title: Magnesium inhibits the calcification of the extracellular matrix in tendon-derived stem cells via the ATP-P2R and mitochondrial pathways.
abstract: Tendon calcification has been widely regarded by researchers to result from the osteogenic differentiation of Tendon-Derived Stem Cells (TDSCs) and ectopic mineralization caused by the calcification of cellular matrix. Recent studies have revealed a correlation between the Mg(2+)/Ca(2+) balance and the degeneration or calcification of tendon tissues. Furthermore, the ATP-P2X/P2Y receptor pathway has been shown to play a decisive role in the process of calcification, with calcium exportation from mitochondria and calcium oscillations potentially representing the cohesive signal produced by this pathway. Our previous study demonstrated that matrix calcification is inhibited by magnesium. In this study, we examined the effects of extracellular Mg(2+) on the deposition of calcium phosphate matrix and cellular pathways in TDSCs. The suppression of the export of calcium from mitochondria has also been detected. We found that a high concentration of extracellular Mg(2+) ([Mg(2+)]e) inhibited the mineralization of the extracellular matrix in TDSCs and that 100 μM ATP reversed this inhibitory effect in vitro. In addition, the spontaneous release of ATP was inhibited by high [Mg(2+)]e levels. A high [Mg(2+)]e suppressed the expression of P2X4, P2X5 and P2X7 and activated the expression of P2Y1, P2Y2, P2Y4 and P2Y14. The interaction between Mg(2+) and Ca(2+) is therefore contradictory, Mg(2+) inhibits mitochondrial calcium concentrations, meanwhile it reverses the opening of mPTP that is induced by Ca(2+). JC-1 staining verified the protective effect of Mg(2+) on mitochondrial membrane potential and the decrease induced by Ca(2+). Taken together, these results indicate that high [Mg(2+)]e interferes with the expression of P2 receptors, resulting in decreased extracellular mineralization. The balance between Mg(2+) and Ca(2+) influences mitochondrial calcium exportation and provides another explanation for the mechanism underlying matrix calcification in TDSCs.
authors.affiliation: Department of Orthopedics, Shanghai Tenth People's Hospital, Tenth People's Hospital of Tongji University, 301 Middle Yanchang Road, Shanghai 200072, People's Republic of China; Tongji University School of Medicine, 1239 Siping Road, Shanghai 200092, People's Republic of China.
authors.forename: Jiaji
authors.initials: J
authors.lastname: Yue
authors.affiliation: Department of Orthopedics, Shanghai Tenth People's Hospital, Tenth People's Hospital of Tongji University, 301 Middle Yanchang Road, Shanghai 200072, People's Republic of China; Tongji University School of Medicine, 1239 Siping Road, Shanghai 200092, People's Republic of China.
authors.forename: Shanzi
authors.initials: S
authors.lastname: Jin
authors.affiliation: Department of Orthopedics, Shanghai Tenth People's Hospital, Tenth People's Hospital of Tongji University, 301 Middle Yanchang Road, Shanghai 200072, People's Republic of China; Tongji University School of Medicine, 1239 Siping Road, Shanghai 200092, People's Republic of China.
authors.forename: Yaqiang
authors.initials: Y
authors.lastname: Li
authors.affiliation: Department of Orthopedics, Shanghai Tenth People's Hospital, Tenth People's Hospital of Tongji University, 301 Middle Yanchang Road, Shanghai 200072, People's Republic of China; Department of Trauma Surgery, Klinikum rechts der lsar, Technical University of Munich, Ismaningerstrasse 22, 81675 Munich, Germany.
authors.forename: Li
authors.initials: L
authors.lastname: Zhang
authors.affiliation: Department of Orthopedics, Shanghai Tenth People's Hospital, Tenth People's Hospital of Tongji University, 301 Middle Yanchang Road, Shanghai 200072, People's Republic of China; The First Clinical Medical College, Nanjing Medical University, 140 Hanzhong Road, Nanjing 210029, People's Republic of China.
authors.forename: Wenwei
authors.initials: W
authors.lastname: Jiang
authors.affiliation: Department of Orthopedics, Shanghai Tenth People's Hospital, Tenth People's Hospital of Tongji University, 301 Middle Yanchang Road, Shanghai 200072, People's Republic of China. Electronic address: chunxi_yang@163.com.
authors.forename: Chunxi
authors.initials: C
authors.lastname: Yang
authors.affiliation: School of Materials Science and Engineering, Tongji University, Shanghai 201804, China. Electronic address: dujiang@tongji.edu.cn.
authors.forename: Jiang
authors.initials: J
authors.lastname: Du
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D002113:Calcification, Physiologic / Q000187:drug effects / Q000502:physiology*; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D005109:Extracellular Matrix / Q000187:drug effects / Q000378:metabolism*; D008274:Magnesium / Q000008:administration & dosage*; D008297:Male; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D015398:Signal Transduction / Q000187:drug effects; D013234:Stem Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D013710:Tendons / Q000166:cytology / Q000378:metabolism*
chemical_list: D000255:Adenosine Triphosphate; D008274:Magnesium
keywords: Magnesium; Mitochondria; P2 receptors; Tendon calcification; Tendon-derived stem cells