title: Enhancement of matrix production and cell proliferation in human annulus cells under bioreactor culture.
abstract: Tissue engineering is a promising approach for treatment of disc degeneration. Herein, we evaluated effects of rotating bioreactor culture on the extracellular matrix production and proliferation of human annulus fibrosus (AF) cells. AF cells were embedded into alginate beads, and then cultured up to 3 weeks in a rotating wall vessel bioreactor or a static vessel. By real-time reverse transcription-polymerase chain reaction, expression of aggrecan, collagen type I and type II, and collagen prolyl 4-hydroxylase II was remarkably elevated, whereas expression of matrix metalloproteinase 3 and a disintegrin and metalloproteinase with thrombospondin motifs 5 was significantly decreased under bioreactor. Biochemical analysis revealed that the levels of the whole cell-associated proteoglycan and collagen were approximately five- and twofolds in rotating bioreactor, respectively, compared to those in static culture. Moreover, AF cell proliferation was augmented in rotating bioreactor. DNA contents were threefolds higher in rotating bioreactor than that in static culture. Expression of the proliferating cell nuclear antigen was robustly enhanced in rotating bioreactor as early as 1 week. Our findings suggested that rotating bioreactor culture would be an effective technique for expansion of human annulus cells for tissue engineering driven treatment of disc degeneration.
authors.affiliation: Orthopaedic Research Lab, University of Virginia, Charlottesville, Virginia 22908, USA.
authors.forename: Xinlin
authors.initials: X
authors.lastname: Yang
authors.forename: Daidong
authors.initials: D
authors.lastname: Wang
authors.forename: Jianrong
authors.initials: J
authors.lastname: Hao
authors.forename: Meiqing
authors.initials: M
authors.lastname: Gong
authors.forename: Vincent
authors.initials: V
authors.lastname: Arlet
authors.forename: Gary
authors.initials: G
authors.lastname: Balian
authors.forename: Francis H
authors.initials: FH
authors.lastname: Shen
authors.forename: Xudong Joshua
authors.initials: XJ
authors.lastname: Li
mesh_terms: D000293:Adolescent; D053674:Aggrecans / Q000378:metabolism; D000464:Alginates / Q000494:pharmacology; D019149:Bioreactors; D018929:Cell Culture Techniques / Q000295:instrumentation* / Q000379:methods*; D049109:Cell Proliferation / Q000187:drug effects; D024043:Collagen Type II / Q000378:metabolism; D005109:Extracellular Matrix / Q000187:drug effects / Q000378:metabolism*; D016326:Extracellular Matrix Proteins / Q000235:genetics / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects; D020723:Glucuronic Acid / Q000494:pharmacology; D006025:Glycosaminoglycans / Q000378:metabolism; D006603:Hexuronic Acids / Q000494:pharmacology; D006801:Humans; D006909:Hydroxyproline / Q000378:metabolism; D007403:Intervertebral Disc / Q000166:cytology*; D008863:Microspheres; D008954:Models, Biological; D010619:Phenazines / Q000378:metabolism; D012399:Rotation
chemical_list: D053674:Aggrecans; D000464:Alginates; D024043:Collagen Type II; D016326:Extracellular Matrix Proteins; D006025:Glycosaminoglycans; D006603:Hexuronic Acids; D010619:Phenazines; D020723:Glucuronic Acid; D006909:Hydroxyproline; C009195:safranine T