title: Cartilage-characteristic matrix reconstruction by sequential addition of soluble factors during expansion of human articular chondrocytes and their cultivation in collagen sponges.
abstract: OBJECTIVE
  Articular cartilage has a poor capacity for spontaneous repair. Tissue engineering approaches using biomaterials and chondrocytes offer hope for treatments. Our goal was to test whether collagen sponges could be used as scaffolds for reconstruction of cartilage with human articular chondrocytes. We investigated the effects on the nature and abundance of cartilage matrix produced of sequential addition of chosen soluble factors during cell amplification on plastic and cultivation in collagen scaffolds.
  
  
  METHODS
  Isolated human articular chondrocytes were amplified for two passages with or without a cocktail of fibroblast growth factor (FGF)-2 and insulin (FI). The cells were then cultured in collagen sponges with or without a cocktail of bone morphogenetic protein (BMP)-2, insulin, and triiodothyronine (BIT). The constructs were cultivated for 36 days in vitro or for another 6-week period in a nude mouse-based contained-defect organ culture model. Gene expression was analyzed using polymerase chain reaction, and protein production was analyzed using Western-blotting and immunohistochemistry.
  
  
  RESULTS
  Dedifferentiation of chondrocytes occurred during cell expansion on plastic, and FI stimulated this dedifferentiation. We found that addition of BIT could trigger chondrocyte redifferentiation and cartilage-characteristic matrix production in the collagen sponges. The presence of FI during cell expansion increased the chondrocyte responsiveness to BIT.
authors.affiliation: Institut de Biologie et Chimie des Protéines, Université de Lyon, Lyon, France.
authors.forename: Stéphanie
authors.initials: S
authors.lastname: Claus
authors.forename: Nathalie
authors.initials: N
authors.lastname: Mayer
authors.forename: Elisabeth
authors.initials: E
authors.lastname: Aubert-Foucher
authors.forename: Hanane
authors.initials: H
authors.lastname: Chajra
authors.forename: Emeline
authors.initials: E
authors.lastname: Perrier-Groult
authors.forename: Jérôme
authors.initials: J
authors.lastname: Lafont
authors.forename: Muriel
authors.initials: M
authors.lastname: Piperno
authors.forename: Odile
authors.initials: O
authors.lastname: Damour
authors.forename: Frédéric
authors.initials: F
authors.lastname: Mallein-Gerin
mesh_terms: D000368:Aged; D000818:Animals; D055396:Bone Morphogenetic Protein 2 / Q000494:pharmacology; D002358:Cartilage, Articular / Q000166:cytology*; D002417:Cattle; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D019902:Chondrocytes / Q000166:cytology* / Q000187:drug effects* / Q000378:metabolism; D020219:Chondrogenesis / Q000187:drug effects / Q000235:genetics; D003094:Collagen / Q000494:pharmacology*; D005109:Extracellular Matrix / Q000187:drug effects / Q000378:metabolism*; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D007150:Immunohistochemistry; D007328:Insulin / Q000494:pharmacology; D036341:Intercellular Signaling Peptides and Proteins / Q000494:pharmacology*; D051379:Mice; D008875:Middle Aged; D014176:Protein Biosynthesis / Q000187:drug effects; D012995:Solubility / Q000187:drug effects; D023822:Tissue Engineering / Q000379:methods*; D054457:Tissue Scaffolds / Q000737:chemistry; D014284:Triiodothyronine / Q000494:pharmacology
chemical_list: D055396:Bone Morphogenetic Protein 2; D007328:Insulin; D036341:Intercellular Signaling Peptides and Proteins; D014284:Triiodothyronine; D016222:Fibroblast Growth Factor 2; D003094:Collagen