WO2008097230A1 - Composition de greffe de tissu comportant une moelle osseuse autologue et une thrombine autologue purifiée - Google Patents

Composition de greffe de tissu comportant une moelle osseuse autologue et une thrombine autologue purifiée Download PDF

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Publication number
WO2008097230A1
WO2008097230A1 PCT/US2007/003502 US2007003502W WO2008097230A1 WO 2008097230 A1 WO2008097230 A1 WO 2008097230A1 US 2007003502 W US2007003502 W US 2007003502W WO 2008097230 A1 WO2008097230 A1 WO 2008097230A1
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Prior art keywords
bone marrow
concentrate
graft material
combining
composition
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PCT/US2007/003502
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English (en)
Inventor
Sherwin Kevy
May Jacobson
James Ellsworth
Kevin Benoit
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Harvest Technologies Corporation
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Priority to PCT/US2007/003502 priority Critical patent/WO2008097230A1/fr
Publication of WO2008097230A1 publication Critical patent/WO2008097230A1/fr

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    • 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

Definitions

  • the present invention relates to a point-of-care method for the preparation of a tissue graft/wound healing composition capable of enhancing tissue regeneration wherein the composition comprises a bone marrow aspirate or bone marrow concentrate and purified autologous thrombin.
  • Bone and soft tissue grafting is a common surgical procedure to achieve wound closure.
  • An example of bone grafting is the fusion of bone tissue for the repair of degenerative, traumatic, oncologic or infectious conditions.
  • An example of soft tissue grafting is the application of autogenous skin recovered from a donor site and applied to a soft tissue injury in order to facilitate healing. The autogenous skin graft creates a biocompatible and bioactive wound covering and wound environment that may enhance wound healing.
  • Osteoconductive bone graft materials typically include synthetic matrices, autogenous bone and matrices of cadaveric origin.
  • Bone graft materials that are osteoinductive include factors, among them bone morphogenetic proteins, which recruit progenitor cells to the graft site and subsequently induce growth and differentiation of the progenitor cells. Osteogenic bone graft materials generally contain the precursor cells that ultimately differentiate to form new bone tissue.
  • the gold standard for graft material is autogenous, that is, tissue taken from the patient, because it possesses all three functional characteristics described above and is inherently non-immunological. Due to the high incidence of donor site morbidity associated with autogenous grafts, however, there has been a great deal of interest in developing other graft substitutes with reduced morbidity.
  • bone marrow In addition to being the repository of precursor cells needed for hematopoiesis, bone marrow also contains a population of mesenchymal stem cells (MSC) which are capable of differentiating into different mesodermal tissues, including bone, muscle, tendon and fat.
  • MSC mesenchymal stem cells
  • Bone marrow has been shown to enhance the rate of bone formation in animal long bone defect models and enhance the rate of healing of soft tissue defects.
  • Graft materials consisting of bone marrow alone or in conjunction with another tissue graft material, have been investigated as potential tissue graft substitutes, with mixed results.
  • the method of the present invention provides for the preparation of an autologous tissue graft material derived from bone marrow and peripheral whole blood harvested from the patient.
  • the bone marrow can be concentrated to yield an increased number of nucleated cells such as osteogenic and hematopoietic progenitors.
  • Purified autologous thrombin is purified from whole blood and combined with the bone marrow or bone marrow cell concentrate to generate a graft material that may be used alone, eliminating the need for harvesting autogenous skin or bone, or in combination with supplemental graft materials including autograft, allograft, xenograft or a synthetic graft composition, prior to application to the graft site.
  • the invention relates to a tissue graft composition
  • a tissue graft composition comprising autologous bone marrow or a concentrate of bone marrow-derived cells and purified autologous thrombin.
  • the composition may further comprise a graft material selected from autograft, allograft, xenograft and a synthetic graft material.
  • the invention relates to a composition
  • a composition comprising autologous bone marrow or bone marrow concentrate, purified autologous thrombin and a platelet concentrate or platelet rich plasma.
  • This composition may further comprise a graft material selected from autograft bone, allograft, xenograft and a synthetic graft material.
  • the invention relates to a method for the preparation of a tissue graft composition comprising a bone marrow concentrate and purified autologous thrombin.
  • the method comprises: a) obtaining a volume of anticoagulated bone marrow from a patient;
  • the invention relates to a method for the preparation of a tissue graft composition wherein the method comprises the further step of mixing the graft composition with a supplemental graft material.
  • suitable graft materials include allograft, autograft, mineralized or demineralized bone, hydroxyapetite or other synthetic matrix material.
  • tissue graft material or "tissue graft composition” refers to a composition applied to a wound site that is the result of injury, disease or surgery and includes sites in bone and cartilage as well as soft tissue.
  • the tissue graft material is used to promote healing and tissue regeneration and can be used therefore, as a wound healing composition.
  • anticoagulant refers to a substance capable of preventing whole blood from clotting. Any anticoagulant capable of inhibiting coagulation of a whole blood or bone marrow specimen is suitable for use in practicing the method of the present invention; examples include, without limitation, ethylenediamine tetraacetic acid (EDTA), heparin and preferably, citrate-based anticoagulants, such as acid-citrate-dextrose (ACD) and citrate-phosphate- dextrose (CPD)
  • EDTA ethylenediamine tetraacetic acid
  • ACD acid-citrate-dextrose
  • CPD citrate-phosphate- dextrose
  • bone marrow concentrate or “concentrate of bone marrow- derived cells” refers to an enriched population of bone-marrow-derived nucleated cells without regard to selection of a subset of nucleated cells.
  • the bone marrow is concentrated by centrifugation of an anticoagulated bone marrow aspirate to generate a product in which nucleated cells and platelets are concentrated and red blood cells and plasma are consequently reduced.
  • the tissue graft material of the present invention derives its benefit from its autologous nature, its rapid preparation and its flexibility to be able to enhance tissue regeneration alone or in combination with other graft materials.
  • the tissue graft material of the present invention combines an autologous bone marrow aspirate or concentrate with a purified autologous thrombin preparation.
  • the bone marrow component is the source of cellular components, both differentiated and undifferentiated
  • the benefits of this particular combination derive from the flexibility associated with first obtaining an anticoagulated autologous bone marrow aspirate, which can then be manipulated, for example, concentrated by reduction of the red blood cell and plasma components and then coagulating the bone marrow preparation in a controlled fashion by the addition of autologous thrombin to the bone marrow aspirate or concentrate.
  • Coagulation of the bone marrow preparation is critical to hemostasis and the stabilization of the graft within the site. Coagulation of the bone marrow also improves handling characteristics and helps the graft adhere to the wound. Coagulation of the bone marrow after processing ensures delivery to the graft site of an optimal amount of the cellular and protein components necessary for improved tissue regeneration. Being able to control the coagulation event, therefore, is an important feature of the present invention.
  • Bone marrow from the patient is harvested in accordance with accepted medical practice, generally, from a site in the iliac crest, tibia, humerus etc. Bone marrow is collected into a syringe (multiple syringes may be required, depending on the amount of bone marrow material needed for the graft) containing an anticoagulant, such as heparin, ACD or CPD to prevent clotting. Optimally, the volume of anticoagulant required is approximately 15-20% of the bone marrow volume, although other anticoagulant volumes may also be effective. Alternatively, the bone marrow is collected in a syringe not containing anticoagulant but is subsequently expressed into a suitable container containing the appropriate amount of anticoagulant for further processing.
  • the anticoagulated bone marrow is then centrifuged at, for example, 1,000 x g for 10 minutes to obtain fractionation of the various bone marrow/whole blood components into discrete regions of the centrifugation vessel.
  • Plasma and red blood cells (RBC) are discarded and the buffy coat, which contains the concentrated bone marrow-derived nucleated cells (BMC) and platelets, is recovered.
  • Bone marrow components may be fractionated using standard centrifugation techniques known to those of skill in the art.
  • a bone marrow concentrate is obtained by placing the bone marrow aspirate in a sterile processing disposable compatible with an automated centrifugation system, such as the SMARTPREP ® system (Harvest Technologies Corp, Madison, MA).
  • SMARTPREP ® system Hardvest Technologies Corp, Madison, MA.
  • This system as described in U.S. Patent No. 5,707,331, consists of an automated microprocessor controlled centrifuge with decanting capability and a swinging bucket designed to allow for rapid automatic separation of plasma and platelets from a sample of whole blood.
  • the system uses a dual chambered processing disposable container of 20ml and 60 ml volume capable of processing any volume of blood or bone marrow from 18ml to 60ml.
  • a unique feature of the disposable is a floating shelf of a specific gravity that rises during the initial centrifugation step and is capable of separating red blood cells from other blood/marrow components.
  • the anticoagulated bone marrow is first placed in one chamber of the container.
  • the centrifuge is then operated to cause the red blood cells to sediment to the bottom of the chamber. Centrifugation is stopped causing the RBC-reduced bone marrow to decant to a second chamber.
  • the container is then centriruged a second time resulting in a density-dependent separation of the nucleated cells of the bone marrow from the plasma component. Most of the plasma is removed.
  • the bone marrow concentrate is then harvested and resuspended to a concentration of approxmately 2 to 4 times baseline levels.
  • a bone marrow concentrate is obtained using the method and apparatus described in U.S. application serial number PCT/US04/15654. Briefly, the bone marrow is aspirated into a syringe which acts as the processing disposable. The anticoagulated bone marrow preparation is then centrifuged at, for example, 1,000 x g for 10 mins. To facilitate recovery of the desired nucleated cell fraction, the syringe, like the processing disposable described above contains a density disk assembly which floats within the syringe such that the assembly encompasses the fractionated nucleated cells. The position of the disc is determined by the patient's hematocrit.
  • the bone marrow concentrate is combined with purified autologous thrombin to form the bone graft material of the invention.
  • autologous thrombin Prior to or concurrently with the preparation of the bone marrow concentrate, autologous thrombin is purified from an aliquot of whole blood taken from the patient.
  • the methodology for the isolation of a purified autologous thrombin preparation is described in co-pending U.S. application serial number 10/765,694. Briefly, purified autologous thrombin is obtained from a whole blood sample taken from the patient.
  • the method comprises the steps of a) obtaining a volume of anticoagulated whole blood from the patient; b) mixing the anticoagulated whole blood with a precipitating agent; c) incubating the mixture of b) for a time sufficient for precipitation of cellular and specific plasma components to occur; d) separating the precipitate obtained in c) from the supernatant (usually by centrifugation and/or filtration); and e) recovering the supernatant wherein the supernatant contains purified autologous thrombin.
  • a small volume of anticoagulated whole blood is obtained by drawing blood from the patient into a blood collection tube or syringe which contains an anticoagulant, for example, acid-citrate-dextrose. After thorough but gentle mixing, the anticoagulated whole blood is transferred to a glass or plastic tube containing a precipitating agent, such as ethanol, and is mixed with the anticoagulated whole blood. The resulting mixture is incubated at room temperature for a period of time sufficient for precipitation of the cellular and specific plasma components of the blood to occur, about 20-60 minutes. Sufficient precipitation will be evidenced by the formation of a viscous precipitate consisting of agglomerized cells and insoluble proteins.
  • an anticoagulant for example, acid-citrate-dextrose.
  • a precipitating agent such as ethanol
  • the mixture is then centrifuged for about 5-30 minutes at 1,000-3,000 x g to pack the precipitate at the bottom of the tube. Finally, the supernatant above the precipitate is removed from the tube; the supernatant being that fraction of the mixture that contains purified autologous thrombin.
  • the volume of whole blood used to prepare the autologous thrombin will be small, for example, as little as 8 to 10 ml.
  • the blood is drawn into a blood collection tube (e.g. a VACUT AINER® tube) or syringe containing a non-hep arm anticoagulant.
  • a blood collection tube e.g. a VACUT AINER® tube
  • syringe containing a non-hep arm anticoagulant.
  • anticoagulants include calcium ion-binding or sequestering anticoagulants, such as, citrate-phosphate-dextrose (CPD) or acid-citrate- dextrose (ACD), sodium citrate, and the like.
  • the preferred anticoagulants are acid-citrate-dextrose (ACD) and ACD/mannitol.
  • Typical precipitating agents will include, for example, polyethylene glycol, ammonium sulfate or ethanol, as well as such components as calcium chloride or magnesium chloride.
  • ethanol is used as a precipitating agent.
  • the final concentration of ethanol will preferably be between 10% and 25%. For an 8 to 10 ml starting whole blood volume, therefore, 1 to 2 ml of 100% or 95% ethanol is added to the whole blood.
  • precipitate may be expected to form in the tube within about 5 to 45 minutes.
  • the initial volume of whole blood may be anticoagulated with a mixture of ACD and mannitol, with the concentration of mannitol being about 5-10 mg/1 ml ACD.
  • the bone marrow concentrate (BMC) is then combined with purified autologous thrombin (AT) in a BMCrAT ratio sufficient to promote clotting of the bone marrow concentrate, preferably 1:1-6:1, and more preferably, 3:1-5:1. Clotting of the BMC may be timed to occur prior to or following insertion of the graft materials into the graft site.
  • BMCrAT ratio sufficient to promote clotting of the bone marrow concentrate, preferably 1:1-6:1, and more preferably, 3:1-5:1. Clotting of the BMC may be timed to occur prior to or following insertion of the graft materials into the graft site.
  • Composite bone graft materials frequently include a porous implantable matrix which provides a scaffolding for the distribution of bone-healing progenitor cells and growth factors. While no matrix is required for the bone graft material of the present invention, if in the surgeon's discretion, a matrix material is desired for the particular indication, the coagulated bone marrow concentrate may be combined with matrices conventionally used to facilitate bone fusion, for example, mineralized and demineralized cancellous bone, and various synthetic matrices including coralline hydroxyapatite.
  • TRP Platelet Rich Plasma
  • TC Platelet Concentrate
  • Platelet rich plasma (PRP) and/or a platelet concentrate (PC) may also be combined with the bone marrow concentrate prior to exposure with purified autologous thrombin to form the bone graft composition of the invention.
  • separation and concentration of bone marrow and peripheral blood platelets and white blood cells is achieved by combining an effective amount of bone marrow aspirate and peripheral blood and processing this composition as described for concentration of bone marrow separately.
  • the combination of purified autologous thrombin with the anticoagulated bone marrow concentrate results in the formation of a coagulated tissue graft composition. Timing of activation of the bone marrow concentrate to form the coagulated tissue graft composition of the present invention varies.
  • the bone marrow concentrate may be injected into the graft site prior to or simultaneously with addition of purified autologous thrombin so that clotting of the bone marrow concentrate occurs in situ.
  • the bone marrow concentrate and purified autologous thrombin may be combined ex vivo and applied directly to the graft site in a coagulated form.
  • supplemental graft material may be combined with the bone marrow concentrate prior to or subsequent to coagulation with purified autologous thrombin.
  • a canine critical bone defect model is used to illustrate the healing ability of a novel mixture of bone marrow-derived cells, autologous thrombin and optimally, platelet concentrate.
  • the critical bone defect is produced by resection of a 21 -mm diaphyseal section of the femur that results in a non-union if left untreated.
  • a duplicate study includes a supplemental graft material, such as tricalcium phosphate (TCP) matrix in groups B-D.
  • a 21mm defect is surgically created at the middiaphysis of the femur, stabilized with a plate and treated. The defect is allowed to heal for 16 weeks. Animals are sacrified and the quality of the bone is analyzed using radiographic and biomechanical techniques.
  • the surgical procedure is described by Kraus et al. and is summarized here.
  • the femur to be operated is chosen in a controlled randomized fashion and prepared for aseptic surgery.
  • a standard lateral approach is made to the femur.
  • An 8-hole, 135mm long, 4.5 mm, leg-lengthening plate (available from Synthes, Paoli, PA) is contoured and applied with bicortical screws to the lateral aspect of the femur.
  • the plate is removed and a 21 -mm cylindrical section of diaphysis and its associated periosteum is removed with an oscillating bone saw.
  • the surgical site is copiously lavaged during the osteotomy procedure to avoid heat necrosis of the bone and to remove all bone debris.
  • the plate is reapplied.
  • the defect is filled with the appropriate graft material according to the treatment randomization.
  • Autologous cancellous bone is harvested from the greater tubercle of the humerus ipsilateral to the femur to be operated.
  • the humerus is prepared for aseptic surgery.
  • a 12-mL syringe is prepared preoperatively by cutting off the end where a hypodermic needle attaches.
  • the inner diameter of the 12mL syringe is 14 mm, similar to the mean diameter of the femoral diaphysis in these dogs.
  • the autologous cancellous bone chips collected with a 7 mm curette is immediately transferred to the 12-mL syringe.
  • the syringe is filled to 21 mm (the length of the defect in the diaphysis).
  • the cancellous bone graft will partially clot during collection forming a cylinder with dimensions 14 mm wide and 21 mm long.
  • the cylindrical graft is transferred to the defect from the syringe.
  • a periosteal elevator is used to mold the graft to completely fill the cavity. Wound closure is routine.
  • the dogs are returned to separate runs after recovery from surgery and an overnight stay in the intensive care unit.
  • PC Platelet Concentrate
  • AT Autologous Thrombin
  • the cervical area is aseptically prepared and jugular venipuncture is performed with a 19 gauge needle and a total of 59 ml of whole blood is collected.
  • 45ml of whole blood is collected in the first syringe containing 5ml of an anticoagulant (anticoagulant citrate dextrose, ACD).
  • 9ml of blood is collected in a second syringe containing ImI of an anticoagulant (mannitol and anticoagulant citrate dextrose, ACD).
  • the 50ml of blood/ACD mixture is centrifuged using the SMARTPREP ® system for separation of the blood into PC. This process will yield a PC volume of 5ml.
  • ImI is used for PC assay and 4ml is placed in a sterile container for mixing with BMC and mixing with AT.
  • the 10ml blood/mannitol ACD mixture is centrifuged in the SMARTPREP ® system and 4ml of AT is produced.
  • the proximal humerus, ipsilateral to the femur that is to be operated, is prepared for aseptic surgery as described above. Twelve, 2ml aspirates are collected from the proximal humerus separated by approximately 1 cm by changing the direction and depth of the needle placement. Two such aspirates are drawn into a 10 ml syringe containing 2ml of ACD as an anticoagulant. This process is repeated to obtain three syringes from the left humerus and three from the right. The total bone marrow volume drawn is 24ml. The total anticoagulated volume is combined in a sterile cup for a total pooled sample volume of 36ml. A ImI sample of the pooled sample is used to assay the number of nucleated cells present as a baseline. The remaining 35ml volume is placed in a separation disposable cartridge and centrifuged using the
  • SMARTPREP ® system for separation of the components of the bone marrow aspirate. This process will yield 4ml of bone marrow concentrate which is placed in a sterile cup to be used in preparing the implantation graft material in Groups B or C.
  • a bone marrow aspiration needle is inserted into the greater tubercle of the proximal humerus.
  • Two aspirates of 2 ml each are drawn, one from the leit humerus and one from the right.
  • Each syringe contains ACD anticoagulant. The contents of both syringes are used in Group D and may be combined with the TCP matrix.
  • Group B implants are prepared as follows: 4 cc of TCP is rehydrated in 4 ml of BMC. The TCP material is placed into a 5 ml syringe and 4 ml of BMC is delivered into the graft material. After 120 seconds, 1.5 ml of AT is delivered into the TCP/BMC material. A 3 ml syringe with a cannula is used to deliver the AT. After delivery, this mixture is allowed to set up for 120 seconds. The resultant formed graft material is expressed from the syringe and placed into the defect.
  • Group C implants are prepared as follows: 4cc of TCP is rehydrated in 4 ml of BMC. The TCP material is placed into a 5 ml syringe and 4 ml BMC is delivered into the graft material. This hydrated material is allowed to stand for 120 seconds. Then a 2ml mixture of PC and AT is delivered in a 3 : 1 ratio. The delivery of these combined autologous materials is accomplished through a dual lumen cannula pushed through the center of the TCP/BMC material to the base of the syringe. The PC/AT mixture is expressed simultaneously as the cannula is withdrawn from the syringe containing the TCP/BMC.
  • Group D implants are prepared as follows: 4 cc of TCP is placed into a 5 ml syringe. 4 ml of BMA is added to the TCP material and allowed to stand for 120 seconds. Then a 2ml mixture of PC and AT is delivered in a 3:1 ratio. The delivery of these combined autologous materials is accomplished through a dual lumen cannula pushed through the center of the TCP/BMC material to the base of the syringe. The PC/AT mixture is expressed simultaneously as the cannula is withdrawn from the syringe containing the TCP/BMC. After delivery, this mixture is allowed to set up for 120 seconds. The resultant formed graft material is expressed from the syringe and placed into the defect. The surface to the graft material and the surrounding tissue is then coated with 2.5 ml PC and 0.7 ml AT expressed simultaneously from a dual lumen cannula to seal the surface of the graft and place an enhanced level of proteins on the adjacent tissue.
  • Body temperature, pulse, and respiration rate is mom ' tored daily. Dogs are evaluated for lameness daily on a 5 point scale (Budsberg, et al. 1993) 64 . The femoral diaphysis is palpated daily after one week and the response to deep palpation is recorded.
  • AU dogs are sedated with acepromazine (0.2 mg/kg) and butorphanol (0.2 mg/kg) intravenously for chemical restraint during radiography.
  • standard cranial-caudal (anterior-posterior) and lateral radiographs of both femora are taken to ensure that all dogs have normal hind limbs.
  • Postoperative cranial-caudal and lateral radiographs are taken to verify implant placement and to serve as a baseline for radiographic evaluation.
  • Serial cranial-caudal radiographs of the operated femur only is taken at 4, 8, 12, and 16 weeks as the leg-lengthening plate used to stabilize the fracture will obscure the bone healing defect in the lateral projection.
  • the lateral radiograph is taken after the plate and screws are removed.
  • radiographic standard is placed at the level of the femur that consists of two radiopaque spheres spaced 100 mm apart. This standard will allow for correction of magnification and reliable measurement of the femoral diameter.
  • An aluminum step wedge phantom will also be included for density quantification.
  • Aradiologist will evaluate the radiographs for evidence of bone healing according to established criteria. Scores are based on mineralization, bony bridging, continuity of bony bridging, loss of cortex at each end of the defect as bone forms, size of extraosseous callus, and integrity of the bone-plate construct, especially signs of screw loosening. The radiologist is blinded to treatment-group assignment.
  • Computed tomography (CT) scans are taken of the excised femora (Picker PQS CT system).
  • a Plexiglass fixture is used to consistently position the bones.
  • 2-mm thick image slices are acquired at 1-mm spacing, producing 1 mm of overlap.
  • a Cann-Genant bone phantom is included in the field ofview to allow conversion of the CT (Hounsfield) units to tissue mineral density.
  • a 3 -cm high volume of interest (VOI) is reconstructed including the defect.
  • the 21-mm defect is selected within the VOI for analysis.
  • the bone mineral density (g/cm 3 ) histogram is determined slice by slice along the defect and for the entire graft VOI by summing across the graft slices.
  • the mineralized graft volume is also determined. If sufficient remodeling has occurred, the polar moment of inertia and section modulus of the newly formed bone tissue is calculated from the CT reconstructions and correlated with the torsional parameters from strength testing.
  • the femora is wrapped in saline-soaked gauze, double bagged, and stored at -20 C until mechanical testing. The whole bone strength of both operated and contralateral non-operated femurs is tested. Only specimens radiographically graded as healed is tested.
  • the prepared specimens are tested in torsion using a servohydraulic biaxial load frame (MiniBionix 858, MTS Systems).
  • the torque is applied at 1 degree/second failure, and no axial load is applied. Torque and angle is sampled at 5 Hz throughout the test. The angular displacement normalized by the gauge length is the twist.
  • the torsional stiffness is determined from the initial linear portion of the torque-twist curve.
  • the torsional strength is the torque at which failure occurred, and the corresponding deformation is the twist at failure.
  • the energy absorbed to failure is calculated from the area beneath the torque-twist curve.
  • the present invention provides a method of preparing an autologous tissue graft material having the following characteristics:
  • It can be prepared at point of care from a bone marrow aspirate and a whole blood sample from the individual receiving the graft during the course of a surgical procedure to implant the graft.
  • the autologous bone marrow graft material can be delivered to the graft site alone or in conjunction with a platelet concentrate or platelet rich plasma by a variety of techniques or devices.
  • the autologous bone marrow graft material of the present invention can be applied directly to a graft site or combined with another graft material.

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Abstract

L'invention concerne un procédé pour la préparation d'une composition de greffe de tissu autologue. De la moelle osseuse est prélevée à partir du patient. Dans un mode de réalisation, les cellules nucléées de la moelle osseuse sont ensuite concentrées. De la thrombine autologue est purifiée à partir d'un volume de sang total prélevé également à partir du patient. La moelle osseuse aspirée ou la moelle osseuse concentrée est ensuite combinée avec la thrombine autologue purifiée pour former un matériau de greffe de tissu coagulé qui peut être utilisé seul ou en association avec d'autres matériaux de greffe.
PCT/US2007/003502 2007-02-09 2007-02-09 Composition de greffe de tissu comportant une moelle osseuse autologue et une thrombine autologue purifiée WO2008097230A1 (fr)

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Cited By (2)

* Cited by examiner, † Cited by third party
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WO2010118979A1 (fr) 2009-04-07 2010-10-21 Velin-Pharma A/S Méthode et dispositif de traitement de pathologies associées à l'inflammation ou à l'activation indésirable du système immunitaire
WO2011029903A1 (fr) 2009-09-10 2011-03-17 Flemming Velin Procédé d'élaboration de micro-arn et application thérapeutique de celui-ci

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US20050101992A9 (en) * 1989-06-23 2005-05-12 Epstein Gordon H. Method of Applying Composition To A Surface
US20050205498A1 (en) * 2003-03-28 2005-09-22 Sowemimo-Coker Samuel O Preparation of a cell concentrate from a physiological solution
US20060246161A1 (en) * 2003-02-21 2006-11-02 Hongtao Xing Method of making medicament for treating anemia

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US20050101992A9 (en) * 1989-06-23 2005-05-12 Epstein Gordon H. Method of Applying Composition To A Surface
US20060246161A1 (en) * 2003-02-21 2006-11-02 Hongtao Xing Method of making medicament for treating anemia
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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2010118979A1 (fr) 2009-04-07 2010-10-21 Velin-Pharma A/S Méthode et dispositif de traitement de pathologies associées à l'inflammation ou à l'activation indésirable du système immunitaire
WO2011029903A1 (fr) 2009-09-10 2011-03-17 Flemming Velin Procédé d'élaboration de micro-arn et application thérapeutique de celui-ci
US9078914B2 (en) 2009-09-10 2015-07-14 Velin-Pharma A/S Method for the preparation of micro-RNA and its therapeutic application

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