10 2023-03-03 奥精医疗科技股份有限公司 一种儿童颅骨修复材料的制备方法
CN116271210A (zh) * 2023-04-21 2023-06-23 国纳之星(上海)纳米科技发展有限公司 一种仿生骨修复涂层改性的钛合金及其制备方法
CN117339016A (zh) * 2023-12-04 2024-01-05 北京大学口腔医学院 一种含纳米金簇的仿生复合材料及其制备方法和应用
RU2822395C2 (ru) * 2019-06-14 2024-07-04 Гайстлих Фарма Аг Коллагеновый матрикс или гранулированная смесь костнозамещающего материала

Families Citing this family (15)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109381745B (zh) * 2018-09-21 2021-03-19 北京市意华健科贸有限责任公司 胶原珊瑚羟基磷灰石复合骨填充支架材料及其材料制备方法
CN109734935A (zh) * 2018-12-29 2019-05-10 广西紫荆生物科技有限公司 一种生物医用可注射凝胶材料及其制备方法和用途
CN110787324B (zh) * 2019-11-03 2021-11-12 太原理工大学 一种药物控释型聚乳酸基骨修复支架材料的制备方法
CN114162799B (zh) * 2021-09-14 2023-04-18 华中科技大学 一种胶原模板仿生矿化锌锶掺杂羟基磷灰石及其制备方法
CN113984853B (zh) * 2021-09-29 2024-03-01 广东省科学院健康医学研究所 一种用于体外降解和模拟矿化的设备及其使用方法
CN114010838B (zh) * 2021-11-15 2022-08-23 中国人民解放军总医院第八医学中心 一种复合材料仿生骨制备方法
CN113913961B (zh) * 2021-11-16 2023-05-16 清华大学 一种活性元素掺杂的矿化胶原纳米纤维及其制备方法
CN114081996A (zh) * 2021-11-16 2022-02-25 清华大学 一种高矿物含量诱导骨再生纤维膜
CN114262374A (zh) * 2021-12-28 2022-04-01 中国科学院烟台海岸带研究所 一种鱼胶原蛋白基仿生骨材料及其制备方法
CN115054743B (zh) * 2022-07-06 2023-12-26 西岭(镇江)医疗科技有限公司 一种可促进牙槽骨再生的屏障膜及其制备方法
CN115444988B (zh) * 2022-08-31 2023-09-29 浙江工业大学 一种高骨活性聚醚醚酮多孔复合材料及其制备方法
CN116570772B (zh) * 2023-06-16 2023-12-19 中山大学附属口腔医院 一种胶原水凝胶及其合成方法和应用
CN117138113A (zh) * 2023-08-25 2023-12-01 重庆生物智能制造研究院 一种矿化胶原材料及其制备方法和应用
CN118141987A (zh) * 2024-03-01 2024-06-07 山西锦波生物医药股份有限公司 一种a型重组人源化胶原蛋白骨修复材料及其制备方法和用途
CN118286511B (zh) * 2024-04-28 2024-10-15 北京工业大学 一种基于动态交联的矿化胶原制备方法及所得矿化胶原

Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6300315B1 (en) * 1999-08-28 2001-10-09 Ceramedical, Inc. Mineralized collagen membrane and method of making same
WO2008076604A1 (fr) * 2006-12-21 2008-06-26 Zimmer Orthobiologics, Inc. Particules de croissance osseuse et composition ostéoinductive de celles-ci
CN101417145A (zh) * 2008-12-04 2009-04-29 山东大学 一种用于骨组织工程的支架材料及其制备方法
CN101791438A (zh) * 2010-03-16 2010-08-04 浙江大学 骨修复用生物活性聚(乳酸-羟基乙酸)/胶原/羟基磷灰石复合纤维膜的制备方法
CN103830775A (zh) * 2012-11-27 2014-06-04 北京奥精医药科技有限公司 高强度胶原基人工骨修复材料
CN104096268A (zh) * 2014-06-19 2014-10-15 北京奥精医药科技有限公司 一种矿化胶原人工骨膜及其制备方法
CN105457097A (zh) * 2015-11-12 2016-04-06 谢宝钢 一种可注射矿化胶原骨修复材料及制备方法

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN100341588C (zh) * 2005-10-10 2007-10-10 胡庆柳 多孔胶原复合纳米羟基磷灰石人工骨的制备方法

Patent Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6300315B1 (en) * 1999-08-28 2001-10-09 Ceramedical, Inc. Mineralized collagen membrane and method of making same
WO2008076604A1 (fr) * 2006-12-21 2008-06-26 Zimmer Orthobiologics, Inc. Particules de croissance osseuse et composition ostéoinductive de celles-ci
CN101417145A (zh) * 2008-12-04 2009-04-29 山东大学 一种用于骨组织工程的支架材料及其制备方法
CN101791438A (zh) * 2010-03-16 2010-08-04 浙江大学 骨修复用生物活性聚(乳酸-羟基乙酸)/胶原/羟基磷灰石复合纤维膜的制备方法
CN103830775A (zh) * 2012-11-27 2014-06-04 北京奥精医药科技有限公司 高强度胶原基人工骨修复材料
CN104096268A (zh) * 2014-06-19 2014-10-15 北京奥精医药科技有限公司 一种矿化胶原人工骨膜及其制备方法
CN105457097A (zh) * 2015-11-12 2016-04-06 谢宝钢 一种可注射矿化胶原骨修复材料及制备方法

Cited By (18)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
RU2822395C2 (ru) * 2019-06-14 2024-07-04 Гайстлих Фарма Аг Коллагеновый матрикс или гранулированная смесь костнозамещающего материала
CN112773938A (zh) * 2019-11-06 2021-05-11 湘潭大学 一种基于马钱子碱的3d打印人工骨组合物和制备基于马钱子碱的3d打印人工骨的方法
WO2022089287A1 (fr) * 2020-10-26 2022-05-05 杜明春 Procédé de préparation et procédé d'utilisation d'une solution de collagène biomimétique pour la surface de la peau
CN114504677B (zh) * 2022-01-11 2023-03-10 武汉亚洲生物材料有限公司 一种3d打印颅骨修复钛网及其制备方法
CN114504677A (zh) * 2022-01-11 2022-05-17 武汉亚洲生物材料有限公司 一种3d打印颅骨修复钛网及其制备方法
CN114404669A (zh) * 2022-02-08 2022-04-29 博纳格科技(天津)有限公司 一种合成矿化胶原骨塞及其制备方法
CN114525630A (zh) * 2022-02-28 2022-05-24 中国人民解放军东部战区总医院 一种复合纤维支架及其制备方法和应用
CN114621484A (zh) * 2022-04-01 2022-06-14 华中科技大学同济医学院附属协和医院 一种仿生矿化骨修复支架的制备方法
CN115120774A (zh) * 2022-06-28 2022-09-30 奥精医疗科技股份有限公司 一种碳纳米管强化矿化胶原材料及其制备方法和应用
CN115120774B (zh) * 2022-06-28 2024-05-28 奥精医疗科技股份有限公司 一种碳纳米管强化矿化胶原材料及其制备方法和应用
CN115105630B (zh) * 2022-08-08 2023-08-18 池州学院 一种内嵌有壳聚糖/明胶复合水凝胶的3d打印材料及其制备方法
CN115105630A (zh) * 2022-08-08 2022-09-27 池州学院 一种内嵌有壳聚糖/明胶复合水凝胶的3d打印材料及其制备方法
CN115607737A (zh) * 2022-09-08 2023-01-17 上海市第六人民医院 一种复合骨仿生支架及其制备方法
CN115721776A (zh) * 2022-11-10 2023-03-03 奥精医疗科技股份有限公司 一种儿童颅骨修复材料的制备方法
CN115721776B (zh) * 2022-11-10 2024-02-20 奥精医疗科技股份有限公司 一种儿童颅骨修复材料的制备方法
CN116271210A (zh) * 2023-04-21 2023-06-23 国纳之星(上海)纳米科技发展有限公司 一种仿生骨修复涂层改性的钛合金及其制备方法
CN117339016A (zh) * 2023-12-04 2024-01-05 北京大学口腔医学院 一种含纳米金簇的仿生复合材料及其制备方法和应用
CN117339016B (zh) * 2023-12-04 2024-03-01 北京大学口腔医学院 一种含纳米金簇的仿生复合材料及其制备方法和应用

Also Published As

Publication number Publication date
ZA201903065B (en) 2020-09-30
CN107952115A (zh) 2018-04-24
CN107952115B (zh) 2021-10-26

Similar Documents

Publication Publication Date Title
WO2018072679A1 (fr) Matériau biomimétique artificiel biominéralisé de réparation osseuse et son procédé de préparation et son utilisation
Sun et al. 3D printed calcium phosphate scaffolds with controlled release of osteogenic drugs for bone regeneration
CN106310380B (zh) 一种纳米纤维化丝素蛋白凝胶及其制备方法
CN111097068B (zh) 一种仿生的羟基磷灰石粉体/明胶/海藻酸钠复合3d打印支架及其制备方法
US20070254007A1 (en) Chitosan/nanocrystalline hydroxyapatite composite microsphere-based scaffolds
Li et al. The degradation regulation of 3D printed scaffolds for promotion of osteogenesis and in vivo tracking
Ghomi et al. A novel investigation on characterization of bioactive glass cement and chitosan-gelatin membrane for jawbone tissue engineering
CN104001208B (zh) 一种生物可降解高分子/甲壳素纳米晶复合支架材料的制备方法
TWI517866B (zh) Porous bone filling material
CN104906637A (zh) 一种可注射-多孔-载药的聚甲基丙烯酸甲酯基复合支架骨移植材料及其制备方法
CN101264341A (zh) 三维多孔组织工程支架材料、其制备及应用
CN111317860A (zh) 一种覆膜生物陶瓷人工骨及制备方法
KR20070010919A (ko) 다공성의 β-인산삼칼슘 과립의 제조 방법
CN105521525A (zh) 一种骨组织工程用多孔复合支架及其制备方法
Van Ho et al. Novel TOCNF reinforced injectable alginate/β-tricalcium phosphate microspheres for bone regeneration
Xie et al. Preparation, in vitro degradability, cytotoxicity, and in vivo biocompatibility of porous hydroxyapatite whisker-reinforced poly (L-lactide) biocomposite scaffolds
CN114504407A (zh) 一种3d打印含生长因子的颅骨修复钛网及其制备方法
Raja et al. Multifunctional calcium-deficient hydroxyl apatite–alginate core–shell-structured bone substitutes as cell and drug delivery vehicles for bone tissue regeneration
CN101249277A (zh) 三维多孔组织工程支架材料、其纤维粘结法制备及应用
CN105412984A (zh) 一种负载蛋白的3d组织工程支架及其制备方法
KR100889593B1 (ko) 생체고분자와 생체세라믹의 이종복합체 및 그 제조 방법
Sun et al. A novel gelatin/carboxymethyl chitosan/nano-hydroxyapatite/β-tricalcium phosphate biomimetic nanocomposite scaffold for bone tissue engineering applications
Tian et al. Nano calcium-deficient hydroxyapatite/O-carboxymethyl chitosan-CaCl2 microspheres loaded with rhein for bone defect repair
CN114732962B (zh) 一种可降解的抗菌引导骨再生膜及其制备方法和应用
Yildirim Preparation and characterization of chitosan/calcium phosphate based composite biomaterials

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: 17861862

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 17861862

Country of ref document: EP

Kind code of ref document: A1