1. Mineralization of bone-related SaOS-2 cells under physiological hypoxic conditions
- Author
-
Xiaohong Wang, Heinz C. Schröder, Werner E. G. Müller, Bärbel Diehl-Seifert, and Emad Tolba
- Subjects
Calcium Phosphates ,0301 basic medicine ,Cell Survival ,Surface Properties ,Bicarbonate ,chemistry.chemical_element ,02 engineering and technology ,Biology ,Calcium ,Biochemistry ,Mineralization (biology) ,Cell Line ,Structure-Activity Relationship ,03 medical and health sciences ,chemistry.chemical_compound ,Calcification, Physiologic ,Antigens, Neoplasm ,Carbonic anhydrase ,Humans ,Lactic Acid ,Particle Size ,Carbonic Anhydrase IX ,Molecular Biology ,Saos-2 cells ,Carbonic Anhydrases ,Cell Proliferation ,Osteoblasts ,Polyphosphate ,Cell Biology ,021001 nanoscience & nanotechnology ,Cell Hypoxia ,Oxygen ,030104 developmental biology ,chemistry ,Anaerobic glycolysis ,Cell culture ,biology.protein ,Biophysics ,Nanoparticles ,0210 nano-technology - Abstract
Inorganic polyphosphate (polyP) is a physiological energy-rich polymer with multiple phosphoric anhydride bonds. In cells such as bone-forming osteoblasts, glycolysis is the main pathway generating metabolic energy in the form of ATP. In the present study, we show that, under hypoxic culture conditions, the growth/viability of osteoblast-like SaOS-2 cells is not impaired. The addition of polyP to those cells, administered as amorphous calcium polyP nanoparticles (aCa-polyP-NP; approximate size 100 nm), significantly increased the proliferation of the cells. In the presence of polyP, the cells produce significant levels of lactate, the end product of anaerobic glycolysis. Under those conditions, an eight-fold increase in the steady-state level of the membrane-associated carbonic anhydrase IX is found, as well as a six-fold induction of the hypoxia-inducible factor 1. Consequently, biomineral formation onto the SaOS-2 cells decreases under low oxygen tension. If the polyP nanoparticles are added to the cells, the degree of mineralization is enhanced. These changes had been measured also in human mesenchymal stem cells. The assumption that the bicarbonate, generated by the carbonic anhydrase in the presence of polyP under low oxygen, is deposited as a constituent of the bioseeds formed during initial hydroxyapatite formation is corroborated by the identification of carbon besides of calcium, oxygen and phosphorus in the initial biomineral deposit onto the cells using the sensitive technology of high-resolution energy dispersive spectrometry mapping. Based on these data, we conclude that polyP is required for the supply of metabolic energy during bone mineral formation under physiological, hypoxic conditions, acting as a 'metabolic fuel' for the cells to grow.
- Published
- 2015