title: Towards biological anulus repair: TGF-β3, FGF-2 and human serum support matrix formation by human anulus fibrosus cells.
abstract: Closure and biological repair of anulus fibrosus (AF) defects in intervertebral disc diseases is a therapeutic challenge. The aim of our study was to evaluate the anabolic properties of bioactive factors on cartilaginous matrix formation by AF cells. Human AF cells were harvested from degenerated lumbar AF tissue and expanded in monolayer culture. AF cell differentiation and matrix formation was initiated by forming pellet cultures and stimulation with hyaluronic acid (HA), human serum (HS), fibroblast growth factor-2 (FGF-2), transforming growth factor-β3 (TGF-β3) and TGF-β3/FGF-2 for up to 4 weeks. Matrix formation was assessed histologically by staining of proteoglycan, type I and type II collagens and by gene expression analysis of typical extracellular matrix molecules and of catabolic matrix metalloproteinases MMP-2 and MMP-13. AF cells, stimulated with HS, FGF-2 and most pronounced with TGF-β3 or TGF-β3/FGF-2 formed a cartilaginous matrix with significantly enhanced expression of matrix molecules and of MMP-13. Stimulation of AF cells with TGF-β3 was accompanied by induction of type X collagen, known to occur in hypertrophic cartilage cells having mineralizing potential. HA did not show any chondro-inductive characteristics. These findings suggest human serum, FGF-2 and TGF-β3 as possible candidates to support biological treatment strategies of AF defects.
authors.affiliation: Department of Neurosurgery, University Medical Center Mannheim, Heidelberg University, Germany.
authors.forename: Aldemar A
authors.initials: AA
authors.lastname: Hegewald
authors.forename: Sabra
authors.initials: S
authors.lastname: Zouhair
authors.forename: Michaela
authors.initials: M
authors.lastname: Endres
authors.forename: Mario
authors.initials: M
authors.lastname: Cabraja
authors.forename: Christian
authors.initials: C
authors.lastname: Woiciechowsky
authors.forename: Claudius
authors.initials: C
authors.lastname: Thomé
authors.forename: Christian
authors.initials: C
authors.lastname: Kaps
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D018929:Cell Culture Techniques; D002478:Cells, Cultured; D005109:Extracellular Matrix / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D005260:Female; D016222:Fibroblast Growth Factor 2 / Q000378:metabolism* / Q000494:pharmacology; D006801:Humans; D007403:Intervertebral Disc / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D055959:Intervertebral Disc Degeneration / Q000378:metabolism / Q000473:pathology; D007405:Intervertebral Disc Displacement / Q000378:metabolism / Q000473:pathology; D012038:Regeneration; D044967:Serum / Q000378:metabolism; D023822:Tissue Engineering; D053782:Transforming Growth Factor beta3 / Q000378:metabolism* / Q000494:pharmacology
chemical_list: D053782:Transforming Growth Factor beta3; D016222:Fibroblast Growth Factor 2