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1,2 Gundul a Schul z e ‐ T anzil , VMD R e s e a r c h g r oup “ Bioreconstruction ”

1,2 Gundul a Schul z e ‐ T anzil , VMD R e s e a r c h g r oup “ Bioreconstruction ”. 1 Institute of Anatom y , Paracelsus Medical Universit y , Salzburg and Nuremberg, Germany

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1,2 Gundul a Schul z e ‐ T anzil , VMD R e s e a r c h g r oup “ Bioreconstruction ”

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  1. 1,2GundulaSchulze‐Tanzil,VMD Researchgroup“Bioreconstruction” 1InstituteofAnatomy,ParacelsusMedicalUniversity,Salzburgand Nuremberg,Germany 2Departmentof Orthopaedic,TraumaandReconstructiveSurgery,Charité- UniversitätsmedizinBerlin,CampusBenjaminFranklin,Germany 1

  2. • • Education: 1990‐1996 1999 1999 2005 Schoolofveterinarymedicine,FreieUniversitätBerlinVMDDegree,FreieUniversitätBerlin PostdoctoralFellow,InstituteofAnatomy,FreieUniversitätBerlinScientificassistant,CentrumofAnatomy,CharitéUniversityof • • • • Medicine,CCM,Berlin Dep.ofTraumaandReconstructiveSurgery,CBF/CentrumofAnatomy,CCM,CharitéUniversityofMedicine,Berlin Leaderoftheresearchlaboratoryofexperimentalsurgery,AGBioreconstruction Dep.ofTraumaandReconstructiveSurgery,CBF,CharitéUniversity 2005‐2008 • 2007 • • 2008‐2014 ofMedicine,Berlin Habilitation:AnatomyandCellBiology ProfessorshipattheInstituteofAnatomy,ParacelsusMedicalUniversity,SalzburgandNuremberg,Germany 2009 2015 • • TeachingActivities: Dissectioncoursesingrossanatomy,coursesinhistology,seminarsandlessionsinanatomyandcellbiology Scholarships: • • • • 1996‐1999 2005‐2008 Doctoralscholarship,FreieUniversitätBerlinRahel‐HirschhabilitationScholarship,CharitéBerlin 2

  3. Adhocreviewerfor ActaBiomaterialia,AnnalsofAnatomy,ArthritisRheumatism,ArthritisResearchTherapy,BerlinerMünchnerTierärztlicheWochenschrift,BMCMusculoskeletalDisorders,Biomaterials,BiotechnologyProgress,BiotechnologyBioengineering,Bone,CellTissueResearch,CellBiologyandToxicology,CellsTissuesOrgans,ClinicalMedicineInsights:ArthritisMusculoskeletalDisorders,ClinicalExperimentalImmunology,ConnectiveTissueResearch,Cytokine,DrugDevreviews,EuropeanCellsMaterials,EuropeanJHistochemistry,ExperimentalBiologyandMedicine,FibrogenesisandTissueRepair,FoliaBiologica,GeneTherapy,HistochemistryCellBiology,HistologyHistopathology,InternationalJMolecularScience,JBiochipsTissueChips,JCellularBiochemistry,JTissueEngineeringRegenerativeMedicine,JBiomedicineandBiotechnology,JBiomedMaterResPartA,JImmunolMethods,JOrthopSurgRes,JNanomedicine,MaterialsLetters,MedicinalChemistry,OsteoarthritisCartilage,OxidAntioxidMedSci,PLOSone,ReviewMaterialLetters,TheLancet,TissueEngineeringPartA.

  4. Scientificinterests • cartilageandtendon/ligamentbiology • osteoarthritis(invitroandinvivomodels) • tendoninjuryandhealing(invitroandinvivomodels) • interplayofanti‐andproinflammatorycytokinesintendonandcartilage,co‐culturemodels • cell‐andbiomaterialbasedcartilageandtendonrepair:cartilageandtendontissueengineering • tendon/ligamentageing • comparativeanatomy/cellbiologyoftendonsandcartilagesubtypes

  5. Techniques • Cellbiology • characterizationofhumanandanimal-derivedprimarycells:chondrocytes,tenocytes,ligamentcells,synovialfibroblasts,PBMCsandneutrophils • three-dimensionalcultures(pellet,organoid,spheroids,hydrogelandscaffoldcultures,staticalanddynamicalseedingofbiomaterialsandselfpreparedcell-freedextracellularmatrices • interactionofmusculoskeletalcellsinco-cultures • immunohistochemistry/-cytochemistry,immunofluorescencemicroscopy • confocallaserscanningmicroscopy • flowcytometry • histology/histopathologyofmusculoskeletaltissues • transmission-/scanningelectronmicroscopy • ELISA,assays(cytokines,proliferation,DMMB-assay,cellvitality,cytotoxicity,caspaseactivity,TUNEL…) • SDS-PAGEandWesternBlotAnalyses • RTPCRandRealTime(RTD)-PCR • transfectionofprimarycells:adenoviral,lipofection,Electroporation • genesilencingusingsiRNA

  6. Techniques • Animalmodels • matrixassistedautologouschondrocytestransplantation(chondralvs.Osteochondraldefects) • LohanAetal.,2014 • partialAchillestendondefectmodelintherabbit(StollCetal.,2011,Biomaterials) • nudemicexenograftmodel:invivo Chondrogenesisandtendogenesis • LohanAetal.,2011:HistochemCellBiol • osteoarthritismodelintherat(surgicallyinduced:MMT) t intheminipigandrabbi grossanatomicalpreparationtechniquesofhumanandanimal-derivedsamples

  7. Somerecentpublications(2014) • MrosewskiI,JorkN,GorteK,ConradC,WiegandE,KohlB,ErtelW,JohnT,OberholzerA,KapsC,Schulze-TanzilG(2014).RegulationofOAassociatedkeymediatorsbyTNFαandIL-10:EffectsofIL-10overexpressioninhumansynovialfibroblastsandasynovialcellline.CellTissueRes357(1):207-23. • ThePhantom5Consortium(2014).Apromoterlevelmammalianexpressionatlas.Nature27;507(7493):462-70. • GrögerD,KerschnitzkiM,WeinhartM,SchneiderT,KohlB,WagermaierW,Schulze-TanzilG,FratzlP,HaagR(2014).SelectivityinBoneTargetingwithDifferentPolyanionicDendriticDyeConjugates.AdvancedHealthcareMaterials3(3):375-85. • LohanA,MarzahnU,ElSayedK,HaischA,MüllerRD,KohlB,StölzelK,ErtelW,JohnT,Schulze-TanzilG(2014).Osteochondralcartilagedefectrepairusingautologousorthotopicandheterotopicchondrocytesintherabbitmodel.AnnAnat.196(5):317-26. • GirkeG,KohlB,BuschC,JohnT,GodkinO,ErtelW,Schulze-TanzilG(2014).Tenocyteactivationandregulationofcomplementfactorsinresponsetoinvitrocellinjury.MolImmunol.60(1):14-22. • StölzelK,Schulze-TanzilG,OlzeH,SchwarzS,FeldmannEM,RotterN(2014).ImmortalisedhumanmesenchymalstemcellsundergochondrogenicdifferentiationinalginateandPGA/PLLAscaffolds.CellTissueBank.2014May16 • HoyerM,MeierC,BreierA,HahnerJ,HeinrichG,DrechselN,MeyerM,RentschC,GarbeLA,ErtelW,LohanA,Schulze-TanzilG.Invitrocharacterizationofself-assembledanteriorcruciateligamentcellspheroidsforligamenttissueengineering.HistochemCellBiol.2014Sep26. • JagielskiM,WolfJ,MarzahnU,VölkerA,LemkeM,MeierC,ErtelW,GodkinO,ArensS,Schulze-TanzilG(2014).TheinfluenceofIL-10andTNFαonchondrogenesisofhumanmesenchymalstromalcellsinthree-dimensionalcultures.IntJMolSci.15(9):15821-44. • HoyerM,DrechselN,MeyerM,MeierC,HinüberC,BreierA,HahnerJ,HeinrichG,RentschC,GarbeLA,ErtelW,Schulze-TanzilG,LohanA.(2014).Embroideredpolymer-collagenhybridscaffoldvariantsfor ligamenttissueengineering.MaterSciEngCMaterBiolAppl.1;43:290-9.

  8. Cartilagetissueengineering tocharacterize/comparethequalityoftissueengineeredcartilageproducedbyheterotopicchondrocytesseededonpolyglycolicacid(PGA)scaffoldsinvitroandinvivo porcineauricular,nasoseptal articularchondrocytes dynamicculture 3weeks→analysis Implantedinonudemice1,6,12weeks → analysis LohanAetal.,2011 HistochemCellBiol.136(1):57-69

  9. Chondrogenesisbyheterotopicchondrocytesinvivo 1week6weeks12weeks explantation articular auricular non-seedednasoseptal resorbed LohanAetal.,2011 HistochemCellBiol.136(1):57-69

  10. Chondrogenesisbyheterotopicchondrocytemono‐ andcocultures articular auriculararticular/auricularnasoseptalarticular/nasoseptal lightmicroscopy 100µm live/deathassay 100µm alcianbluestaining 20µm articular/nasoseptal articular/auricular celltracking ElSayedKetal.,2013 JTissueEng RegenMed7,61-72. 100µm

  11. Cartilagerepairinvivo intactarticularcartilage chondralfullthicknessdefect repairtissueafter6month HE glycosaminoglycanstaining(red) LohanAetal.,2013,195(5):488-97.

  12. influenceofautologousleukocytesontenocytes? PBMCs inflammation tenocyteresponse • tocytokines? • toleukocytes? • tocomplementactivation? • tocellinjury? transwellsystem:indirectco-culture neutrophils Achillestenocytes 10ng/mLTNFα,24h4x106rabbitleukocytes 105rabbittenocytes MMP-1 100µm relativegeneexpression TNFα IL-6 IL-1β 30 30 10 8 2.5 2.0 * * * 20 20 6 4 2 0 1.5 1.0 0.5 0.0 ** 10 10 0 0 co co co co TNF TNF PBMC TNF PBMC TNF neutro. PBMC neutro. PBMC neutro. neutro. Al-SadiOetal.,2012.MusclesLigaments TendonsJ.20121(3):68-76.

  13. tenocyteresponsetocellinjury scratchassay computerguidedreproducibleplottingsystem „healing“afterinjury 4h8h 16h 24h green:vital,red:deadcells humanHamstringtenocytes GirkeGetal.,2014MolImmunol60(1):14-22.

  14. tenocyteresponsetocellinjury tenocyteresponse 10 • tocytokines? • toleukocytes? • tocomplementactivation? • tocellinjury? * 8 relativegeneexpression 6 4 * * * 2 * * * 0 control 4+24h 4h 4h 4h 4h 4h 4h 4h 24h 24h 24h 24h 24h 24h 24h MMP-1 C3aR CD46 CD55 C5aR TNFα IL-1β humanHamstringtenocytes GirkeGetal.,2014MolImmunol

  15. decellularizedtendonECMscaffolds humantenocytes recellularization decellularization porcineAStendon decellularized ECM reseededtendonECM tendonrepair? HE Lohanetal.,2013,Schulze-TanzilGetal.,2012 ConnectTissueRes.54(4-5):305-12.Cells1:1010.

  16. tenocyteimplantationinpartialtendondefects(rabbit) rabbitAchilles Achillestenocytes tendon rabbittenocytes expansionin monolayer PGA-culture PGA-culture cellvitality medialM. gastrocnemiustendon 1week emptydefect implantedPGA implantationin tendondefect PGA-transplant 6weeks healing? green=vitalred=dead macroscopicalandhistologicalscoring StollCetal.,2011Biomaterials32:4806

  17. tenocyteimplantationinpartialtendondefects(rabbit) coemptydefecttenocytes+PGA PGA HE macroscopicalscore 15 healthy=17points * 10 sulphatedGAGs:blue PGA+ 5 PGA cells empty 0 histologicalscore,6weeks 15 healthy=20points elasticfibers 10 PGA+ cells 5 PGA empty 0 StollCetal.,2011Biomaterials32:4806

  18. Lapineanteriorcruciateligament(ACL) meansubstance posteriorcruciateligament enthesis Dimensions: anterior cruciateligament 15mmx3mmx4mm Hoyeretal.,2014.MaterialsScienceandEngineering1;43:290-9.

  19. embroideredscaffoldswithcollagen • anteriorcruciateligament(ACL)tissueengineeringusingembroideredscaffoldssupplementedwithcollagen embroiderypattern PLA-CLPLA+PLA-CLPLA stitchlength stitchangle loadingaxis spheroid-basedseeding P(LA-CL) +collagenfoam • Poly(lactic-co-ε-caprolacton)=PLA-CL,monofilament • Poly-L-lacticacid=PLA,multifilament • bothmaterialscombined • DimensionsofthelapineACL(15mmx3mmx4mm) vitality Hoyeretal.,2014.MaterialsScienceandEngineering1;43:290-9.

  20. M Cellularity 60 ACLspheroidcharacterization [nuclei/µm2] 40 20 HE SafraninO AzAn 0 days 7 7 0 0 A B C 14 [µg/106cellsperspheroid] 0days N TotalsGAG *** 4 200µm 200µm 200µm 3 sy da E F D 2 1 7daysstatical 7 200µm 200µm 200µm Ø Ø 0 H I G 7daysdynamical days 0 7 0 7 14 Totalcollagen O*** * [µg/106cellsperspheroid] 200µm 200µm 200µm 100 80 60 s y J K L 14daysstatical da14 40 20 200µm 200µm 200µm 4 3 2 1 0 days dynamical statical 0 7 0 7 14 Hoyeretal.,2014.HistochemCellBiol.In press

  21. Collagenscaffoldsseededwithligamentcells 7d green:vital,red:deadcells 21 Hoyeretal.,2014.MaterialsScienceandEngineering1;43:290-9.

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