Abstract
Oral cancer disease is among the most common cancers in the world and are associated with mortality and morbidity. The characterization of saliva samples may help to distinguish patients with oral cancer disease from normal subjects. To characterize spectra of saliva samples from normal subjects and oral cancer patients by use of fluorescence, absorption, and 1H-NMR spectroscopy. Whole unstimulated saliva samples were collected from patients with oral cancer disease and normal subjects. The saliva samples were analyzed by absorption, fluorescence and 1H-NMR spectroscopic techniques. The characteristic spectra of saliva samples from patients with oral cancer disease and normal subjects were compared. For fluorescence spectroscopic studies, six fluorophores were found in saliva samples. Autofluorescence emission spectra and synchronous spectra of saliva were different between normal subjects and oral cancer patients. For absorption spectroscopic studies, the typical absorption spectra of saliva samples from normal subjects and oral cancer patients were also different in absorption intensity, 1st and 2nd derivative of absorption spectra values. For 1H-NMR studies, nine metabolites and four metabolites were found in saliva samples taken from normal subjects and oral cancer patients, respectively. The metabolic profiles of saliva samples from normal subjects and oral cancer patients were not similar. The characteristic spectra of saliva samples from normal subjects and oral cancer patients were found. These results showed differences in the spectra of saliva samples between both that groups. The spectra from each spectroscopic techniques could determine a candidate saliva biomarkers for distinguishing patients with oral cancer disease from normal subjects.
Similar content being viewed by others
Data Availability
Not applicable.
Code Availability
Not applicable.
References
Chaturvedi P, Majumder S, Krishna H, Muttagi S, Gupta P (2010) Fluorescence spectroscopy for noninvasive early diagnosis of oral mucosal malignant and potentially malignant lesions. J Cancer Res Ther 6(4):497–502. https://doi.org/10.4103/0973-1482.77097
Chandra S, Sah K (2010) Spectroscopy: a new diagnostic technique for detection of potentially malignant oral lesions. J Indian Acad Oral Med Radiol 22(3):144–146
Ingrams DR, Dhingra JK, Roy K, Perrault DF Jr, Bottrill ID, Kabani S, Rebeiz EE, Pankratov MM, Shapshay SM, Manoharan R, Itzkan I, Feld MS (1997) Autofluorescence characteristics of oral mucosa. Head Neck 19(1):27–32. https://doi.org/10.1002/(SICI)1097-0347(199701)19:1<27::AID-HED5>3.0.CO;2-X
Scully C, Bagan JV, Hopper C, Epstein JB (2008) Oral cancer: current and future diagnostic techniques. Am J Dent 21(4):199–209
Fedele S (2009) Diagnostic aids in the screening of oral cancer. Head Neck Oncol 1:5. https://doi.org/10.1186/1758-3284-1-5
Rajasekaran R, Aruna PR, Koteeswaran D, Bharanidharan G, Baludavid M, Ganesan S (2014) Steady-state and time-resolved fluorescence spectroscopic characterization of urine of healthy subjects and cervical cancer patients. J Biomed Opt 19(3):37003. https://doi.org/10.1117/1.Jbo.19.3.037003
Madhuri S, Vengadesan N, Aruna P, Koteeswaran D, Venkatesan P, Ganesan S (2003) Native fluorescence spectroscopy of blood plasma in the characterization of oral malignancy. Photochem Photobiol 78(2):197–204. https://doi.org/10.1562/0031-8655(2003)078<0197:nfsobp>2.0.co;2
Shpitzer T, Hamzany Y, Bahar G, Feinmesser R, Savulescu D, Borovoi I, Gavish M, Nagler RM (2009) Salivary analysis of oral cancer biomarkers. Br J Cancer 101(7):1194–1198. https://doi.org/10.1038/sj.bjc.6605290
Bahar G, Feinmesser R, Shpitzer T, Popovtzer A, Nagler RM (2007) Salivary analysis in oral cancer patients: DNA and protein oxidation, reactive nitrogen species, and antioxidant profile. Cancer 109(1):54–59. https://doi.org/10.1002/cncr.22386
Yuvaraj M, Udayakumar K, Jayanth V, Prakasa Rao A, Bharanidharan G, Koteeswaran D, Munusamy BD, Murali Krishna C, Ganesan S (2014) Fluorescence spectroscopic characterization of salivary metabolites of oral cancer patients. J Photochem Photobiol B 130:153–160. https://doi.org/10.1016/j.jphotobiol.2013.11.006
Yuvaraj M, Aruna P, Koteeswaran D, Tamilkumar P, Ganesan S (2015) Rapid fluorescence spectroscopic characterization of salivary DNA of normal subjects and OSCC patients using ethidium bromide. J Fluoresc 25(1):79–85. https://doi.org/10.1007/s10895-014-1482-0
Pobozy E, Czarkowska W, Trojanowicz M (2006) Determination of amino acids in saliva using capillary electrophoresis with fluorimetric detection. J Biochem Biophys Methods 67(1):37–47. https://doi.org/10.1016/j.jbbm.2006.01.001
Pobozy E, Czarkowska W, Trojanowicz M (2007) Determination of amino acids in saliva using capillary electrophoresis with fluorimetric detection. J Biochem Biophys Methods 69(3):Xiii-xxiii
Venkatakrishna K, Kartha VB, Pai KM, Krishna CM, Ravikiran O, Kurian J, Alexander M, Ullas G (2003) HPLC-LIF for early detection of oral cancer. Curr Sci 84:551–557
Silwood CJ, Lynch E, Claxson AW, Grootveld MC (2002) 1H and (13)C NMR spectroscopic analysis of human saliva. J Dent Res 81(6):422–427. https://doi.org/10.1177/154405910208100613
Harris AT, Lungari A, Needham CJ, Smith SL, Lones MA, Fisher SE, Yang XB, Cooper N, Kirkham J, Smith DA, Martin-Hirsch DP, High AS (2009) Potential for Raman spectroscopy to provide cancer screening using a peripheral blood sample. Head Neck Oncol 1:34. https://doi.org/10.1186/1758-3284-1-34
Kuznetsov A, Frorip A, Maiste A, Ots-Rosenberg M, Sünter A (2014) Visible auto-fluorescence in biological fluids as biomarker of pathological processes and new monitoring tool. J Innov Opt Health Sci 08:140601175004007. https://doi.org/10.1142/S1793545815410035
Markopoulos AK, Michailidou EZ, Tzimagiorgis G (2010) Salivary markers for oral cancer detection. Open Dent J 4:172–178. https://doi.org/10.2174/1874210601004010172
Wu JY, Yi C, Chung HR, Wang DJ, Chang WC, Lee SY, Lin CT, Yang YC, Yang WC (2010) Potential biomarkers in saliva for oral squamous cell carcinoma. Oral Oncol 46(4):226–231. https://doi.org/10.1016/j.oraloncology.2010.01.007
Sohn M, Himmelsbach DS, Barton FE II, Fedorka-Cray PJ (2009) Fluorescence spectroscopy for rapid detection and classification of bacterial pathogens. Appl Spectrosc 63(11):1251–1255. https://doi.org/10.1366/000370209789806993
Ramanujam N (2000) Fluorescence spectroscopy of neoplastic and non-neoplastic tissues. Neoplasia 2(1–2):89–117. https://doi.org/10.1038/sj.neo.7900077
Gillenwater A, Jacob R, Ganeshappa R, Kemp B, El-Naggar AK, Palmer JL, Clayman G, Mitchell MF, Richards-Kortum R (1998) Noninvasive diagnosis of oral neoplasia based on fluorescence spectroscopy and native tissue autofluorescence. Arch Otolaryngol Head Neck Surg 124(11):1251–1258. https://doi.org/10.1001/archotol.124.11.1251
Lingen MW, Kalmar JR, Karrison T, Speight PM (2008) Critical evaluation of diagnostic aids for the detection of oral cancer. Oral Oncol 44(1):10–22. https://doi.org/10.1016/j.oraloncology.2007.06.011
Patton LL, Epstein JB, Kerr AR (2008) Adjunctive techniques for oral cancer examination and lesion diagnosis: a systematic review of the literature. J Am Dent Assoc 139(7):896–905. https://doi.org/10.14219/jada.archive.2008.0276 (quiz 993 – 894)
De Veld DC, Witjes MJ, Sterenborg HJ, Roodenburg JL (2005) The status of in vivo autofluorescence spectroscopy and imaging for oral oncology. Oral Oncol 41(2):117–131. https://doi.org/10.1016/j.oraloncology.2004.07.007
Inaguma M, Hashimoto K (1999) Porphyrin-like fluorescence in oral cancer: In vivo fluorescence spectral characterization of lesions by use of a near-ultraviolet excited autofluorescence diagnosis system and separation of fluorescent extracts by capillary electrophoresis. Cancer 86 (11):2201–2211. https://doi.org/10.1002/(sici)1097-0142(19991201)86:11<2201::aid-cncr5>3.0.co;2-9
Kumar P, Singh A, Kumar Kanaujia S, Pradhan A (2018) Human saliva for oral precancer detection: a comparison of fluorescence & stokes shift spectroscopy. J Fluoresc 28(1):419–426. https://doi.org/10.1007/s10895-017-2203-2
Kumari S, Goyal V, Kumaran SS, Dwivedi SN, Srivastava A, Jagannathan NR (2020) Quantitative metabolomics of saliva using proton NMR spectroscopy in patients with Parkinson’s disease and healthy controls. Neurol Sci 41(5):1201–1210. https://doi.org/10.1007/s10072-019-04143-4
Dacosta RS, Wilson BC, Marcon NE (2002) New optical technologies for earlier endoscopic diagnosis of premalignant gastrointestinal lesions. J Gastroenterol Hepatol 17 Suppl:S85–104. https://doi.org/10.1046/j.1440-1746.17.s1.8.x
Nagler RM (2009) Saliva as a tool for oral cancer diagnosis and prognosis. Oral Oncol 45(12):1006–1010. https://doi.org/10.1016/j.oraloncology.2009.07.005
Purushothama P, Neethi H, Indu M, Babu N, Juneja S (2016) Autofluorescence spectroscopy of blood plasma in characterization of oral malignancy - a pilot study. J Dent Specialities 4(2):174–177. https://doi.org/10.18231/2393-9834.2016.0015
Kaur J, Jacobs R (2015) Combination of Autofluorescence imaging and salivary protoporphyrin in Oral precancerous and cancerous lesions: Non-invasive tools. J Clin Exp Dent 7(2):e187–e191. https://doi.org/10.4317/jced.52100
Raja P, Yuvaraj M, Einstein G, Sangeetha R, Prakasarao A, Jayachandran S, Singaravelu G (2020) Correlation of metabolites in saliva and in vivo tissue of oral cancer patients based on fluorescence spectral deconvolution. In: Proc. SPIE. https://doi.org/10.1117/12.2545662
Jayanthi JL, Mallia RJ, Shiny ST, Baiju KV, Mathews A, Kumar R, Sebastian P, Madhavan J, Aparna GN, Subhash N (2009) Discriminant analysis of autofluorescence spectra for classification of oral lesions in vivo. Lasers Surg Med 41(5):345–352. https://doi.org/10.1002/lsm.20771
Yan SK, Wei BJ, Lin ZY, Yang Y, Zhou ZT, Zhang WD (2008) A metabonomic approach to the diagnosis of oral squamous cell carcinoma, oral lichen planus and oral leukoplakia. Oral Oncol 44(5):477–483. https://doi.org/10.1016/j.oraloncology.2007.06.007
Wei J, Xie G, Zhou Z, Shi P, Qiu Y, Zheng X, Chen T, Su M, Zhao A, Jia W (2011) Salivary metabolite signatures of oral cancer and leukoplakia. Int J Cancer 129(9):2207–2217. https://doi.org/10.1002/ijc.25881
Ishikawa S, Sugimoto M, Edamatsu K, Sugano A, Kitabatake K, Iino M (2020) Discrimination of oral squamous cell carcinoma from oral lichen planus by salivary metabolomics. Oral Dis 26(1):35–42. https://doi.org/10.1111/odi.13209
Lohavanichbutr P, Zhang Y, Wang P, Gu H, Nagana Gowda GA, Djukovic D, Buas MF, Raftery D, Chen C (2018) Salivary metabolite profiling distinguishes patients with oral cavity squamous cell carcinoma from normal controls. PLoS One 13(9):e0204249. https://doi.org/10.1371/journal.pone.0204249
Acknowledgements
Khin TheNu Aye would like to thank the Ph.D. degree program in biomedical sciences, Faculty of Associated Medical Sciences, Chiang Mai University, under the CMU Presidential Scholarship. The authors would like to thank the Faculty of Associated Medical Sciences, Chiang Mai University and Lampang Cancer Hospital, Lampang, Thailand for their support. The authors also would like to thank Pranruethai Chaimongkol and Chadaporn Thaithiang for their participation.
Funding
This research was partially supported by Chiang Mai University. This study was also supported by the Faculty of Associated Medical Sciences, Chiang Mai University.
Author information
Authors and Affiliations
Contributions
BS. and KTNA. ; acquisition and analysis of data. JR. and WN. ; sample collection., SK.; acquisition, analysis, interpretation of data and provided critical discussions. JP.; interpretation of data and provided critical discussions. MT.; study conception and design, acquisition, analysis, and interpretation of data, provided critical discussions, drafting the manuscript and writing of the manuscript. All authors have read and agreed to the published version of the manuscript.
Corresponding author
Ethics declarations
Conflicts of Interest/Competing Interests
The authors declare no conflicts of interest/competing interests.
Ethics Approval
The present study was approved by the Ethics Committee of the Faculty of Associated Medical Sciences, Chiang Mai University, Chaing Mai, Thailand (No. AMSEC-63EX-012).
Consent to Participate
Not applicable.
Consent for Publication
Not applicable.
Additional information
Publisher’s Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
Supawat, B., Aye, K.T., Ritpanja, J. et al. Differences in Spectroscopic Properties of Saliva Taken From Normal Subjects and Oral Cancer Patients: Comparison Studies. J Fluoresc 31, 747–754 (2021). https://doi.org/10.1007/s10895-021-02707-2
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10895-021-02707-2