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An Atypical Degenerative Osteoarthropathy in Hyp Mice is Characterized by a Loss in the Mineralized Zone of Articular Cartilage

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Abstract

Patients with X-linked hypophosphatemia (XLH) develop enthesophytes and osteophytes secondary to articular cartilage degeneration and together are the primary cause of morbidity in adult patients so afflicted. We have previously characterized the enthesopathy in Hyp mice, a murine model of XLH. We now extend these studies to the synovial joint in order to characterize potential cellular changes in articular cartilage that may predispose patients to the osteoarthropathy of XLH. We report that, despite highly elevated levels of alkaline phosphatase activity throughout articular cartilage, there is a complete loss in the mineralized zone of articular cartilage as assessed by von Kossa staining of mineral and as quantified by EPIC-microCT analysis and evidence of vascular invasion. We also identify the downregulation of extracellular matrix (ECM) factors identified as regulators of terminally differentiated mineralizing articular chondrocytes. There is also a striking increase in the histochemical staining of sulfated proteoglycans, a change that may reflect the loss of a transitional tissue that reduces mechanical stress at the interface between cartilage and subchondral bone. The failure of mineralizing articular chondrocytes to develop in the hypophosphatemic state suggests that phosphate may be a key regulator of chondrocyte mineralization. Accordingly, we find that the appropriate zonal arrangement and phenotypic markers of articular cartilage are significantly reestablished by phosphate-replacement therapy. Given the turnover and maintenance of articular cartilage ECM, the identification of early and abnormal cellular changes unique to XLH will undoubtedly aid in a more effective management of this disease to minimize the onset of degenerative osteoarthropathy.

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Acknowledgments

The authors thank Drs. Thomas Carpenter, Arthur Broadus, and Pete Amos for helpful discussion. We also thank Ali Nasiri, Nancy Troiano, and Christiane Coady for excellent technical assistance.

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  1. Section of Endocrinology and Metabolism, Department of Internal Medicine, School of Medicine, Yale University, New Haven, CT, 06520, USA

    Guoying Liang, Joshua VanHouten & Carolyn M. Macica

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  1. Guoying Liang
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Correspondence to Carolyn M. Macica.

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223_2011_9502_MOESM1_ESM.tif

Supplementary material 1: Schematic of EPIC-microCT strategy (shown is a 12 wk old wildtype mouse) showing the discrimination of specific regions of interest to segregate unmineralized and mineralized cartilage and underlying subchondral bone; arrow showing cement line (the interface between calcified articular cartilage and subchondral bone). See Materials and Methods for experimental detail (TIFF 2184 kb)

223_2011_9502_MOESM2_ESM.tif

Supplementary material 2: Localization of alkaline phosphatase and markers of mineralizing growth plate chondrocytes of representative 12 wk old control, Hyp and Hyp mice treated with oral phosphate and calcitriol. (A), Tibia of 12 wk old control mouse; shown at low power; (B-E), Control mouse, alkaline phosphatase activity localized to hypertrophic growth plate [GP] chondrocytes (B); Immunoreactive OPN secreted into the matrix of hypertrophic GP chondrocytes (C); Immunoreactive MMP13 secreted into the matrix of GP chondrocytes (D); Histochemical staining of sulfated proteoglycans by safranin O/fast green staining (E). (F), Tibia of 12 wk old Hyp mouse; shown at low power, note widened growth plate (rickets), metaphyseal splaying and high levels of tissue alkaline phosphatase levels; (G-J), alkaline phosphatase activity localized to hypertrophic GP chondrocytes (G); Absence of immunoreactive OPN secreted into the matrix of expanded hypertrophic GP chondrocytes (H); Absence of immunoreactive MMP13 secreted into the matrix of GP chondrocytes (I); Histochemical staining of sulfated proteoglycans by safranin O/fast green staining, note significant induction of proteoglycan synthesis (J); (K), Tibia of 12 wk old Hyp mouse treated with oral phosphate and calcitriol; shown at low power, note loss of rachitic lesions; (L-O), alkaline phosphatase activity localized to hypertrophic GP chondrocytes (L); Recovery of immunoreactive OPN secreted into the matrix of hypertrophic GP chondrocytes (M); Recovery of immunoreactive MMP13 secreted into the matrix of GP chondrocytes (N); Histochemical staining of sulfated proteoglycans by safranin O/fast green staining (O) showing significant downregulation of proteoglycans (TIFF 26078 kb)

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Liang, G., VanHouten, J. & Macica, C.M. An Atypical Degenerative Osteoarthropathy in Hyp Mice is Characterized by a Loss in the Mineralized Zone of Articular Cartilage. Calcif Tissue Int 89, 151–162 (2011). https://doi.org/10.1007/s00223-011-9502-4

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