title: Cotransfected human chondrocytes: over-expression of IGF-I and SOX9 enhances the synthesis of cartilage matrix components collagen-II and glycosaminoglycans.
abstract: 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.
authors.affiliation: Department of Biochemistry and Molecular Medicine, Faculty of Medicine, Autonomous University of Nuevo León, Monterrey, NL, Mexico.
authors.forename: M
authors.initials: M
authors.lastname: Simental-Mendía
authors.affiliation: Autonomous University of Nuevo León, Laboratory of Tissue Engineering, Bone and Tissue Bank, Universitary Hospital, Monterrey, NL, Mexico.
authors.forename: J
authors.initials: J
authors.lastname: Lara-Arias
authors.affiliation: Autonomous University of Nuevo León, Laboratory of Tissue Engineering, Bone and Tissue Bank, Universitary Hospital, Monterrey, NL, Mexico.
authors.forename: E
authors.initials: E
authors.lastname: Álvarez-Lozano
authors.affiliation: Department of Biochemistry and Molecular Medicine, Faculty of Medicine, Autonomous University of Nuevo León, Monterrey, NL, Mexico.
authors.forename: S
authors.initials: S
authors.lastname: Said-Fernández
authors.affiliation: Department of Histology, Faculty of Medicine, Autonomous University of Nuevo León, Monterrey, NL, Mexico.
authors.forename: A
authors.initials: A
authors.lastname: Soto-Domínguez
authors.affiliation: Department of Biochemistry and Molecular Medicine, Faculty of Medicine, Autonomous University of Nuevo León, Monterrey, NL, Mexico.
authors.forename: G R
authors.initials: GR
authors.lastname: Padilla-Rivas
authors.affiliation: Department of Biochemistry and Molecular Medicine, Faculty of Medicine, Autonomous University of Nuevo León, Monterrey, NL, Mexico.
authors.forename: H G
authors.initials: HG
authors.lastname: Martínez-Rodríguez
mesh_terms: D002358:Cartilage, Articular / Q000293:injuries / Q000378:metabolism; D019902:Chondrocytes / Q000378:metabolism*; D024043:Collagen Type II / Q000032:analysis / Q000096:biosynthesis*; D005109:Extracellular Matrix / Q000737:chemistry; D015870:Gene Expression; D006025:Glycosaminoglycans / Q000032:analysis / Q000096:biosynthesis*; D006801:Humans; D007334:Insulin-Like Growth Factor I / Q000235:genetics / Q000378:metabolism*; D064235:Matrilin Proteins / Q000096:biosynthesis* / Q000235:genetics; D061251:Primary Cell Culture; D012333:RNA, Messenger / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D055758:SOX9 Transcription Factor / Q000235:genetics / Q000378:metabolism*; D013053:Spectrophotometry; D014162:Transfection / Q000379:methods*
chemical_list: D024043:Collagen Type II; D006025:Glycosaminoglycans; D064235:Matrilin Proteins; D012333:RNA, Messenger; D055758:SOX9 Transcription Factor; D007334:Insulin-Like Growth Factor I