title: Variations in gene and protein expression in canine chondrodystrophic nucleus pulposus cells following long-term three-dimensional culture.
abstract: Intervertebral disc (IVD) degeneration greatly affects quality of life. The nucleus pulposus (NP) of chondrodystrophic dog breeds (CDBs) is similar to the human NP, because the cells disappear with age and are replaced by fibrochondrocyte-like cells. However, because IVD develops as early as within the first year of life, we used canines as a model to investigate in vitro the mechanisms underlying IVD degeneration. Specifically, we evaluated the potential of a three-dimensional (3D) culture of healthy NP as an in vitro model system to investigate the mechanisms of IVD degeneration. Agarose hydrogels were populated with healthy NP cells from beagles after performing magnetic resonance imaging, and mRNA expression profiles and pericellular extracellular matrix (ECM) protein distribution were determined. After 25 days of 3D culture, there was a tendency for redifferentiation into the native NP phenotype, and mRNA levels of Col2A1, COMP, and CK18 were not significantly different from those of freshly isolated cells. Our findings suggest that long-term 3D culture promoted chondrodystrophic NP redifferentiation through reconstruction of the pericellular microenvironment. Further, lipopolysaccharide (LPS) induced expression of TNF-α, MMP3, MMP13, VEGF, and PGES mRNA in the 3D cultures, creating a molecular milieu that mimics that of degenerated NP. These results suggest that this in vitro model represents a reliable and cost-effective tool for evaluating new therapies for disc degeneration.
authors: Division of Veterinary Surgery, Department of Veterinary Science, Faculty of Veterinary Medicine, Nippon Veterinary and Life Science University, 1-7-1 Kyonan-cho, Musashino, Tokyo, Japan.
authors: Munetaka
authors: M
authors: Iwata
authors: Hiroki
authors: H
authors: Ochi
authors: Yoshinori
authors: Y
authors: Asou
authors: Hirotaka
authors: H
authors: Haro
authors: Takeshi
authors: T
authors: Aikawa
authors: Yasuji
authors: Y
authors: Harada
authors: Yoshinori
authors: Y
authors: Nezu
authors: Takuya
authors: T
authors: Yogo
authors: Masahiro
authors: M
authors: Tagawa
authors: Yasushi
authors: Y
authors: Hara
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D049109:Cell Proliferation / Q000187:drug effects; D024043:Collagen Type II / Q000235:genetics; D016207:Cytokines / Q000235:genetics; D004285:Dogs; D005347:Fibroblasts / Q000473:pathology; D005819:Genetic Markers / Q000235:genetics; D006025:Glycosaminoglycans / Q000235:genetics; D006820:Hyaluronic Acid / Q000235:genetics; D007403:Intervertebral Disc / Q000166:cytology / Q000378:metabolism / Q000473:pathology*; D007405:Intervertebral Disc Displacement / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D008070:Lipopolysaccharides / Q000494:pharmacology; D010641:Phenotype; D013122:Spinal Diseases / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D013997:Time Factors; D015854:Up-Regulation / Q000187:drug effects
chemical_list: D024043:Collagen Type II; D016207:Cytokines; D005819:Genetic Markers; D006025:Glycosaminoglycans; D008070:Lipopolysaccharides; D006820:Hyaluronic Acid
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