EP0413794A1 - Facteur de regeneration osseuse et peridentaire avec matieres et procedes correspondants - Google Patents

Facteur de regeneration osseuse et peridentaire avec matieres et procedes correspondants

Info

Publication number
EP0413794A1
EP0413794A1 EP90904015A EP90904015A EP0413794A1 EP 0413794 A1 EP0413794 A1 EP 0413794A1 EP 90904015 A EP90904015 A EP 90904015A EP 90904015 A EP90904015 A EP 90904015A EP 0413794 A1 EP0413794 A1 EP 0413794A1
Authority
EP
European Patent Office
Prior art keywords
factor
growth factor
cells
periodontal ligament
pdl
Prior art date
Legal status (The legal status 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 status listed.)
Withdrawn
Application number
EP90904015A
Other languages
German (de)
English (en)
Other versions
EP0413794A4 (en
Inventor
Victor P. Terranova
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
CYTOTAXIS Inc
Original Assignee
CYTOTAXIS Inc
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 CYTOTAXIS Inc filed Critical CYTOTAXIS Inc
Publication of EP0413794A1 publication Critical patent/EP0413794A1/fr
Publication of EP0413794A4 publication Critical patent/EP0413794A4/en
Withdrawn legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/475—Growth factors; Growth regulators
    • C07K14/51—Bone morphogenetic factor; Osteogenins; Osteogenic factor; Bone-inducing factor
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/02—Stomatological preparations, e.g. drugs for caries, aphtae, periodontitis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides

Definitions

  • This invention relates to a periodontal ligament cell-attractant factor useful for periodontal and bone regeneration.
  • the invention also relates to regeneration methods, compositions and materials.
  • tetracycline HCl and/or citric acid is given in:
  • a method for promoting periodontal regeneration which includes exposing an area of tooth surface, preconditioning the area with a tetracycline salt, applying fibronectin (FN) and optionally
  • ECGF endothelial cell growth factor
  • Chemotaxis is an essential feature of many biological processes in health and disease.
  • a specific example of chemotaxis is the movement of endothelial cells in the process of neovascularization as disclosed in:
  • laminin The chemotactic properties of laminin, fibronectin and various polypeptides are described, and laminin is shown to promote chemotaxis and growth of human gingival epithelial cells, in:
  • Periodontology Vol. 58, No. 4, April 1987, pp. 247257, the disclosure of which is incorporated herein by reference, describes a new assay system that tests the ability of a number of proteins and polypeptide growth factors applied on dentin to stimulate a chemotactic and proliferative response from various cell types.
  • the invention relates to an isolated
  • periodontal ligament cell-attractant factor which comprises a protein obtainable from periodontal ligament (PDL) cells, said factor having chemoattractant activity to periodontal ligament cells.
  • the invention also relates to a composition useful for periodontal regeneration which comprises a pharmaceutically acceptable amount of said PDL-CTX factor and a pharmaceutically acceptable medium.
  • the invention further relates to a kit useful for periodontal regeneration, which includes said PDL-CTX factor.
  • the invention relates to improvements in a method of periodontal regeneration or of bone regeneration in which said PDL-CTX factor is applied to the surface to be regenerated.
  • the invention relates to improvements in a method for inducing periodontal cell migration on dentin, in which said PDL-CTX factor is applied to the dentin.
  • PDL periodontal ligament
  • the invention relates to improvements in a method of periodontal regeneration in which periodontal ligament cells are applied to the root surface of a tooth, which comprises covering the treated surface with an artificial basement membrane comprised of collagen.
  • the invention also relates to a method of periodontal regeneration, which comprises:
  • Figs. 1 to 5 are graphical representations of experimental data illustrating the chemotactic attraction of PDL cells to the PDL-CTX factor.
  • Fig. 6 is a graphical representation of experimental data illustrating the chemotactic attraction of bone cells to the PDL-CTX factor.
  • Fig. 7 is a graphical representation of experimental data illustrating the chemotactic attraction of parent and selected PDL cells to the PDL-CTX factor.
  • Fig. 8 is a graphical representation of experimental data illustrating 3 H-thymidine
  • Fig. 9 is a chromatogram of a proteincontaining concentrate of PDL cell conditioned media precipitated with ammonium sulfate.
  • PDGF platelet derived growth factor
  • TGF-ß transforming growth factor-ß
  • PDGF transforming growth factor- ⁇
  • EGF epidermal growth factor
  • TGF-ß transforming growth factor-ß appears to have a particularly important role in the repair process. This peptide is found in relatively high concentrations in platelets, in activated T lymphocytes, as well as in macrophages.
  • TGF-ß stimulates the formation of collagen in human or rodent fibroblasts and when injected subcutaneously in newborn mice causes rapid fibrotic and angiogenic response at the site of injection.
  • Another recently discovered source of TGF-ß is bone.
  • TGF-ß is present in bone in amount of almost 100 fold greater than found in other soft tissues. In vitro studies indicate that TGF-ß can control the effects of several other polypeptide growth factors including PDGF, TGF- ⁇ , EGF, AFGF
  • BFGF basic fibroblast growth factor
  • fibroblast growth factor fibroblast growth factor
  • IL-2 interleukin-2
  • AMF autocrine motility factor
  • hepatocytes and bladder epithelial cells hepatocytes and bladder epithelial cells.
  • Terranova et al. enables testing of the potential activity of various biological response modifiers on a dentin substrate.
  • the assay systems is divided into two parts.
  • ASSAY I allows the measurement of the chemotactic activity of the test substance bound to dentin.
  • the cells must actively move through a filter ("Nuclepore” [registered trademark, 100 ⁇ m thick, 8 ⁇ m pore diameter) in response to biological response modifiers bound to dentin.
  • Nuclepore registered trademark, 100 ⁇ m thick, 8 ⁇ m pore diameter
  • ASSAY II the ability of the dentin-bound factors to stimulate directed movement and proliferation of periodontal tissue cells on dentin surfaces is measured. Using these assays it was demonstrated that PDL cells migrate to FN, ECGF and AFGF.
  • FN, ECGF and FGF induce a proliferative response in PDL cells grown on surface-demineralized dentin.
  • Gingival epithelial cells were shown to migrate to LM (laminin) and LM fragments.
  • LM was also shown to stimulate gingival epithelial cell proliferation on native dentin surfaces.
  • the PDL-CTX factor of the present invention is a potent new PDL cell and bone cell attractant factor which is a protein isolated from, derived from or obtainable from PDL cells. It may also be obtained by recombinant DNA methodology or lay peptide
  • the protein factor may be recovered from a medium conditioned by PDL cells (the medium obtained after being used in the tissue culture of the PDL cells).
  • a concentrate of the protein may be recovered as a precipitate by treatment of said medium with a salt such as a
  • An especially useful PDL-CTX factor was derived from periodontal ligament cells which were selected from a plurality of periodontal ligament cells, the selected periodontal ligament cells having been selected on the basis of an increased chemotactic response to said factor relative to the chemotactic response of the other periodontal ligament cells of said plurality of cells.
  • the useful factor may comprise a monomer which forms said tetramer, said monomer having a molecular weight of about 12,500 daltons.
  • Fig. 1 The graphical representation of data in Fig. 1 shows the directed migration of PDL cells to
  • fibroblasts to various concentrations (dilutions) of PDL cell conditioned media where migration is assayed using ASSAY I and each point represents the mean +/- s.d. of triplicate samples.
  • Figure 4 is a graphical representation of data which shows the effect of various concentrations of TGF- ⁇ and TGF-ß on the chemotaxis of PDL cells where migration is assayed using ASSAY 1, all factors are allowed to bind to TTC conditioned dentin (dentin preconditioned in 100 mg/ml of tetracycline HCl) for 30 minutes at 37oC in 100% humidity, PDL cell
  • each assay point represents the mean +/- s.d. of triplicate samples.
  • FIG 5 Data represented in Fig 5 show the effect of various antibodies against extracellular matrix components on migration of PDL cells to 50% PDL cell conditioned media where antibody dilutions are
  • the graphical representation of data in Fig. 6 shows directed migration of human bone cells (ATCC 7009) to 50% PDL cell conditioned media or TGF-ß where migration is assayed using ASSAY I, both factors are allowed to bind to previously TTC conditioned dentin for 30 minutes at 37°C in 100% humidity and each assay point represents the mean of duplicate samples not differing by more than 10%.
  • Fig. 7 the graphical representation of data shows directed migration of parent and selected PDL cells to various dilutions of conditioned media where selection of PDL cells is accomplished by the methodology established in the above mentioned
  • EDTA ethylenediaminetetraacetic acid
  • EGTA ethylene glycol bis(ß-ami ⁇ oethyl ether)-N,N,N',N'-teteraacetic acid
  • 3 H-radioactivity incorporated into the cell is 0.1% ethylenediaminetetraacetic acid (EDTA), 0.1% ethylene glycol bis(ß-ami ⁇ oethyl ether)-N,N,N',N'-teteraacetic acid (EGTA) in a divalent cation free PBS (phosphate-buffered saline), 3 H-radioactivity incorporated into the cell is
  • EDTA ethylenediaminetetraacetic acid
  • EGTA ethylene glycol bis(ß-ami ⁇ oethyl ether)-N,N,N',N'-teteraacetic acid
  • the chromatogram reproduced in Fig. 9 is a profile resulting from PDL cell conditioned media being subjected to (NH 4 ) 2 SO 4 salting out procedure, subjecting a concentrate containing the precipitated protein to TSK55F (molecular sieve column) open column chromatography, and then subjecting the eluted protein having increased activity to reverse-phase high performance liquid chromatography (HPLC).
  • NH 4 ) 2 SO 4 salting out procedure subjecting a concentrate containing the precipitated protein to TSK55F (molecular sieve column) open column chromatography
  • HPLC reverse-phase high performance liquid chromatography
  • epithelial cells did not respond to various conditions
  • concentrations of the PDL cell conditioned media indicating the response is most likely not due to other polypeptide growth factors acting on populations of these cells that may be present in the PDL
  • TGF-ß enhances the activity of the PDL cell conditioned media but
  • NH 2 terminal sequence data indicate a unique peptide.
  • Information from the NIH (U.S. National Institutes of Health) data banks indicates that there is no
  • a chemotactic factor for PDL cells is
  • Antibody against FN or LM does not inhibit the chemotactic properties of this factor.
  • Antibody against ECGF, FGF or EGF does not inhibit the chemotactic properties of this factor.
  • TGF- ⁇ and TGF- ⁇ do not stimulate chemotaxis of PDL cells to the same degree as does the factor.
  • a subpopulation of PDL cells can be isolated that are more chemotactically responsive to the factor.
  • This peptide has high biological activity for PDL cells. Both chemotaxis and
  • the connective tissue of the periodonitium is a complex structure consisting of fibroblasts
  • cementoblasts with associated extracellular matrix, alveolar bone and an extensive extracellular matrix comprised of collagen, glycoproteins and
  • PDL-CTX is believed to have an amino acid sequence from the N-terminal as shown in the following Table A which also, indicate sequence data of known polypeptide growth factors for comparison.
  • the present invention further relates to an isolated periodontal ligament cellattractant protein obtainable from human periodontal ligament cells, said protein comprising an initial amino acid sequence, from the N-terminal, as follows: Val Pro Asp Ser Ser Ala His Lys Lys Ala ... .
  • the present invention further relates to an isolated periodontal ligament cell-attractant protein obtainable from human periodontal ligament cells, said protein comprising an amino acid sequence, ending at the C-terminal, as follows: . . . Pro Val Val Pro Ala Tyr Ala Pro Pro .
  • the present invention further relates to an isolated periodontal ligament cell-attractant protein obtainable from human periodontal ligament cells , said protein having an amino acid sequence, from the N-terminal to the C-terminal , substantially as follows :
  • the PDL-CTX factor of the invention is isolated by being separated from a natural or
  • the PDL-CTX factor of the invention can be in the form of a concentrate of said factor, e.g. a concentrated aqueous solution or dispersion of said factor.
  • the PDL-CTX factor can suitably be obtained by tissue culture of PDL cells in a customary medium, following which the medium contains the PDL-CTX factor.
  • Media containing the factor are referred to as PDL cell conditioned media.
  • separation media may be concentrated, for example by chromatography,
  • a preferred procedure employs a sequence of (1) salting out with ammonium sulfate; (2) centrifugation and discarding the supernatant; repeating (1) and (2); (3) resuspending sediment; (4) fractionating by chromatography; (5) fractionating the eluate
  • the separated or PDL-CTX factor or concentrate thereof is a component of the composition of the invention.
  • a pharmaceutically acceptable amount of the PDL-CTX factor will vary depending upon the intended use, and depending upon the activity of the concentrate. The acceptable amount also may be in an acceptable dosage form for application to teeth or to bone. The acceptable amount of the factor is combined with a pharmaceutically acceptable medium to form the composition of the invention.
  • the growth factors may be selected from the ones described where they enhance regeneration in combination with the PDL-CTX factor.
  • the growth factor suitably may consist of transforming growth factor-ß (TGF-ß), platelet derived growth factor (PDGF), and mixtures thereof.
  • the medium of the composition is aqueous.
  • a suitable amount of the PDL-CTX factor is an amount of from about 10 picograms to about 10 micrograms per ml of the composition.
  • the composition may further comprise TGF-ß in an amount of from about 10 picograms to about 10 micrograms per ml of said composition.
  • the composition may comprise both TGF-ß and PDGF, with TGF-ß in an amount of from about 10 picograms to about 10 micrograms per ml of said composition, and PDGF in an amount of about 10 picograms to about 10 micrograms per ml of said composition.
  • a kit in accordance with the present invention comprises as individual components:
  • the polypeptide growth factor of component (b) may be selected form the group consisting of TGF- ⁇ , TGF-ß, PDGF, and mixtures thereof.
  • the kit may further include as an individual component:
  • polypeptide growth factor of component (b) in a pharmacologically acceptable medium.
  • Component (c) may be selected from the same growth factors as is component (b), but will differ from the particular growth factor which was selected from component (b).
  • the media of components (a), (b) and (c) preferably is aqueous.
  • the active factors of (a), (b) and (c) may be present in the same amounts as specified above in relation to the composition of the present invention.
  • the kit may include an agent which demineralizes enamel and/or dentin, to pre-condition the tooth surface and to provide enhanced binding of components (a) to (c), and improved regeneration.
  • (d-1) an aqueous solution of citric acid.
  • the further agent is tetracycline HCl.
  • the kit may also include, if desired, directions for use, and appropriate materials for the use, such as syringes and materials for suturing and dress the treated area.
  • kit of the invention containing components (a)-(c) is also useful for bone regeneration.
  • Another embodiment of the present invention resides in the use of an artificial basement membrane comprised of collagen to cover the treated surface of a tooth in a method of periodontal regeneration.
  • the treated surface is covered with the membrane.
  • This membrane acts as a selective barrier which inhibits cells which do not display a strong chemotactic response to the factor from
  • a preferred artificial basement membrane comprises type I collagen overlayered with type IV collagen and laminin.
  • the root surface is treated with the PDL-CTX factor of the present invention, and desirably said root surface is additionally treated with a polypeptide growth factor selected from the group consisting of TGF- ⁇ , TGF-ß, PDGF and mixtures thereof.
  • the artificial basement membrane also may be used in the kit of the present invention as a further component.
  • the PDL-CTX factor of the present invention is generally useful in a method of periodontal or bone regeneration. It is particularly useful for applying to dentin in a method directed at inducing PDL cell migration to the dentin.
  • a particularly preferred technique involves treatment of the diseased root with PDL cells selected from the
  • Another embodiment of the present invention relates to a technique for selection of PDL cells in connection with a method of periodontal regeneration.
  • the steps comprise:
  • PDL periodontal ligament
  • step (c) an enriched
  • step (b) population of PDL cells of the culture of step (b) is obtained by incubating the PDL cells of said culture in a first compartment of a chamber having a second compartment containing a solution comprising the PDL-CTX factor, the first and second chambers being separated by a semipermeable membrane, selecting an enriched sub-population of cells which migrate by chemotaxis directed against said PDL-CTX factor, and using said enriched sub-population to further select therefrom cells which migrate through the collagen barrier in step (c).
  • a preferred therapeutic procedure for periodontal regeneration comprises:
  • step (b) before or simultaneously with step (b) there is additionally applied to said surface a pharmaceutically acceptable amount of polypeptide growth factor selected from the group consisting of TGF- ⁇ , TGF-ß, PDGF, and mixtures thereof.
  • a pharmaceutically acceptable amount of polypeptide growth factor selected from the group consisting of TGF- ⁇ , TGF-ß, PDGF, and mixtures thereof.
  • step (b) there is applied to said surface an aqueous solution of tetracycline HCl or an aqueous, saturated solution of citric acid, the application of tetracycline HCl being the preferred one.
  • Assay conditions are 37oC, 100% humidity and a 6 hour incubation using the modified Boyden Chamber Assay System.
  • the barrier in this assay is an 8 ⁇ m pore
  • Nuclepore filter polyvinyl parolodine free.
  • the filters are precoated with gelatin at a concentration of 5 ⁇ g/filter.
  • ASSAY I assay for specific cell migration
  • conditioned media adsorbed to dentin produces a dose dependent response for both PDL cells and bone cells, as shown in the following Table II.
  • Assaying conditions were TTC conditioned dentin (50 mg/block for 5 minutes at 37oC) with 5 ⁇ l application of conditioned media for 30 minutes at 37°C and 100% humidity.
  • TTC conditioned dentin blocks are further conditioned by the application of FN (50 ⁇ g/block) a small but not
  • trypsin sensitivity of the conditioned media is tested. Fifty percent conditioned media is incubated with purified trypsin 10 ⁇ g/ml for various time points. Incubation is stopped by the addition of SBTI (soybean trypsin inhibitor) at a 10 fold excess. Chemoattractant activity is decreased over a 60 minute incubation with trypsin (Fig. 2). SBTI by itself had no effect (either positive or negative) on PDL cell
  • LM is tested at 10 ⁇ g/block for gingival epithelial cells and FN is tested at 100 g/block for gingival fibroblasts. Neither gingival epithelial cells nor gingival fibroblasts respond to conditioned media of PDL cells.
  • PDL-CTX of the present invention is a unique PDL cell synthesized factor
  • the ability of anti-ECGF, anti-FGF and anti-EGF to inhibit 50% conditioned media in a dose dependent manner is examined. No effect of any antibodies on PDL cell movement is altered.
  • Non-specific IgG (NSIgG) is also evaluated, see the following Table III.
  • concentrations all adjusted to start with 100 mg protein per ml media) are incubated with the PDL cells above the Nuclepore filter. Migration is assayed using ASSAY 1. Each assay point is the mean +/- s.d. of triplicate samples.
  • TGF-ß is found in high level in bone and since both TGF- ⁇ and TGF-ß are implicated in both fibroblast proliferation and chemotaxis, the ability of TGF- ⁇ and TGF-ß and 50% conditioned media to generate a chemotactic response in PDL cells is compared. A dose dependent chemotactic response of PDL cells to 50% PDL cell conditioned media, TGF- ⁇ and TGF-ß is observed. PDL cell conditioned media is significantly more effective as a chemoattractant (Fig. 4).
  • TGF-ß is believed to be a "panregulin" the ability of 1 and 10 ng/block TGF/ß ti augment 50% PDL cell conditioned media (CM) is evaluated.
  • CM PDL cell conditioned media
  • dentin blocks are conditioned with 50 mg/ml TTC for 5 minutes at 37oC, rinsed in cold PBS (6 ⁇ 1 minute each rinse) then both TGF-ß and 50% conditioned media applied in 5 ⁇ l aliquotes.
  • TGF-ß 50% conditioned media applied in 5 ⁇ l aliquotes.
  • 1 and 10 ng/block of added TGF-ß an increase in PDL cell migration is observed, as shown in the following Table IV.
  • TGF-ß induced chemotactic response but not the conditioned media induced chemotactic response.
  • TGF-ß fragments of LM or FN.
  • An assay system that can be utilized to separate subpopulations of cells based on their migratory
  • a collagen barrier separates the upper and lower chambers of a Modified Boyden Chamber.
  • a collagen barrier is placed directly over a type IV collagen coated Nuclepore filter (1 ⁇ m pore diameter). Cells that traverse the collagen barrier attach to the coated filter. After various times of incubation the filters are removed and the attached cells subcultured.
  • Type I collagen is cross-linked such that on formation of a 13 mm disc the pore size is no more than 5 ym. This type I collagen matrix is then overlayered with 200 ⁇ g of type IV collagen and 200 ⁇ g of laminin dispersed in 0.05 M Tris, 0.15 M NaCl, pH 7.4. After lyophilization, the
  • artificial basement membranes are sterilized (gas sterilization) and packaged in sterile air tight plastic wrap for subsequent use.
  • a suspension of PDL cells in PBS is applied to teeth using extracted human teeth as a model. 5 ⁇ 10 6 selected PDL cells in PBS are applied by use of a pipet in a drop-wise fashion. The cells are allowed to attach to the root for 30 minutes. After this time period, the teeth are trypsinized and the number of attached cell quantitated by the use of a cell particles counter.
  • Widman-type flaps are raised in the gums of a patient and scrapings from around the junction of the tooth and bone are taken.
  • the scrapings contain PDL cells.
  • scrapings are cultured for growth of PDL cells as follows.
  • the scrapings are added to media containing collagenase-dispase at 100 mg/ml in 10 ml of an isotonic salt solution (ISS) containing 100 mM NaCl, 60 mM mannitol, 25 mM Hepes, 10 mM NaHCO 3 , 6 mM K 2 HPO 4 , lmM CaCl 2 , pH 7.4.
  • ISS isotonic salt solution
  • the contents are vigorously vortexed for two minutes. The medium is removed and centrifuged at 500 x g at 4oC for 3 minutes.
  • the resulting cell pellet is resuspended and washed 3 times in DMEM Dulbecco modified minimal essential melin) with 500 ⁇ g/ml gentamycin.
  • the resulting cells ar then added to type I collagen and FN coated (300 ⁇ g/dish respectively) 35mm tissue culture dishes.
  • the culture medium consists of Media NCTC 109 supplemented with 15% fetal bovine serum (FBS), 1% sodium pyruvate, 1% nonessential amino acid and 25 ⁇ g/ml gentamycin.
  • the most responsive cells are isolated utilizing the following selection technique.
  • the Nuclepore filters are carefully removed and placed top-side down on sterile glass slides. The under-surface is then carefully scraped using a sterile rubber policeman. Ten ⁇ l of NCTC 109 with 10% FBS is next applied to the surface and gently aspirated into a pasteur pipette. These 10 ⁇ l samples with removed PDL cells are transferred to a 35 mm tissue culture dish which has previously been coated with 10 ⁇ l of type I collagen and 10 ⁇ g of FN. The media volume is brought up to 5 ml and the dish immediately anchored at 37o C, 5% CO 2 for 120 minutes. After this time, the
  • unattracted cells are removed by decanting the medium, fresh media added, and the dish returned to the
  • porous type 1 collagen barriers (100 ⁇ m pore size) are overlayed on gelatin-coated 1 ⁇ m pore Nuclepore filters and the selection procedure repeated.
  • the cells that migrate through the collagen barriers attach to the collagen-coated
  • TGF-ß in concentration ranges between 10 picograms per ml PBS and 10 micrograms per ml PBS
  • PDL-CTX in concentrations between 10 picograms and 10 micrograms per ml PBS.
  • Application is accomplished by dripping the material onto the teeth by means of a pasteur pipette.
  • the area treated as described above is then overlayed with an artificial basement membrane.
  • This basement membrane is placed such that the type I collagen side is next to the tooth-bone surface while the type IV collagen-laminin side is next to the soft tissue.
  • the membrane is placed such that it extends 10 mm below the tooth-bone interface, 10 mm to either side (right and left) of the area treated and 10 mm above the crest of the soft tissue flap.
  • Soft tissue flaps are then sutured such that the artificial basement membrane is folded over the crest of the soft tissue flap and secured to the soft tissue with methylmethacrylate.

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Animal Behavior & Ethology (AREA)
  • Engineering & Computer Science (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Orthopedic Medicine & Surgery (AREA)
  • Toxicology (AREA)
  • Zoology (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Biochemistry (AREA)
  • Biophysics (AREA)
  • Genetics & Genomics (AREA)
  • Molecular Biology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
  • Peptides Or Proteins (AREA)

Abstract

Facteur isolé d'attraction des cellules de ligaments péridentaires (PDL-CTX), utile pour la régénération péridentaire et osseuse, comprenant une protéine tirée des cellules de ligaments péridentaires. Ce facteur exerce une attraction chimique sur les cellules de ligaments péridentaires (PDL-). Il peut être combiné avec des facteurs de croissance polypeptidique dans des composés et des kits. Les procédés de régénération péridentaire comprennent l'utilisation du facteur PDL-CTX, l'utilisation de cellules PDL pour une meilleure réaction chimiotactique au facteur PDL-CTX, et/ou l'utilisation d'une membrane basale artificielle à sélection cellulaire.
EP19900904015 1989-02-23 1990-02-22 Periodontal and bone regeneration factor, materials and methods Withdrawn EP0413794A4 (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US31449789A 1989-02-23 1989-02-23
US314497 1999-05-19

Publications (2)

Publication Number Publication Date
EP0413794A1 true EP0413794A1 (fr) 1991-02-27
EP0413794A4 EP0413794A4 (en) 1992-02-26

Family

ID=23220194

Family Applications (1)

Application Number Title Priority Date Filing Date
EP19900904015 Withdrawn EP0413794A4 (en) 1989-02-23 1990-02-22 Periodontal and bone regeneration factor, materials and methods

Country Status (5)

Country Link
EP (1) EP0413794A4 (fr)
JP (1) JPH03505218A (fr)
AU (2) AU5197790A (fr)
CA (1) CA2027583A1 (fr)
WO (1) WO1990010017A1 (fr)

Families Citing this family (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5206023A (en) * 1991-01-31 1993-04-27 Robert F. Shaw Method and compositions for the treatment and repair of defects or lesions in cartilage
US5656593A (en) * 1991-03-11 1997-08-12 Creative Biomolecules, Inc. Morphogen induced periodontal tissue regeneration
CA2363965C (fr) 1991-03-11 2010-05-18 Curis, Inc. Morphogenese induite par des proteines
US5270300A (en) * 1991-09-06 1993-12-14 Robert Francis Shaw Methods and compositions for the treatment and repair of defects or lesions in cartilage or bone
WO1994006399A1 (fr) * 1992-09-15 1994-03-31 Creative Biomolecules, Inc. Regeneration tissulaire periodontique induite par des morphogenes

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0105014A2 (fr) * 1982-09-24 1984-04-04 THE UNITED STATES OF AMERICA as represented by the Secretary United States Department of Commerce Rétablissement de tissu chez les animaux

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0105014A2 (fr) * 1982-09-24 1984-04-04 THE UNITED STATES OF AMERICA as represented by the Secretary United States Department of Commerce Rétablissement de tissu chez les animaux

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
JOURNAL OF DENTAL RESEARCH, vol. 67, special issue, 9th - 13th March 1988, Montreal PO, page 186, abstract no. 584, Washington, D.C., US; V. P. TERRANOVA et al.: "Periodontal ligament cells synthesize an autocrine chemotactic and mitogenic factor" *
See also references of WO9010017A1 *

Also Published As

Publication number Publication date
JPH03505218A (ja) 1991-11-14
AU5197790A (en) 1990-09-26
WO1990010017A1 (fr) 1990-09-07
AU4610993A (en) 1993-12-02
CA2027583A1 (fr) 1990-08-24
EP0413794A4 (en) 1992-02-26

Similar Documents

Publication Publication Date Title
Terranova et al. Repopulation of dentin surfaces by periodontal ligament cells and endothelial cells: effect of basic fibroblast growth factor
EP0148155B1 (fr) Facteurs ostéogéniques
JP3159705B2 (ja) 血管形成ペプチド
Tsukamoto et al. Macrophage production of fibronectin, a chemoattractant for fibroblasts.
Gospodarowicz et al. Isolation of fibroblast growth factor from bovine adrenal gland: physicochemical and biological characterization
Terranova et al. A biochemical approach to periodontal regeneration: Tetracycline treatment of dentin promotes fibroblast adhesion and growth
Terranova et al. Chemotaxis of human gingival epithelial cells to laminin: a mechanism for epithelial cell apical migration
Graf et al. A pentapeptide from the laminin B1 chain mediates cell adhesion and binds to 67000 laminin receptor
AU654938B2 (en) Inhibiting transforming growth factor beta to prevent accumulation of extracellular matrix
AU599647B2 (en) Wound healing composition
US4863899A (en) Biologically active polypeptides
EP0362278A1 (fr) Facteurs de motilite de l'autocrine dans le diagnostic et le traitement du cancer
JPH07504650A (ja) 細胞外マトリックスの蓄積を防止するためのトランスフォーミング増殖因子βの阻害
JP3159216B2 (ja) 白血球由来の成長因子
EP0278781B1 (fr) Peptides à activité de laminine
Yaoi et al. Insulin binds to type V collagen with retention of mitogenic activity
EP0413794A1 (fr) Facteur de regeneration osseuse et peridentaire avec matieres et procedes correspondants
Tweden et al. Neovascularization of surface demineralized dentin
US4935497A (en) Dentin chondrogenic inductive agent
Terranova et al. Chemotaxis of cells isolated from periodontal tissues to different biological response modifiers
Larjava et al. Enhanced synthesis of hyaluronic acid by human gingival fibroblasts exposed to human dental bacterial extract
US4702734A (en) Method of promoting periodontal regeneration and fibroblast bonding
EP0572573A1 (fr) Facteur de stimulation du chondrocyte derive de la thyroide
Ogata Demonstration of pancreatic secretory trypsin inhibitor in serum-free culture medium conditioned by the human pancreatic carcinoma cell line CAPAN-1.
Singh et al. Purification and biochemical properties of a human monocyte-derived growth factor.

Legal Events

Date Code Title Description
PUAI Public reference made under article 153(3) epc to a published international application that has entered the european phase

Free format text: ORIGINAL CODE: 0009012

AK Designated contracting states

Kind code of ref document: A1

Designated state(s): AT BE CH DE DK ES FR GB IT LI LU NL SE

17P Request for examination filed

Effective date: 19910307

A4 Supplementary search report drawn up and despatched

Effective date: 19920107

AK Designated contracting states

Kind code of ref document: A4

Designated state(s): AT BE CH DE DK ES FR GB IT LI LU NL SE

17Q First examination report despatched

Effective date: 19940207

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: THE APPLICATION IS DEEMED TO BE WITHDRAWN

18D Application deemed to be withdrawn

Effective date: 19940818