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The role of fibroblast in ectopic calcification
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The role of fibroblast in ectopic calcification

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The role of fibroblast in ectopic calcification The role of fibroblast in ectopic calcification Presentation Transcript

  • The Role of Dermal Fibroblasts inthe Development of EctopicCalcificationsGiulia AnnoviDepartment of Life SciencesUniversity of Modena and Reggio Emilia
  • CalcificationBiologicalmineralizationEctopiccalcificationPhysiological processBone - TeethCell-mediated processRegulated by proteins, ions,functional moleculesPathological processSoft connective tissuesCell-mediated processInduced by several mechanisms
  • 5. Loss of “inhibitors”4. Presence of “Bone Proteins”1. Circulating nucleational complexes (high Ca / P)2. Cell death3. Proteolysis and ECM degradation
  • CellInhibitors:ANKHPC-1MGPOsteopontinPPiStimulators:Matrix vesiclesANKHPC-1AnnexinsApoptotic bodiesTNAPType I, II, X collagenPiMineral formation(Kirsch, 2006)
  • OsteoarthritisAnkylosing SpondylitisKeutel SyndromeAgingAtherosclerosisAgingChronic renal failureDiabetesBone formationOsteopetrosisAgingOsteoblastsVascular SmoothMuscle CellsChondrocytesFibroblasts?Mesenchymal cells which are resident in a tissue that is only rarely affected bymineralization process.
  • Angioid streaksHaemorragiesCentral vision lossPapulesSkin laxity and redundancyCardiovascularcomplicationsPseudoxanthoma elasticumas soft connective tissue calcification model
  • SEManalysis
  • Calcifying MediumAscorbic Acidβ glicerophosphateDexamethasone
  • Day10Day40Day30Day20DMEM CMHumanDermalFibroblasts
  • Day20Day30Day40
  • The calcification process in a fibroblast cell culturesystem is a slowly progressive event.Fibroblasts phenotype could be different from that ofother mesenchymal cells.
  • Control PXE CM P CM C CM 20GG 6HPXE CM 20GG 6H01234567ControlPXEANKHgeneexpression6h6h20dDMEM CM CMControl6 PXE6 Control6cm PXE6cm C XXg CM P xxg CM0.00.51.01.52.02.5ControlPXEANKH(Relativebandintensity)DMEM CM CM6h 6h 20d
  • Control PXE CM P CM C Cm 20 GG 6HPXE CM 6H 20GG01020PXEControlNPP1geneexpressionDMEM CM CM6h 6h20dc 6h P 6h C 6h cm P 6h CM C XXg cm P XXg CM01234ControlPXEPC1(Relativebandintensity)DMEM CM CM6h 6h20d
  • C CM PXE CM01234ControlPXE4142434ALPactivity(arbitraryunit)DMEM CM CM CM6h6h10d20dTNAP determins higher Pi levels.
  • DMEM CMCM +100 uMLevamisoleControlPXE
  • # Genetic studies have implicated PPi metabolism in the development ofcalcification, but this finding sustains the hypothesis that the calcificationprocess could be due to alterations in local PPi metabolism.# Results indicate that NPP1, being capable to produce PPi, is normallyexpressed by human dermal fibroblasts and is significantly upregulated in acalcifying environment.# However, the potent inhibitory action of PPi is abolished by thecontemporary hyperactivation of TNAP, which is consistent with its critical rolein mineralization process.# The higher upregulation of TNAP in PXE could be a condition associatedwith ECM calcification.# Restoration of PPi extracellular pool, by inhibiting TNAP activity, is able tocounteract mineral deposits formation.Conclusions
  • Federica BoraldiRoberta TiozzoIvonne RonchettiDaniela QuaglinoUniversity of ModenaMaria I. Garcia FernandezUniversity of MalagaThanks toAnne de PaepeOlivier VanakkerUniversity of GhentLeon J.SchurgersCees VermeerUniversity of Maastricht
  • Is it TNAP sufficient to induce ectopic calcification in PXE?No,it is NECESSARY but NOT SUFFICIENT50kDa37kDaActin Control3-10NL50kDa37kDaActin PXE3-10NLGla-MGP Control15kDa10kDaGla-MGP PXE15kDa10kDaMGP
  • Control PXE C 20GG 6h PXE 20GG 6H01234ControlPXEBMP2GENEexpression6h20dControl6 PXE60.00.51.01.5 ControlPXEBMP2(Relativebandintensity)6h20dBMP2