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Braz. j. med. biol. res ; 48(12): 1063-1070, Dec. 2015. tab, graf
Article Dans Anglais | LILACS | ID: lil-762916

Résumé

Damage to cartilage causes a loss of type II collagen (Col-II) and glycosaminoglycans (GAG). To restore the original cartilage architecture, cell factors that stimulate Col-II and GAG production are needed. Insulin-like growth factor I (IGF-I) and transcription factor SOX9are essential for the synthesis of cartilage matrix, chondrocyte proliferation, and phenotype maintenance. We evaluated the combined effect of IGF-I and SOX9 transgene expression on Col-II and GAG production by cultured human articular chondrocytes. Transient transfection and cotransfection were performed using two mammalian expression plasmids (pCMV-SPORT6), one for each transgene. At day 9 post-transfection, the chondrocytes that were over-expressing IGF-I/SOX9 showed 2-fold increased mRNA expression of the Col-II gene, as well as a 57% increase in Col-II protein, whereas type I collagen expression (Col-I) was decreased by 59.3% compared with controls. The production of GAG by these cells increased significantly compared with the controls at day 9 (3.3- vs 1.8-times, an increase of almost 83%). Thus, IGF-I/SOX9 cotransfected chondrocytes may be useful for cell-based articular cartilage therapies.


Sujets)
Humains , Chondrocytes/métabolisme , Collagène de type II/biosynthèse , Glycosaminoglycanes/biosynthèse , Facteur de croissance IGF-I/métabolisme , Matrilines/biosynthèse , Facteur de transcription SOX-9/métabolisme , Transfection/méthodes , Cartilage articulaire/traumatismes , Cartilage articulaire/métabolisme , Collagène de type II/analyse , Matrice extracellulaire/composition chimique , Expression des gènes , Glycosaminoglycanes/analyse , Facteur de croissance IGF-I/génétique , Matrilines/génétique , Culture de cellules primaires , Réaction de polymérisation en chaine en temps réel , ARN messager/métabolisme , Facteur de transcription SOX-9/génétique , Spectrophotométrie
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