Chemical analysis of in vivo–irradiated dentine of head and neck cancer patients by ATR-FTIR and Raman spectroscopy

  • Rafael Resende de Miranda
  • Anielle Christine Almeida Silva
  • Noelio Oliveira Dantas
  • Carlos José Soares
  • Veridiana Resende NovaisEmail author
Original Article



To evaluate the effect of in vivo radiotherapy on the chemical properties of human dentine by Fourier-transform infrared spectroscopy (FTIR) and Raman analysis.

Materials and methods

Chemical composition was evaluated comparing control and irradiated group (n = 8). Irradiated teeth were obtained from radiotherapy patients subjected to fractionated X-ray radiation of 1.8 Gy daily totaling 72 Gy. The teeth were sectioned according to the type of dentine (crown or root dentine), obtaining 3-mm dentine cervical slices. The analyzed parameters by FTIR and Raman spectroscopies were mineral/matrix ratio (M:M), carbonate/mineral ratio (C:M), amide I/amide III ratio, and amide I/CH2 ratio. Raman also calculated the phosphate and carbonate crystallinity.


FTIR revealed that M:M had a decrease in both factors (p = 0.008; p = 0.043, respectively) and root dentine showed a lower C:M in the irradiated group (p = 0.003). Raman revealed a higher phosphate crystallinity and a lower carbonate crystallinity in crown dentine of irradiated group (p = 0.021; p = 0.039). For amide I/amide III, the irradiated showed a lower ratio when compared to the control group (FTIR p = 0.002; Raman p = 0.017). For amide I/CH2, the root dentine showed a higher ratio than the crown dentine in both methods (p < 0.001).


Radiotherapy altered the chemical composition of human dentine. The exchange of phosphate-carbonate ions in the hydroxyapatite and higher concentration of organic components was found after radiotherapy.

Clinical relevance

The increased risk of radiation-related caries in patients undergoing head and neck radiotherapy is due not only to salivary, dietary, and microbiological changes but also to changes in tooth chemical composition.


Dentine Fourier-transform infrared spectroscopy Head and neck cancer Radiotherapy Raman spectroscopy 



The authors thank FAPEMIG, CNPq and CAPES (Brazil) for the financial support. The authors are also grateful to the Laboratory of New Insulating Materials and Semiconductors (LNMIS) at Institute of Physics, Federal University of Uberlândia, Minas Gerais, Brazil.


The study was supported by FAPEMIG, CNPq and CAPES for the granting of scholarship and resource for the purchase of consumer material.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.

Informed consent

Informed consent was obtained from all individual participants included in the study. Additional informed consent was obtained from all individual participants for whom identifying information is included in this article.


  1. 1.
    Ferlay J, Soerjomataram I, Dikshit R, Eser S, Mathers C, Rebelo M, Parkin DM, Forman D, Bray F (2015) Cancer incidence and mortality worldwide: sources, methods and major patterns in GLOBOCAN 2012. Int J Cancer 136(5):E359–E386CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Tolentino ES, Centurion BS, Ferreira LHC, Souza AP, Damante JH, Rubira-Bullen IRF (2011) Oral adverse effects of head and neck radiotherapy: literature review and suggestion of a clinical oral care guideline for irradiated patients. J Appl Oral Sci 19(5):448–454CrossRefPubMedCentralGoogle Scholar
  3. 3.
    Reed R, Xu C, Liu Y, Gorski JP, Wang Y, Walker MP (2015) Radiotherapy effect on nano-mechanical properties and chemical composition of enamel and dentine. Arch Oral Biol 60(5):690–697CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    Lieshout HFJ, Bots CP (2014) The effect of radiotherapy on dental hard tissue – a systematic review. Clin Oral Invest 18(1):17–24CrossRefGoogle Scholar
  5. 5.
    Gonçalves LM, Palma-Dibb RG, Paula-Silva FW, Oliveira HF, Nelson-Filho P, Silva LA, Queiroz AM (2014) Radiation therapy alters microhardness and microstructure of enamel and dentin of permanent human teeth. J Dent 42(8):986–992CrossRefPubMedGoogle Scholar
  6. 6.
    de Siqueira Mellara T, Palma-Dibb RG, de Oliveira HF, Garcia Paula-Silva FW, Nelson-Filho P, da Silva RA, da Silva LA, de Queiroz AM (2014) The effect of radiation therapy on the mechanical and morphological properties of the enamel and dentin of deciduous teeth – an in vitro study. Radiat Oncol 9:30. CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Qing P, Huang S, Gao S, Qian L, Yu H (2016) Effect of gamma irradiation on the wear behavior of human tooth dentin. Clin Oral Invest 20(9):2379–2386CrossRefGoogle Scholar
  8. 8.
    Liang X, Zhang JY, Cheng IK, Li JK (2016) Effect of high X-ray irradiation on the nano-mechanical properties of human enamel and dentine. Braz Oral Res 30:e9. CrossRefGoogle Scholar
  9. 9.
    Novais VR, Soares PB, Guimarães CM, Schliebe LR, Braga SS, Soares CJ (2016) Effect of gamma radiation and endodontic treatment on mechanical properties of human and bovine root dentin. Braz Oral Res 27(6):670–674. CrossRefGoogle Scholar
  10. 10.
    Rodrigues RB, Soares CJ, Junior PCS, Lara VC, Arana-Chavez VE, Novais VR (2018) Influence of radiotherapy on the dentin properties and bond strength. Clin Oral Investig 22(2):875–883CrossRefPubMedGoogle Scholar
  11. 11.
    Ferreira EMS, Soares LES, Antunes HE, Uemura ST, Barbosa PS, Salmon HA Jr, Sant’Anna GR (2016) Effect of therapeutic doses of radiotherapy on the organic and inorganic contents of the deciduous enamel: an in vitro study. Clin Oral Invest 20(8):1953–1961CrossRefGoogle Scholar
  12. 12.
    Fang YZ, Yang S, Wu G (2002) Free radicals, antioxidants and nutrition. Nutrition 18(10):872–879CrossRefPubMedGoogle Scholar
  13. 13.
    Pioch T, Golfels D, Staehle HJ (1992) An experimental study of the stability of irradiated teeth in the region of the dentinoenamel junction. Endod Dent Traumatol 8(6):241–244CrossRefPubMedGoogle Scholar
  14. 14.
    Walker MP, Wichman B, Cheng A, Coster J, Williams K (2011) Impact of radiotherapy dose on dentition breakdown in head and neck cancer patients. Pract Radiat Oncol 1(3):142–148CrossRefPubMedPubMedCentralGoogle Scholar
  15. 15.
    Kielbassa AM, Beetz J, Schendera A, Hellwing E (1997) Irradiation effects on microhardness of fluoridated and non-fluoridated bovine dentin. Eur J Oral Sci 105(5 Pt1):444–447CrossRefPubMedGoogle Scholar
  16. 16.
    McGuire JD, Mousa AA, Zhang BJ, Todoki LS, Huffman NT, Chandrababu KB, Moradian-Oldak J, Keightley A, Wang Y, Walker MP, Gorski JP (2014) Extracts of irradiated mature human tooth crowns contain MMP-20 protein and activity. J Dent 42(5):626–635CrossRefPubMedPubMedCentralGoogle Scholar
  17. 17.
    Liu Y, Yao X, Liu YW, Wang Y (2014) A Fourier transform infrared spectroscopy analysis of carious dentin from transparent zone to normal zone. Caries Res 48(4):320–329CrossRefPubMedPubMedCentralGoogle Scholar
  18. 18.
    Lopes CCA, Limirio PHJO, Novais VR, Dechichi P (2018) Fourier transform infrared spectroscopy (FTIR) application chemical characterization of enamel, dentin and bone. Appl Spectrosc Rev 53:747–769. CrossRefGoogle Scholar
  19. 19.
    Xu C, Wang Y (2011) Cross-linked demineralized dentin maintains its mechanical stability when challenged by bacterial collagenase. J Biomed Mater Res B Appl Biomater 96(2):242–248CrossRefPubMedGoogle Scholar
  20. 20.
    Toledano M, Aguilera FS, Osorio E, Cabello I, Toledano-Osorio M, Osorio R (2015) Functional and molecular structural analysis of dentine interfaces promoted by a Zn-doped self-etching adhesive and an in vitro load cycling model. J Mech Behav Biomed Mater 50:131–149CrossRefPubMedGoogle Scholar
  21. 21.
    Pucéat E, Reynard B, Lécuyer C (2004) Can crystallinity be used to determine the degree of chemical alteration of biogenic apatites? Chem Geol 205(1–2):83–97CrossRefGoogle Scholar
  22. 22.
    Salehi H, Terrer E, Panayotov I, Levallois B, Jacquot B, Tassery H, Cuisinier F (2013) Functional mapping of human sound and carious enamel and dentin with Raman spectroscopy. J Biophotonics 6(10):765–774PubMedGoogle Scholar
  23. 23.
    Seredin P, Goloshchapov D, Prutskij T, Ippolitov Y (2015) Phase transformations in a human tooth tissue at the initial stage of caries. PLoS One 10(4):e0124008. CrossRefPubMedPubMedCentralGoogle Scholar
  24. 24.
    Zieba-Palus J, Kunicki M (2006) Application of the micro-FTIR spectroscopy, Raman spectroscopy and XRF method examination of inks. Forensic Sci Int 158(2–3):164–172CrossRefPubMedGoogle Scholar
  25. 25.
    Cui Y, Fung KH, Xu J, Ma H, Jin Y, He S, Fang NX (2012) Ultrabroadband light absorption by a sawtooth anisotropic metamaterial slab. Nano Lett 12(3):1443–1447CrossRefPubMedGoogle Scholar
  26. 26.
    Andrew Chan KL, Kazarian SG (2016) Attenuated total reflection Fourier-transform infrared (ATR-FTIR) imaging of tissues and live cells. Chem Soc Rev 45(7):1850–1864CrossRefPubMedGoogle Scholar
  27. 27.
    Duke ES, Lindemuth J (1991) Variability of clinical dentin substrates. Am J Dent 4(5):241–246PubMedGoogle Scholar
  28. 28.
    da Cunha SRB, Fonseca FP, Ramos PAMM, Haddad CMK, Fregnani ER, Aranha ACC (2017) Effects of different radiation doses on the microhardness, superficial morphology, and mineral components of human enamel. Arch Oral Biol 80:130–135CrossRefGoogle Scholar
  29. 29.
    Jiang T, Ma X, Wang Y, Zhu Z, Tong H, Hu J (2007) Effects of hydrogen peroxide on human dentin structure. J Dent Res 86(11):1040–1045CrossRefPubMedGoogle Scholar
  30. 30.
    Hannig M, Dounis E, Henning T, Apitz N, Stösser L (2006) Does irradiation affect the protein composition of saliva? Clin Oral Investig 10(1):61–65CrossRefPubMedGoogle Scholar
  31. 31.
    Shellis RP, Featherstone JDB, Lussi A (2014) Understanding the chemistry of dental erosion. Monogr Oral Sci 25:163–179CrossRefPubMedGoogle Scholar
  32. 32.
    Leventouri T, Antonakos A, Kyriacou A, Venturelli R, Liarokapis E, Perdikatsis V (2009) Crystal structure studies of human dental apatite as a function of age. Int J Biomater 2009:1–6. CrossRefGoogle Scholar
  33. 33.
    Rey C, Collins B, Goehl T, Dickson IR, Glimcher MJ (1989) The carbonate environment in bone mineral: a resolution-enhanced Fourier transform infrared spectroscopy study. Calcif Tissue Int 45(3):157–164CrossRefPubMedGoogle Scholar
  34. 34.
    Liu Y, Hsu CY (2007) Laser-induced compositional changes on enamel: a FT-Raman study. J Dent 35(3):226–230CrossRefPubMedGoogle Scholar
  35. 35.
    Soares CJ, Castro CG, Neiva NA, Soares PV, Santos-Filho PCF, Naves LZ, Pereira PNR (2010) Effect of gamma irradiation on ultimate tensile strength of enamel and dentin. J Dent Res 89(2):159–164CrossRefPubMedGoogle Scholar
  36. 36.
    Xu C, Wang Y (2012) Chemical composition and structure of peritubular and intertubular human dentine revisited. Arch Oral Biol 57(4):383–391CrossRefPubMedGoogle Scholar
  37. 37.
    Yamauchi M, Sricholpech M (2012) Lysine post-translational modifications of collagen. Essays Biochem 52:113–133CrossRefPubMedPubMedCentralGoogle Scholar
  38. 38.
    Cheung DT, Perelman N, Tong D, Nimni ME (1990) The effect of gamma irradiation on collagen molecules, isolated alpha-chains and crosslinked native fibers. J Biomed Mater Res 24(5):581–589CrossRefGoogle Scholar
  39. 39.
    Naves LZ, Novais VR, Armstrong SR, Correr-Sobrinho L, Soares CJ (2012) Effect of gamma radiation on bonding to human enamel and dentin. Support Care Cancer 20(11):2873–2878CrossRefPubMedGoogle Scholar
  40. 40.
    Yadav S, Yadav H (2013) Ionizing irradiation affects the microtensile resin dentin bond strength under simulated clinical conditions. J Conserv Dent 16(2):148–151CrossRefPubMedPubMedCentralGoogle Scholar
  41. 41.
    Springer IN, Niehoff P, Warnke PH, Böcek G, Kovács G, Suhr M, Wiltfang J, Açil Y (2005) Radiation caries - radiogenic destruction of dental collagen. Oral Oncol 41(7):723–728CrossRefPubMedGoogle Scholar
  42. 42.
    da Cunha SR, Ramos PA, Haddad CM, da Silva JL, Fregnani ER, Aranha ACC (2016) Effects of different radiation doses on the bond strengths of two different adhesive systems to enamel and dentin. J Adhes Dent 18(2):151–156PubMedGoogle Scholar
  43. 43.
    Marshall GW Jr, Marshall SJ, Kinney JH, Balooch M (1997) The dentin substrate: structure and properties related to bonding. J Dent 25(6):441–458CrossRefPubMedGoogle Scholar
  44. 44.
    Lussi A, Schlueter N, Rakhmatullina E, Ganss C (2011) Dental erosion – an overview with emphasis on chemical and histopathological aspects. Caries Res 45(suppl 1):2–12CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of Operative Dentistry and Dental Materials, School of DentistryFederal University of UberlândiaUberlândiaBrazil
  2. 2.Laboratory of New Nanostructured and Functional Materials, Institute of PhysicsFederal University of AlagoasMaceióBrazil

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