Genes expressed during amelogenesis are candidates to increase the risk of dental fluorosis (DF). Thus, this study aimed to evaluate the association between polymorphisms in enamel development genes and susceptibility to DF in mice. Mice of both sexes, representing strains 129P3/J (n = 20; resistant to DF) and A/J (n = 20; susceptible to DF), were divided into 2 groups. Each strain received a diet with a low concentration of fluoride (F) and drinking water containing 0 or 50 mg/L of F for 6 weeks. Clinical evaluation and analysis of Vickers enamel microhardness of the incisors were performed. Livers were collected for genomic DNA extraction. Seventeen genetic polymorphisms in Amelx, Ambn, Ambn, Col14a1, Col1a1, Col5a2, Enam, Fam20a, Fam83h, Foxo1, Klk4, Mmp20, Serpinf1, Serpinh1, Smad3, Tuft1, and Wdr72 were genotyped by real-time PCR using Taqman chemistry. Overrepresentation of alleles and genotypes in DF was evaluated using the χ2 test with an alpha of 5%. The clinical aspects of the enamel and the surface enamel microhardness confirmed the DF condition. In the polymorphisms rs29569969, rs13482592, and rs13480057 in Ambn, Col14a1, and Mmp20, respectively, genotype and allele distributions were statistically significantly different between A/J and 129P3/J strains (p < 0.05). In conclusion, polymorphisms in Ambn, Col14a1, and Mmp20 are associated with the susceptibility to DF.

Keywords:
Polymorphisms, Gene, Dental fluorosis
1.
Aoba T, Fejerskov O: Dental fluorosis: chemistry and biology. Crit Rev Oral Biol Med 2002; 13:155–170.
2.
Bartlett JD, Skobe Z, Nanci A, Smith CE: Matrix metalloproteinase 20 promotes a smooth enamel surface, a strong dentino-enamel junction, and a decussating enamel rod pattern. Eur J Oral Sci 2011; 119(suppl 1): 199–205.
3.
Bartlett JD, Smith CE: Modulation of cell-cell junctional complexes by matrix metalloproteinases. J Dent Res 2013; 92: 10–17.
4.
Bhussry BR: Chemical and physical studies of enamel from human teeth. J Dent Res 1959; 38: 369–373.
5.
Bronckers AL, Lyaruu DM, DenBesten PK: The impact of fluoride on ameloblasts and the mechanisms of enamel fluorosis. J Dent Res 2009; 88: 877–893.
6.
Buzalaf MA, Levy SM: Fluoride intake of children: considerations for dental caries and dental fluorosis. Monogr Oral Sci 2011; 22: 1–19.
7.
Buzalaf MA, Whitford GM: Fluoride metabolism. Monogr Oral Sci 2011; 22: 20–36.
8.
Carvalho JG, Leite Ade L, Peres-Buzalaf C, Salvato F, Labate CA, Everett ET, Whitford GM, Buzalaf MA: Renal proteome in mice with different susceptibilities to fluorosis. PLoS One 2013; 8:e53261.
9.
Carvalho JG, Leite AL, Yan D, Everett ET, Whitford GM, Buzalaf MA: Influence of genetic background on fluoride metabolism in mice. J Dent Res 2009; 88: 1054–1058.
10.
Chan HC, Estrella NM, Milkovich RN, Kim JW, Simmer JP, Hu JC: Target gene analyses of 39 amelogenesis imperfecta kindreds. Eur J Oral Sci 2011; 119(suppl 1): 311–323.
11.
Charone S, De Lima Leite A, Peres-Buzalaf C, Silva Fernandes M, Ferreira de Almeida L, Zardin Graeff MS, Cardoso de Oliveira R, Campanelli AP, Groisman S, Whitford GM, Everett ET, Buzalaf MA: Proteomics of secretory-stage and maturation-stage enamel of genetically distinct mice. Caries Res 2016; 50: 24–31.
12.
Chun YH, Lu Y, Hu Y, Krebsbach PH, Yamada Y, Hu JC, Simmer JP: Transgenic rescue of enamel phenotype in Ambn null mice. J Dent Res 2010; 89: 1414–1420.
13.
DenBesten PK, Yan Y, Featherstone JD, Hilton JF, Smith CE, Li W: Effects of fluoride on rat dental enamel matrix proteinases. Arch Oral Biol 2002; 47: 763–770.
14.
Denbesten P, Li W: Chronic fluoride toxicity: dental fluorosis. Monogr Oral Sci 2011; 22: 81–96.
15.
Dunipace AJ, Brizendine EJ, Zhang W, Wilson ME, Miller LL, Katz BP, Warrick JM, Stookey GK: Effect of aging on animal response to chronic fluoride exposure. J Dent Res 1995; 74: 358–368.
16.
Everett ET: Fluoride’s effects on the formation of teeth and bones, and the influence of genetics. J Dent Res 2011; 90: 552–560.
17.
Everett ET, McHenry MA, Reynolds N, Eggertsson H, Sullivan J, Kantmann C, Martinez-Mier EA, Warrick JM, Stookey GK: Dental fluorosis: variability among different inbred mouse strains. J Dent Res 2002; 81: 794–798.
18.
Everett ET, Yan D, Weaver M, Liu L, Foroud T, Martinez-Mier EA: Detection of dental fluorosis-associated quantitative trait loci on mouse chromosomes 2 and 11. Cells Tissues Organs 2009; 189: 212–218.
19.
Everett ET, Yin Z, Yan D, Zou F: Fine mapping of dental fluorosis quantitative trait loci in mice. Eur J Oral Sci 2011; 119(suppl 1): 8–12.
20.
Huang H, Ba Y, Cui L, Cheng X, Zhu J, Zhang Y, Yan P, Zhu C, Kilfoy B, Zhang Y: COL1A2 gene polymorphisms (Pvu II and Rsa I), serum calciotropic hormone levels, and dental fluorosis. Community Dent Oral Epidemiol 2008; 36: 517–522.
21.
Iwata T, Yamakoshi Y, Hu JC, Ishikawa I, Bartlett JD, Krebsbach PH, Simmer JP: Processing of ameloblastin by Mmp-20. J Dent Res 2007; 86: 153–157.
22.
Krebsbach PH, Lee SK, Matsuki Y, Kozak CA, Yamada KM, Yamada Y: Full-length sequence, localization, and chromosomal mapping of ameloblastin. A novel tooth-specific gene. J Biol Chem 1996; 271: 4431–4435.
23.
Küchler EC, Dea Bruzamolin C, Ayumi Omori M, Costa MC, Antunes LS, Pecharki GD, Trevilatto PC, Vieira AR, Brancher JA: Polymorphisms in nonamelogenin enamel matrix genes are associated with dental fluorosis. Caries Res 2017a; 52: 1–6.
24.
Küchler EC, Tannure PN, Oliveira DS, Charone S, Nelson-Filho P, Silva RA, Costa MC, Antunes LS, Calasans Maia MD, Antunes LA: Polymorphisms in genes involved in enamel development are associated with dental fluorosis. Arch Oral Biol 2017b; 76: 66–69.
25.
Lee KE, Lee SK, Jung SE, Song SJ, Cho SH, Lee ZH, Kim JW: A novel mutation in the AMELX gene and multiple crown resorptions. Eur J Oral Sci 2011; 119(suppl 1): 324–328.
26.
Nagano T, Kakegawa A, Yamakoshi Y, Tsuchiya S, Hu JC, Gomi K, Arai T, Bartlett JD, Simmer JP: Mmp-20 and KLK-4 cleavage site preferences for amelogenin sequences. J Dent Res 2009; 88: 823–828.
27.
Papagerakis P, Lin HK, Lee KY, Hu Y, Simmer JP, Bartlett JD, Hu JC: Premature stop codon in Mmp-20 causing amelogenesis imperfecta. J Dent Res 2008; 87: 56–59.
28.
Ryu OH, Fincham AG, Hu CC, Zhang C, Qian Q, Bartlett JD, Simmer JP: Characterization of recombinant pig enamelyzin activity and cleavage of recombinant pig and mouse amelogenins. J Dent Res 1999; 78: 743–750.
29.
Shinoda H: Effect of long term administration of fluoride on physic-chemical properties of the rat incisor enamel. Calcif Tissue Res 1975; 18: 91–100.
30.
Simmer JP, Hu Y, Richardson AS, Bartlett JD, Hu JC: Why does enamel in Klk4-null mice break above the dentino-enamel junction? Cells Tissues Organs 2011; 194: 211–215.
31.
Vieira AP, Hanocock R, Eggertsson H, Everett ET, Grynpas MD: Tooth quality in dental fluorosis genetic and environmental factors. Calcif Tissue Int 2005; 76: 17–25.
32.
Yan D, Willett TL, Gu XM, Martinez-Mier EA, Sardone L, McShane L, Grynpas M, Everett ET: Phenotypic variation of fluoride responses between inbred strains of mice. Cells Tissues Organs 2011; 194: 261–267.
© 2018 S. Karger AG, Basel
2018
Copyright / Drug Dosage / Disclaimer
Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher.
Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in government regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug.
Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.
You do not currently have access to this content.