title: Effects of low molecular weight hyaluronan combined with carprofen on canine osteoarthritis articular chondrocytes and cartilage explants in vitro.
abstract: Intra-articular injection with non-steroidal anti-inflammatory drugs (NSAIDs) is used to treat inflammatory joint disease, but the side effects of NSAIDs include chondrotoxicity. Hyaluronan has shown positive effects on chondrocytes by reducing apoptosis and increasing proteoglycan synthesis. The purposes of this study were to evaluate the effects of low molecular weight hyaluronan (low MW HA), carprofen 25 mg/ml, carprofen 12.5 mg/ml, and a combination of HA and carprofen on canine osteoarthritis (OA) articular chondrocytes and a cartilage explant model in terms of cell viability, extracellular matrix remaining, and gene expression after exposure. In chondrocyte culture, MTT assay was used to evaluate the chondrotoxicity of IC50 and IC80 of carprofen with HA. In cartilage explant culture, two kinds of extracellular matrix (uronic acid and collagen) remaining in cartilage were used to evaluate cartilage damage for 14 d after treatment. Expression of COL2A1, AGG, and MMP3 was used to evaluate the synthesis and degradation of the matrix for 7 d after treatment. In chondrocyte culture, low MW HA could preserve OA chondrocyte viability but could not reduce the chondrotoxicity level of carprofen (P < 0.05). In explant culture, low MW HA combined with 12.5 mg/ml carprofen caused less destruction of uronic acid and collagen structure when compared with the control (P < 0.05). Low MW HA caused high expression levels of COL2A1 and AGG in OA cartilage (P < 0.05); HA combined with carprofen resulted in higher COL2A1 and AGG expression levels than carprofen alone.
authors.affiliation: Animal Bone and Joint Research Laboratory, Department of Veterinary Biosciences and Public Health, Faculty of Veterinary Medicine, Chiang Mai University, Chiang Mai, 50100, Thailand.
authors.forename: Thippaporn
authors.initials: T
authors.lastname: Euppayo
authors.affiliation: Animal Bone and Joint Research Laboratory, Department of Veterinary Biosciences and Public Health, Faculty of Veterinary Medicine, Chiang Mai University, Chiang Mai, 50100, Thailand.
authors.forename: Puntita
authors.initials: P
authors.lastname: Siengdee
authors.affiliation: Department of Biology, Faculty of Science, Chiang Mai University, Chiang Mai, 50200, Thailand.
authors.forename: Kittisak
authors.initials: K
authors.lastname: Buddhachat
authors.affiliation: Department of Biology, Faculty of Science, Chiang Mai University, Chiang Mai, 50200, Thailand.
authors.forename: Waranee
authors.initials: W
authors.lastname: Pradit
authors.affiliation: Department of Biochemistry, Faculty of Medicine, Chiang Mai University, Chiang Mai, 50100, Thailand.
authors.forename: Nawarat
authors.initials: N
authors.lastname: Viriyakhasem
authors.affiliation: Department of Biology, Faculty of Science, Chiang Mai University, Chiang Mai, 50200, Thailand.
authors.forename: Siriwadee
authors.initials: S
authors.lastname: Chomdej
authors.affiliation: Department of Biochemistry, Faculty of Medicine, Chiang Mai University, Chiang Mai, 50100, Thailand.
authors.forename: Siriwan
authors.initials: S
authors.lastname: Ongchai
authors.affiliation: Divisions of Veterinary Surgery, Nippon Veterinary and Life Science University, 1-7-1, Kyonann-cho, Musashinoshi, Tokyo, 180-8602, Japan.
authors.forename: Yasuji
authors.initials: Y
authors.lastname: Harada
authors.affiliation: Animal Bone and Joint Research Laboratory, Department of Veterinary Biosciences and Public Health, Faculty of Veterinary Medicine, Chiang Mai University, Chiang Mai, 50100, Thailand. korakot.n@cmu.ac.th.|Excellence Center in Osteology Research and Training Center, Chiang Mai University, Chiang Mai, 50200, Thailand. korakot.n@cmu.ac.th.
authors.forename: Korakot
authors.initials: K
authors.lastname: Nganvongpanit
mesh_terms: D000276:Adjuvants, Immunologic / Q000008:administration & dosage / Q000494:pharmacology / Q000627:therapeutic use*; D000818:Animals; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000008:administration & dosage / Q000494:pharmacology / Q000627:therapeutic use*; D002227:Carbazoles / Q000494:pharmacology*; D002358:Cartilage, Articular / Q000187:drug effects*; D019902:Chondrocytes / Q000187:drug effects*; D004283:Dog Diseases / Q000188:drug therapy*; D004285:Dogs; D004359:Drug Therapy, Combination; D005786:Gene Expression Regulation / Q000187:drug effects / Q000502:physiology; D006820:Hyaluronic Acid / Q000494:pharmacology*; D066298:In Vitro Techniques; D010003:Osteoarthritis / Q000188:drug therapy / Q000662:veterinary*
chemical_list: D000276:Adjuvants, Immunologic; D000894:Anti-Inflammatory Agents, Non-Steroidal; D002227:Carbazoles; D006820:Hyaluronic Acid; C007005:carprofen
keywords: Articular cartilage; Carprofen; Cartilage explant; Hyaluronan; Osteoarthritis