title: BMP-2, hypoxia, and COL1A1/HtrA1 siRNAs favor neo-cartilage hyaline matrix formation in chondrocytes.
abstract: Osteoarthritis (OA) is an irreversible pathology that causes a decrease in articular cartilage thickness, leading finally to the complete degradation of the affected joint. The low spontaneous repair capacity of cartilage prevents any restoration of the joint surface, making OA a major public health issue. Here, we developed an innovative combination of treatment conditions to improve the human chondrocyte phenotype before autologous chondrocyte implantation. First, we seeded human dedifferentiated chondrocytes into a collagen sponge as a scaffold, cultured them in hypoxia in the presence of a bone morphogenetic protein (BMP), BMP-2, and transfected them with small interfering RNAs targeting two markers overexpressed in OA dedifferentiated chondrocytes, that is, type I collagen and/or HtrA1 serine protease. This strategy significantly decreased mRNA and protein expression of type I collagen and HtrA1, and led to an improvement in the chondrocyte phenotype index of differentiation. The effectiveness of our in vitro culture process was also demonstrated in the nude mouse model in vivo after subcutaneous implantation. We, thus, provide here a new protocol able to favor human hyaline chondrocyte phenotype in primarily dedifferentiated cells, both in vitro and in vivo. Our study also offers an innovative strategy for chondrocyte redifferentiation and opens new opportunities for developing therapeutic targets.
authors: 1 Microenvironnement Cellulaire et Pathologies Laboratory (MILPAT) EA 4652, Federative Research Structure (FRS) 146 ICORE, Faculty of Medicine, IBFA, University of Caen/Lower-Normandy (UCLN) , Caen, France .
authors: David
authors: D
authors: Ollitrault
authors: Florence
authors: F
authors: Legendre
authors: Carole
authors: C
authors: Drougard
authors: Mélanie
authors: M
authors: Briand
authors: Hervé
authors: H
authors: Benateau
authors: Didier
authors: D
authors: Goux
authors: Hanane
authors: H
authors: Chajra
authors: Laurent
authors: L
authors: Poulain
authors: Daniel
authors: D
authors: Hartmann
authors: Denis
authors: D
authors: Vivien
authors: Vijayalakshmi
authors: V
authors: Shridhar
authors: Alfonso
authors: A
authors: Baldi
authors: Frédéric
authors: F
authors: Mallein-Gerin
authors: Karim
authors: K
authors: Boumediene
authors: Magali
authors: M
authors: Demoor
authors: Philippe
authors: P
authors: Galera
mesh_terms: D000368:Aged; D000369:Aged, 80 and over; D000818:Animals; D055396:Bone Morphogenetic Protein 2 / Q000494:pharmacology*; D002358:Cartilage, Articular / Q000166:cytology*; D002417:Cattle; D015687:Cell Hypoxia / Q000187:drug effects; D002478:Cells, Cultured; D019902:Chondrocytes; D020219:Chondrogenesis / Q000187:drug effects; D024042:Collagen Type I / Q000378:metabolism*; D000090602:Collagen Type I, alpha 1 Chain; D005109:Extracellular Matrix / Q000187:drug effects / Q000378:metabolism*; D000074289:High-Temperature Requirement A Serine Peptidase 1; D006801:Humans; D006818:Hyalin / Q000378:metabolism*; D006984:Hypertrophy; D007700:Kinetics; D008819:Mice, Nude; D008875:Middle Aged; D010012:Osteogenesis / Q000187:drug effects; D010641:Phenotype; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000378:metabolism*; D012697:Serine Endopeptidases / Q000378:metabolism*
chemical_list: D055396:Bone Morphogenetic Protein 2; D024042:Collagen Type I; D000090602:Collagen Type I, alpha 1 Chain; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D000074289:High-Temperature Requirement A Serine Peptidase 1; C474012:HTRA1 protein, human; D012697:Serine Endopeptidases