Manuel Scimeca1, Lucia Anemona2, Annarita Granaglia3, Rita Bonfiglio4, Nicoletta Urbano5, Nicola Toschi6, Giuseppe Santeusanio7, Stefania Schiaroli8, Silvestro Mauriello9, Virginia Tancredi10, Orazio Schillaci11, Elena Bonanno12, Alessandro Mauriello13. 1. Department of Biomedicine and Prevention, University of Rome "Tor Vergata", Via Montpellier 1, Rome, 00133, Italy; San Raffaele University, Via di Val Cannuta 247, 00166, Rome, Italy; Fondazione Umberto Veronesi (FUV), Piazza Velasca 5, 20122, Milano, Italy; Saint Camillus International University of Health Sciences, Via di Sant'Alessandro, 8, 00131 Rome, Italy. Electronic address: manuel.scimeca@uniroma2.it. 2. Department of Experimental Medicine, University of Rome "Tor Vergata", Rome, Italy. Electronic address: anemona@uniroma2.it. 3. Department of Experimental Medicine, University of Rome "Tor Vergata", Rome, Italy. Electronic address: a.granaglia@gmail.com. 4. Department of Experimental Medicine, University of Rome "Tor Vergata", Rome, Italy. Electronic address: bonfiglio.rita@gmail.com. 5. Nuclear Medicine, Policlinico "Tor Vergata", Rome, Italy. Electronic address: n.urbano@virgilio.it. 6. Department of Biomedicine and Prevention, University of Rome "Tor Vergata", Via Montpellier 1, Rome, 00133, Italy; Department of Radiology, Athinoula A. Martinos Center for Biomedical Imaging and Harvard Medical School, Massachusetts General Hospital, Boston, MA, USA. Electronic address: toschi@med.uniroma2.it. 7. Department of Experimental Medicine, University of Rome "Tor Vergata", Rome, Italy. Electronic address: santeusa@uniroma2.it. 8. Department of Experimental Medicine, University of Rome "Tor Vergata", Rome, Italy. Electronic address: schiaroli@yahoo.com. 9. Department of Biomedicine and Prevention, University of Rome "Tor Vergata", Via Montpellier 1, Rome, 00133, Italy. Electronic address: mauriello.silvestro@gmail.com. 10. San Raffaele University, Via di Val Cannuta 247, 00166, Rome, Italy; Department of Systems Medicine, School of Sport and Exercise Sciences, University of Rome Tor Vergata, Rome, Italy; Centre of Space Biomedicine, University of Rome Tor Vergata, Rome, Italy. Electronic address: tancredi@uniroma2.it. 11. Department of Biomedicine and Prevention, University of Rome "Tor Vergata", Via Montpellier 1, Rome, 00133, Italy; IRCCS Neuromed, Pozzilli, Italy. Electronic address: orazio.schillaci@uniroma2.it. 12. Saint Camillus International University of Health Sciences, Via di Sant'Alessandro, 8, 00131 Rome, Italy; IRCCS Neuromed Lab. "Diagnostica Medica"; and "Villa dei Platani", Avellino, Italy. Electronic address: elena.bonanno@uniroma2.it. 13. Saint Camillus International University of Health Sciences, Via di Sant'Alessandro, 8, 00131 Rome, Italy; TorVergata Oncoscience Research (TOR), University of Rome "Tor Vergata", Rome, Italy. Electronic address: alessandro.mauriello@uniroma2.it.
Abstract
BACKGROUND AND AIMS: The aim of this study was to investigate possible associations among markers of mineralization, plaque instability and the main risk factors of atherosclerosis. METHODS AND RESULTS: A Tissue MicroArray containing 52 samples of calcified carotid plaques from 52 symptomatic and asymptomatic patients were built. TMA serial sections were used to study the expression of inflammatory and mineralization markers (BMP-2, BMP-4, VDR, RANKL, Osteopontin, Sclerostin, β-catenin and calmodulin) by immunohistochemistry. Our data clearly demonstrated the expression of mineralization markers in atheromatic plaques. Indeed, with the exception of RANKL, all investigated markers were expressed in at least 60% of cases. Specifically, multivariate analysis displayed significant associations between both the expression of BMP-2 and the presence of unstable plaques as well as between the expression of β-catenin and the presence of stable plaques. We also found a significant inverse association between both a) the presence of hypertension and VDR and b) smoking habits and calmodulin expression. Finally, we noted a higher density of RANKL positive cells in plaques from diabetic patients as compared to non-diabetic ones and a significant positive association between hypertriglyceridemia and BMP-4 expression. CONCLUSION: Our results support the hypothesis that the process of atherosclerotic plaque calcification presents a number of similarities with the physiological processes that occur in bone, involving both osteoblasts- and osteoclasts-like arterial cells. Finally, the present study suggests that risk factors, such as hypertension, cigarette smoke and diabetes, can cause the destabilization of the atheromatic plaque acting on calcification process as well as inflammation.
BACKGROUND AND AIMS: The aim of this study was to investigate possible associations among markers of mineralization, plaque instability and the main risk factors of atherosclerosis. METHODS AND RESULTS: A Tissue MicroArray containing 52 samples of calcified carotid plaques from 52 symptomatic and asymptomatic patients were built. TMA serial sections were used to study the expression of inflammatory and mineralization markers (BMP-2, BMP-4, VDR, RANKL, Osteopontin, Sclerostin, β-catenin and calmodulin) by immunohistochemistry. Our data clearly demonstrated the expression of mineralization markers in atheromatic plaques. Indeed, with the exception of RANKL, all investigated markers were expressed in at least 60% of cases. Specifically, multivariate analysis displayed significant associations between both the expression of BMP-2 and the presence of unstable plaques as well as between the expression of β-catenin and the presence of stable plaques. We also found a significant inverse association between both a) the presence of hypertension and VDR and b) smoking habits and calmodulin expression. Finally, we noted a higher density of RANKL positive cells in plaques from diabeticpatients as compared to non-diabetic ones and a significant positive association between hypertriglyceridemia and BMP-4 expression. CONCLUSION: Our results support the hypothesis that the process of atherosclerotic plaque calcification presents a number of similarities with the physiological processes that occur in bone, involving both osteoblasts- and osteoclasts-like arterial cells. Finally, the present study suggests that risk factors, such as hypertension, cigarette smoke and diabetes, can cause the destabilization of the atheromatic plaque acting on calcification process as well as inflammation.