N-methyl-N-nitrosourea-induced changes in epithelial rests of Malassez and the development of odontomas in rats

Kimura,Ayako; Yoshizawa,Katsuhiko; Sasaki,Tomo; Uehara,Norihisa; Kinoshita,Yuichi; Miki,Hisanori; Yuri,Takashi; Uchida,Takashi; Tsubura,Airo

doi:10.3892/etm.2012.559

N-methyl-N-nitrosourea-induced changes in epithelial rests of Malassez and the development of odontomas in rats

Authors:
- Ayako Kimura
- Katsuhiko Yoshizawa
- Tomo Sasaki
- Norihisa Uehara
- Yuichi Kinoshita
- Hisanori Miki
- Takashi Yuri
- Takashi Uchida
- Airo Tsubura
View Affiliations

Affiliations: Department of Pathology II, Kansai Medical University, Morguchi, Osaka 570-8506, Japan, Department of Oral Biology, Hiroshima University Graduate School of Biomedical Science, Hiroshima 734-8553, Japan
Published online on: April 23, 2012 https://doi.org/10.3892/etm.2012.559
Pages: 15-20

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Morphological changes in the epithelial rests of Malassez (ERM) and the development of odontogenic tumors in the molars of female Lewis rats treated at 4 weeks of age with a single intraperitoneal injection of 50 mg/kg of N-methyl-N-nitrosourea (MNU) were examined at 12, 18 and 30 weeks of age. Following MNU exposure, the total number and average area of ERM in the cervical and furcational regions of the first, second and third molars of the mandible and maxilla were compared with age-matched control animals. The number of ERM at each time point was significantly greater in the MNU-treated group compared to the control group, but there was no time-dependent increase in the number of ERM in either group. The area of ERM was significantly larger in the MNU-treated group compared to the control group at each time point, and it increased in a time-dependent manner in the MNU-treated group. No increases in the number or area of ERM were observed in the control group. At 30 weeks of age, 23% of the MNU-treated rats had developed odontomas (complex type) in the molar region as well as in the incisor region. Immunohistochemically, the expression of tyrosine receptor kinase A (TrkA) and cytokeratin 14 (CK14) decreased, whereas p63 expression remained high during ERM enlargement. In tumors, ameloblast-like cells were positive for amelogenin, TrkA and CK14 but negative for p63, whereas odontoblast-like cells were negative for all antigens examined. In conclusion, a single intraperitoneal injection of MNU caused the development of odontomas in the molar region; these tumors were possibly derived from ERM.

View Figures

View References

1	Brown HR and Hardisty JF: Oral cavity, esophagus, and stomach. Pathology of the Fischer Rat: Reference and Atlas. Boorman GA, Montgomery CA Jr and MacKenzie WF: Academic Press; San Diego: pp. 9–30. 1990
2	Long PH and Leininger JR: Teeth. Pathology of the Mouse: Reference and Atlas. Maronpot RR, Boorman GA and Gaul BW: Cache River Press; Vienna: pp. 13–28. 1999
3	Abe T and Miyajima H: Effect of drugs on developing enamel in rat incisors. J Toxicol Pathol. 3:245–256. 1990.(Abstract in English, text in Japanese).
4	Kuijpers MH, van de Kooij AJ and Slootweg PJ: The rat incisor in toxicologic pathology. Toxicol Pathol. 24:346–360. 1996. View Article : Google Scholar : PubMed/NCBI
5	Weber K: Induced and spontaneous lesions in teeth of laboratory animals. J Toxicol Pathol. 20:203–213. 2007. View Article : Google Scholar
6	Kumamoto H: Molecular pathology of odontogenic tumors. J Oral Pathol Med. 35:65–74. 2006. View Article : Google Scholar : PubMed/NCBI
7	Takeda Y: Histogenesis and clinical pathology of odontogenic tumors. Dent J Iwate Med Univ. 31:143–152. 2006.(In Japanese).
8	Edwards MB: Some effects of N-methylnitrosourea on the oral and odontogenic tissues of the Syrian golden hamster. Arch Oral Biol. 23:515–524. 1978. View Article : Google Scholar : PubMed/NCBI
9	Maeda H, Kameyama Y, Fujita K, Sato E and Mizutani M: Experimental odontogenic tumors produced by ethylnitrosourea injections and mechanical injuries. J Oral Pathol Med. 20:296–299. 1991. View Article : Google Scholar : PubMed/NCBI
10	Satoh H, Usegi Y, Kawabata T, Mori K, Fujii F and Kashimoto Y: Morphological classification of dental lesions induced by various antitumor drugs in mice. Toxicol Pathol. 29:292–299. 2001. View Article : Google Scholar : PubMed/NCBI
11	Azuma T and Komori A: An experimental study of odontogenic tumors in rats. Induction of tumors by gastric intubation of MNU. Jpn J Oral Biol. 27:631–639. 1985. View Article : Google Scholar
12	Berman JJ and Rice JM: Odontogenic tumours produced in Fischer rats by a single intraportal injection of methylnitrosourea. Arch Oral Biol. 25:213–220. 1980. View Article : Google Scholar : PubMed/NCBI
13	Eblin H, Barbachan JJD, Do Valle JGC and De Oliveira LY: N-Methyl-N-nitrosourea-induced odontogenic neoplasms in rats. J Dent Res. 52:1771973. View Article : Google Scholar : PubMed/NCBI
14	Eisenberg E, Murthy AS, Vawter GF and Krutchkoff DJ: Odontogenic neoplasms in Wistar rats treated with N-methylnitrosourea. Oral Surg Oral Med Oral Pathol. 55:481–486. 1983. View Article : Google Scholar : PubMed/NCBI
15	Gardner DG: Experimentally induced ameloblastomas: a critical review. J Oral Pathol Med. 21:337–339. 1992. View Article : Google Scholar : PubMed/NCBI
16	Leaver DD, Swann PF and Magee PN: The induction of tumors in the rat by a single oral dose of N-nitrosomethylurea. Br J Cancer. 223:177–187. 1969. View Article : Google Scholar
17	Smulow J B, Konstantinidis A and Sonnenschein C: Age-dependent odontogenic lesions in rats after a single i.p. injection of N-nitroso-N-methylurea. Carcinogenesis. 4:1085–1088. 1983.PubMed/NCBI
18	Herrold KM: Odontogenic tumors and epidermoid carcinomas of the oral cavity. An experimental study in Syrian hamsters. Oral Surg Oral Med Oral Pathol. 25:262–272. 1968. View Article : Google Scholar : PubMed/NCBI
19	Kohgo T: An experimental study of odontogenic tumors in hamsters by N-nitrosomethylurea. J Stomatol Soc Jpn. 39:191–212. 1972. View Article : Google Scholar : PubMed/NCBI
20	Kohgo T, Iizuka T and Shindoh M: Pathological evaluation of the effects of intentional disocclusion and overloading occlusion in odontogenesis disorders in N-methylnitrosourea-treated hamsters. Toxicol Pathol. 27:226–232. 1999. View Article : Google Scholar : PubMed/NCBI
21	Wentz FM, Weinmann JP and Schour I: The prevalence, distribution, and morphologic changes of the epithelial remnants in the molar region of the rat. J Dent Res. 29:637–646. 1950. View Article : Google Scholar : PubMed/NCBI
22	Bykov VL: Epithelial cell rests of Malassez: tissue, cell, and molecular biology. Morfologiia. 124:95–103. 2003.(In Russian).
23	Kat PS, Sampson WJ, Wilson DF and Wiebkin OW: Distribution of the epithelial rests of Malassez and their relationship to blood vessels of the periodontal ligament during rat tooth development. Aust Orthod J. 19:77–86. 2003.PubMed/NCBI
24	Huang X, Bringas P, Slavkin HC and Chai Y: Fate of HERS during tooth root development. Dev Biol. 334:22–30. 2009. View Article : Google Scholar : PubMed/NCBI
25	Shinmura Y, Tsuchiya S, Hata K and Honda MJ: Quiescent epithelial cell rests of Malassez can differentiate into ameloblast-like cells. J Cell Physiol. 217:728–738. 2008. View Article : Google Scholar : PubMed/NCBI
26	Hamamoto Y, Hamamoto N, Nakajima T and Ozawa H: Morphological changes of epithelial rests of Malassez in rat molars induced by local administration of N-methylnitrosourea. Arch Oral Biol. 43:899–906. 1998. View Article : Google Scholar : PubMed/NCBI
27	Rincon JC, Young WG and Bartold PM: The epithelial cell rests of Malassez - a role in periodontal regeneration? J Periodontal Res. 41:245–252. 2006. View Article : Google Scholar : PubMed/NCBI
28	Yagoto M, Shimojyo Y, Uotani Y, et al: Methodology of histological preparation of rat teeth tissue including incisor and molar. Jpn J Histotech. 9:29–32. 2000.(In Japanese).
29	Long PH, Leininger JR and Lieuallen WG: Non-proliferative lesions of bone, cartilage, tooth, and synovium in rats. Guides for Toxicologic Pathology STP/ARP/AFIP Washington, D.C.: 1996
30	Long PH, Leininger JR, Nold JB and Lieuallen WG: Proliferative lesions of bone, cartilage, tooth, and synovium in rats. Guides for Toxicologic Pathology STP/ARP/AFIP Washington, D.C.: 1996
31	Kurihara H, Shinohara H, Yoshino H, Takeda K and Shiba H: Neurotrophins in cultured cells from periodontal tissues. J Periodontol. 74:76–84. 2003. View Article : Google Scholar : PubMed/NCBI
32	Woodnutt DA and Byers MR: Morphological variation in the tyrosine receptor kinase A- immunoreactive periodontal ligament. Arch Oral Biol. 46:163–171. 2001. View Article : Google Scholar : PubMed/NCBI
33	Brikic A, Mutlu S, Kocak-Berberoglu H and Olgac V: Pathological changes and immunoexpression of p63 gene in dental follicles of asymptomatic impacted lower third molars: an immunohistochemical study. J Craniofac Surg. 21:854–857. 2010. View Article : Google Scholar : PubMed/NCBI
34	Casasco M, Cornaglia AI, Riva F, Calligaro A and Casasco A: Expression of p63 transcription factor in ectoderm-derived oral tissues. Int J Anat Embryol. 111:125–131. 2006.PubMed/NCBI
35	Kock M, Nolting D, Kjaer KW, Hansen BF and Kjaer I: Immunohistochemical expression of p63 in human prenatal tooth primordia. Acta Odontol Scand. 63:253–257. 2005. View Article : Google Scholar : PubMed/NCBI
36	Nam H, Kim J, Park J, et al: Expression profile of the stem cell markers in human Hertwig’s epithelial root sheath/epithelial rests of Malassez cells. Mol Cells. 31:355–360. 2011.PubMed/NCBI
37	Rufini A, Weil M, McKeon F, Barlattani A, Melino G and Candi E: p63 protein is essential for the embryonic development of vibrissae and teeth. Biochem Biophys Res Commun. 340:737–741. 2006. View Article : Google Scholar : PubMed/NCBI
38	Xiong J, Gronthos S, Krzysztof M and Bartold PM: Characterisation and stem-cell properties of epithelial cell rests of Malassez. In: Abstract in IADR/AADR/CADR 89th General Session and Exhibition; San Diego, USA. 2011
39	Uchida T, Tanabe T and Fukae M: Immunocytochemical localization of amelogenins in the deciduous tooth germs of the human fetus. Arch Histol Cytol. 52:543–552. 1989. View Article : Google Scholar : PubMed/NCBI
40	Uchida T, Tanabe T, Fukae M, et al: Immunochemical and immunohistochemical studies, using antisera against porcine 25 kDa amelogenin, 89 kDa enamelin and the 13–17 kDa nonamelogenins, on immature enamel of the pig and rat. Histochemistry. 96:129–138. 1991.PubMed/NCBI
41	Hasegawa N, Kawaguchi H, Ogawa T, Uchida T and Kurihara H: Immunohistochemical characteristics of epithelial cell rests of Malassez during cementum repair. J Periodont Res. 38:51–56. 2003. View Article : Google Scholar : PubMed/NCBI
42	Nishio C, Wazen R, Kuroda S, Moffatt P and Nanci A: Disruption of periodontal integrity induces expression of apin by epithelial cell rests of Malassez. J Periodont Res. 45:709–713. 2010. View Article : Google Scholar : PubMed/NCBI
43	Lewis DJ, Cherry CP and Gibson WA: Ameloblastoma (adamantinoma) of the mandible in the rat. J Comp Pathol. 90:379–384. 1980. View Article : Google Scholar : PubMed/NCBI
44	Wright JT, Hansen L, Mahler J, Szczesniak C and Spalding JW: Odontogenic tumors in the v-Ha-ras (TGAC) transgenic mouse. Archs Oral Biol. 40:631–638. 1995. View Article : Google Scholar : PubMed/NCBI
45	Grachtchouk M, Liu J, Wang A, et al: Odontogenic keratocysts arise from quiescent epithelial rests and are associated with deregulated hedgehog signaling in mice and humans. Am J Pathol. 169:806–814. 2006. View Article : Google Scholar : PubMed/NCBI
46	Gao Y, Yang G, Weng T, et al: Disruption of Smad4 in odontoblasts causes multiple keratocytic odontogenic tumors and tooth malformation in mice. Mol Cell Biol. 29:5941–5951. 2009. View Article : Google Scholar : PubMed/NCBI
47	Yamashiro T, Fujiyama K, Fukunaga T, Wang Y and Takano-Yamamoto T: Epithelial rests of Malassez express immunoreactivity of TrkA and its distribution is regulated by sensory nerve innervation. J Histochem Cytochem. 48:979–984. 2000. View Article : Google Scholar : PubMed/NCBI
48	Lo Muzio L, Santarelli A, Caltabiano R, et al: p63 expression correlates with pathological features and biological behaviour of odontogenic tumours. Histopathology. 49:211–214. 2006.PubMed/NCBI
49	Hamamoto Y, Nakajima T, Ozawa H and Uchida T: Production of amelogenin by enamel epithelium of Hertwig’s root sheath. Oral Surg Oral Med Oral Pathol Oral Radiol Endod. 81:703–709. 1996.PubMed/NCBI
50	Kumamoto H, Yoshida M and Ooya K: Immunohistochemical detection of amelogenin and cytokeratin 19 in epithelial odontogenic tumors. Oral Dis. 7:171–176. 2001. View Article : Google Scholar : PubMed/NCBI
51	Ravindranath RM, Tam WY, Bringas P Jr, Santos V and Fincham AG: Amelogenin-cytokeratin 14 interaction in ameloblasts during enamel formation. J Biol Chem. 276:36586–36597. 2001. View Article : Google Scholar : PubMed/NCBI
52	Crivelini MM, de Araújo VC, de Sousa SO and de Araújo NS: Cytokeratins in epithelia of odontogenic neoplasms. Oral Dis. 9:1–6. 2003. View Article : Google Scholar : PubMed/NCBI
53	Rufini A, Barlattani A, Docimo R, et al: p63 in tooth development. Biochem Pharmacol. 82:1256–1261. 2011. View Article : Google Scholar
54	Kumamoto H, Ohki K and Ooya K: Expression of p63 and p73 in ameloblastomas. J Oral Pathol Med. 34:220–226. 2005. View Article : Google Scholar : PubMed/NCBI