WO2012097506A1 - 一种关节软骨修复再生用支架及其制备方法 - Google Patents

一种关节软骨修复再生用支架及其制备方法 Download PDF

Info

Publication number
WO2012097506A1
WO2012097506A1 PCT/CN2011/070377 CN2011070377W WO2012097506A1 WO 2012097506 A1 WO2012097506 A1 WO 2012097506A1 CN 2011070377 W CN2011070377 W CN 2011070377W WO 2012097506 A1 WO2012097506 A1 WO 2012097506A1
Authority
WO
WIPO (PCT)
Prior art keywords
bone
decalcified
cancellous
cortical bone
stent
Prior art date
Application number
PCT/CN2011/070377
Other languages
English (en)
French (fr)
Inventor
敖英芳
张辛
何震明
马勇
周春燕
Original Assignee
北京大学第三医院
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 北京大学第三医院 filed Critical 北京大学第三医院
Priority to PCT/CN2011/070377 priority Critical patent/WO2012097506A1/zh
Publication of WO2012097506A1 publication Critical patent/WO2012097506A1/zh

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61F—FILTERS IMPLANTABLE INTO BLOOD VESSELS; PROSTHESES; DEVICES PROVIDING PATENCY TO, OR PREVENTING COLLAPSING OF, TUBULAR STRUCTURES OF THE BODY, e.g. STENTS; ORTHOPAEDIC, NURSING OR CONTRACEPTIVE DEVICES; FOMENTATION; TREATMENT OR PROTECTION OF EYES OR EARS; BANDAGES, DRESSINGS OR ABSORBENT PADS; FIRST-AID KITS
    • A61F2/00—Filters implantable into blood vessels; Prostheses, i.e. artificial substitutes or replacements for parts of the body; Appliances for connecting them with the body; Devices providing patency to, or preventing collapsing of, tubular structures of the body, e.g. stents
    • A61F2/02—Prostheses implantable into the body
    • A61F2/30—Joints
    • A61F2/30756—Cartilage endoprostheses
    • 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
    • A61L27/34—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
    • A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
    • A61L27/3604—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix characterised by the human or animal origin of the biological material, e.g. hair, fascia, fish scales, silk, shellac, pericardium, pleura, renal tissue, amniotic membrane, parenchymal tissue, fetal tissue, muscle tissue, fat tissue, enamel
    • A61L27/3608—Bone, e.g. demineralised bone matrix [DBM], bone powder
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
    • A61L27/3641—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix characterised by the site of application in the body
    • A61L27/3645—Connective tissue
    • A61L27/365—Bones
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
    • A61L27/3683—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix subjected to a specific treatment prior to implantation, e.g. decellularising, demineralising, grinding, cellular disruption/non-collagenous protein removal, anti-calcification, crosslinking, supercritical fluid extraction, enzyme treatment
    • A61L27/3687—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix subjected to a specific treatment prior to implantation, e.g. decellularising, demineralising, grinding, cellular disruption/non-collagenous protein removal, anti-calcification, crosslinking, supercritical fluid extraction, enzyme treatment characterised by the use of chemical agents in the treatment, e.g. specific enzymes, detergents, capping agents, crosslinkers, anticalcification agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61F—FILTERS IMPLANTABLE INTO BLOOD VESSELS; PROSTHESES; DEVICES PROVIDING PATENCY TO, OR PREVENTING COLLAPSING OF, TUBULAR STRUCTURES OF THE BODY, e.g. STENTS; ORTHOPAEDIC, NURSING OR CONTRACEPTIVE DEVICES; FOMENTATION; TREATMENT OR PROTECTION OF EYES OR EARS; BANDAGES, DRESSINGS OR ABSORBENT PADS; FIRST-AID KITS
    • A61F2/00—Filters implantable into blood vessels; Prostheses, i.e. artificial substitutes or replacements for parts of the body; Appliances for connecting them with the body; Devices providing patency to, or preventing collapsing of, tubular structures of the body, e.g. stents
    • A61F2/02—Prostheses implantable into the body
    • A61F2/30—Joints
    • A61F2/3094—Designing or manufacturing processes
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61F—FILTERS IMPLANTABLE INTO BLOOD VESSELS; PROSTHESES; DEVICES PROVIDING PATENCY TO, OR PREVENTING COLLAPSING OF, TUBULAR STRUCTURES OF THE BODY, e.g. STENTS; ORTHOPAEDIC, NURSING OR CONTRACEPTIVE DEVICES; FOMENTATION; TREATMENT OR PROTECTION OF EYES OR EARS; BANDAGES, DRESSINGS OR ABSORBENT PADS; FIRST-AID KITS
    • A61F2/00—Filters implantable into blood vessels; Prostheses, i.e. artificial substitutes or replacements for parts of the body; Appliances for connecting them with the body; Devices providing patency to, or preventing collapsing of, tubular structures of the body, e.g. stents
    • A61F2/02—Prostheses implantable into the body
    • A61F2/30—Joints
    • A61F2002/30001—Additional features of subject-matter classified in A61F2/28, A61F2/30 and subgroups thereof
    • A61F2002/30003—Material related properties of the prosthesis or of a coating on the prosthesis
    • A61F2002/30004—Material related properties of the prosthesis or of a coating on the prosthesis the prosthesis being made from materials having different values of a given property at different locations within the same prosthesis
    • A61F2002/30057—Material related properties of the prosthesis or of a coating on the prosthesis the prosthesis being made from materials having different values of a given property at different locations within the same prosthesis made from both cortical and cancellous adjacent parts
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61F—FILTERS IMPLANTABLE INTO BLOOD VESSELS; PROSTHESES; DEVICES PROVIDING PATENCY TO, OR PREVENTING COLLAPSING OF, TUBULAR STRUCTURES OF THE BODY, e.g. STENTS; ORTHOPAEDIC, NURSING OR CONTRACEPTIVE DEVICES; FOMENTATION; TREATMENT OR PROTECTION OF EYES OR EARS; BANDAGES, DRESSINGS OR ABSORBENT PADS; FIRST-AID KITS
    • A61F2/00—Filters implantable into blood vessels; Prostheses, i.e. artificial substitutes or replacements for parts of the body; Appliances for connecting them with the body; Devices providing patency to, or preventing collapsing of, tubular structures of the body, e.g. stents
    • A61F2/02—Prostheses implantable into the body
    • A61F2/30—Joints
    • A61F2/30756—Cartilage endoprostheses
    • A61F2002/30766—Scaffolds for cartilage ingrowth and regeneration
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61F—FILTERS IMPLANTABLE INTO BLOOD VESSELS; PROSTHESES; DEVICES PROVIDING PATENCY TO, OR PREVENTING COLLAPSING OF, TUBULAR STRUCTURES OF THE BODY, e.g. STENTS; ORTHOPAEDIC, NURSING OR CONTRACEPTIVE DEVICES; FOMENTATION; TREATMENT OR PROTECTION OF EYES OR EARS; BANDAGES, DRESSINGS OR ABSORBENT PADS; FIRST-AID KITS
    • A61F2/00—Filters implantable into blood vessels; Prostheses, i.e. artificial substitutes or replacements for parts of the body; Appliances for connecting them with the body; Devices providing patency to, or preventing collapsing of, tubular structures of the body, e.g. stents
    • A61F2/02—Prostheses implantable into the body
    • A61F2/30—Joints
    • A61F2/30767—Special external or bone-contacting surface, e.g. coating for improving bone ingrowth
    • A61F2/30771—Special external or bone-contacting surface, e.g. coating for improving bone ingrowth applied in original prostheses, e.g. holes or grooves
    • A61F2002/30772—Apertures or holes, e.g. of circular cross section
    • A61F2002/30784—Plurality of holes
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61F—FILTERS IMPLANTABLE INTO BLOOD VESSELS; PROSTHESES; DEVICES PROVIDING PATENCY TO, OR PREVENTING COLLAPSING OF, TUBULAR STRUCTURES OF THE BODY, e.g. STENTS; ORTHOPAEDIC, NURSING OR CONTRACEPTIVE DEVICES; FOMENTATION; TREATMENT OR PROTECTION OF EYES OR EARS; BANDAGES, DRESSINGS OR ABSORBENT PADS; FIRST-AID KITS
    • A61F2310/00—Prostheses classified in A61F2/28 or A61F2/30 - A61F2/44 being constructed from or coated with a particular material
    • A61F2310/00005—The prosthesis being constructed from a particular material
    • A61F2310/00359—Bone or bony tissue
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61F—FILTERS IMPLANTABLE INTO BLOOD VESSELS; PROSTHESES; DEVICES PROVIDING PATENCY TO, OR PREVENTING COLLAPSING OF, TUBULAR STRUCTURES OF THE BODY, e.g. STENTS; ORTHOPAEDIC, NURSING OR CONTRACEPTIVE DEVICES; FOMENTATION; TREATMENT OR PROTECTION OF EYES OR EARS; BANDAGES, DRESSINGS OR ABSORBENT PADS; FIRST-AID KITS
    • A61F2310/00—Prostheses classified in A61F2/28 or A61F2/30 - A61F2/44 being constructed from or coated with a particular material
    • A61F2310/00005—The prosthesis being constructed from a particular material
    • A61F2310/00365—Proteins; Polypeptides; Degradation products thereof
    • A61F2310/00371—Collagen

Definitions

  • the invention relates to the field of medical materials, in particular to a bracket for repairing and regenerating articular cartilage and a preparation method thereof.
  • the currently used cartilage repair scaffold materials can be broadly divided into two types: natural scaffold materials and artificial scaffold materials.
  • the natural support material is derived from macromolecules naturally occurring in animals, plants or human bodies, and has good biocompatibility and tissue affinity.
  • Artificial scaffold materials are currently studied more materials, mainly polyglycolic acid, polylactic acid, and polyanhydride. They can be adjusted according to their physical, chemical, biomechanical and degradation properties, easy to form and form, and have good production repeatability.
  • the natural scaffold material has problems in that the material properties cannot be changed, and the mechanical strength and the degradation rate are difficult to control.
  • Commonly used natural scaffold materials are collagen, hydroxyapatite, etc.
  • the biomechanical properties of natural scaffolds are difficult to adjust, and it is difficult to find biomechanical properties similar to those of cartilage.
  • Some natural biomaterial scaffolds have poor mechanical mechanics and are not resistant to the mechanical stress generated during joint movement. They are prone to collapse of the repair site and also affect the growth of cells on the material, such as hydroxyapatite scaffold elasticity and creep than cartilage. Poor, too high hardness can cause the corresponding cartilage joint surface wear.
  • the biocompatibility of artificial scaffold materials is relatively poor, and there are different processes. Degree of inflammatory response, as well as immunogenicity problems, and even carcinogenic problems. Summary of the invention
  • An object of the present invention is to provide a stent for repairing and regenerating articular cartilage which has both good biocompatibility and mechanical strength in view of the above-mentioned drawbacks of the prior art.
  • Another object of the present invention is to provide a method for preparing a stent for articular cartilage repair and regeneration.
  • the material of the scaffold is decalcified cancellous-cortical bone, and the decalcified cancellous-cortical bone is provided with small holes extending through the decalcified cancellous bone and the decalcified cortical bone.
  • the small hole is perpendicular to the horizontal surface of the decalcified cortical bone, the diameter of the small hole is 0. l_lmm.
  • the pores have a porosity of 5% to 50%.
  • a method for preparing a stent for repairing and regenerating articular cartilage comprising the following steps:
  • the decalcification solution is 0.1 M-0. 7 M ethylenediaminetetraacetic acid, pH 6-10, temperature: 2 _10 °C.
  • the biological scaffold is subjected to cobalt 60 irradiation for 12-72 hours before transplanting the biological scaffold.
  • the bone is a cryopreserved human humerus, vertebral body or long bone metaphysis.
  • the bone library bone is the raw material that has been approved by the Food and Drug Administration and applied to the clinical for many years.
  • the donor source is sufficient and safe, and it is easy to be clinically popularized.
  • Decalcification of many animal cancellous-cortical bones can also be used for the repair of human cartilage damage by the method of the present invention.
  • the number of cattle and sheep is large, and the materials used for bones are more extensive.
  • the present invention utilizes a new method for decalcifying the biological characteristics of decalcified bone scaffolds, and at the same time solves the problems of decalcification with ethylenediaminetetraacetic acid (EDTA).
  • EDTA ethylenediaminetetraacetic acid
  • Commonly used acids hydroochloric acid, nitric acid and formic acid, etc.
  • decalcification of human bone is performed using EDTA (neutral calcium ion chelating agent) (such a neutral decalcifying agent has minimal damage to the collagen scaffold).
  • the present invention abandons the simple decalcified cortical bone scaffold and selects the cortical-cancellous bone.
  • the thickness of the cortical bone in the scaffold is only 0. l-lmm, and the remaining thickness of the cancellous bone complements the depth of the defect, due to the porous structure of the cancellous bone. , making the decalcifying solution easy to enter the center of the material. Not only can it completely decalcify, but it also significantly shortens the decalcification time and meets clinical applications.
  • the fixing material can be used to fix the repairing material to the cartilage defect, increase the firmness of the cartilage repairing stent, and shorten the time for the patient to carry weight and walk back and resume exercise function.
  • the loose porous structure of the cancellous bone in the stent can accommodate more bone marrow and bone marrow-derived stem cells released from the subchondral bone, and has a wider space for the bone marrow stem cells to adhere, migrate and exchange nutrients, accelerate the proliferation of stem cells and Differentiation, repair of cartilage defects.
  • the natural collagen scaffold material used in the invention has no problem of immune rejection, is easy to adsorb by cells, and has good biocompatibility.
  • the decalcified cancellous-cortical bone scaffold itself is rich in bone morphogenetic proteins (BMPs), which can promote the transformation of stem cells or bone marrow stromal cells in the middle of the bone marrow into chondrocytes or bone cells, and promote stem cell secretion, cartilage or bone. Secretion of the matrix. It plays an important role in the initiation, development, regulation and shaping of cartilage repair.
  • BMPs bone morphogenetic proteins
  • the longitudinally drilled decalcified cancellous-cortical bone articular cartilage regeneration stent of the present invention uses the microfracture technique to release its own bone marrow stem cells, and spreads through the longitudinal borehole on the stent to the interior of the decalcified cancellous bone stent.
  • the decalcified cancellous bone scaffold supports the three-dimensional growth of autologous bone marrow stem cells, and the bone morphological proteins (BMPs) on the scaffold induce bone marrow stem cells to grow and differentiate into bone and cartilage.
  • BMPs bone morphological proteins
  • FIG. 1 is a schematic view of a stent for repairing and regenerating articular cartilage according to an embodiment of the present invention
  • Figure 2 is a cross-sectional view of Figure 1;
  • FIG. 3 is a flow chart of preparing a stent for repairing and regenerating articular cartilage according to an embodiment of the present invention
  • FIG. 4 is a structural view of a scanning electron microscope of a stent for repairing and regenerating articular cartilage according to an embodiment of the present invention
  • 5 is a surgical diagram of a human decalcified cortex-cancellous bone scaffold for repairing knee articular cartilage defects according to an embodiment of the present invention
  • 6 is a schematic view showing the repair of the knee articular cartilage defect at 12 weeks after repairing the knee articular cartilage defect by the human decalcified cortex-cancellous bone scaffold provided by the embodiment of the present invention
  • FIG. 7 is a 12-week HE staining diagram of a human decalcified cortex-cancellous bone scaffold for repairing a knee articular cartilage defect according to an embodiment of the present invention
  • Figure 8 is a diagram showing the specific staining of toluidine blue cartilage in a 12-week for the treatment of knee articular cartilage defects by a decalcified cortex-cancellous bone scaffold provided by an embodiment of the present invention
  • Figure 9 is a diagram showing the immunohistochemical staining of type I collagen I for 12 weeks after repairing knee articular cartilage defects with a human decalcified cortex-cancellous bone scaffold provided by an embodiment of the present invention. detailed description
  • a stent for repairing and regenerating articular cartilage the material of the stent is decalcified cancellous-cortical bone, and the decalcified cortical bone 1 is connected to decalcified cancellous bone 3, the decalcified cancellous substance-
  • the cortical bone is provided with a small hole 2 extending through the decalcified cancellous bone 3 and the decalcified cortical bone 1.
  • the diameter of the small hole 2 is 0. lmm, 0. 5mm, lmm.
  • the distance between the center of the hole of the adjacent small hole 2 is 0. 5 ⁇ 5mm, and the distance between the centers of the holes may be 0. 5 hidden, 1. 2 hidden, 3. 4 hidden, 5 hidden.
  • the porosity of the small holes 2 is 5% - 20%, and the overall thickness of the stent is greater than 2 mm.
  • a preparation method of a stent for repairing and regenerating articular cartilage includes the following steps:
  • Step 102 soaking the bone in the decalcifying solution for decalcification
  • Step 103 monitoring the decalcification process by an atomic absorption spectrophotometer
  • Step 104 X-ray and CT examination are performed on the decalcified specimen to prove that the bone is completely decalcified, and decalcified cancellous-cortical bone is obtained;
  • Step 105 laser drilling on the surface of the decalcified cortical bone with a diameter of 0.1-lmm;
  • Step 106 The nanoelastic modulus of each part of the decalcified cancellous-cortical bone scaffold is detected by a nanoindentation biomechanical tester, and the decalcified cancellous-cortical bone similar to the normal cartilage elastic modulus is selected for repair and regeneration of articular cartilage. Bracket, will take off Calcareous cortical-cortical bone is cut into the required specifications for use, frozen storage;
  • Step 107 The biological scaffold is subjected to cobalt 60 irradiation for 12-24 hours before transplanting the biological scaffold.
  • shaving is performed in the osteochondral defect area to remove fibrotic, irregular lesion cartilage and subchondral bone.
  • subchondral bone grinding is performed to shape the osteochondral defect into a relatively regular shape. Holes in the subchondral bone are then made to release the bone marrow of itself.
  • the laser-drilled decalcified bone scaffold is trimmed into a shape conforming to the osteochondral defect, and the defect is filled in, and the cortical bone portion is the bottom layer.
  • the bone marrow released by the subchondral bone due to drilling will penetrate into the decalcified cancellous bone through the small hole in the cortical bone of the stent, thereby repairing the osteochondral defect.
  • the released self-bone marrow stem cells enhance the repair ability.
  • the bone marrow released by the pure subchondral bone drill can not bear the stress and can not be repaired according to specific structural specifications.
  • the resulting articular cartilage lacks normal structure.
  • the stent for regenerating decalcified bone and articular cartilage of the present invention is used to infiltrate the released bone marrow into the decalcified cancellous bone through the decalcified cortical bone hole in the stent, thereby increasing the mechanical strength of the repaired portion at the early stage of repairing, and repairing the site.
  • the mechanical stress has a three-dimensional regulation of bone marrow repair in the shape of the defect.
  • the preparation flow chart is shown in Figure 3.
  • the cryopreserved human tibia or vertebral body bone was purchased and immersed in 0. 1M-1M EDTA decalcifying solution.
  • the temperature of the decalcifying solution was 6 ° C and the pH was 8.
  • the calcium in the cortical bone is removed by chelation, and the decalcifying solution is replaced daily.
  • the EDTA decalcifying solution was prepared in deionized water and contained in a plastic container.
  • the daily decalcification solution is measured by an atomic absorption spectrophotometer to detect the calcium ion concentration to monitor the decalcification process. When the concentration of Ca chelate in the decalcification solution is ⁇ 0.5 g/L, it is complete decalcification, decalcification.
  • the present invention uses a laser drilling technique as shown in FIG. 2, the horizontal surface of the decalcified cortical bone is vertically drilled with a diameter of 0.5 mm, and the borehole is laser drilled.
  • the experimental rabbit animal model has a pore diameter applied to the clinical patient. The difference is that the size of the joints of the human and the rabbit are different, so the aperture and the hole spacing are different. After the comparison, the interval between the holes is set to 2 mm.
  • the laser-drilled decalcified cancellous-cortical bone stent is used. Cut to a certain size and size, frozen storage. Prior to transplantation, the prepared laser-drilled decalcified cancellous-cortical bone scaffold was subjected to cobalt 60 irradiation for 18 hours.
  • FIG. 4 a scanning electron micrograph of the structure of the cancellous bone portion of the scaffold for decalcified cancellous-cortical osteochondral joint regeneration. It can be seen that the collagen fibers in the decalcified cancellous bone are loosely staggered, the collagen fibers are filled with voids, and the pore structure is interlaced, which is beneficial to the infiltration of the bone marrow matrix and factors and the attachment of cells.
  • Decalcified cortical bone natural layer there are several advantages: 1. Collagen structure can resist vertical traction and pressure, which is conducive to cartilage regeneration stent fixation in cartilage defects. 2. The interlamellar gap facilitates the infiltration of bone marrow matrix and factors and the attachment of cells; facilitates the exchange of cellular nutrients. Surgical procedure for repairing cartilage defects in animal models
  • Figure 6 is a general view of the New Zealand white rabbits using laser-drilled decalcified cancellous-cortical bone articular cartilage repair and regeneration stent for soft bone repair after 12 weeks.
  • the repaired area indicated by the arrow on the gross specimen showed a cartilage-like change, which was very similar to the surrounding normal tissue.
  • the junction with the normal cartilage was not obvious, and no gap appeared. No obvious immune inflammatory response was observed.
  • the HE staining map of the new tissue is shown in Fig. 7; the neonatal tissue toluidine blue staining (specifically for the mucopolysaccharide component in the cartilage matrix) is positive as shown in Fig. 8; the neonatal tissue type II collagen immunohistochemical staining is positive as shown in Fig. 9, which proves that the new tissue is rich.
  • Type II collagen is hyaline cartilage similar to normal cartilage tissue.
  • the laser-drilled decalcified bone and articular cartilage regeneration stent of the present invention is prepared by using decalcified cortical bone and cancellous bone of allogeneic humans, and using laser-drilled decalcified cancellous-cortical bone articular cartilage regeneration.
  • autologous bone marrow not only provides autologous stem cells, but also provides a microenvironment for cell survival, and has higher cell survival than chondrocytes or stem cell transplantation alone. rate.
  • the method of the present invention utilizes allogeneic decalcified human bone scaffolds to solve the biocompatibility problem of synthetic scaffolds.
  • Biomechanical tests show that the mechanical stress and elastic modulus of the prepared decalcified bone scaffold are closer to that of cartilage, which solves the poor mechanical mechanics of natural biomaterials such as silk, collagen, fibrin and human amnion, and is incapable of resisting joint movement.
  • the problem of mechanical stress It does not cause collapse of the repair site, and it also supports the growth of cells on the material.
  • the decalcified cancellous-cortical bone articular cartilage regeneration stent is a human decalcified cancellous-cortical bone, which itself is rich in bone morphogenetic proteins (BMPs), which can promote the transformation of stem cells or bone marrow stromal cells in the middle of bone marrow in the induced scaffold.
  • BMPs bone morphogenetic proteins
  • Chondrocytes or bone cells promote the secretion of cartilage or bone matrix by stem cells. It plays an important role in the initiation, development, regulation and shaping of cartilage repair.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Biomedical Technology (AREA)
  • Oral & Maxillofacial Surgery (AREA)
  • Transplantation (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Epidemiology (AREA)
  • Medicinal Chemistry (AREA)
  • Dermatology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Botany (AREA)
  • Orthopedic Medicine & Surgery (AREA)
  • Molecular Biology (AREA)
  • Vascular Medicine (AREA)
  • Urology & Nephrology (AREA)
  • Zoology (AREA)
  • Rheumatology (AREA)
  • Heart & Thoracic Surgery (AREA)
  • Cardiology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Prostheses (AREA)

Description

一种关节软骨修复再生用支架及其制备方法
技术领域
本发明涉及医用材料领域， 特别涉及一种关节软骨修复再生用支架及其制备方法。 背景技术 说
关节软骨损伤是运动医学领域的难治性损伤。 运动引起的关节软骨损伤发生率很高， 北 京大学运动医学研究所的调查结果显示软骨损伤占膝关节运动创伤的 40. 5%以上。 软骨损伤 后常会引起关节肿痛等症状。 由于软骨组织结构特殊， 其内无神经、 血管和淋巴管， 软骨的 营养供给是通过关节活动产生的压力变化， 使滑液在关书节腔与软骨基质之间进行交流， 故损 伤后的关节软骨不能修复与再生， 且易导致明显的功能障碍。 国内、 外软骨修复方法很多， 但是尚无一种公认的完善修复方法。 目前， 利用自身骨髓修复软骨缺损， 如： 软骨下骨微骨 折技术成为了软骨缺损修复的重要方法之一。 由于它操作简单， 对于小面积的软骨缺损取得 良好结果， 在临床软骨修复上已经广泛应用。但由于微骨折是通过骨髓形成的瘢块填充缺损， 瘢块的机械应力太低， 无法承受关节活动产生的压力。 因此微骨折术后较长时间 6周不能负 重； 修复位置可能出现塌陷或过度增生； 微骨折方法对于大面积和非水平位置的软骨微骨折 效果并不理想， 而且新生软骨多为纤维软骨。 因此， 寻找适合的软骨缺损填充支架也成为软 骨修复的主要研究方向之一。
目前使用的软骨修复支架材料可大体分为天然支架材料和人工支架材料两大类。 天然支 架材料是来源于动、 植物或者人体内天然存在的大分子材料， 具有良好的生物相容性、 组织 亲合性。 人工支架材料是目前研究较多的材料， 主要有聚羟基乙酸、 聚乳酸、 以及聚酸酐等， 可根据需要调整其物理、 化学、 生物力学和降解性能， 容易加工成形， 生产重复性好。
在实现本发明的过程中， 发明人发现现有技术至少存在以下问题： 天然支架材料存在材 料性质无法改变， 机械强度及降解速度难于控制等问题。 常用的天然支架材料有胶原蛋白、 羟基磷灰石等， 天然支架生物力学性质难以调整， 很难找到和软骨生物力学性质相似的。 有 些天然生物材料支架的机械力学差， 无法抵抗关节运动时产生的机械应力， 容易造成修补处 的塌陷， 也同时影响材料上细胞的生长， 而如羟基磷灰石支架弹性和蠕变性较软骨差， 硬度 过高易造成相对应的软骨关节面的磨损。 人工支架材料的生物相容性相对较差， 存在不同程 度的炎性反应， 以及免疫原性问题， 甚至致癌性问题。 发明内容
本发明的目的是针对上述现有技术的缺陷， 提供一种兼有良好的生物相容性和机械强度 的关节软骨修复再生用支架。
本发明的另一个目的是提供关节软骨修复再生用支架的制备方法。
为了实现上述目的本发明采取的技术方案是：
所述支架的材料为脱钙松质-皮质骨， 所述脱钙松质-皮质骨上设有贯穿脱钙松质骨和脱 钙皮质骨的小孔。
所述小孔垂直于所述的脱钙皮质骨水平表面设置， 所述小孔的直径为 0. l_lmm。
所述小孔为两个以上， 相邻所述小孔的中心距离为 0. 5〜5mm。
所述小孔的孔隙率为 5%_50%。
本发明提供的另一个技术方案是： 一种关节软骨修复再生用支架的制备方法， 包括以下 步骤：
( 1 ) 选择包括皮质骨和松质骨双层结构的骨骼；
( 2) 将所述骨骼浸泡于脱钙液中进行脱钙处理；
( 3) 用原子吸收分光光度计监测脱钙进程；
(4) 脱钙后的标本进行 X线和 CT检测， 当骨骼完全脱钙后， 得到脱钙松质-皮质骨；
( 5)垂直于脱钙皮质骨水平表面以 0. 1-lmm的直径激光打孔， 钻孔贯穿脱钙皮质骨和脱 钙松质骨两层， 穿透整个支架；
(6) 将脱钙松质 -皮质骨剪裁成使用需要的规格， 冷冻贮存。
所述脱钙液为 0. 1M-0. 7M乙二胺四乙酸， pH值 6-10， 温度： 2 _10°C。
进一步地， 在移植所述生物支架前， 所述生物支架进行钴 60照射 12-72小时。
所述骨骼为低温冷冻的人髂骨、 椎体骨或长骨干骺端。
本发明实施例提供的技术方案带来的有益效果是：
1. 运用成熟的商业骨库中的人骨为原料， 骨库骨是已经通过药监局批准， 并应用于临床 多年的原料， 供体来源充分且安全， 易于临床普及。 很多动物松质-皮质骨 （牛、 羊等）脱钙 经本发明的方法处理也可以用于人软骨损伤的修复。 牛、 羊养殖数量多， 取骨做材料更为广 泛。
2. 本发明运用了保护脱钙骨支架生物特性脱钙新方法， 同时解决了用乙二胺四乙酸 (EDTA) 脱钙所存在的问题。 普遍运用的酸 （盐酸、 硝酸和甲酸等） 脱钙方法， 对骨的胶原 支架损伤很大， 造成支架变脆， 弹性模量下降等生物学和生物力学性质的改变， 无法再用于 软骨修复。 本发明中运用 EDTA (中性钙离子螯合剂） （此种中性脱钙剂对骨胶原支架损伤最 小）进行人骨的脱钙。 但由于 EDTA脱钙剂只能渗透人皮质骨表面 2mm， 无法完全脱去皮质骨 中心的钙质， 而且时间过长， 无法满足临床应用需要。 因此本发明放弃单纯脱钙皮质骨支架 而选用皮质-松质骨， 支架中皮质骨厚度只取 0. l-lmm, 其余用松质骨的厚度补足缺损深度， 由于松质骨的多孔隙结构， 使脱钙液很容易进入材料中心。 不仅能完全脱钙， 而且显著縮短 了脱钙时间， 满足临床应用。
3.利用材料中的薄层皮质骨， 可以运用固定钉将修复材料固定于软骨缺损处， 增加软骨 修复支架的牢固程度， 縮短病人术后负重行走、 恢复运动功能的时间。
4.支架中松质骨的疏松多孔结构，可以容纳更多软骨下骨释放的骨髓及骨髓来源干细胞， 有更加宽阔的空间让骨髓干细胞进行粘附、 迁移和营养物质的交换， 加速干细胞的增殖和分 化， 软骨缺损的修复。
5. 本发明采用的天然胶原支架材料， 无免疫排斥反应的问题， 易于细胞吸附， 生物相容 性好。
6. 在脱钙松质 -皮质骨支架自身富含骨形态蛋白（BMPs ), 可促进诱导支架中的骨髓中间 的干细胞或骨髓基质细胞转化成软骨细胞或骨细胞， 促进干细胞分泌、软骨或骨基质的分泌。 对软骨修复的启动、 发育、 调控及塑形均有重要作用。
7.操作简便， 不需二次手术， 节省了手术中取材时间， 易于临床应用。 本发明的纵向钻 孔的脱钙松质-皮质骨关节软骨再生用支架， 使用时利用微骨折技术释放自身的骨髓干细胞， 使其通过支架上的纵向钻孔扩散至脱钙松质骨支架内部， 脱钙松质骨支架支撑自体骨髓干细 胞三维立体生长， 同时支架上的骨形态蛋白 （BMPs )诱导骨髓干细胞向骨、 软骨生长、 分化。 本发明避开了组织工程中体外支架培养细胞， 外源因子诱导， 生成物回植等复杂的步骤， 无 需二次手术， 更适用于临床应用。 附图说明
图 1 是本发明实施例提供的关节软骨修复再生用支架示意图；
图 2是图 1的剖视图；
图 3 是本发明实施例提供的关节软骨修复再生用支架的制备流程图；
图 4是本发明实施例提供的关节软骨修复再生用支架扫描电镜结构图；
图 5是本发明实施例提供的人脱钙皮质-松质骨支架修复膝关节软骨缺损手术图； 图 6是本发明实施例提供的人脱钙皮质-松质骨支架修复膝关节软骨缺损后 12周取材观 察膝关节软骨缺损修复处大体图；
图 7是本发明实施例提供的人脱钙皮质 -松质骨支架修复膝关节软骨缺损 12周 HE染色 图；
图 8是本发明实施例提供的人脱钙皮质 -松质骨支架修复膝关节软骨缺损 12周甲苯胺蓝 软骨特异性染色图；
图 9是本发明实施例提供的人脱钙皮质 -松质骨支架修复膝关节软骨缺损 12周 I I型胶原 免疫组化染色图。 具体实施方式
为使本发明的目的、 技术方案和优点更加清楚， 下面将结合附图对本发明实施方式作进 一步地详细描述。
参见图 1和图 2， 一种关节软骨修复再生用支架， 所述支架的材料为脱钙松质-皮质骨， 脱钙皮质骨 1连接着脱钙松质骨 3， 所述脱钙松质-皮质骨上设有贯穿脱钙松质骨 3和脱钙皮 质骨 1的小孔 2。
作为一种优选， 小孔 2垂直于脱钙皮质骨 1水平表面设置， 小孔 2的直径为 0. 1-lmm, 可以为 0. lmm、 0. 5mm, lmm。 小孔 2为两个以上， 相邻小孔 2的孔中心的距离为 0. 5〜5mm， 孔中心距离可以为 0. 5隱、 1. 2隱、 3. 4隱、 5隱。 小孔 2的孔隙率为 5%- 20%， 支架总体厚度大 于 2mm。
实施例 1
参见图 3， 一种关节软骨修复再生用支架的制备方法， 包括以下步骤：
步骤 101 : 选择皮质骨下带有松质骨的骨骼， 将皮质骨厚度修整在 0. 5-3mm， 脱钙液为 0. 1M-0. 7M乙二胺四乙酸， pH值 6-10， 温度： 2 _10°C ;
步骤 102: 将骨骼浸泡于脱钙液中进行脱钙处理；
步骤 103: 用原子吸收分光光度计监测脱钙进程；
步骤 104: 脱钙后的标本进行 X线和 CT检测， 以证明骨骼完全脱钙， 得到脱钙松质-皮 质骨；
步骤 105: 在脱钙皮质骨水平表面以 0. 1-lmm的直径激光打孔；
步骤 106: 通过纳米压痕生物力学测试仪检测脱钙松质-皮质骨支架各部分纳米弹性模 量， 筛选出和正常软骨弹性模量相似的脱钙松质 -皮质骨作为关节软骨修复再生用支架， 将脱 钙松质 -皮质骨剪裁成使用需要的规格， 冷冻贮存；
步骤 107 : 在移植所述生物支架前， 所述生物支架进行钴 60照射 12-24小时。
采用本实施例的关节软骨修复再生用支架的修复方法主要步骤为：
首先， 在骨软骨缺损区域进行刨刮去除纤维变、 不规则的病变软骨和软骨下骨。 继而， 进行软骨下骨磨削， 将骨软骨缺损修成比较规则的形状。 然后进行软骨下骨的钻孔使能够释 放自身的骨髓。
接着， 将激光钻孔处理的脱钙骨支架， 修整成符合骨软骨缺损的形状， 填入缺损处， 皮 质骨部分为底层。 软骨下骨由于钻孔而释放的自身的骨髓， 会通过支架皮质骨上的小孔渗透 入脱钙松质骨中， 进而修复骨软骨缺损处。 释放出的自身骨髓干细胞， 增强了修复能力。 单 纯软骨下骨钻孔释放出的自身骨髓， 无支架不能承担应力且无法按照特定结构规范修复， 生 成的关节软骨缺乏正常的结构。 而采用了本发明的脱钙骨关节软骨再生用支架， 使释放的骨 髓经过支架中的脱钙皮质骨孔道渗入脱钙松质骨中，既增加了修复早期的修补处的机械强度， 修补处的机械应力有可按缺损形状三维立体的调控骨髓修复作用。
实施例 2
关节软骨修复再生用支架的制备方法：
制备流程图如图 3所示， 购置深低温冻存人髂骨或椎体骨， 浸泡于 0. 1M-1M EDTA脱钙液 中， 脱钙液的温度为 6 ° C内， pH为 8， 利用螯合脱去皮质骨中的钙， 脱钙液每日更换。 用去 离子水配制 EDTA脱钙液， 用塑料容器盛载。每日换下的脱钙液用原子吸收分光光度计检测钙 离子浓度以监测脱钙进程， 当脱钙液中的 Ca螯合物浓度〈0. 5g/L， 即为完全脱钙， 脱钙后的 标本进行 X线和 CT检测， 证明完全脱钙。 由于脱钙皮质骨表面结构过于致密， 不宜于细胞的 长入。 因此本发明通过激光钻孔技术如图 2所示， 将脱钙皮质骨水平表面以 0. 5mm的直径垂 直钻孔， 钻孔采用激光钻孔， （实验兔动物模型上与临床病人应用孔径有所不同， 因为人、 兔 的关节大小不同， 所以孔径和孔距都不同）， 比较后设定孔间间隔 2mm， 然后， 如图 3所示， 将激光钻孔脱钙松质-皮质骨支架剪裁成一定的大小规格， 冷冻贮存。在移植前， 制备的激光 钻孔的脱钙松质 -皮质骨支架进行钴 60照射 18小时消毒。
参见图 4，激光钻孔脱钙松质 -皮质骨软骨关节再生用支架松质骨部分的结构扫描电镜图。 可见脱钙松质骨中胶原纤维疏松交错排列， 胶原纤维间充满空隙， 孔隙结构交错相通， 有利 于骨髓基质和因子的渗入及细胞的附着。 脱钙皮质骨天然片层样， 有几点优势： 1.胶原结构 可对抗铅垂方向牵引力和压力， 有利于软骨再生支架固定于软骨缺损。 2 .片层间的间隙有利 于骨髓基质和因子的渗入及细胞的附着； 有利于细胞营养物质的交换。 在动物模型上进行修补软骨缺损实验的手术过程
为验证本发明的关节软骨修复再生用支架在修补方法中的效果， 在动物模型上进行了模 拟手术实验。 选择 2. 5-3. 0kg的新西兰大耳白兔， 麻醉， 备皮， 消毒， 如图 5所示， 切口膝 关节露出股骨滑车， 在股骨滑车上做 4mm直径， 1. 5mm深的缺损， 在软骨下骨钻孔微骨折处 理后， 在已消毒好的激光钻孔脱钙松质-皮质骨支架修成 4mm直径， 高度 1. 5mm的圆柱体， 填 充入缺损。 分层缝合伤口， 术后 12周取材， 进行组织学的观察。
图 6 为新西兰大耳白兔使用激光钻孔脱钙松质 -皮质骨关节软骨修复再生用支架进行软 骨修复 12周后的大体图。在大体标本上可见箭头所指的修复处， 已呈现软骨样改变， 和周围 正常组织极为相似， 与正常软骨的交界处已不明显， 未出现缝隙。 未见明显免疫炎症反应。
参见图 7、 图 8和图 9的新西兰大耳白兔激光钻孔脱钙松质-皮质骨关节软骨再生用支架 进行软骨修复 12周后的组织学切片染色。 可见箭头所指的修复处的脱钙松质-皮质骨支架已 完全被新生的透明软骨所替代， 修复处未出现塌陷， 与图右侧的正常软骨的交界处紧密接合， 界线模糊。 由于早期观察软骨下骨还未被骨替代， 呈现软骨样组织。 未见巨噬细胞等炎症细 胞浸润， 未见明显免疫炎症反应。 新生组织 HE染色图如图 7; 新生组织甲苯胺蓝染色 （特异 性针对软骨基质中粘多糖成分） 阳性如图 8; 新生组织 II型胶原免疫组织化学染色阳性如图 9， 证明新生组织富含 II型胶原为类似正常软骨组织的透明软骨。
本发明的激光钻孔的脱钙骨关节软骨再生用支架是运用同种异体人的脱钙的皮质骨和松 质骨制备的， 采用激光钻孔的脱钙松质-皮质骨关节软骨再生用支架修复时， 操作简便， 易于 临床应用。 而且使用本发明的关节软骨修复再生用支架可以解决异体移植排斥反应的缺点， 修复时自体骨髓不仅提供自体干细胞， 同时提供细胞生存的微环境， 比单纯软骨细胞或干细 胞移植有更高的细胞生存率。 本发明方法运用同种异体的脱钙人骨支架解决人工合成支架的 生物相容性问题。 通过生物力学测试表明制备的脱钙骨支架的机械应力和弹性模量更接近于 软骨， 解决了蚕丝、 胶原蛋白、 纤维蛋白、 人羊膜等天然生物材料支架机械力学差， 无法抵 抗关节运动时产生的机械应力的问题。 不会造成修补处的塌陷， 也同时可支持材料上细胞的 生长。 而且脱钙松质 -皮质骨关节软骨再生用支架是使用人的脱钙松质-皮质骨， 本身富含骨 形态蛋白（BMPs ), 可促进诱导支架中的骨髓中间的干细胞或骨髓基质细胞转化成软骨细胞或 骨细胞， 促进干细胞分泌软骨或骨基质的分泌。 对软骨修复的启动、 发育、 调控及塑形均有 重要作用。
以上所述仅为本发明的较佳实施例， 并不用以限制本发明， 凡在本发明的精神和原则之 内， 所作的任何修改、 等同替换、 改进等， 均应包含在本发明的保护范围之内。

Claims

权 利 要 求 书
1、 一种关节软骨修复再生用支架， 其特征在于， 所述支架的材料为脱钙松质-皮质骨， 所述脱钙松质 -皮质骨上设有贯穿脱钙松质骨和脱钙皮质骨的小孔。
2、根据权利要求 1所述的支架， 其特征在于， 所述小孔垂直于所述脱钙皮质骨水平表面 设置， 所述小孔的直径为 0. 1-1隱。
3、 根据权利要求 2所述的支架， 其特征在于， 所述小孔为两个以上， 相邻所述小孔的中 心距离为 0. 5〜5mm。
4、 根据权利要求 1所述的支架， 其特征在于， 所述小孔的孔隙率为 5%_50%。
5、 一种关节软骨修复再生用支架的制备方法， 包括以下步骤：
( 1 ) 选择包括皮质骨和松质骨双层结构的骨骼；
( 2 ) 将所述骨骼浸泡于脱钙液中进行脱钙处理；
( 3 ) 用原子吸收分光光度计监测脱钙进程；
( 4) 脱钙后的标本进行 X线和 CT检测， 当骨骼完全脱钙后， 得到脱钙松质-皮质骨；
( 5 )垂直于脱钙皮质骨水平表面以 0. 1-lmm的直径激光打孔， 钻孔贯穿脱钙皮质骨和脱 钙松质骨两层， 穿透整个支架；
( 6 ) 将脱钙松质 -皮质骨剪裁成使用需要的规格， 冷冻贮存。
6、根据权利要求 5所述的制备方法， 其特征在于， 所述脱钙液为 0. 1M-0. 7M乙二胺四乙 酸， pH值 6-10， 温度： 2 -10°C。
7、 根据权利要求 5或 6所述的制备方法， 其特征在于， 在移植所述生物支架前， 所述 生物支架进行钴 60照射 12-72小时。
8、 根据权利要求 5所述的制备方法， 其特征在于， 所述骨骼为低温冷冻的人髂骨、 椎 体骨或长骨干骺端。
PCT/CN2011/070377 2011-01-19 2011-01-19 一种关节软骨修复再生用支架及其制备方法 WO2012097506A1 (zh)

Priority Applications (1)

Application Number Priority Date Filing Date Title
PCT/CN2011/070377 WO2012097506A1 (zh) 2011-01-19 2011-01-19 一种关节软骨修复再生用支架及其制备方法

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
PCT/CN2011/070377 WO2012097506A1 (zh) 2011-01-19 2011-01-19 一种关节软骨修复再生用支架及其制备方法

Publications (1)

Publication Number Publication Date
WO2012097506A1 true WO2012097506A1 (zh) 2012-07-26

Family

ID=46515086

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2011/070377 WO2012097506A1 (zh) 2011-01-19 2011-01-19 一种关节软骨修复再生用支架及其制备方法

Country Status (1)

Country Link
WO (1) WO2012097506A1 (zh)

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20150132152A (ko) * 2013-03-15 2015-11-25 알로소스 천공된 골연골 동종이식 조성물
EP2958523A4 (en) * 2013-02-22 2016-11-30 Allosource CARTEL MUSAIC COMPOSITIONS AND METHODS
CN110743039A (zh) * 2019-12-11 2020-02-04 成都奇璞生物科技有限公司 一种自体颅骨用于回植材料的制备方法
US11202674B2 (en) 2018-04-03 2021-12-21 Convergent Dental, Inc. Laser system for surgical applications

Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4950296A (en) * 1988-04-07 1990-08-21 Mcintyre Jonathan L Bone grafting units
US6096081A (en) * 1996-01-16 2000-08-01 University Of Florida Tissue Bank, Inc. Diaphysial cortical dowel
CN101332313A (zh) * 2008-08-06 2008-12-31 北京大学第三医院 纵向钻孔的脱钙皮质骨关节软骨再生用支架及其制备方法
CN101530634A (zh) * 2009-04-21 2009-09-16 昆明医学院第一附属医院 一种仿生双相陶瓷样生物活性骨及其制备方法
US20090312842A1 (en) * 2008-06-16 2009-12-17 Predrag Bursac Assembled Cartilage Repair Graft
US7837740B2 (en) * 2007-01-24 2010-11-23 Musculoskeletal Transplant Foundation Two piece cancellous construct for cartilage repair

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4950296A (en) * 1988-04-07 1990-08-21 Mcintyre Jonathan L Bone grafting units
US6096081A (en) * 1996-01-16 2000-08-01 University Of Florida Tissue Bank, Inc. Diaphysial cortical dowel
US7837740B2 (en) * 2007-01-24 2010-11-23 Musculoskeletal Transplant Foundation Two piece cancellous construct for cartilage repair
US20090312842A1 (en) * 2008-06-16 2009-12-17 Predrag Bursac Assembled Cartilage Repair Graft
CN101332313A (zh) * 2008-08-06 2008-12-31 北京大学第三医院 纵向钻孔的脱钙皮质骨关节软骨再生用支架及其制备方法
CN101530634A (zh) * 2009-04-21 2009-09-16 昆明医学院第一附属医院 一种仿生双相陶瓷样生物活性骨及其制备方法

Cited By (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP2958523A4 (en) * 2013-02-22 2016-11-30 Allosource CARTEL MUSAIC COMPOSITIONS AND METHODS
US9700415B2 (en) 2013-02-22 2017-07-11 Allosource Cartilage mosaic compositions and methods
US10335281B2 (en) 2013-02-22 2019-07-02 Allosource Cartilage mosaic compositions and methods
US11123193B2 (en) 2013-02-22 2021-09-21 Allosource Cartilage mosaic compositions and methods
KR20150132152A (ko) * 2013-03-15 2015-11-25 알로소스 천공된 골연골 동종이식 조성물
EP2967874A4 (en) * 2013-03-15 2016-11-16 Allosource PERFORATED OSTEOCHONDRAL ALLOGRAFT COMPOSITIONS
US9603710B2 (en) 2013-03-15 2017-03-28 Allosource Methods of manufacturing perforated osteochondral allograft compositions
KR102138399B1 (ko) 2013-03-15 2020-07-27 알로소스 천공된 골연골 동종이식 조성물
US11202674B2 (en) 2018-04-03 2021-12-21 Convergent Dental, Inc. Laser system for surgical applications
CN110743039A (zh) * 2019-12-11 2020-02-04 成都奇璞生物科技有限公司 一种自体颅骨用于回植材料的制备方法

Similar Documents

Publication Publication Date Title
US6884621B2 (en) Method and carrier for culturing multi-layer tissue in vitro
TWI394597B (zh) 骨科用生物可分解性補綴物
KR100839875B1 (ko) 관절연골 재생용 지지체
CN101979105A (zh) 一种用于软骨缺损修复的组织工程支架材料及其制备方法
KR20090010607A (ko) 콜라겐을 함유한 관절연골 치료용 이중 지지체
Marmotti et al. Autologous cartilage fragments in a composite scaffold for one stage osteochondral repair in a goat model
CN106075582A (zh) 一种组织工程血管化支架及其构建方法
EP3785740B1 (en) Gradient mineralized bone extracellular matrix material and preparation method therefor
CN104606713B (zh) 三维平行胶原纤维－蚕丝支架及其制备方法与应用
CN101332313B (zh) 纵向钻孔的脱钙皮质骨关节软骨再生用支架及其制备方法
CN102078642B (zh) 一种关节软骨修复再生用支架及其制备方法
Qiang et al. Xenoimplantation of an extracellular-matrix-derived, biphasic, cell-scaffold construct for repairing a large femoral-head high-load-bearing osteochondral defect in a canine model
WO2012097506A1 (zh) 一种关节软骨修复再生用支架及其制备方法
CN110152067B (zh) 组织工程骨支架及其制备方法
CN106421917A (zh) 制备用于软骨损伤修复的组合物的方法
CN102178981B (zh) 一种软骨修复支架材料的制备方法
CN110368529B (zh) 软骨或软骨下骨全层修复支架及其制备方法
CN111514374A (zh) 一种基于共培养系统构建的组织工程软骨促进软骨修复的方法
CN204601181U (zh) 一种半月板支架
Cai et al. A Col I and BCP ceramic bi-layer scaffold implant promotes regeneration in osteochondral defects
CN210843633U (zh) 一种关节软骨修复支架
CN201949388U (zh) 一种关节软骨修复再生用支架
CN106552288A (zh) 制备软骨细胞组合物的方法
CN106421918A (zh) 软骨细胞组合物
CN109876187B (zh) 球状蛋白作致孔剂的组织工程软骨修复支架及其制备方法

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: 11855933

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: 11855933

Country of ref document: EP

Kind code of ref document: A1