Original Article

Cytotoxic Effect of Addition of Different Concentrations of Nanohydroxyapatite to Resin Modified and Conventional Glass Ionomer Cements on L929 Murine Fibroblasts


Objectives: In this study we assessed the cytotoxic effect of nanohydroxyapatite (NHA) incorporated into resin modified and conventional glass ionomer cements (RMGICs and CGICs) on L929 murine fibroblasts.
Materials and Methods: In this in vitro study, 0wt%, 1wt%, 2wt%, 5wt%, 7wt% and 10wt% concentrations of NHA were added to Fuji II LC RMGIC and Fuji IX CGIC powders. Eighteen samples (5×3mm) were fabricated from each type of glass ionomer, in six experimental groups (n=3): CG0, CG1, CG2, CG5, CG7, CG10, RMG0, RMG1, RMG2, RMG5, RMG7, and RMG10. Samples were incubated for 72h. The overlaying solution was removed and added to L929 fibroblasts. The methyl thiazolyl tetrazolium bromide (MTT) assay was performed at 24, 48 and 72h. The wavelength was read by a spectrophotometer. Data were analyzed by ANOVA and Tukey’s test.
Results: There was no significant difference in cytotoxicity of the two types of glass ionomers, with and without NHA, except for CG0 and RMG0 groups after 72h. RMG0 group was significantly more cytotoxic than the CG0 group (P<0.05). In CG groups during the first 24h, the cytotoxicity of CG5 and CG7 groups was significantly higher than that of CG1; while, there was no significant difference between the RMG groups. Cytotoxicity significantly decreased in all groups after 24h (P<0.05).
Conclusion: Incorporation of NHA into Fuji II LC RMGIC and Fuji IX CGIC did not affect their biocompatibility and therefore its addition to these materials can provide favorable biological properties, especially considering its beneficial effects on the other properties of GICs.

1. Sidhu SK, Nicholson JW. A review of glass-ionomer cements for clinical dentistry. J Funct. 2016 Sep;7(3):16.
2. Garoushi S, Vallittu PK, Lassila L. Characterization of fluoride releasing restorative dental materials. Dent Mater. 2018 Jan; 30:2017-161.
3. Genaro LE, Anovazzi G, Hebling J, Zuanon AC. Glass ionomer cement modified by resin with incorporation of nanohydroxyapatite: In vitro evaluation of physical-biological properties. Nanomaterials. 2020 Jul;10(7):1412.
4. Şuhani MF, Băciuţ G, Băciuţ M, Şuhani R, Bran S. Current perspectives regarding the application and incorporation of silver nanoparticles into dental biomaterials. Clujul medical. 2018 Jul;91(3):274.
5. Alatawi RA, Elsayed NH, Mohamed WS. Influence of hydroxyapatite nanoparticles on the properties of glass ionomer cement. J Mater Res Technol. 2019 Jan;8(1):344-9.
6. Munguía-Moreno S, Martínez-Castañón GA, Patiño-Marín N, Cabral-Romero C, Zavala-Alonso NV. Biocompatibility and surface characteristics of resin-modified glass ionomer cements with ammonium quaternary compounds or silver nanoparticles: An in vitro study. J Nanomater. 2018;2018.
7. Bilić-Prcić M, Rajić VB, Ivanišević A, Pilipović A, Gurgan S, Miletić I. Mechanical Properties of Glass Ionomer Cements after Incorporation of Marine Derived Porous Cuttlefish Bone Hydroxyapatite. Materials. 2020 Jan;13(16):3542.
8. Mohammadi Basir M, Ataei M, Rezvani MB, Golkar P. Effect of incorporation of various amounts of nano-sized Hydroxyapatite on the mechanical properties of a resin modified glass ionomer. J Dent Sch. 2013 Mar 15;30(4):216-23.
9. Moshaverinia A, Ansari S, Moshaverinia M, Roohpour N, Darr JA, Rehman I. Effects of incorporation of hydroxyapatite and fluoroapatite nanobioceramics into resin cements (GIC). Acta Biomater. 2008 Mar 31;4(2):432-40.
10. Poorzandpoush K, Omrani LR, Jafarnia SH, Golkar P, Atai M. Effect of addition of Nano hydroxyapatite particles on wear of resin modified glass ionomer by tooth brushing simulation. J Clin Exp Dent. 2017 Mar;9(3):e372.
11. Koohpeima F, Mokhtari MJ, Doozandeh M, Jowkar Z, Yazdanshenas F. Comparison of Cytotoxicity of New Nanohybrid Composite, Giomer, Glass Ionomer and Silver Reinforced Glass Ionomer using Human Gingival Fibroblast Cell Line. J Clin Pediatr Dent. 2017;41(5):368-73.
12. Zarrabian M, Ostad SN, Abbasi MA, Mohseni ME. In vitro evaluation of mutagenicity and cytotoxicity of four root canal sealers. J Dent Med. 2007 Jun 15;20(3):205-11.
13. Wataha JC. Predicting clinical biological responses to dental materials. Dent Mater. 2012 Jan;28(1):23-40.
14. Aydın N, Karaoğlanoğlu S, Oktay EA, Süloğlu AK. Evaluating Cytotoxic Effects of Highly Esthetic Dental Composites. Braz Dent Sci. 2020 Jan;23(1):8.
15. Motskin M, Wright DM, Muller K, Kyle N, Gard TG, Porter AE, et al. Hydroxyapatite nano and microparticles: correlation of particle properties with cytotoxicity and biostability. Biomaterials. 2009 Jul;30(19):3307-17.
16. Coelho CC, Grenho L, Gomes PS, Quadros PA, Fernandes MH. Nano-hydroxyapatite in oral care cosmetics: Characterization and cytotoxicity assessment. Sci Rep .2019 Jul;9(1):1-0.
17. Dahl JE, Frangou‐Polyzois MJ, Polyzois GL. In vitro biocompatibility of denture relining
materials. Gerodontology. 2006 Mar;23(1):17-22.
18. Naguib GH, Al-Hazmi FE, Kurakula M, Al-dharrab AA, Hosny KM, Alkhalidi HM, et al. Zein Coated zinc oxide nanoparticles: Fabrication and antimicrobial evaluation as dental aid. Pharmacology. 2018 Jan;14(8):1051-9.
19. Davidson CL. Advances in glass-ionomer cements. J Appl Oral Sci. 2006,14 (SPE):3-9.
20. Sasanaluckit P, Albustany KR, Doherty PJ, Williams DF. Biocompatibility of glass ionomer cements. Biomaterials. 1993 Oct;14(12):906-16.
21. Six N, Lasfargues JJ, Goldberg M. In vivo study of the pulp reaction to Fuji IX, a glass ionomer cement. J Dent. 2000 Aug;28(6):413-22.
22. Hii SC, Luddin N, Kannan TP, Ab Rahman I, Ghani NR. The biological evaluation of conventional and nano-hydroxyapatite-silica glass ionomer cement on dental pulp stem cells: A comparative study. Contemp Clin Dent. 2019 Apr;10(2):324.
23. Shi Z, Huang X, Cai Y, Tang R, Yang D. Size effect of hydroxyapatite nanoparticles on proliferation and apoptosis of osteoblast-like cells. Acta Biomater. 2009 Jan;5(1):338-45.
24. Xie D, Yang Y, Zhao J, Park JG, Zhang JT. A novel comonomer-free light-cured glass-ionomer cement for reduced cytotoxicity and enhanced mechanical strength Dent Mater. 2007 Aug;23(8):994-1003.
25. Noorani TY, Luddin N, Rahman IA, Masudi SM. In vitro cytotoxicity evaluation of novel nano-hydroxyapatite-silica incorporated glass ionomer cement. J Clin Diagn Res. 2017 Apr;11(4):ZC105.
26. Aoki H. Science and medical applications of hydroxyapatite. J Anal At Spectrom.1991;1991:123-34.
27. Moshaverinia A, Ansari S, Movasaghi Z, Billington RW, Darr JA, Rehman IU. Modification of conventional glass-ionomer cements with N-vinylpyrrolidone containing polyacids, nano-hydroxy and fluorapatite to improve mechanical properties. Dent Mater. 2008 Oct;24(10):1381-90.
28. Najeeb S, Khurshid Z, Zafar MS, Khan AS, Zohaib S, Martí JM, Sauro S, Matinlinna JP, Rehman IU. Modifications in glass ionomer cements: Nano-sized fillers and bioactive nanoceramics. Int J Mol Sci 2016 Jul;17(7):1134.
29. Goenka S, Balu R, Kumar TS. Effects of nanocrystalline calcium deficient hydroxyapatite incorporation in glass ionomer cements. J Mech Behav Biomed 2012 Mar;7:69-76.
30. Sharafeddin F, Shoale S, Kowkabi M. Effects of different percentages of microhydroxyapatite on microhardness of resin-modified glass-ionomer and zirconomer. J Clin Exp Dent. 2017 Jun;9(6):e805.
IssueVolume 18 QRcode
SectionOriginal Article
DOI https://doi.org/10.18502/fid.v18i17.6248
Hydroxyapatites Glass Ionomer Cements Fibroblasts Toxicity Tests

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Golkar P, Ranjbar Omrani L, Zohourinia S, Ahmadi E, Asadian F. Cytotoxic Effect of Addition of Different Concentrations of Nanohydroxyapatite to Resin Modified and Conventional Glass Ionomer Cements on L929 Murine Fibroblasts. Front Dent. 2021;18.