Cytotoxic Effects of Zinc Oxide on Human Periodontal Ligament Fibroblasts In Vitro

Abstract

Zinc oxide ZnO has been widely used in wound dressing, food additive and root canal filling material owing to its antibacterial and antifungal properties. This study describes potential cytotoxic effect of ZnO on human periodontal ligament fibroblasts hPDLFs in vitro. ZnO was characterized using BET Braun-Emmet-Teller and DLS dynamic light scattering analyses. The hPDLFs were treated with 0.1, 1, 10, 50 and 100 μg/mL of ZnO for 6 h, 24 h and 48 h durations. After exposure to ZnO, phase contrast microscopy, transmission electron microscopy TEM , mitochondrial function MTT and lactate dehydrogenase LDH assays were used to evaluate cell morphology, particle uptake, cell viability and membrane leakage. In addition, real time cell responses to ZnO exposure were monitored by impedance measurements. Results indicated that ZnO exposure caused toxic effect in a dose- and timedependent manner when compared to control group, especially at 50 and 100 μg/mL.

Keywords

• Cytotoxicity,Zinc oxide,Periodontal ligament fibroblasts,Impedance,In vitro toxicity assays

References

1. Dönmez Güngüneş Ç, Şeker Ş, Elçin AE, Elçin YM. A comparative study on the in vitro cytotoxic responses of two mammalian cell types to fullerenes, carbon nanotubes and iron oxide nanoparticles. Drug and Chemical Toxicology 40(2) (2017) 215-227.
2. Martinez-Serra J, Gutierrez A, Muñoz-Capó S, Navarro- Palou M, Ros T, Amat JC, Lopez B, Marcus TF, Fueyo L, Suquia AG, Gines J, Rubio F, Ramos R, Besalduch J. xCELLigence system for real-time label-free monitoring of growth and viability of cell lines from hematological malignancies. OncoTargets and Therapy 7 (2014) 985-994.
3. Urcan E, Haertel U, Styllou M, Hickel R, Scherthan H, Reichl FX. Real-time xCELLigence impedance analysis of the cytotoxicity of dental composite components on human gingival fibroblasts. Dental Materials 26(1) (2010) 51-58.
4. Şeker Ş, Elçin AE, Yumak T, Sınağ A, Elçin YM. In vitro cytotoxicity of hydrothermally synthesized ZnO nanoparticles on human periodontal ligament fibroblast and mouse dermal fibroblast cells. Toxicology in Vitro 28(8) (2014) 1349-1358.
5. Gatoo MA, Naseem S, Arfat MY, Mahmood Dar A, Qasim K, Zubair S. Physicochemical properties of nanomaterials: implication in associated toxic manifestations. BioMed Research International 2014 (2014) 1-8.
6. Pan CH, Liu WT, Bien MY, Lin IC, Hsiao TC, Ma CM, Lai CH, Chen MC, Chuang KJ, Chuang HC. Effects of size and surface of zinc oxide and aluminum-doped zinc oxide nanoparticles on cell viability inferred by proteomic analyses. International Journal of Nanomedicine 9 (2014) 3631-3643.
7. Milić M, Leitinger G, Pavičić I, Zebić Avdičević M, Dobrović S, Goessler W, Vinković Vrček I. Cellular uptake and toxicity effects of silver nanoparticles in mammalian kidney cells. Journal of Applied Toxicology 35(6) (2015) 581-592.
8. Lin W, Xu Y, Huang CC, Ma Y, Shannon KB, Chen DR, Huang YW. Toxicity of nano-and micro-sized ZnO particles in human lung epithelial cells. Journal of Nanoparticle Research 11(1) (2009) 25-39.

9. Condello M, De Berardis B, Ammendolia MG, Barone F, Condello G, Degan P, Meschini S. ZnO nanoparticle tracking from uptake to genotoxic damage in human colon carcinoma cells. Toxicology In Vitro 35 (2016) 169-179.
10. Niles AL, Moravec RA, Riss TL. In vitro viability and cytotoxicity testing and same-well multi-parametric combinations for high throughput screening. Current Chemical Genomics 3 (2009) 33-41.
11. Ng CT, Yong LQ, Hande MP, Ong CN, Yu LE, Bay BH, Baeg GH. Zinc oxide nanoparticles exhibit cytotoxicity and genotoxicity through oxidative stress responses in human lung fibroblasts and Drosophila melanogaster. International Journal of Nanomedicine 12 (2017) 1621-1637.
12. Silva EJNLD, Santos CC, Zaia AA. Long-term cytotoxic effects of contemporary root canal sealers. Journal of Applied Oral Science 21(1) (2013) 43-47.
13. Ashraf H, MoradiMajd N, Mozayeni MA, Dianat O, Mahjour F, Yadegari Z. Cytotoxicity evaluation of three resin-based sealers on an L929 cell line. Dental Research Journal 9(5) (2012) 549-553.
14. Chakar S, Changotade S, Osta N, Khalil I. Cytotoxic evaluation of a new ceramic-based root canal sealer on human fibroblasts. European Journal of Dentistry 11(2) (2017) 141-148.