title: FGF2 and dexamethasone increase the production of hyaluronan in two-dimensional culture of elastic cartilage-derived cells: in vitro analyses and in vivo cartilage formation.
abstract: Elastic cartilage-derived cells cultured two-dimensionally with FGF2 and corticosteroid produce gel-type masses that become mature cartilage when injected into a subcutaneous pocket. This unique method has previously been clinically applied for treatments of nasal augmentation. However, the components of the gel-type mass and the mechanism of its synthesis remain unknown. Here, we have investigated the components of the gel-type mass produced by elastic cartilage-derived cells, and whether this gel-type mass can be produced by using other cell sources or other media. Human elastic cartilage-derived cells from auricular cartilage, hyaline cartilage-derived cells from articular cartilage, and mesenchymal stem cells from synovium were cultured in three media: "redifferentiation medium" containing FGF2 and dexamethasone; "chondrogenic medium" containing bone morphogenetic protein-2, transforming growth factor-beta3, and dexamethasone specific for in vitro chondrogenesis of mesenchymal stem cells; control medium. The elastic cartilage-derived cells cultured in redifferentiation medium produced a gelatinous matrix positive for Alcian blue. During culture, the amount of chondroitin 4-sulfate, chondroitin 6-sulfate, and especially hyaluronan increased. However, the expression of RNAs for most chondrogenic genes did not increase. We also reproduced cartilage tissue formation by the injection of elastic cartilage-derived cells with the gelatinous mass into the subcutaneous space of the nude mouse. The synthesis of gelatinous matrix in vitro and the formation of cartilage tissue in vivo could be obtained only for the combination of elastic cartilage-derived cells with redifferentiation medium.
authors.affiliation: Section of Orthopaedic Surgery, Graduate School, Tokyo Medical and Dental University, Tokyo, Japan.
authors.forename: Akiko
authors.initials: A
authors.lastname: Yokoyama
authors.forename: Takeshi
authors.initials: T
authors.lastname: Muneta
authors.forename: Akimoto
authors.initials: A
authors.lastname: Nimura
authors.forename: Hideyuki
authors.initials: H
authors.lastname: Koga
authors.forename: Tomoyuki
authors.initials: T
authors.lastname: Mochizuki
authors.forename: Yuiro
authors.initials: Y
authors.lastname: Hata
authors.forename: Ichiro
authors.initials: I
authors.lastname: Sekiya
mesh_terms: D000368:Aged; D000818:Animals; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000187:drug effects; D049109:Cell Proliferation; D002648:Child; D020219:Chondrogenesis / Q000187:drug effects; D003907:Dexamethasone / Q000494:pharmacology*; D051472:Elastic Cartilage / Q000166:cytology / Q000378:metabolism*; D005109:Extracellular Matrix / Q000378:metabolism; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology*; D006025:Glycosaminoglycans / Q000096:biosynthesis; D006801:Humans; D051457:Hyaline Cartilage / Q000166:cytology / Q000378:metabolism*; D006820:Hyaluronic Acid / Q000096:biosynthesis*; D007223:Infant; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000378:metabolism*; D051379:Mice; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D006025:Glycosaminoglycans; D016222:Fibroblast Growth Factor 2; D003907:Dexamethasone; D006820:Hyaluronic Acid