Kensuke Ochi1, Assia Derfoul, Rocky S Tuan. 1. Cartilage Biology and Orthopaedics Branch, National Institute of Arthritis, and Musculoskeletal and Skin Diseases, National Institutes of Health, Department of Health and Human Services, Bethesda, MD 20892-8022, USA.
Abstract
OBJECTIVE: Cartilage tissue engineering using multipotential human mesenchymal stem cells (hMSCs) is a promising approach to develop treatment for degenerative joint diseases. A key requirement is that the engineered tissues maintain their hyaline articular cartilage phenotype and not proceed towards hypertrophy. It is noteworthy that osteoarthritic articular cartilage frequently contains limited regions of reparative cartilage, suggesting the presence of bioactive factors with regenerative activity. Based on this idea, we recently performed cDNA microarray analysis to identify genes that are strongly expressed only in articular cartilage and encode secreted gene products. One of the genes that met our criteria was SCRG1. This study aims to analyze SCRG1 function in cartilage development using an in vitro mesenchymal chondrogenesis system. METHODS: Full-length SCRG1 cDNA was subcloned into pcDNA5 vector, and transfected into hMSCs and murine C3H10T1/2 mesenchymal cells, placed in pellet cultures and micromass cultures, respectively. The cultures were analyzed by reverse transcription-polymerase chain reaction for the expression of SCRG1 and cartilage marker genes, and by histological staining for cartilage phenotype. RESULTS: Induction of SCRG1 expression was seen during in vitro chondrogenesis, and was dependent on dexamethasone (DEX) known to promote chondrogenesis. Immunohistochemistry revealed that SCRG1 protein was localized to the extracellular matrix. Forced expression of SCRG1 in hMSCs suppressed their proliferation, and stimulated chondrogenesis in C3H10T1/2 cells, confirmed by reduced collagen type I and elevated collagen type IIB expression. CONCLUSION: These results suggest that SCRG1 is involved in cell growth suppression and differentiation during DEX-dependent chondrogenesis. SCRG1 may be of utility in gene-mediated cartilage tissue engineering.
OBJECTIVE:Cartilage tissue engineering using multipotential human mesenchymal stem cells (hMSCs) is a promising approach to develop treatment for degenerative joint diseases. A key requirement is that the engineered tissues maintain their hyaline articular cartilage phenotype and not proceed towards hypertrophy. It is noteworthy that osteoarthritic articular cartilage frequently contains limited regions of reparative cartilage, suggesting the presence of bioactive factors with regenerative activity. Based on this idea, we recently performed cDNA microarray analysis to identify genes that are strongly expressed only in articular cartilage and encode secreted gene products. One of the genes that met our criteria was SCRG1. This study aims to analyze SCRG1 function in cartilage development using an in vitro mesenchymal chondrogenesis system. METHODS: Full-length SCRG1 cDNA was subcloned into pcDNA5 vector, and transfected into hMSCs and murine C3H10T1/2 mesenchymal cells, placed in pellet cultures and micromass cultures, respectively. The cultures were analyzed by reverse transcription-polymerase chain reaction for the expression of SCRG1 and cartilage marker genes, and by histological staining for cartilage phenotype. RESULTS: Induction of SCRG1 expression was seen during in vitro chondrogenesis, and was dependent on dexamethasone (DEX) known to promote chondrogenesis. Immunohistochemistry revealed that SCRG1 protein was localized to the extracellular matrix. Forced expression of SCRG1 in hMSCs suppressed their proliferation, and stimulated chondrogenesis in C3H10T1/2 cells, confirmed by reduced collagen type I and elevated collagen type IIB expression. CONCLUSION: These results suggest that SCRG1 is involved in cell growth suppression and differentiation during DEX-dependent chondrogenesis. SCRG1 may be of utility in gene-mediated cartilage tissue engineering.
Authors: Arif Kaya; Murat Kara; Tülay Tiftik; Mehmet Engin Tezcan; Mehmet Akif Öztürk; Ayşen Akıncı; Levent Özçakar Journal: Rheumatol Int Date: 2012-07-21 Impact factor: 2.631
Authors: Kosei Ijiri; Luiz F Zerbini; Haibing Peng; Hasan H Otu; Kaneyuki Tsuchimochi; Miguel Otero; Cecilia Dragomir; Nicole Walsh; Benjamin E Bierbaum; David Mattingly; Geoff van Flandern; Setsuro Komiya; Thomas Aigner; Towia A Libermann; Mary B Goldring Journal: Arthritis Rheum Date: 2008-07
Authors: Arthur M Mandelin; Philip J Homan; Alexander M Shaffer; Carla M Cuda; Salina T Dominguez; Emily Bacalao; Mary Carns; Monique Hinchcliff; Jungwha Lee; Kathleen Aren; Anjali Thakrar; Anna B Montgomery; S Louis Bridges; Joan M Bathon; John P Atkinson; David A Fox; Eric L Matteson; Christopher D Buckley; Costantino Pitzalis; Deborah Parks; Laura B Hughes; Laura Geraldino-Pardilla; Robert Ike; Kristine Phillips; Kerry Wright; Andrew Filer; Stephen Kelly; Eric M Ruderman; Vince Morgan; Hiam Abdala-Valencia; Alexander V Misharin; G Scott Budinger; Elizabeth T Bartom; Richard M Pope; Harris Perlman; Deborah R Winter Journal: Arthritis Rheumatol Date: 2018-05-03 Impact factor: 10.995
Authors: Louise Bundgaard; Allan Stensballe; Kirstine Juul Elbæk; Lise Charlotte Berg Journal: Stem Cell Res Ther Date: 2020-05-20 Impact factor: 6.832