This study was designed to investigate the odontogenic effects of Silicon, Calcium and Phosphorous on human dental pulp cells. Human dental pulp cells derived from extracted pristine teeth were cultured in growth media with supplements of Si 25ppm, Si 25ppm+Ca 8.3ppm, Si 25ppm+Ca 8.3ppm+P 4.16ppm, Si 50ppm, Si 50ppm+Ca 16.7ppm, Si 50ppm+Ca 16.7ppm+P 8.3ppm and media without additional supplement as control, for the time intervals of 16 hours, 7, 12, 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. Mineralization of cultures was measured by Alizarin Red staining. The data were presented as the mean of triplicates and normalized on a per million cell basis. Statistical analysis was conducted using ANOVA and Tukey HSD post-hoc tests. Culture with 50ppm supplemental Si at day 21 yield significantly higher levels of ALP activity, DSP expression and mineralization (P<0.05) compared to the control group and other supplemented groups. Cultures with Si 25ppm+Ca 8.3ppm supplemental and Si 50ppm+Ca 16.7ppm supplemental displayed significantly higher cell proliferation rates compared to the control group at day 12 (P<0.05) and at day 21 (P<0.05). Supplemental silicon in concentration of 50 ppm could significantly induce differentiation and mineralization of normal human dental pulp cells. Calcium has a synergetic effect in up-regulating the proliferation rates. This is the first report to demonstrate the silicon- and calcium-induced mineralized tissue formation of human dental pulp cell cultures, leading to the potential development and clinical application of a future novel dental pulp capping material.
Effect of Silicon and Calcium on Human Dental Pulp Cell Cultures, International Journal of Materials Science and Applications.
Vol. 6, No. 6,
2017, pp. 290-296.
Ricketts, D., Restorative dentistry: Management of the deep carious lesion and the vital pulp dentine complex. Br Dent J, 2001. 191(11): p. 606-610.
Yu, F., et al., Effect of an Experimental Direct Pulp-capping Material on the Properties and Osteogenic Differentiation of Human Dental Pulp Stem Cells. Scientific Reports, 2016. 6: p. 34713.
Cho, S.-Y., et al., Prognostic Factors for Clinical Outcomes According to Time after Direct Pulp Capping. Journal of Endodontics, 2013. 39(3): p. 327-331.
Komabayashi, T., et al., Current status of direct pulp-capping materials for permanent teeth. Dental Materials Journal, 2016. 35(1): p. 1-12.
Hilton, T. J., Keys to Clinical Success with Pulp Capping: A Review of the Literature. Operative dentistry, 2009. 34(5): p. 615-625.
Nowicka, A., et al., Tomographic Evaluation of Reparative Dentin Formation after Direct Pulp Capping with Ca(OH)2, MTA, Biodentine, and Dentin Bonding System in Human Teeth. Journal of Endodontics, 2015. 41(8): p. 1234-1240.
Holland, R., et al., Reaction of rat connective tissue to implanted dentin tubes filled with mineral trioxide aggregate or calcium hydroxide. Journal of Endodontics, 1999. 25(3): p. 161-166.
Christodoulou, I., et al., Dose- and time-dependent effect of bioactive gel-glass ionic-dissolution products on human fetal osteoblast-specific gene expression. Journal of Biomedical Materials Research Part B: Applied Biomaterials, 2005. 74B (1): p. 529-537.
Tsigkou, O., et al., Differentiation of fetal osteoblasts and formation of mineralized bone nodules by 45S5 Bioglass® conditioned medium in the absence of osteogenic supplements. Biomaterials, 2009. 30(21): p. 3542-3550.
Gough, J. E., J. R. Jones, and L. L. Hench, Nodule formation and mineralisation of human primary osteoblasts cultured on a porous bioactive glass scaffold. Biomaterials, 2004. 25(11): p. 2039-2046.
Eklou-Kalonji, E., et al., Effects of extracellular calcium on the proliferation and differentiation of porcine osteoblasts in vitro. Cell and Tissue Research, 1998. 292(1): p. 163-171.
Catauro, M., et al., Biological influence of Ca/P ratio on calcium phosphate coatings by sol-gel processing. Materials Science and Engineering: C, 2016. 65: p. 188-193.
Galliano, P., et al., Sol-gel coatings on 316L steel for clinical applications. Journal of sol-gel science and technology, 1998. 13(1-3): p. 723-727.
Hench, L. L., The story of Bioglass®. Journal of Materials Science: Materials in Medicine, 2006. 17(11): p. 967-978.
Stanislawski, L., et al., In vitro culture of human dental pulp cells: some aspects of cells emerging early from the explant. Clinical Oral Investigations, 1997. 1(3): p. 131-140.
Freshney, R. I., Culture of animal cells. A Manual of Basic, 1994.
Chou, L., et al., Effects of titanium on transcriptional and post-transcriptional regulation of fibronectin in human fibroblasts. Journal of Biomedical Materials Research, 1996. 31(2): p. 209-217.
Wang, W. and T. Kirsch, Retinoic acid stimulates annexin-mediated growth plate chondrocyte mineralization. The Journal of Cell Biology, 2002. 157(6): p. 1061-1070.
Nakashima, M. and K. Iohara, Mobilized Dental Pulp Stem Cells for Pulp Regeneration: Initiation of Clinical Trial. Journal of Endodontics, 2014. 40(4, Supplement): p. S26-S32.
Barthel, C. R., et al., TNF-α release in monocytes after exposure to calcium hydroxide treated Escherichia coli LPS. International Endodontic Journal, 1997. 30(3): p. 155-159.
Modena, K. C. d. S., et al., cytotoxicity and biocompatibility of direct and indirect pulp capping materials. Journal of Applied Oral Science, 2009. 17(6): p. 544-554.
Fava, L. R. G. and W. P. Saunders, Calcium hydroxide pastes: classification and clinical indications. International Endodontic Journal, 1999. 32(4): p. 257-282.
Accorinte, M. L. R., et al., Response of human pulps capped with different self-etch adhesive systems. Clinical Oral Investigations, 2008. 12(2): p. 119-127.
Hebling, J., E. M. A. Giro, and C. A. de Souza Costa, Biocompatibility of an adhesive system applied to exposed human dental pulp. Journal of Endodontics, 1999. 25(10): p. 676-682.
Schuurs, A. H. B., R. J. M. Gruythuysen, and P. R. Wesselink, Pulp capping with adhesive resin-based composite vs. calcium hydroxide: a review. Dental Traumatology, 2000. 16(6): p. 240-250.
Dominguez, M. S., et al., Histological and Scanning Electron Microscopy Assessment of Various Vital Pulp-Therapy Materials. Journal of Endodontics, 2003. 29(5): p. 324-333.
Asgary, S., et al., A comparative study of histologic response to different pulp capping materials and a novel endodontic cement. Oral Surgery, Oral Medicine, Oral Pathology, Oral Radiology, and Endodontology, 2008. 106(4): p. 609-614.
Hørsted, P., K. El Attar, and K. Langeland, Capping of monkey pulps with Dycal and a Ca-eugenol cement. Oral Surgery, Oral Medicine, Oral Pathology, 1981. 52(5): p. 531-553.
Estrela, C. and R. Holland, Calcium hydroxide: study based on scientific evidences. Journal of Applied Oral Science, 2003. 11: p. 269-282.
Torabinejad, M., T. F. Watson, and T. R. Pitt Ford, Sealing ability of a mineral trioxide aggregate when used as a root end filling material. Journal of Endodontics, 1993. 19(12): p. 591-595.
McNamara, R. P., et al., Biocompatibility of Accelerated Mineral Trioxide Aggregate in a Rat Model. Journal of Endodontics, 2010. 36(11): p. 1851-1855.
Zhu, C., B. Ju, and R. Ni, Clinical outcome of direct pulp capping with MTA or calcium hydroxide: a systematic review and meta-analysis. International Journal of Clinical and Experimental Medicine, 2015. 8(10): p. 17055-17060.
Roberts, H. W., et al., Mineral trioxide aggregate material use in endodontic treatment: A review of the literature. Dental Materials, 2008. 24(2): p. 149-164.
Parirokh, M. and M. Torabinejad, Mineral Trioxide Aggregate: A Comprehensive Literature Review—Part I: Chemical, Physical, and Antibacterial Properties. Journal of Endodontics, 2010. 36(1): p. 16-27.
Zhai, W., et al., Silicate bioceramics induce angiogenesis during bone regeneration. Acta Biomaterialia, 2012. 8(1): p. 341-349.
Huang, Y., et al., In vitro and in vivo evaluation of akermanite bioceramics for bone regeneration. Biomaterials, 2009. 30(28): p. 5041-5048.
Chou, L., et al. Atomic and Molecular Mechanisms Underlying the Osteogenic Effects of Bioglass [R] Materials. in bioceramics-conference-. 1998.
Keeting, P. E., et al., Zeolite a increases proliferation, differentiation, and transforming growth factor β production in normal adult human osteoblast-like cells in vitro. Journal of Bone and Mineral Research, 1992. 7(11): p. 1281-1289.
Matsuoka, H., et al., In vitro analysis of the stimulation of bone formation by highly bioactive apatite- and wollastonite-containing glass-ceramic: Released calcium ions promote osteogenic differentiation in osteoblastic ROS17/2.8 cells. Journal of Biomedical Materials Research, 1999. 47(2): p. 176-188.
Dvorak, M. M., et al., Physiological changes in extracellular calcium concentration directly control osteoblast function in the absence of calciotropic hormones. Proceedings of the National Academy of Sciences of the United States of America, 2004. 101(14): p. 5140-5145.
Knabe, C., et al., Morphological evaluation of osteoblasts cultured on different calcium phosphate ceramics. Biomaterials, 1997. 18(20): p. 1339-1347.
Vrouwenvelder, W. C. A., C. G. Groot, and K. de Groot, Behaviour of fetal rat osteoblasts cultured in vitro on bioactive glass and nonreactive glasses. Biomaterials, 1992. 13(6): p. 382-392.
Li, P., et al., The role of hydrated silica, titania, and alumina in inducing apatite on implants. Journal of Biomedical Materials Research, 1994. 28(1): p. 7-15.
Sautier, J. M., et al., Bioactive glass-ceramic containing crystalline apatite and wollastonite initiates biomineralization in bone cell cultures. Calcified Tissue International, 1994. 55(6): p. 458-466.
Loty, C., et al., Bioactive Glass Stimulates In Vitro Osteoblast Differentiation and Creates a Favorable Template for Bone Tissue Formation. Journal of Bone and Mineral Research, 2001. 16(2): p. 231-239.
Yang, X., 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(1): p. 66-73.
Väkevä, L., et al., 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(1): p. 69-76.
Butler, W. T., et al., Isolation, Characterization and Immunolocalization of a 53-kDal Dentin Sialoprotein (DSP). Matrix, 1992. 12(5): p. 343-351.
Couble, M.-L., et al., Odontoblast Differentiation of Human Dental Pulp Cells in Explant Cultures. Calcified Tissue International, 2000. 66(2): p. 129-138.
Kim, J., et al., Evaluation of reparative dentin formation of ProRoot MTA, Biodentine and Bio Aggregate using micro-CT and immunohistochemistry. Restorative Dentistry & Endodontics, 2016. 41(1): p. 29-36.
Chaudhary, S. C., et al., Phosphate induces formation of matrix vesicles during odontoblast-initiated mineralization in vitro. Matrix Biology, 2016. 52–54: p. 284-300.
Montazerolghaem, M., H. Engqvist, and M. Karlsson Ott, Sustained release of simvastatin from premixed injectable calcium phosphate cement. Journal of Biomedical Materials Research Part A, 2014. 102(2): p. 340-347.
Lian, J. B. and G. S. Stein, Concepts of osteoblast growth and differentiation: basis for modulation of bone cell development and tissue formation. Critical Reviews in Oral Biology & Medicine, 1992. 3(3): p. 269-305.