International Journal of Materials Science and Applications
Volume 8, Issue 3, May 2019, Pages: 40-46
Received: Jul. 1, 2019;
Accepted: Aug. 5, 2019;
Published: Aug. 23, 2019
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Jomana Alsenan, Department of Restorative Sciences & Biomaterials, Goldman School of Dental Medicine, Boston University, Boston, USA
Laisheng Chou, Department of Restorative Sciences & Biomaterials, Goldman School of Dental Medicine, Boston University, Boston, USA
This study was designed to investigate the effect of inorganic phosphate (Pi) at different concentrations on odontogenesis of the normal human dental pulp cells (hDPCs). Normal human dental pulp cells derived from extracted pristine teeth were cultured in growth medium with supplements of inorganic phosphate (Pi) in 0 ppm, 2 ppm, 4 ppm, 5 ppm and 8 ppm, for the time intervals of 16 hours, 7, 14, and 21 days. Cell proliferation rates were measured by the optical density of crystal violet dye stained cells. ALP activity was measured by fluorometric assay. Expression of Dentin Sialoprotein (DSP) was measured by ELISA. The data were presented as the mean of triplicates. Statistical analysis was conducted using JMP Pro 12 (ver. 12.1.0) in one-way ANOVA and Tukey HSD post-hoc tests. Cell attachment efficiency was reduced significantly by additional Pi of 2, 4 and 5 ppm (P<0.05). At 21 days, cultures with 2, 4 and 5 ppm supplemental Pi displayed significantly higher cell proliferation rates compared to the control group at day 14 (P<0.05) and at day 21 (P<0.05). At day 7, cultures with 2, 4, 5 and 8 ppm supplemental Pi yield significantly higher levels of ALP activity (P<0.05) compared to the control group. At day 7, cultures with 5 ppm Pi supplement showed significantly higher levels of DSP expression (P<0.05) compared to the control group and the rest of the other groups. Supplemental Pi in concentration of 5 ppm could significantly induce proliferation and odontogenesis of hDPCs. This is the first report to demonstrate Pi-induced odontogenesis, leading to potential development and clinical application of future Pi containing dental pulp capping or root canal filling materials.
Inorganic Phosphate Effect of on Human Dental Pulp Cell Cultures, International Journal of Materials Science and Applications.
Vol. 8, No. 3,
2019, pp. 40-46.
Williams R. Phosphorus biochemistry. 1978.
Del Valle HB, Yaktine AL, Taylor CL, Ross AC. Dietary reference intakes for calcium and vitamin D: National Academies Press; 2011.
Iheagwara OS, Ing TS, Kjellstrand CM, Lew SQ. Phosphorus, phosphorous, and phosphate. Hemodialysis International. 2013; 17: 479-82.
Hong S-H, Park S-J, Lee S, Kim S, Cho M-H. Biological effects of inorganic phosphate: potential signal of toxicity. The Journal of Toxicological Sciences. 2015; 40: 55-69.
Zhang R, Lu Y, Ye L, Yuan B, Yu S, Qin C, et al. Unique roles of phosphorus in endochondral bone formation and osteocyte maturation. Journal of Bone and Mineral Research. 2011; 26: 1047-56.
Hsu Y-J, Hsu S-C, Huang S-M, Lee H-S, Lin S-H, Tsai C-S, et al. Hyperphosphatemia induces protective autophagy in endothelial cells through the inhibition of Akt/mTOR signaling. Journal of Vascular Surgery. 2015; 62: 210-21.e2.
Nadkarni GN, Uribarri J. Phosphorus and the Kidney: What Is Known and What Is Needed. Advances in Nutrition. 2014; 5: 98-103.
Beck GR, Zerler B, Moran E. Phosphate is a specific signal for induction of osteopontin gene expression. Proceedings of the National Academy of Sciences. 2000; 97: 8352-7.
Beck GR, Moran E, Knecht N. Inorganic phosphate regulates multiple genes during osteoblast differentiation, including Nrf2. Experimental cell research. 2003; 288: 288-300.
Beck GR. Inorganic phosphate as a signaling molecule in osteoblast differentiation. Journal of cellular biochemistry. 2003; 90: 234-43.
Meleti Z, Shapiro I, Adams CS. Inorganic phosphate induces apoptosis of osteoblast-like cells in culture. Bone. 2000; 27: 359-66.
Adams CS, Shapiro I. Mechanisms by which extracellular matrix components induce osteoblast apoptosis. Connective tissue research. 2003; 44: 230-9.
Adams CS, Mansfield K, Perlot RL, Shapiro IM. Matrix regulation of skeletal cell apoptosis role of calcium and phosphate ions. Journal of Biological Chemistry. 2001; 276: 20316-22.
Gupta G, Kirakodu S, El-Ghannam A. Effects of exogenous phosphorus and silicon on osteoblast differentiation at the interface with bioactive ceramics. Journal of Biomedical Materials Research Part A. 2010; 95A: 882-90.
Stanislawski L, Carreau JP, Pouchelet M, Chen ZHJ, Goldberg M. In vitro culture of human dental pulp cells: some aspects of cells emerging early from the explant. Clinical Oral Investigations. 1997; 1: 131-40.
Freshney RI. Culture ofanimal cells. A Manual ofBasic. 1994.
Chou L, Firth JD, Nathanson D, Uitto V-J, Brunette DM. Effects of titanium on transcriptional and post-transcriptional regulation of fibronectin in human fibroblasts. Journal of Biomedical Materials Research. 1996; 31: 209-17.
Takeda E, Yamamoto H, Nashiki K, Sato T, Arai H, Taketani Y. Inorganic phosphate homeostasis and the role of dietary phosphorus. Journal of Cellular and Molecular Medicine. 2004; 8: 191-200.
Ariffin S, Manogaran T, Abidin I, Wahab R, Senafi S. A Perspective On Stem Cells As Biological Systems That Produce Differentiated Osteoblasts And Odontoblasts. Current stem cell research & therapy. 2016.
Chou L, Firth JD, Uitto V-J, Brunette DM. Substratum surface topography alters cell shape and regulates fibronectin mRNA level, mRNA stability, secretion and assembly in human fibroblasts. Journal of Cell Science. 1995; 108: 1563-73.
Chou L, Firth JD, Uitto VJ, Brunette DM. Effects of titanium substratum and grooved surface topography on metalloproteinase‐2 expression in human fibroblasts. Journal of biomedical materials research. 1998; 39: 437-45.
Keller JC, Collins JG, Niederauer GG, McGee TD. In vitro attachment of osteoblast-like cells to osteoceramic materials. Dental Materials. 1997; 13: 62-8.
Knabe C, Gildenhaar R, Berger G, Ostapowicz W, Fitzner R, Radlanski R, et al. Morphological evaluation of osteoblasts cultured on different calcium phosphate ceramics. Biomaterials. 1997; 18: 1339-47.
Stanford CM, Jacobson PA, Eanes ED, Lembke LA, Midura RJ. Rapidly forming apatitic mineral in an osteoblastic cell line (UMR 10601 BSP). Journal of Biological Chemistry. 1995; 270: 9420-8.
Addison WN, Azari F, Sørensen ES, Kaartinen MT, McKee MD. Pyrophosphate inhibits mineralization of osteoblast cultures by binding to mineral, up-regulating osteopontin, and inhibiting alkaline phosphatase activity. Journal of Biological Chemistry. 2007; 282: 15872-83.
Fleish H, Neuman WF. Mechanisms of calcification: role of collagen, polyphosphates, and phosphatase. American Journal of Physiology--Legacy Content. 1961; 200: 1296-300.
Boskey A, Posner A. Optimal conditions for Ca-acidic phospholipid-PO4 formation. Calcified tissue international. 1981; 34: S1-7.
Yang X, van den Dolder J, Walboomers XF, Zhang W, Bian Z, Fan M, et al. The odontogenic potential of STRO-1 sorted rat dental pulp stem cells in vitro. Journal of Tissue Engineering and Regenerative Medicine. 2007; 1: 66-73.
Väkevä L, Mackie E, Kantomaa T, Thesleff I. Comparison of the distribution patterns of tenascin and alkaline phosphatase in developing teeth, cartilage, and bone of rats and mice. The Anatomical Record. 1990; 228: 69-76.
Kim J, Song Y-S, Min K-S, Kim S-H, Koh J-T, Lee B-N, et al. Evaluation of reparative dentin formation of ProRoot MTA, Biodentine and BioAggregate using micro-CT and immunohistochemistry. Restorative Dentistry & Endodontics. 2016; 41: 29-36.
Butler WT, Bhown M, Brunn JC, D'Souza RN, Farach-Carson MC, Happonen R-P, et al. Isolation, Characterization and Immunolocalization of a 53-kDal Dentin Sialoprotein (DSP). Matrix. 1992; 12: 343-51.
Couble M-L, Farges J-C, Bleicher F, Perrat-Mabillon B, Boudeulle M, Magloire H. Odontoblast Differentiation of Human Dental Pulp Cells in Explant Cultures. Calcified Tissue International. 2000; 66: 129-38.
Owen TA, Aronow M, Shalhoub V, Barone LM, Wilming L, Tassinari MS, et al. Progressive development of the rat osteoblast phenotype in vitro: reciprocal relationships in expression of genes associated with osteoblast proliferation and differentiation during formation of the bone extracellular matrix. Journal of cellular physiology. 1990; 143: 420-30.