Abstract
The growing interest of nanotechnology in dentistry has sparked various applications of biomaterials in nanoscale to be developed. The aim of this study was to evaluate the genotoxicity effect of locally produced hydroxyapatite-silica nanocomposite (School of Dental Sciences, Universiti Sains Malaysia, Malaysia) using Comet assay on human lung fibroblast cell line, MRC-5. Extraction of this test material was prepared and the concentrations which produced IC10, IC25 and IC50 in cytotoxicity tests (MTT assay) were recorded. Three specific concentrations, 0.00005 g/mL, 0.0009 g/mL and 0.1 g/mL for IC10, 1C25 and IC50 respectively were further used to evaluate the genotoxicity effect along with concurrent positive (hydrogen peroxide) and negative (Eagle’s Minimum Essential Medium) controls. There was no significant difference in the tail moments between negative control and treatment groups (0.00005 g/mL). Dose-dependent relationship was observed, where significant difference was noticed in the tail moments between 0.0009 g/mL and 0.1 g/mL groups with that of the negative control. However, since the values were still less than 5, it can be considered as non-genotoxic. The tail moments between different concentrations of hydroxyapatite-silica nanocomposite and positive control differed significantly (P/0.05). This concludes that the locally produced HAsilica nanocomposite is non-genotoxic by Comet assay under the present test conditions.
Similar content being viewed by others
References
Mitra, S. B., Wu, D. & Holmes, B. N. An application of nanotechnology in advanced dental materials. J Am Dent Assoc 134:1382–1390 (2003).
Terry, D. A. Direct applications of a nanocomposite resin system: Part 1-the evolution of contemporary composite materials. Pract Proced Aesthet Dent 16: 1–7 (2004).
Saunders, S. A. Current practicality of nanotechnology in dentistry. Part 1: Focus on nanocomposite restoratives and biomimetics. Clin Cosmet Investig Dent 1: 47–61 (2009).
Dee, K. C., Puleo, D. A. & Bizios, R. Biomaterials, John Wiley and Sons, New Jersey, pp. 1–2 (2002). 5. IS
International Organization of Standardization (ISO), Biological evaluation of medical devices (10993-3) (1999).
Burhans, W. C. et al. Apoptosis-like yeast cell death in response to DNA damage and replication defects. Mutat Res 532:227–243 (2003).
Fang, B., Wan, Y. Z., Tang, T. T., Gao, C. & Dai, K. R. Proliferation and osteoblastic differentiation of human bone marrow stromal cells on hydroxyapatite/bacterial cellulose nanocomposite scaffolds. Tissue Eng Part A 15:1091–1098 (2009).
Diaz, M. et al. Synthesis and antimicrobial activity of a silver-hydroxyapatite nanocomposite. Nanomaterials 10:498–505 (2009).
Jantová, S., Letasiová, M., Birosová, L. & Palou T. M. In vitro effects of fluor-hydroxyapatite, fluorapatite and hydroxyapatite on colony formation, DNA damage and mutagenicity. Mutat Res 652:139–144 (2008).
Xu, J. L. & Khor, K. A. Chemical analysis of silica doped hydroxyapatite biomaterials consolidated by a spark plasma sintering method. J Inorg Biochem 101: 187–195 (2007).
Ravarian, R. et al. Synthesis, characterization and bioactivity investigation of bioglass/hydroxyapatite composite. Ceramics International 36:291–297 (2010).
Chen, L., Mccrate, J. M., Lee, J. C. & Li, H. The role of surface charge on the uptake and biocompatibility of hydroxyapatite nanoparticles with osteoblast cells. Nanotechnology 22:105–115 (2011).
Zhang, Z., Yang, Y. & Ong, J. L. Bionanotechnology: Global Aspect, Taylor & Francis Group, London, p. 249 (2008).
Sharma, V. et al. DNA damaging potential of zinc oxide nanoparticles in human epidermal cells. Toxicol Lett 185:211–218 (2009).
Collins, A. R. The Comet assay for DNA damage and repair: principles, applications, and limitations. Mol Biotechnol 26:249–261 (2004).
Tice, R. et al. Single cell gel/Comet assay: guidelines for in vitro and in vivo genetic toxicology testing. Environ Mol Mutagen 35:206–221 (2000).
Park, S. Y. & Choi, J. H. Cytotoxicity, genotoxicity and ecotoxicity assay using human cell and environmental species for the screening of the risk from pollutant exposure. Environ Int 33:817–822 (2007).
Venturi, M. et al. Genotoxic activity in human faecal water and the role of bile acids: a study using the alkaline comet assay. Carcinogenesis 18:2353–2359 (1997).
Zeni, O. et al. Cytotoxicity investigation on cultured human blood cells treated with single-wall carbon nanotubes. Sensors 8:488–499 (2008).
Mohammed, N., Kannan, T. P., Adam, H., Haswati, A. & Abdul, R. I. Genotoxicity evaluation of locally produced dental porcelain-an in vitro study using the ames and comet assays. Toxicol In Vitro 23:1145–1150 (2009).
Katzer, A., Marquardt, H., Westendorf, J., Wening, J. V. & von Foerster, G. Polyetheretherketone-Cytotoxicity and mutagenicity in vitro. Biomaterials 23: 1749–1759 (2002).
Madle, S., Korte, A. & Ball, R. Experience with mutagenicity testing of new drugs: view point of a regulatory agency. Mutat Res 182:187–192 (1987).
ICH. International Conference on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use (2008).
Theiszovaa, M., Jantovaa, S., Draguňovab, J., Grznarovaa, P. & Palouc, M. Comparison the cytotoxicity of hydroxyapatite measured by direct cell counting and MTT test in murine fibroblast NIH-3T3 cells. Biomed Pap Med Fac Univ Palacky Olomouc Czech Repub 149:393–396 (2005).
Napierska, D. et al. Size-dependent cytotoxicity of monodisperse silica nanoparticles in human endothelial cells. Small 5:846–853 (2009).
Horváthová, E. et al. The nature and origin of DNA single-strand breaks determined with the comet assay. Mutat Res 409:163–171 (1998).
Ponti, J. et al. Genotoxicity and morphological transformation induced by cobalt nanoparticles and cobalt chloride: an in vitro study in Balb/3T3 mouse fibroblasts. Mutagenesis 24:439–445 (2009).
Trouiller, B., Reliene, R., Westbrook, A., Solaimani, P. & Schiestl, R. H. Titanium dioxide nanoparticles induce DNA damage and genetic instability in vivo in mice. Cancer Res 69:8784–8789 (2009).
Nabeshi, H. et al. Amorphous nanosilica induce endocytosis-dependent ROS generation and DNA damage in human keratinocytes. Part Fibre Toxicol 8:1 (2011).
Beckerman, M. Cellular Signaling in Health and Disease, Springer, New York, p. 149 (2009).
Cemeli, E. et al. Antigenotoxic properties of selenium compounds on potassium dichromate and hydrogen peroxide. Teratog Carcinog Mutagen 2:53–67 (2003).
Hassan, A. & Swaminathan, D. An in vitro study to evaluate the genotoxicity of value added hydroxyapatite as a bone replacement material. Sains Malaysiana 40:139–147 (2011).
Rajab, N. F. et al. DNA damage evaluation of hydroxyapatite on fibroblast cell L929 using the single cell gel electrophoresis assay. Med J Malaysia 59:170–171 (2004).
Barnes, A. C. et al. Reproducible comet assay of amorphous silica nanoparticles detects no genotoxicity. Nano Lett 8: 3069–3074 (2008).
Sayes, C. M. et al. Changing the dose metric for inhalation toxicity studies: short-term study in rats with engineered aerosolized amorphous silica nanoparticles. Inhal Toxicol 22:348–354 (2010).
ISO. International Organization of Standardization (ISO), Biological evaluation of medical devices (10993-5) (1999).
Mosmann, T. Rapid colorimetric assay for cellular growth and survival: Application to proliferation and cytotoxicity assays. J Immunol Method 65:55–63 (1983).
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Musa, M., Kannan, T.P., Masudi, S.M. et al. Assessment of DNA damage caused by locally produced hydroxyapatite-silica nanocomposite using Comet assay on human lung fibroblast cell line. Mol. Cell. Toxicol. 8, 53–60 (2012). https://doi.org/10.1007/s13273-012-0007-7
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s13273-012-0007-7