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01-11-2010 | Original Article

Identification of oxytetracycline as a chondrogenic compound using a cell-based screening system

Authors: Hironori Hojo, Fumiko Yano, Shinsuke Ohba, Kazuyo Igawa, Keiji Nakajima, Yuske Komiyama, Akinori Kan, Toshiyuki Ikeda, Takayuki Yonezawa, Je-Tae Woo, Tsuyoshi Takato, Kozo Nakamura, Hiroshi Kawaguchi, Ung-il Chung

Published in: Journal of Bone and Mineral Metabolism | Issue 6/2010

Abstract

To effectively treat degenerative joint diseases including osteoarthritis (OA), small chemical compounds need to be developed that can potently induce chondrogenic differentiation without promoting terminal differentiation. For this purpose, we screened natural and synthetic compound libraries using a Col2GFP-ATDC5 system and identified oxytetracycline (Oxy) as a chondrogenic compound. Oxy induced cartilaginous matrix synthesis and mRNA expressions of chondrocyte markers in ATDC5 cells. In addition, Oxy suppressed mineralization and mRNA expressions of terminal chondrocyte differentiation markers in ATDC5 cells, primary chondrocytes, and cultured metatarsal bones. Oxy’s induction of Col2 mRNA expression was decreased by the addition of Noggin and was increased by the addition of BMP2. Furthermore, Oxy increased mRNA expression of Id1, Bmp2, Bmp4, and Bmp6. These data suggest that Oxy induces chondrogenic differentiation in a BMP-dependent manner and suppresses terminal differentiation. Oxy may be useful for treatment of OA and also for regeneration of cartilage tissue.

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Hollander AP, Pidoux I, Reiner A, Rorabeck C, Bourne R, Poole AR (1995) Damage to type II collagen in aging and osteoarthritis starts at the articular surface, originates around chondrocytes, and extends into the cartilage with progressive degeneration. J Clin Invest 96:2859–2869CrossRefPubMed

Kronenberg HM (2003) Developmental regulation of the growth plate. Nature 423:332–336CrossRefPubMed

Lefebvre V, Smits P (2005) Transcriptional control of chondrocyte fate and differentiation. Birth Defects Res C Embryo Today 75:200–212CrossRefPubMed

Billinghurst RC, Dahlberg L, Ionescu M, Reiner A, Bourne R, Rorabeck C, Mitchell P, Hambor J, Diekmann O, Tschesche H, Chen J, Van Wart H, Poole AR (1997) Enhanced cleavage of type II collagen by collagenases in osteoarthritic articular cartilage. J Clin Invest 99:1534–1545CrossRefPubMed

Pullig O, Weseloh G, Gauer S, Swoboda B (2000) Osteopontin is expressed by adult human osteoarthritic chondrocytes: protein and mRNA analysis of normal and osteoarthritic cartilage. Matrix Biol 19:245–255CrossRefPubMed

Mitchell PG, Magna HA, Reeves LM, Lopresti-Morrow LL, Yocum SA, Rosner PJ, Geoghegan KF, Hambor JE (1996) Cloning, expression, and type II collagenolytic activity of matrix metalloproteinase-13 from human osteoarthritic cartilage. J Clin Invest 97:761–768CrossRefPubMed

Ikeda T, Kamekura S, Mabuchi A, Kou I, Seki S, Takato T, Nakamura K, Kawaguchi H, Ikegawa S, Chung UI (2004) The combination of SOX5, SOX6, and SOX9 (the SOX trio) provides signals sufficient for induction of permanent cartilage. Arthritis Rheum 50:3561–3573CrossRefPubMed

Kolettas E, Muir HI, Barrett JC, Hardingham TE (2001) Chondrocyte phenotype and cell survival are regulated by culture conditions and by specific cytokines through the expression of Sox-9 transcription factor. Rheumatology (Oxford) 40:1146–1156CrossRef

Zehentner BK, Dony C, Burtscher H (1999) The transcription factor Sox9 is involved in BMP-2 signaling. J Bone Miner Res 14:1734–1741CrossRefPubMed

10.

Tuli R, Tuli S, Nandi S, Huang X, Manner PA, Hozack WJ, Danielson KG, Hall DJ, Tuan RS (2003) Transforming growth factor-beta-mediated chondrogenesis of human mesenchymal progenitor cells involves N-cadherin and mitogen-activated protein kinase and Wnt signaling cross-talk. J Biol Chem 278:41227–41236CrossRefPubMed

11.

Church VL, Francis-West P (2002) Wnt signalling during limb development. Int J Dev Biol 46:927–936PubMed

12.

Fang J, Zhu YY, Smiley E, Bonadio J, Rouleau JP, Goldstein SA, McCauley LK, Davidson BL, Roessler BJ (1996) Stimulation of new bone formation by direct transfer of osteogenic plasmid genes. Proc Natl Acad Sci USA 93:5753–5758CrossRefPubMed

13.

Kan A, Ikeda T, Saito T, Yano F, Fukai A, Hojo H, Ogasawara T, Ogata N, Nakamura K, Chung UI, Kawaguchi H (2009) Screening of chondrogenic factors with a real-time fluorescence-monitoring cell line ATDC5–C2ER: identification of sorting nexin 19 as a novel factor. Arthritis Rheum 60:3314–3323CrossRefPubMed

14.

Yano F, Kugimiya F, Ohba S, Ikeda T, Chikuda H, Ogasawara T, Ogata N, Takato T, Nakamura K, Kawaguchi H, Chung UI (2005) The canonical Wnt signaling pathway promotes chondrocyte differentiation in a Sox9-dependent manner. Biochem Biophys Res Commun 333:1300–1308CrossRefPubMed

15.

Saito T, Ikeda T, Nakamura K, Chung UI, Kawaguchi H (2007) S100A1 and S100B, transcriptional targets of SOX trio, inhibit terminal differentiation of chondrocytes. EMBO Rep 8:504–509CrossRefPubMed

16.

Ohba S, Ikeda T, Kugimiya F, Yano F, Lichtler AC, Nakamura K, Takato T, Kawaguchi H, Chung UI (2007) Identification of a potent combination of osteogenic genes for bone regeneration using embryonic stem (ES) cell-based sensor. FASEB J 21:1777–1787CrossRefPubMed

17.

Hojo H, Igawa K, Ohba S, Yano F, Nakajima K, Komiyama Y, Ikeda T, Lichtler AC, Woo JT, Yonezawa T, Takato T, Chung UI (2008) Development of high-throughput screening system for osteogenic drugs using a cell-based sensor. Biochem Biophys Res Commun 376:375–379CrossRefPubMed

18.

Wang L, Hinoi E, Takemori A, Nakamichi N, Yoneda Y (2006) Glutamate inhibits chondral mineralization through apoptotic cell death mediated by retrograde operation of the cystine/glutamate antiporter. J Biol Chem 281:24553–24565CrossRefPubMed

19.

Ohba S, Kawaguchi H, Kugimiya F, Ogasawara T, Kawamura N, Saito T, Ikeda T, Fujii K, Miyajima T, Kuramochi A, Miyashita T, Oda H, Nakamura K, Takato T, Chung UI (2008) Patched1 haploinsufficiency increases adult bone mass and modulates Gli3 repressor activity. Dev Cell 14:689–699CrossRefPubMed

20.

Yu LP Jr, Smith GN Jr, Hasty KA, Brandt KD (1991) Doxycycline inhibits type XI collagenolytic activity of extracts from human osteoarthritic cartilage and of gelatinase. J Rheumatol 18:1450–1452PubMed

21.

Nganvongpanit K, Pothacharoen P, Suwankong N, Ong-Chai S, Kongtawelert P (2009) The effect of doxycycline on canine hip osteoarthritis: design of a 6-months clinical trial. J Vet Sci 10(3):239–247CrossRefPubMed

22.

Yu LP Jr, Smith GN Jr, Brandt KD, Myers SL, O’Connor BL, Brandt DA (1992) Reduction of the severity of canine osteoarthritis by prophylactic treatment with oral doxycycline. Arthritis Rheum 35:1150–1159CrossRefPubMed

23.

Smith GN Jr, Yu LP Jr, Brandt KD, Capello WN (1998) Oral administration of doxycycline reduces collagenase and gelatinase activities in extracts of human osteoarthritic cartilage. J Rheumatol 25:532–535PubMed

24.

Yoon BS, Ovchinnikov DA, Yoshii I, Mishina Y, Behringer RR, Lyons KM (2005) Bmpr1a and Bmpr1b have overlapping functions and are essential for chondrogenesis in vivo. Proc Natl Acad Sci USA 102:5062–5067CrossRefPubMed

25.

Kobayashi T, Lyons KM, McMahon AP, Kronenberg HM (2005) BMP signaling stimulates cellular differentiation at multiple steps during cartilage development. Proc Natl Acad Sci USA 102:18023–18027CrossRefPubMed

Title: Identification of oxytetracycline as a chondrogenic compound using a cell-based screening system
Authors: Hironori Hojo
Fumiko Yano
Shinsuke Ohba
Kazuyo Igawa
Keiji Nakajima
Yuske Komiyama
Akinori Kan
Toshiyuki Ikeda
Takayuki Yonezawa
Je-Tae Woo
Tsuyoshi Takato
Kozo Nakamura
Hiroshi Kawaguchi
Ung-il Chung
Publication date: 01-11-2010
Publisher: Springer Japan
Published in: Journal of Bone and Mineral Metabolism / Issue 6/2010
Print ISSN: 0914-8779
Electronic ISSN: 1435-5604
DOI: https://doi.org/10.1007/s00774-010-0179-y

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