Characterization of an in vitro model of calcification in retinal pigmented epithelial cells.

Little is known about the relationship at the molecular and cellular levels between vascular calcification and elastic fibers essential for elasticity. To gain a better understanding of the physiological function of elastin in vascular calcification, we developed a calcification model on cultured bovine retinal-pigmented-epithelial cells (RPEs) that do not express endogenous tropoelastin. The addition of inorganic phosphate (NaH2PO4; Pi) induced calcium deposition in RPEs. The Pi-induced calcification, as assessed by the o-cresolphthalein complexone method, Goldenbergs method, and von Kossa staining, was completely inhibited by treatment with clodronate (DMDP) and phosphonoformic acid (PFA) and was weakly suppressed by treatment with levamisole. Moreover, the osteopontin mRNA expression was upregulated in the Pi-induced calcification of RPEs. These reactions in RPEs were characteristically consistent with those already established in cultured bovine aortic smooth muscle cells (BASMCs). Furthermore, bacterially expressed tropoelastin inhibited calcium deposition in RPEs as well as in BASMCs. Finally, Pi-induced calcification was partially suppressed after the addition of tropoelastin due to elastic fiber formation. In conclusion, we suggest that this calcification model in RPEs is useful for analyzing the relation between elastic fibers and vascular calcification, and that tropoelastin and elastic fibers may contribute to the inhibition of vascular calcification.

[1]  R. Mecham,et al.  Accelerated calcification represses the expression of elastic fiber components and lysyl oxidase in cultured bovine aortic smooth muscle cells. , 2002, Journal of atherosclerosis and thrombosis.

[2]  R. Aebersold,et al.  Smooth Muscle Cell Phenotypic Transition Associated With Calcification: Upregulation of Cbfa1 and Downregulation of Smooth Muscle Lineage Markers , 2001, Circulation research.

[3]  H. Koyama,et al.  Mönckeberg's medial sclerosis and inorganic phosphate in uremia. , 2001, American journal of kidney diseases : the official journal of the National Kidney Foundation.

[4]  D. Sane,et al.  Arterial calcification: A review of mechanisms, animal models, and the prospects for therapy , 2001, Medicinal research reviews.

[5]  S. Tajima,et al.  Nitric oxide stimulates elastin expression in chick aortic smooth muscle cells. , 2001, Biological & pharmaceutical bulletin.

[6]  H. Dietz,et al.  Phenotypic Alteration of Vascular Smooth Muscle Cells Precedes Elastolysis in a Mouse Model of Marfan Syndrome , 2001, Circulation research.

[7]  M. McKee,et al.  Phosphate regulation of vascular smooth muscle cell calcification. , 2000, Circulation research.

[8]  R. Mecham,et al.  Characterization of an in vitro model of elastic fiber assembly. , 1999, Molecular biology of the cell.

[9]  Frederick J. Schoen,et al.  Elastin calcification and its prevention with aluminum chloride pretreatment. , 1999, The American journal of pathology.

[10]  T. Osakabe,et al.  [Qualitative change in the elastin from the calcified portion of human artery]. , 1999, Nihon Ronen Igakkai zasshi. Japanese journal of geriatrics.

[11]  J. Wallach,et al.  Biological effects of elastin peptides. , 1998, Pathologie-biologie.

[12]  YoshikiNishizawa,et al.  Parathyroid Hormone–Related Peptide as a Local Regulator of Vascular Calcification , 1997 .

[13]  Y. Nishizawa,et al.  Parathyroid hormone-related peptide as a local regulator of vascular calcification. Its inhibitory action on in vitro calcification by bovine vascular smooth muscle cells. , 1997, Arteriosclerosis, thrombosis, and vascular biology.

[14]  J. Atkinson,et al.  Calcification of medial elastic fibers and aortic elasticity. , 1997, Hypertension.

[15]  M. Loghman-Adham,et al.  Use of phosphonocarboxylic acids as inhibitors of sodium-phosphate cotransport. , 1996, General pharmacology.

[16]  Y. Bobryshev,et al.  Calcified deposit formation in intimal thickenings of the human aorta. , 1995, Atherosclerosis.

[17]  H. Koyama,et al.  β-Glycerophosphate Accelerates Calcification in Cultured Bovine Vascular Smooth Muscle Cells , 1995 .

[18]  S. Tajima,et al.  Modulation of elastin expression by heparin is dependent on the growth condition of vascular smooth muscle cells: up-regulation of elastin expression by heparin in the proliferating cells is mediated by the inhibition of protein kinase C activity. , 1995, Journal of biochemistry.

[19]  T. Nishikawa,et al.  Minoxidil stimulates elastin expression in aortic smooth muscle cells. , 1994, Archives of biochemistry and biophysics.

[20]  J. Womack,et al.  Sequence of the coding region of the bovine fibrillin cDNA and localization to bovine chromosome 10. , 1994, Genomics.

[21]  S. Tajima,et al.  Elastin synthesis is inhibited by angiotensin II but not by platelet-derived growth factor in arterial smooth muscle cells. , 1994, Biochimica et biophysica acta.

[22]  K. Watson,et al.  Bone morphogenetic protein expression in human atherosclerotic lesions. , 1993, The Journal of clinical investigation.

[23]  W. Paule,et al.  Calcification of implanted vascular tissues associated with elastin in an experimental animal model. , 1992, Journal of biomedical materials research.

[24]  R. Levy,et al.  Calcification of allograft aortic wall in a rat subdermal model. Pathophysiology and inhibition by Al3+ and aminodiphosphonate preincubations. , 1992, The American journal of pathology.

[25]  M. Young,et al.  The cDNA cloning and RNA distribution of bovine osteopontin. , 1991, Gene.

[26]  P. Trackman,et al.  Cloning of rat aorta lysyl oxidase cDNA: complete codons and predicted amino acid sequence. , 1991, Biochemistry.

[27]  J. Tímár,et al.  Interaction between elastin and tumor cell lines with different metastatic potential; in vitro and in vivo studies , 1991, Journal of Cancer Research and Clinical Oncology.

[28]  P. Trackman,et al.  Cloning of rat aorta lysyl oxidase cDNA: complete codons and predicted amino acid sequence. , 1990, Biochemistry.

[29]  K. Raju,et al.  Primary structures of bovine elastin a, b, and c deduced from the sequences of cDNA clones. , 1987, The Journal of biological chemistry.

[30]  W. Hornebeck,et al.  Inducible adhesion of mesenchymal cells to elastic fibers: elastonectin. , 1986, Proceedings of the National Academy of Sciences of the United States of America.

[31]  J. Rosenbloom,et al.  Elastin: relation of protein and gene structure to disease. , 1984, Laboratory investigation; a journal of technical methods and pathology.

[32]  R. Mecham,et al.  Chemotactic responses of fibroblasts to tropoelastin and elastin-derived peptides. , 1982, The Journal of clinical investigation.

[33]  S. Teitelbaum,et al.  Stereospecific inhibition of alkaline phosphatase by L-tetramisole prevents in vitro cartilage calcification. , 1980, Laboratory investigation; a journal of technical methods and pathology.

[34]  R. Mecham,et al.  Chemotactic activity of elastin-derived peptides. , 1980, The Journal of clinical investigation.

[35]  R. Cortivo,et al.  Calcium ion-binding study on α-elastin , 1974 .

[36]  D. Urry.  Neutral sites for calcium ion binding to elastin and collagen: a charge neutralization theory for calcification and its relationship to atherosclerosis. , 1971, Proceedings of the National Academy of Sciences of the United States of America.

[37]  A. L. Brown,et al.  Significance of calcification of the coronary arteries. , 1970, The American journal of cardiology.

[38]  Beadenkopf Wg,et al.  CALCIFICATION IN THE CORONARY ARTERIES AND ITS RELATIONSHIP TO ARTERIOSCLEROSIS AND MYOCARDIAL INFARCTION. , 1964 .

[39]  S. Tajima,et al.  Endothelin-1 Down-Regulates Expression of Tropoelastin and Lysyl Oxidase mRNA in Cultured Chick Aortic Smooth Muscle Cells , 2001 .

[40]  L. Robert,et al.  Interaction between cells and elastin, the elastin-receptor. , 1999, Connective tissue research.

[41]  M. Hayashi,et al.  Comparative effects of vitamin K2 and vitamin E on experimental arteriosclerosis. , 1999, International journal for vitamin and nutrition research. Internationale Zeitschrift fur Vitamin- und Ernahrungsforschung. Journal international de vitaminologie et de nutrition.

[42]  V. Velebný,et al.  Biological effects of elastin peptides. , 1988, Sbornik vedeckych praci Lekarske fakulty Karlovy university v Hradci Kralove.

[43]  R. Wuthier,et al.  Effect of pyrophosphate and two diphosphonates on 45Ca and 32Pi uptake and mineralization by matrix vesicle-enriched fractions and by hydroxyapatite. , 1985, Bone.

[44]  D. Jackson,et al.  Carbohydrate content of insoluble elastins prepared from adult bovine and calf ligamentum nuchae and tropoelastin isolated from copper-deficient porcine aorta. , 1971, The Biochemical journal.