Objective: Recent in vitro studies have shown that chitosan nanoparticles in several root canal sealers, intracanal medicament, and irrigation solutions could enhance the antimicrobial activity. However, the nanotoxicity of chitosan has not been fully studied. The aim of this study was to evaluate cellular uptake and genotoxicity of various sizes and concentrations of chitosan nanoparticles cultured with human dental pulp cells. Methods: Human dental pulp cells were derived from human dental pulp tissues and cultured for 24 hours with 50 nm and 318 nm FITC-tagged chitosan nanoparticles in concentrations: 0.1 mg/mL, 0.5 mg/mL, and 2 mg/mL as study groups, and 0 mg/mL as a control. The fluorescence intensity of the FITC tagged chitosan nanoparticles was measured using a spectrophotometer to determine the cellular uptake. Genotoxicity was assessed by the Cytokinesis-block micronucleus method and by measuring the fluorescent intensity of the phosphorylated H2AX nuclear foci. Statistical analysis was performed using One-Way ANOVA, post-hoc Tukey, and Chi-square tests. Results: Chitosan nanoparticles were able to internalize the human dental pulp cells and significantly induced micronuclei, nuclear buds, and pH2AX foci at concentrations of 0.5 mg/mL and 2 mg/mL as compared to 0.1 mg/mL (P < 0.01) and control group (P < 0.01). At both concentrations, 0.5 mg/mL and 2 mg/mL, 50 nm chitosan significantly induced higher proportions of micronuclei (P=0.001), nuclear buds (P=0.009), and pH2AX nuclear foci (P=0.00004) as compared to 318 nm chitosan. Conclusion: 50 nm and 318 nm chitosan nanoparticles at concentrations 0.5 mg/mL and 2 mg/mL penetrated human dental pulp cells and induced genotoxicity in dose-dependent and size-associated manners.
Published in | International Journal of Materials Science and Applications (Volume 10, Issue 4) |
DOI | 10.11648/j.ijmsa.20211004.11 |
Page(s) | 79-86 |
Creative Commons |
This is an Open Access article, distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution and reproduction in any medium or format, provided the original work is properly cited. |
Copyright |
Copyright © The Author(s), 2021. Published by Science Publishing Group |
Chitosan, Nanoparticles, Dental Pulp, Cellular Uptake, Genotoxicity
[1] | Divya K., Vijayan Smitha, George Tijith K., Jisha M. S. Antimicrobial properties of chitosan nanoparticles: Mode of action and factors affecting activity. Fibers Polym 2017; 18 (2): 221–30. Doi: 10.1007/s12221-017-6690-1. |
[2] | Kishen Anil, Shi Zhilong, Shrestha Annie, Neoh Koon Gee. An Investigation on the Antibacterial and Antibiofilm Efficacy of Cationic Nanoparticulates for Root Canal Disinfection. J Endod 2008; 34 (12): 1515–20. Doi: 10.1016/j.joen.2008.08.035. |
[3] | Chavez de Paz L. E., Resin A., Howard K. A., Sutherland D. S., Wejse P. L. Antimicrobial Effect of Chitosan Nanoparticles on Streptococcus mutans Biofilms. Appl Environ Microbiol 2011; 77 (11): 3892–5. Doi: 10.1128/AEM.02941-10. |
[4] | Del Carpio-Perochena Aldo, Kishen Anil, Shrestha Annie, Bramante Clovis Monteiro. Antibacterial Properties Associated with Chitosan Nanoparticle Treatment on Root Dentin and 2 Types of Endodontic Sealers. J Endod 2015; 41 (8): 1353–8. Doi: 10.1016/j.joen.2015.03.020. |
[5] | del Carpio-Perochena Aldo, Kishen Anil, Felitti Rafael, et al. Antibacterial Properties of Chitosan Nanoparticles and Propolis Associated with Calcium Hydroxide against Single- and Multispecies Biofilms: An In Vitro and In Situ Study. J Endod 2017; 43 (8): 1332–6. Doi: 10.1016/j.joen.2017.03.017. |
[6] | Travan Andrea, Marsich Eleonora, Donati Ivan, et al. Silver-polysaccharide nanocomposite antimicrobial coatings for methacrylic thermosets. Acta Biomater 2011; 7 (1): 337–46. Doi: 10.1016/j.actbio.2010.07.024. |
[7] | Jiang Li Qun, Wang Ting Yu, Webster Thomas J., et al. Intracellular disposition of chitosan nanoparticles in macrophages: Intracellular uptake, exocytosis, and intercellular transport. Int J Nanomedicine 2017; 12: 6383–98. Doi: 10.2147/IJN.S142060. |
[8] | Gigault Julien, Halle Alexandra ter, Baudrimont Magalie, et al. Current opinion: What is a nanoplastic? Environ Pollut 2018; 235: 1030–4. Doi: 10.1016/j.envpol.2018.01.024. |
[9] | Zhang Xu, Li Yanqiu, Sun Xiaoxi, et al. Biomimetic remineralization of demineralized enamel with nano-complexes of phosphorylated chitosan and amorphous calcium phosphate. J Mater Sci Mater Med 2014; 25 (12): 2619–28. Doi: 10.1007/s10856-014-5285-2. |
[10] | Stanislawski L, Carreau J P, Pouchelet M, Chen Z H, Goldberg M. In vitro culture of human dental pulp cells: some aspects of cells emerging early from the explant. Clin Oral Investig 1997; 1 (3): 131–40. Doi: 10.1007/s007840050024. |
[11] | Alonso M J, Calvo P, Remun C. Novel Hydrophilic Chitosan – Polyethylene Oxide Nanoparticles as Protein Carriers n. d.: 125–32. |
[12] | Gan Quan, Wang Tao, Cochrane Colette, McCarron Paul. Modulation of surface charge, particle size and morphological properties of chitosan-TPP nanoparticles intended for gene delivery. Colloids Surfaces B Biointerfaces 2005; 44 (2–3): 65–73. Doi: 10.1016/j.colsurfb.2005.06.001. |
[13] | Alhomrany Rami, Zhang Chang, Chou Laisheng. Cytotoxic effect of chitosan nanoparticles on normal human dental pulp cells 2019; 3 (1): 1–9. |
[14] | Qaqish Roula B, Amiji Mansoor M. Synthesis of a - uorescent chitosan derivative and its application for the study of chitosan ± mucin interactions 1999; 38: 99–107. |
[15] | Patiño Tania, Soriano Jorge, Barrios Lleonard, Ibáñez Elena, Nogués Carme. Surface modification of microparticles causes differential uptake responses in normal and tumoral human breast epithelial cells. Nat Publ Gr 2015; (December 2014): 1–12. Doi: 10.1038/srep11371. |
[16] | Landuyt Kirsten Van, Styllou Panorea, Rothmund Lena, et al. Cytotoxicity and induction of DNA double-strand breaks by components leached from dental composites in primary human gingival fibroblasts. Dent Mater 2013; 29 (9): 971–9. Doi: 10.1016/j.dental.2013.07.007. |
[17] | Thomas Philip, Fenech Michael. Cytokinesis-block micronucleus cytome assay in lymphocytes. Methods Mol Biol 2011; 682: 217–34. Doi: 10.1007/978-1-60327-409-8_16. |
[18] | HAMIDA R. S.;, ALBASTER G.;, BIN-MEFERIJ M. M. Oxidative Stress and Apoptotic Responses Elicited by. Cancers (Basel) 2020; 12 (8): 1–25. |
[19] | Azouz Rehab A., Korany Reda M. S. Toxic Impacts of Amorphous Silica Nanoparticles on Liver and Kidney of Male Adult Rats: an In Vivo Study. Biol Trace Elem Res 2020. Doi: 10.1007/s12011-020-02386-3. |
[20] | Shrestha Suja, Diogenes Anibal, Kishen Anil. Temporal-controlled Dexamethasone Releasing Chitosan Nanoparticle System Enhances Odontogenic Differentiation of Stem Cells from Apical Papilla. J Endod 2015; 41 (8): 1253–8. Doi: 10.1016/j.joen.2015.03.024. |
[21] | Del Carpio-Perochena A, Bramante C M, Duarte M A, de Moura M R, Aouada F A, Kishen A. Chelating and antibacterial properties of chitosan nanoparticles on dentin. Restor Dent Endod 2015; 40 (3): 195–201. Doi: 10.5395/rde.2015.40.3.195. |
[22] | Shrestha Suja, Diogenes Anibal, Kishen Anil. Temporal-controlled release of bovine serum albumin from chitosan nanoparticles: effect on the regulation of alkaline phosphatase activity in stem cells from apical papilla. J Endod 2014; 40 (9): 1349–54. Doi: 10.1016/j.joen.2014.02.018. |
[23] | Dasilva Luis, Finer Yoav, Friedman Shimon, Basrani Bettina, Kishen Anil. Biofilm formation within the interface of bovine root dentin treated with conjugated chitosan and sealer containing chitosan nanoparticles. J Endod 2013; 39 (2): 249–53. Doi: 10.1016/j.joen.2012.11.008. |
[24] | Bruinink Arie, Wang Jing, Wick Peter. Effect of particle agglomeration in nanotoxicology. Arch Toxicol 2015; 89 (5): 659–75. Doi: 10.1007/s00204-015-1460-6. |
[25] | Allouni Zouhir E., Cimpan Mihaela R., Høl Paul J., Skodvin Tore, Gjerdet Nils R. Agglomeration and sedimentation of TiO2 nanoparticles in cell culture medium. Colloids Surfaces B Biointerfaces 2009; 68 (1): 83–7. Doi: 10.1016/j.colsurfb.2008.09.014. |
[26] | Bae Eunjoo, Park Hee Jin, Lee Jeongjin, et al. Bacterial cytotoxicity of the silver nanoparticle related to physicochemical metrics and agglomeration properties. Environ Toxicol Chem 2010; 29 (10): 2154–60. Doi: 10.1002/etc.278. |
[27] | Andersson Per Ola, Lejon Christian, Ekstrand-Hammarström Barbro, et al. Polymorph- and size-dependent uptake and toxicity of TiO2 nanoparticles in living lung epithelial cells. Small 2011; 7 (4): 514–23. Doi: 10.1002/smll.201001832. |
[28] | Ahamed Maqusood, Karns Michael, Goodson Michael, et al. DNA damage response to different surface chemistry of silver nanoparticles in mammalian cells. Toxicol Appl Pharmacol 2008; 233 (3): 404–10. Doi: 10.1016/j.taap.2008.09.015. |
[29] | Huang Min, Ma Zengshuan, Khor Eugene, Lim Lee Yong. Uptake of FITC-chitosan nanoparticles by A549 cells. Pharm Res 2002; 19 (10): 1488–94. Doi: 10.1023/A:1020404615898. |
[30] | Kuo Linda J., Yang Li Xi. γ-H2AX- A novel biomaker for DNA double-strand breaks. In Vivo (Brooklyn) 2008; 22 (3): 305–10. |
[31] | Sharma Arishya, Singh Kamini, Almasan Alexandru. Histone H2AX Phosphorylation: A Marker for DNA Damage 2012; 920: 613–26. Doi: 10.1007/978-1-61779-998-3_40. |
[32] | Shuga J., Zhang J., Samson L. D., Lodish H. F., Griffiths L. G. In vitro erythropoiesis from bone marrow-derived progenitors provides a physiological assay for toxic and mutagenic compounds. Proc Natl Acad Sci U S A 2007; 104 (21): 8737–42. Doi: 10.1073/pnas.0701829104. |
[33] | Xu Liming, Li Xuefei, Takemura Taro, Hanagata Nobutaka, Wu Gang, Chou Laisheng L. Genotoxicity and molecular response of silver nanoparticle (NP)-based hydrogel. J Nanobiotechnology 2012; 10: 1–11. Doi: 10.1186/1477-3155-10-16. |
[34] | Zhou Furong, Liao Fen, Chen Lingying, Liu Yuanfeng, Wang Wuxiang, Feng Shaolong. The size-dependent genotoxicity and oxidative stress of silica nanoparticles on endothelial cells. Environ Sci Pollut Res 2019; 26 (2): 1911–20. Doi: 10.1007/s11356-018-3695-2. |
[35] | Arancibia R., Maturana C., Silva D., et al. Effects of chitosan particles in periodontal pathogens and gingival fibroblasts. J Dent Res 2013; 92 (8): 740–5. Doi: 10.1177/0022034513494816. |
[36] | Aliasghari Azam, Khorasgani Mohammad Rabbani, Vaezifar Sedigheh, Rahimi Fateh, Younesi Habibollah, Khoroushi Maryam. Evaluation of antibacterial efficiency of chitosan and chitosan nanoparticles on cariogenic streptococci: An in vitro study. Iran J Microbiol 2016; 8 (2): 93–100. Doi: 10.5281/zenodo.3342597. |
[37] | Covarrubias Cristian, Trepiana Diego, Corral Camila. Synthesis of hybrid copper-chitosan nanoparticles with antibacterial activity against cariogenic Streptococcus mutans. Dent Mater J 2018; 37 (3): 379–84. Doi: 10.4012/dmj.2017-195. |
[38] | Chen Chueh Pin, Chen Chin Tin, Tsai Tsuimin. Chitosan nanoparticles for antimicrobial photodynamic inactivation: Characterization and in vitro investigation. Photochem Photobiol 2012; 88 (3): 570–6. Doi: 10.1111/j.1751-1097.2012.01101.x. |
APA Style
Rami Alhomrany, Chang Zhang, Laisheng Chou. (2021). Genotoxicity Induced by Cellular Uptake of Chitosan Nanoparticles in Human Dental Pulp Cells. International Journal of Materials Science and Applications, 10(4), 79-86. https://doi.org/10.11648/j.ijmsa.20211004.11
ACS Style
Rami Alhomrany; Chang Zhang; Laisheng Chou. Genotoxicity Induced by Cellular Uptake of Chitosan Nanoparticles in Human Dental Pulp Cells. Int. J. Mater. Sci. Appl. 2021, 10(4), 79-86. doi: 10.11648/j.ijmsa.20211004.11
AMA Style
Rami Alhomrany, Chang Zhang, Laisheng Chou. Genotoxicity Induced by Cellular Uptake of Chitosan Nanoparticles in Human Dental Pulp Cells. Int J Mater Sci Appl. 2021;10(4):79-86. doi: 10.11648/j.ijmsa.20211004.11
@article{10.11648/j.ijmsa.20211004.11, author = {Rami Alhomrany and Chang Zhang and Laisheng Chou}, title = {Genotoxicity Induced by Cellular Uptake of Chitosan Nanoparticles in Human Dental Pulp Cells}, journal = {International Journal of Materials Science and Applications}, volume = {10}, number = {4}, pages = {79-86}, doi = {10.11648/j.ijmsa.20211004.11}, url = {https://doi.org/10.11648/j.ijmsa.20211004.11}, eprint = {https://article.sciencepublishinggroup.com/pdf/10.11648.j.ijmsa.20211004.11}, abstract = {Objective: Recent in vitro studies have shown that chitosan nanoparticles in several root canal sealers, intracanal medicament, and irrigation solutions could enhance the antimicrobial activity. However, the nanotoxicity of chitosan has not been fully studied. The aim of this study was to evaluate cellular uptake and genotoxicity of various sizes and concentrations of chitosan nanoparticles cultured with human dental pulp cells. Methods: Human dental pulp cells were derived from human dental pulp tissues and cultured for 24 hours with 50 nm and 318 nm FITC-tagged chitosan nanoparticles in concentrations: 0.1 mg/mL, 0.5 mg/mL, and 2 mg/mL as study groups, and 0 mg/mL as a control. The fluorescence intensity of the FITC tagged chitosan nanoparticles was measured using a spectrophotometer to determine the cellular uptake. Genotoxicity was assessed by the Cytokinesis-block micronucleus method and by measuring the fluorescent intensity of the phosphorylated H2AX nuclear foci. Statistical analysis was performed using One-Way ANOVA, post-hoc Tukey, and Chi-square tests. Results: Chitosan nanoparticles were able to internalize the human dental pulp cells and significantly induced micronuclei, nuclear buds, and pH2AX foci at concentrations of 0.5 mg/mL and 2 mg/mL as compared to 0.1 mg/mL (P P P=0.001), nuclear buds (P=0.009), and pH2AX nuclear foci (P=0.00004) as compared to 318 nm chitosan. Conclusion: 50 nm and 318 nm chitosan nanoparticles at concentrations 0.5 mg/mL and 2 mg/mL penetrated human dental pulp cells and induced genotoxicity in dose-dependent and size-associated manners.}, year = {2021} }
TY - JOUR T1 - Genotoxicity Induced by Cellular Uptake of Chitosan Nanoparticles in Human Dental Pulp Cells AU - Rami Alhomrany AU - Chang Zhang AU - Laisheng Chou Y1 - 2021/07/15 PY - 2021 N1 - https://doi.org/10.11648/j.ijmsa.20211004.11 DO - 10.11648/j.ijmsa.20211004.11 T2 - International Journal of Materials Science and Applications JF - International Journal of Materials Science and Applications JO - International Journal of Materials Science and Applications SP - 79 EP - 86 PB - Science Publishing Group SN - 2327-2643 UR - https://doi.org/10.11648/j.ijmsa.20211004.11 AB - Objective: Recent in vitro studies have shown that chitosan nanoparticles in several root canal sealers, intracanal medicament, and irrigation solutions could enhance the antimicrobial activity. However, the nanotoxicity of chitosan has not been fully studied. The aim of this study was to evaluate cellular uptake and genotoxicity of various sizes and concentrations of chitosan nanoparticles cultured with human dental pulp cells. Methods: Human dental pulp cells were derived from human dental pulp tissues and cultured for 24 hours with 50 nm and 318 nm FITC-tagged chitosan nanoparticles in concentrations: 0.1 mg/mL, 0.5 mg/mL, and 2 mg/mL as study groups, and 0 mg/mL as a control. The fluorescence intensity of the FITC tagged chitosan nanoparticles was measured using a spectrophotometer to determine the cellular uptake. Genotoxicity was assessed by the Cytokinesis-block micronucleus method and by measuring the fluorescent intensity of the phosphorylated H2AX nuclear foci. Statistical analysis was performed using One-Way ANOVA, post-hoc Tukey, and Chi-square tests. Results: Chitosan nanoparticles were able to internalize the human dental pulp cells and significantly induced micronuclei, nuclear buds, and pH2AX foci at concentrations of 0.5 mg/mL and 2 mg/mL as compared to 0.1 mg/mL (P P P=0.001), nuclear buds (P=0.009), and pH2AX nuclear foci (P=0.00004) as compared to 318 nm chitosan. Conclusion: 50 nm and 318 nm chitosan nanoparticles at concentrations 0.5 mg/mL and 2 mg/mL penetrated human dental pulp cells and induced genotoxicity in dose-dependent and size-associated manners. VL - 10 IS - 4 ER -