title: Fibrin sealant promotes migration and proliferation of human articular chondrocytes: possible involvement of thrombin and protease-activated receptors.
abstract: Fibrin sealant (FS), a biological adhesive material, has been recently recommended as an adjunct in autologous chondrocyte implantation (ACI). While FS has been shown to possess osteoinductive potential, little is known about its effects on chondrogenic cells. In this study, we assessed the bioactivity of FS (Tisseel) on the migration and proliferation of human articular chondrocytes in vitro. Using a co-culture assay to mimic matrix-induced ACI (MACI), chondrocytes were found to migrate from collagen membranes towards FS within 12 h of culture, with significant migratory activity evident by 24 h. In addition, 5-bromo-2'-deoxyuridine (BrdU) incorporation experiments revealed that thrombin, the active component of the tissue glue, stimulated chondrocyte proliferation, with maximal efficacy observed at 48 h post-stimulation (1-10 U/ml). In an effort to elucidate the molecular mechanisms underlying these thrombin-induced effects, we examined the expression and activation of protease-activated receptors (PARs), established thrombin receptors. Using a combination of RT-PCR and immunohistochemistry, all four PARs were detected in human chondrocytes, with PAR-1 being the major isoform expressed. Moreover, thrombin and a PAR-1, but not other PAR-isotype-specific peptide agonists, were found to induce rapid intracellular Ca2+ responses in human chondrocytes in calcium mobilization assays. Together, these data demonstrate that FS supports both the migration and proliferation of human chondrocytes. We propose that these effects are mediated, at least in part, via thrombin-induced PAR-1 signalling in human chondrocytes.
authors.affiliation: School of Surgery and Pathology (Orthopaedics), The University of Western Australia, Crawley WA 6009, Australia.
authors.forename: Yaowanuj
authors.initials: Y
authors.lastname: Kirilak
authors.forename: Nathan J
authors.initials: NJ
authors.lastname: Pavlos
authors.forename: Craig R
authors.initials: CR
authors.lastname: Willers
authors.forename: Renzhi
authors.initials: R
authors.lastname: Han
authors.forename: Haotian
authors.initials: H
authors.lastname: Feng
authors.forename: Jiake
authors.initials: J
authors.lastname: Xu
authors.forename: Nithiananthan
authors.initials: N
authors.lastname: Asokananthan
authors.forename: Geoffrey A
authors.initials: GA
authors.lastname: Stewart
authors.forename: Peter
authors.initials: P
authors.lastname: Henry
authors.forename: David
authors.initials: D
authors.lastname: Wood
authors.forename: Ming H
authors.initials: MH
authors.lastname: Zheng
mesh_terms: D002118:Calcium / Q000378:metabolism; D002358:Cartilage, Articular / Q000166:cytology; D002465:Cell Movement / Q000187:drug effects*; D049109:Cell Proliferation / Q000187:drug effects*; D002478:Cells, Cultured; D019902:Chondrocytes / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D015718:Fibrin Tissue Adhesive / Q000494:pharmacology*; D006801:Humans; D007150:Immunohistochemistry; D007424:Intracellular Fluid / Q000187:drug effects / Q000378:metabolism; D018613:Microscopy, Confocal; D010446:Peptide Fragments / Q000494:pharmacology; D020033:Protein Isoforms / Q000819:agonists / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D044463:Receptor, PAR-1 / Q000819:agonists / Q000235:genetics / Q000378:metabolism; D044464:Receptor, PAR-2 / Q000819:agonists / Q000235:genetics / Q000378:metabolism; D044462:Receptors, Proteinase-Activated / Q000819:agonists / Q000235:genetics / Q000378:metabolism; D018179:Receptors, Thrombin / Q000819:agonists / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013917:Thrombin / Q000494:pharmacology
chemical_list: D015718:Fibrin Tissue Adhesive; D010446:Peptide Fragments; D020033:Protein Isoforms; D012333:RNA, Messenger; D044463:Receptor, PAR-1; D044464:Receptor, PAR-2; D044462:Receptors, Proteinase-Activated; D018179:Receptors, Thrombin; C082835:thrombin receptor peptide (42-47); D013917:Thrombin; D002118:Calcium