WO1996003160A1 - Suspension de cellules de fibrine pour la creation de nouveaux tissus - Google Patents

Suspension de cellules de fibrine pour la creation de nouveaux tissus Download PDF

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Publication number
WO1996003160A1
WO1996003160A1 PCT/US1995/009374 US9509374W WO9603160A1 WO 1996003160 A1 WO1996003160 A1 WO 1996003160A1 US 9509374 W US9509374 W US 9509374W WO 9603160 A1 WO9603160 A1 WO 9603160A1
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Prior art keywords
cells
fibrinogen
cell
fibrin
patient
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PCT/US1995/009374
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English (en)
Inventor
Charles A. Vacanti
Mark A. Randolph
C. Derek Sims
Peter E. M. Butler
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Children's Medical Center Corporation
The General Hospital Corporation
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Application filed by Children's Medical Center Corporation, The General Hospital Corporation filed Critical Children's Medical Center Corporation
Priority to AU31457/95A priority Critical patent/AU3145795A/en
Publication of WO1996003160A1 publication Critical patent/WO1996003160A1/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
    • A61L27/38—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix containing added animal cells
    • A61L27/3804—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix containing added animal cells characterised by specific cells or progenitors thereof, e.g. fibroblasts, connective tissue cells, kidney cells
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L24/00—Surgical adhesives or cements; Adhesives for colostomy devices
    • A61L24/0005—Ingredients of undetermined constitution or reaction products thereof
    • 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
    • A61L26/00—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form
    • A61L26/0009—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form containing macromolecular materials
    • A61L26/0028—Polypeptides; Proteins; Degradation products thereof
    • A61L26/0042—Fibrin; Fibrinogen
    • 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
    • A61L26/00—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form
    • A61L26/0057—Ingredients of undetermined constitution or reaction products thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
    • A61L27/38—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix containing added animal cells
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
    • A61L27/38—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix containing added animal cells
    • A61L27/3839—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix containing added animal cells characterised by the site of application in the body
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • C12N5/0068—General culture methods using substrates
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2533/00—Supports or coatings for cell culture, characterised by material
    • C12N2533/50—Proteins
    • C12N2533/56—Fibrin; Thrombin

Definitions

  • the present invention is generally in the area of medical treatments, and specifically relates to an method for creating new tissue, especially cartilage, bone and endothelial tissue.
  • Cartilage is a specialized type of dense connective tissue consisting of cells embedded in a matrix. There are several kinds of cartilage.
  • Translucent cartilage having a homogeneous matrix containing collagenous fibers is found in articular cartilage, in costal cartilages, in the septum of the nose, in larynx and trachea.
  • Articular cartilage is hyaline cartilage covering the articular surfaces of bones.
  • Costal cartilage connects the true ribs and the sternum.
  • Fibrous cartilage contains collagen fibers.
  • Yellow cartilage is a network of elastic fibers holding cartilage cells which is primarily found in the epiglottis, the external ear, and the auditory tube.
  • U.S. Patent No. 5,041,138 to Vacanti, et al. discloses the use of an open synthetic polymeric matrix which is seeded with cells for implantation into a patient to create new cartilage.
  • a fibrous structure is preferred to allow maximum surface area for attachment of cells at the same time as allowing for sufficient diffusion of nutrients and gases to maintain cell viability in the absence of vascularization.
  • dissociated cells are seeded onto matrices which are then implanted and evolve into new tissue. This technique has now been used extensively in animals and a human, with promising results. This technique is limited however, by the availability of suitable polymeric support materials.
  • a method for creating new tissue wherein cells are suspended in cryoprecipitate or fibrinogen (jointly referred to herein as "fibrinogen") which is clotted by exposure to thrombin or other serine esterase.
  • the cells and fibrinogen are autologous, derived by biopsy and cryoprecipitation of a plasma sample, respectively.
  • Other protein matrices can be substituted for the fibrin, where the protein has similar characteristics and can be crosslinked easily by addition of an agent such as calcium.
  • the method is particularly useful for creation of new cartilage in the repair of defects such as worn or torn joint linings or construction of ears and noses.
  • the method is also particularly useful where tissue such as endothelial tissue or cartilage is used to oblate a structure, such as fallopian tubes or the vas deferens, or in the repair of defects such as incomplete closure of structures, for example, the heart. Examples demonstrate formation of new cartilage in mice.
  • the method described herein generally consists of making a suspension of dissociated cells that are to be implanted to form new tissue with an aqueous solution of a crosslinkable endogenous protein, implanting the suspension into the site where tissue is to be formed, and crosslinking the protein.
  • the protein solution is cryoprecipitate or fibrinogen derived from the patient in which the tissue is to be implanted, which is clotted with human thrombin, and the cells are autologous cells derived from the patient by biopsy.
  • cryoprecipitate is prepared from a plasma sample obtained directly from the patient into which the cell suspension is to be implanted.
  • One unit of human blood consists of approximately 350 to 450 mis, which is preferably collected in ACD (citric acid-dextrose) anticoagulant, although other acceptable anticoagulants can be used such as ethylene diamine tetraacetate.
  • ACD citric acid-dextrose
  • the red blood cells are removed by centrifugation or filtration, and the separated plasma chilled at 4°C until cryoprecipitate is formed, typically about three days.
  • Cryoprecipitate consists predominantly of fibrinogen. Fresh frozen plasma can also be used.
  • fibrinogen is also available from commercial suppliers such as Sigma Chemical Co., Baxter Diagnostics, and Ortho Pharmaceuticals. As used herein, the term "fibrinogen” is intended to encompass either cryoprecipitate or purified fibrinogen, unless specifically stated otherwise.
  • Other materials that can be used as a source of fibrinogen besides cryoprecipitate, fresh frozen plasma, and purified fibrinogen include factor VIII concentrate, platelet concentrate, and platelet rich plasma.
  • Proteins other than fibrinogen can be substituted for the fibrinogen where the protein can be prepared and crosslinked using a physiologically acceptable crosslinker such as calcium.
  • a physiologically acceptable crosslinker such as calcium.
  • cells of the same species and preferably immunological profile are obtained by biopsy, either from the patient or a close relative, which are then grown to confluence in culture using standard techniques known for cell culture and used as needed. If cells that may elicit an immune reaction are used, such as human chondrocytes from an immunologically distinct individual, then the recipient can be immunosuppressed as needed, for example, using a schedule of steroids and other immunosuppressant drugs such as cyclosporine A, Imuran, or prednisone, using the dosages recommended by the manufacturer for immunosuppression of transplants, and/or the cells irradiated prior to transplantation.
  • the cells are autologous. Cultured cells of some cell types are available from sources such as the American Type Culture Collection, Rockville, MD. Only normal, non-transformed cells which exhibit contact inhibition characteristic of normal cells should be implanted.
  • Examples of cell types that can be implanted to form cartilage include chondrocytes, subchondral cells and fibroblasts.
  • Examples of cell types that can be implanted to form bone include osteocytes, periosteocytes and fibroblasts, under appropriate conditions.
  • Examples of parenchymal cell types include many types of cells present in tissues such as intestine, pancreas, liver. Examples of other cell types that can be implanted include muscle cells, Dermafibroblasts, mesenchymal cells and cells forming tendons and ligaments.
  • cartilage can be obtained from another site in the patient or from autopsy, using for example, cartilage obtained from joints or rib regions.
  • the cartilage is sterilized, for example, by washing in Povidone-Iodine 10% solution (Betadine, Purdue
  • the muscle attachments are dissected from the underlying bone to expose the joint surfaces.
  • the cartilage from the articulating surfaces of the joint is then sharply dissected from the underlying bone.
  • the cartilage is cut into pieces with dimensions of less than 5 mm per side and washed twice in Phosphate Buffered Saline (PBS) with electrolytes and adjusted to neutral pH.
  • PBS Phosphate Buffered Saline
  • the minced cartilage is then incubated at 37°C in a solution of 0.2% clostridial collagenase (Worthington CLS II, 140 U/mg) and agitated overnight as described by Klagsbrun, (Methods in Enzvmolocrv. Vol. VIII). This suspension is then filtered using a 153 ⁇ g nylon sieve (Tetko,
  • the cells are washed with PBS, counted using a hemocytometer, and concentrated to yield a chondrocyte concentration of between approximately 10 7 to 10 8 cells/cc, most preferably 5 x 10 7 cells/cc.
  • the isolated chondrocytes can be cultured in Hamm's F-12 culture media and 10% fetal calf serum with L-glutamine (292 ⁇ g/cc) , penicillin (100 U/cc) , streptomycin (100 ⁇ g/cc) and ascorbic acid (5 ⁇ g/cc) , at 37°C.
  • chondrocytes Precursor cells of chondrocytes can also be used in place of the chondrocytes.
  • An example is fibroblasts which differentiate to form chondrocytes.
  • chondrocytes includes chondrocyte precursor cells. Other cell types are similarly dispersed.
  • muscle cells can be obtained by mincing small detrusor muscle fragments to approximately 0.5 mm diameter and using these as explants in 100 mm tissue culture dishes containing 10 mL of DMEM supplemented with 10% fetal calf serum. Outgrowth is routinely observed at 72 hr after explants are placed in culture. When cultures are 80% confluent, the cells were trypsinized and passaged. Cell populations highly enriched in elongated, striated cells are routinely obtained using this method.
  • Cells are preferably suspended in nutrient media, such as Ham F12 or MEM 199, or in saline or buffer, for mixing with fibrinogen and implantation.
  • nutrient media such as Ham F12 or MEM 199
  • saline or buffer for mixing with fibrinogen and implantation.
  • the fibrinogen is clotted by exposure to an enzyme such as thrombin in the presence of physiological calcium immediately prior to or at the time of implantation. Additional crosslinking is obtained by exposure to factor Xllla.
  • thrombin is commercially available from the same suppliers as purified human fibrinogen, for example, Parke- Davis or Johnson and Johnson; it appears to be irrelevant whether it is human or bovine. In the preferred embodiment, 10,000 units of thrombin is added to the cryoprecipitate from one unit of whole blood.
  • Other serine esterases are known to those skilled in the art, such as factor Xa, which can be used instead of thrombin to cleave the fibrinogen into fibrin. Other esterases such as factor Xllla can also be used.
  • Thrombin and factor Xllla cleave fibrinogen or fibrin monomers, respectively, during the normal clotting process.
  • the cells are mixed with the fibrinogen, thrombin and optionally factor Xllla, in the presence of calcium (typically, 80 mM) , to form a cell-hydrogel matrix which is implanted.
  • the fibrinogen from one unit of blood is mixed with approximately 25 x 10 6 cells/ml.
  • the cell- fibrin/fibrinogen or cryoprecipitate mixture is then implanted at the site where tissue is to be created. As the cells proliferate in si tu, new tissue forms to the extent of the implanted material, i.e., the fibrinogen-cell mixture, but not beyond.
  • tissue formation is effective to treat the disorder or defect.
  • cartilage is formed that serves as a joint lining.
  • the cell solution is implanted into the joint, and cartilage forms over a period of a few weeks.
  • the joint is immobilized while new.cartilage is formed.
  • cartilage is formed that serves as a structure such as a nose or ear.
  • the fibrinogen-cell suspension is implanted at the site and immediately shaped, preferably simultaneously or immediately prior to crosslinking.
  • tissue which can be cartilage or other cell types is formed by implantation of the appropriate fibrinogen-cell suspension at the site where tissue formation is desired. In some cases this will be to obstruct a lumen or close an opening resulting from a congenital defect, trauma, or disease.
  • alginate As reported by Atala et al, "Injectable alginate seeded with chondrocyte as a potential treatment for vesicoureteral reflux" J. Urol. 150:745 (1993), alginate, a biodegradable polymer, embedded with chondrocytes, can serve as a synthetic substrate for the injectable delivery and maintenance of cartilage architecture in humans.
  • a biopsy of the symphysis pubis can be easily and quickly performed using a biopsy gun followed by chondrocyte processing and endoscopic injection of the autologous chondrocyte/alginate suspension for the treatment reflux.
  • the alginate can be replaced with fibrinogen clotted with thrombin. It is preferable to implant rather than inject the fibrinogen-cell mixture, however, and the degradation profile of the totally endogenous fibrinogen is distinct from that of alginate or synthetic polymers.
  • chondrocytes can be easily harvested and combined with a matrix in vi tro, implanted cystoscopically and the elastic cartilage tissue formed is able to correct vesicoureteral reflux without any evidence of obstruction.
  • the resulting autologous cartilage is non-antigenic, non-migratory, and volume stable. Since the chondrocyte are autologous, this method of treatment does not require FDA approval.
  • the procedure can be performed under 15 minutes, with a short period of a mask anesthetic, in the outpatient unit, without any need for a hospital stay. Neither vesical nor perivesical drainage is required. Since the whole procedure is done endoscopically and the bladder is not entered surgically, there is no postoperative discomfort. The patient can return to a normal level of activity almost immediately.
  • the cells to be implanted are obtained, for example, by biopsy from the patient who is also to be the recipient, the cells are dissociated and proliferated, a unit of blood is drawn from the patient and cryoprecipitate made, the cryoprecipitate is mixed with the cells and thrombin in the presence of calcium, and the mixture implanted at the site where tissue is to be formed: in a lumen, in a joint lining, in the mesentery or adjacent an adequate blood supply or highly vascularized area, optionally with a portocaval shunt in the case of hepatocytes and other cells benefitting from the addition of hepatotropic factors.
  • the cell-matrix suspensions could be used to obliterate fistula related to the sinus tract, nasal-cutaneous, bronchopleural and intraabdominal fistulae, by endoscopic injection of the suspension into the cavity.
  • Heart valves, trachea, inner ear structures, and implants could be formed by using molds seeded with cells in a fibrin matrix allowed to grow in vi tro, then implanted subcutaneously.
  • Example 1 Preparation of Fibrinogen-Cell Matrix.
  • the twice frozen and thawed plasma was centrifuged at 4°C at 4200 rpm for 12 minutes. Following centrifugation the bag containing the cryoprecipitated fibrinogen was inverted and the plasma drained into the attached transfer bag, leaving 3 to 5 cc of concentrated fibrinogen.
  • the product for clinical use routinely contains 39.8 to 58.9 mg/ml fibrinogen.
  • the autologous fibrinogen concentrate can be frozen at less than -18°C for storage of up to one year from the collection date of the whole blood. The range of concentrations obtainable is from 20 to 77 mg/ml.
  • the autologous fibrinogen concentration Prior to surgical application, the autologous fibrinogen concentration is thawed at 30 to 37°C in a water bath.
  • the thawed fibrinogen concentrate may be stored at 20 to 24°C for up to six hours before preparing the fibrin matrix.
  • Preparation of Fibrin Matrix Four ml of calcium chloride solution (40 ⁇ iM/liter) was added to a vial of thrombin (1000 U) and swirled until completely dissolved, for a final concentration of 250 U/ml. Equal volumes of calcium chloride/thrombin and fibrinogen concentrate were mixed together simultaneously according to the surgical protocol for application of the fibrin suspension. Fibrinogen gels into a fibrin clot within 20 to 30 seconds with a thrombin mixture concentrate of 250 U/cc, which is preferred for injection. Higher concentrations of thrombin
  • Example 2 Neocartilage formation in a fibrin glue matrix.
  • Bovine chondrocytes were isolated by enzymatic digestion from the glenohumeral joint surface of calf forelimbs. The isolated cells were combined with 1 ml of cryoprecipitate to create a final cell density of 12.5 x 10 6 cells/ml. Bovine thrombin was added to the mixture to produce a polymerizing gel. The resulting fibrin glue-chondrocyte admixture was placed subcutaneously in eighteen sites in nude mice. Separate control implants consisting of chondrocyte suspension or fibrin glue alone were also implanted. All specimens were weighed and processed for histological, total DNA, and glycoaminoglycan (GAG) analysis to demonstrate production of neocartilage.
  • GAG glycoaminoglycan
  • Histology and histomorphometry Specimens from each concentration and time point were collected. One half of each specimen was placed in 10% buffered formalin for histological preparation (the remaining half was used for GAG and DNA analysis described below) . The tissue was fixed for a minimum of 24 hours and dehydrated in increasing concentrations of ethanol and embedded in paraffin. Five micron sections were made and stained with hematoxylin and eosin, toluidine blue, safranin-O. Cartilage samples from donor articular cartilage processed in an identical manner served as one control. The histological sections were examined in a blinded fashion by three independent investigators and scored.
  • Glycosaminoglycan content Total glycosaminoglycan (GAG) content of the cell/polymer constructs was determined using a dimethylmethylene blue (DMB) dye method. DMB stock solution was made using DMB, NaCl, Glycine, Na azide, 1 N HC1, and water. The samples were digested with papain (125 mg/ml buffer of 0.1 M NaH 2 P0 4 , 5 mM EDTA, 5 mM cysteine HCl at pH 7.0 overnight at 60°C) before adding the dye. GAG was to be reported as a percent of wet weight.
  • DMB dimethylmethylene blue
  • Total DNA content was determined using one-fourth of an implant specimen using the bisbenzimidazol fluorescent dye method.
  • the dye (Hoechst 33258) was used in a concentration of 0.2 mg/ml in a 0.01 M Tris, 1 mM EDTA and 0.1 M NaCl.
  • the samples were digested with papain (125 mg/ml in buffer of 0.1 M NaH 2 P0 4 , 5 mM EDTA, 5 mM cysteine HC1 at pH 7.0 overnight at 60°C) before adding the dye.
  • a spectrofluorometer was used to estimate the fluorescence of the digested samples, emission was measured at 400-550 nm for an excitation wavelength of 365 nm.
  • Uncultured chondrocytes in a solution of calf thymus DNA was used to generate standardized curves.
  • the fibrin glue-chondrocyte implant specimens had the gross appearance of cartilage; whereas the control specimens were entirely absorbed and unretrievable.
  • the presence of cartilage was confirmed histological by a blinded histopathological observer.
  • DNA/GAG analysis confirmed the presence of actively proliferating chondrocytes with production of a well-formed cartilagenous matrix.
  • the mean GAG content in the neomatrix was 473 ⁇ g/ml (SD ⁇ 240) .
  • the mean amount of GAG in relation to wet weight was 1.9% (SD + 0.8%) .
  • the mean DNA content was estimated at 40.7 ⁇ g/ml (SD + 12.4).

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Abstract

Procédé de création de nouveaux tissus dans lequel des cellules sont mises en suspension dans un cryoprécipité ou dans du fibrinogène (tous deux désignés ici par fibrinogène) qui est coagulé en présence de thrombine ou d'une autre sérine estérase. Dans le mode de réalisation préféré, les cellules et le fibrinogène sont autologues, et dérivés respectivement par biopsie et cryoprécipitation d'un échantillon de plasma. Ce procédé s'avère particulièrement utile pour créer de nouveaux cartilages dans la réparation de défectuosités telles que des ligaments usés ou déchirés, ou la reconstitution de nez ou d'oreilles. Il est particulièrement utile lorsque des tissus tels que des cartilages ou de l'endothélium sont utilisés pour obturer une structure telle que les trompes de Fallope ou le canal déférent, ou pour réparer des malformations telles que la fermeture incomplète de structures, telles que le c÷ur. Des exemples décrivent la formation de nouveaux cartilages chez les souris.
PCT/US1995/009374 1994-07-26 1995-07-26 Suspension de cellules de fibrine pour la creation de nouveaux tissus WO1996003160A1 (fr)

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Cited By (14)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1996040304A1 (fr) * 1995-06-07 1996-12-19 Reprogenesis, Inc. Compositions d'hydrogels injectables
WO1999015209A2 (fr) * 1997-09-19 1999-04-01 Baxter Aktiengesellschaft Eponge a base de fibrine
US5912177A (en) * 1994-06-29 1999-06-15 Common Services Agency Stem cell immobilization
US6139578A (en) * 1995-05-19 2000-10-31 Etex Corporation Preparation of cell seeded ceramic compositions
DE19926083A1 (de) * 1999-06-08 2000-12-14 Universitaetsklinikum Freiburg Biologisches Gelenkkonstrukt
WO2001059068A2 (fr) * 2000-02-08 2001-08-16 Schmitt-Waldburg Gbr Copeaux osseux artificiels, procedes permettant de les produire et utilisation
ES2221770A1 (es) * 2002-04-19 2005-01-01 Eduardo Anitua Aldecoa Metodo de preparacion de un compuesto para la regeneracion de tejidos.
EP2316462A1 (fr) * 2002-09-07 2011-05-04 Quy Biosciences Limited Compositions comprenant des cellules souches mésenchymateuses
US8480757B2 (en) 2005-08-26 2013-07-09 Zimmer, Inc. Implants and methods for repair, replacement and treatment of disease
US8518433B2 (en) 2003-12-11 2013-08-27 Zimmer, Inc. Method of treating an osteochondral defect
US9138318B2 (en) 2007-04-12 2015-09-22 Zimmer, Inc. Apparatus for forming an implant
US9314420B2 (en) 2005-10-26 2016-04-19 Jan-Eric Ahlfors Acellular bioabsorbable tissue regeneration matrices
US10167447B2 (en) 2012-12-21 2019-01-01 Zimmer, Inc. Supports and methods for promoting integration of cartilage tissue explants
US11918608B2 (en) 2014-07-25 2024-03-05 Recellerate, Inc. Methods of treating exercise-induced pulmonary hemorrhage

Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
GB2137209A (en) * 1983-03-23 1984-10-03 Univ Ramot Repair of cartilage and of bones
EP0339607A2 (fr) * 1988-04-29 1989-11-02 Samuel Dr. Itay Composition pour la réparation de cartilage et d'os et procédé pour sa préparation comme implant de tissu du squelette
WO1992009301A1 (fr) * 1990-11-27 1992-06-11 The American National Red Cross Compositions contenant un agent de fermeture des tissus et des facteurs de croissance, qui accelerent la cicatrisation
WO1992022312A1 (fr) * 1991-06-17 1992-12-23 Wadstroem Jonas Composition pour le traitement de tissus corporels, contenant de la fibrine ou du fibrogene et un polymere biodegradable et biocompatible
WO1994002182A1 (fr) * 1992-07-18 1994-02-03 Opperbas Holding B.V. Composition de colle fibrine a deux composants servant a ameliorer la fertilisation in vitro
WO1994025080A1 (fr) * 1993-04-30 1994-11-10 Massachusetts Institute Of Technology Compositions de cellules de polysaccharide injectables

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
GB2137209A (en) * 1983-03-23 1984-10-03 Univ Ramot Repair of cartilage and of bones
EP0339607A2 (fr) * 1988-04-29 1989-11-02 Samuel Dr. Itay Composition pour la réparation de cartilage et d'os et procédé pour sa préparation comme implant de tissu du squelette
WO1992009301A1 (fr) * 1990-11-27 1992-06-11 The American National Red Cross Compositions contenant un agent de fermeture des tissus et des facteurs de croissance, qui accelerent la cicatrisation
WO1992022312A1 (fr) * 1991-06-17 1992-12-23 Wadstroem Jonas Composition pour le traitement de tissus corporels, contenant de la fibrine ou du fibrogene et un polymere biodegradable et biocompatible
WO1994002182A1 (fr) * 1992-07-18 1994-02-03 Opperbas Holding B.V. Composition de colle fibrine a deux composants servant a ameliorer la fertilisation in vitro
WO1994025080A1 (fr) * 1993-04-30 1994-11-10 Massachusetts Institute Of Technology Compositions de cellules de polysaccharide injectables

Cited By (27)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5912177A (en) * 1994-06-29 1999-06-15 Common Services Agency Stem cell immobilization
US6277151B1 (en) 1995-05-19 2001-08-21 Etex Corporation Cartilage growth from cell seeded ceramic compositions
US6139578A (en) * 1995-05-19 2000-10-31 Etex Corporation Preparation of cell seeded ceramic compositions
US6544290B1 (en) 1995-05-19 2003-04-08 Etex Corporation Cell seeding of ceramic compositions
AU720569B2 (en) * 1995-06-07 2000-06-08 Curis, Inc. Injectable hydrogel compositions
WO1996040304A1 (fr) * 1995-06-07 1996-12-19 Reprogenesis, Inc. Compositions d'hydrogels injectables
WO1999015209A2 (fr) * 1997-09-19 1999-04-01 Baxter Aktiengesellschaft Eponge a base de fibrine
WO1999015209A3 (fr) * 1997-09-19 1999-06-17 Immuno Ag Eponge a base de fibrine
US6548729B1 (en) 1997-09-19 2003-04-15 Baxter Aktiengesellschaft Fibrin sponge
DE19926083A1 (de) * 1999-06-08 2000-12-14 Universitaetsklinikum Freiburg Biologisches Gelenkkonstrukt
WO2000074741A3 (fr) * 1999-06-08 2002-07-18 Universitaetsklinikum Freiburg Produit de synthese biologique pour articulation
WO2000074741A2 (fr) * 1999-06-08 2000-12-14 Universitätsklinikum Freiburg Produit de synthese biologique pour articulation
WO2001059068A3 (fr) * 2000-02-08 2002-02-14 Michael Sittinger Copeaux osseux artificiels, procedes permettant de les produire et utilisation
WO2001059068A2 (fr) * 2000-02-08 2001-08-16 Schmitt-Waldburg Gbr Copeaux osseux artificiels, procedes permettant de les produire et utilisation
ES2221770A1 (es) * 2002-04-19 2005-01-01 Eduardo Anitua Aldecoa Metodo de preparacion de un compuesto para la regeneracion de tejidos.
EP2316462A1 (fr) * 2002-09-07 2011-05-04 Quy Biosciences Limited Compositions comprenant des cellules souches mésenchymateuses
US8524268B2 (en) 2003-12-11 2013-09-03 Zimmer, Inc. Cadaveric allogenic human juvenile cartilage implant
US8518433B2 (en) 2003-12-11 2013-08-27 Zimmer, Inc. Method of treating an osteochondral defect
US8652507B2 (en) 2003-12-11 2014-02-18 Zimmer, Inc. Juvenile cartilage composition
US8765165B2 (en) 2003-12-11 2014-07-01 Zimmer, Inc. Particulate cartilage system
US8784863B2 (en) 2003-12-11 2014-07-22 Zimmer, Inc. Particulate cadaveric allogenic cartilage system
US8834914B2 (en) 2003-12-11 2014-09-16 Zimmer, Inc. Treatment methods using a particulate cadaveric allogenic juvenile cartilage particles
US8480757B2 (en) 2005-08-26 2013-07-09 Zimmer, Inc. Implants and methods for repair, replacement and treatment of disease
US9314420B2 (en) 2005-10-26 2016-04-19 Jan-Eric Ahlfors Acellular bioabsorbable tissue regeneration matrices
US9138318B2 (en) 2007-04-12 2015-09-22 Zimmer, Inc. Apparatus for forming an implant
US10167447B2 (en) 2012-12-21 2019-01-01 Zimmer, Inc. Supports and methods for promoting integration of cartilage tissue explants
US11918608B2 (en) 2014-07-25 2024-03-05 Recellerate, Inc. Methods of treating exercise-induced pulmonary hemorrhage

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