A pilot study for early detection of oral premalignant diseases using oral cytology and Raman micro-spectroscopy: Assessment of confounding factors
Corresponding Author
Isha Behl
Centre for Radiation and Environmental Science, FOCAS Research Institute, Technological University Dublin. City Campus, Dublin, Ireland
School of Physics & Clinical & Optometric Sciences, Technological University Dublin, City Campus, Dublin, Ireland
Correspondence
Dr Isha Behl, Centre for Radiation and Environmental Science, FOCAS Research Institute, Technological University Dublin, City Campus, Dublin, Ireland.
Email: [email protected]
Search for more papers by this authorGenecy Calado
Centre for Radiation and Environmental Science, FOCAS Research Institute, Technological University Dublin. City Campus, Dublin, Ireland
School of Physics & Clinical & Optometric Sciences, Technological University Dublin, City Campus, Dublin, Ireland
Search for more papers by this authorAlison Malkin
School of Biological and Health Sciences, Technological University Dublin, City Campus, Dublin, Ireland
Search for more papers by this authorStephen Flint
Oral Medicine Unit, Dublin Dental University Hospital, Trinity College Dublin, Dublin, Ireland
Search for more papers by this authorSheila Galvin
Oral Medicine Unit, Dublin Dental University Hospital, Trinity College Dublin, Dublin, Ireland
Search for more papers by this authorClaire M. Healy
Oral Medicine Unit, Dublin Dental University Hospital, Trinity College Dublin, Dublin, Ireland
Search for more papers by this authorMarina Leite Pimentel
Division of Restorative Dentistry and Periodontology, Dublin Dental University Hospital, Trinity College Dublin, Dublin, Ireland
Search for more papers by this authorHugh J. Byrne
FOCAS Research Institute, Technological University Dublin, City Campus, Dublin, Ireland
Search for more papers by this authorFiona M. Lyng
Centre for Radiation and Environmental Science, FOCAS Research Institute, Technological University Dublin. City Campus, Dublin, Ireland
School of Physics & Clinical & Optometric Sciences, Technological University Dublin, City Campus, Dublin, Ireland
Search for more papers by this authorCorresponding Author
Isha Behl
Centre for Radiation and Environmental Science, FOCAS Research Institute, Technological University Dublin. City Campus, Dublin, Ireland
School of Physics & Clinical & Optometric Sciences, Technological University Dublin, City Campus, Dublin, Ireland
Correspondence
Dr Isha Behl, Centre for Radiation and Environmental Science, FOCAS Research Institute, Technological University Dublin, City Campus, Dublin, Ireland.
Email: [email protected]
Search for more papers by this authorGenecy Calado
Centre for Radiation and Environmental Science, FOCAS Research Institute, Technological University Dublin. City Campus, Dublin, Ireland
School of Physics & Clinical & Optometric Sciences, Technological University Dublin, City Campus, Dublin, Ireland
Search for more papers by this authorAlison Malkin
School of Biological and Health Sciences, Technological University Dublin, City Campus, Dublin, Ireland
Search for more papers by this authorStephen Flint
Oral Medicine Unit, Dublin Dental University Hospital, Trinity College Dublin, Dublin, Ireland
Search for more papers by this authorSheila Galvin
Oral Medicine Unit, Dublin Dental University Hospital, Trinity College Dublin, Dublin, Ireland
Search for more papers by this authorClaire M. Healy
Oral Medicine Unit, Dublin Dental University Hospital, Trinity College Dublin, Dublin, Ireland
Search for more papers by this authorMarina Leite Pimentel
Division of Restorative Dentistry and Periodontology, Dublin Dental University Hospital, Trinity College Dublin, Dublin, Ireland
Search for more papers by this authorHugh J. Byrne
FOCAS Research Institute, Technological University Dublin, City Campus, Dublin, Ireland
Search for more papers by this authorFiona M. Lyng
Centre for Radiation and Environmental Science, FOCAS Research Institute, Technological University Dublin. City Campus, Dublin, Ireland
School of Physics & Clinical & Optometric Sciences, Technological University Dublin, City Campus, Dublin, Ireland
Search for more papers by this authorFunding information: Fiosraigh Dean of Graduate Studies Award; Science Foundation Ireland, Grant/Award Number: 12/IP/1494; Technological University Dublin
Abstract
This study demonstrates the efficacy of Raman micro-spectroscopy of oral cytological samples for differentiating dysplastic, potentially malignant lesions from those of normal, healthy donors. Cells were collected using brush biopsy from healthy donors (n = 20) and patients attending a Dysplasia Clinic (n = 20). Donors were sampled at four different sites (buccal mucosa, tongue, alveolus, gingiva), to ensure matched normal sites for all lesions, while patient samples were taken from clinically evident, histologically verified dysplastic lesions. Spectra were acquired from the nucleus and cytoplasm of individual cells of all samples and subjected to partial least squares-discriminant analysis. Discriminative sensitivities of 94% and 86% and specificity of 85% were achieved for the cytoplasm and nucleus, respectively, largely based on lipidic contributions of dysplastic cells. Alveolar/gingival samples were differentiated from tongue/buccal samples, indicating that anatomical site is potentially a confounding factor, while age, gender, smoking and alcohol consumption were confirmed not to be.
CONFLICT OF INTEREST
The authors declare no conflicts of interest.
Supporting Information
| Filename | Description |
|---|---|
| jbio202000079-sup-0001-FigureS1.jpgJPEG image, 849.4 KB | Figure S1 PLS-DA predictive model for cytoplasm (normal vs lesion). (a) Scores plot, (b) latent variable 1, (c) latent variable 2, (d) predictive model and (e) confusion matrix. (One-way ANOVA, P < .00001) |
| jbio202000079-sup-0002-FigureS2.jpgJPEG image, 718.5 KB | Figure S2 (a) PLS-DA score plot for cytoplasm spectra from healthy donors (normal) vs patients (lesions). (b) Depicting anatomical sites |
| jbio202000079-sup-0003-FigureS3.jpgJPEG image, 745.1 KB | Figure S3 PLS-DA predictive model for cytoplasm spectra from buccal mucosa/tongue (normal vs lesion). (a) Score plot, (b) latent variable 1, (c) latent variable 2, (d) predictive model and (e) confusion matrix |
| jbio202000079-sup-0004-FigureS4.jpgJPEG image, 671.3 KB | Figure S4 PLS-DA predictive model for nucleus spectra for healthy donors(normal) vs patient samples (lesion). (a) Score plot gender, (b) score plot age, (c) score plot smoking, (d) score plot alcohol consumption. (One-way ANOVA P < .00001 |
Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.
REFERENCES
- 1GLOBOCAN: Estimated Cancer Incidence, Mortality and Prevalence Worldwide in 2018, International Agency for Research on Cancer, 2018, (online), http://globocan.iarc.fr/Pages/fact_sheets_population.aspx (accessed: March 2020).
- 2C. F. Poh, C. E. MacAulay, D. M. Laronde, P. M. Williams, L. Zhang, M. P. Rosin, Periodontol 2000 2011, 57(1), 73.
- 3B. W. Neville, T. a Day, Ca-Cancer J. Clin. 2002, 52(4), 195.
- 4P. V. Centelles, J. M. Seoane-romero, I. Gómez, P. Diz-dios, N. S. De Melo, J. Seoane, Timing of Oral Cancer Diagnosis: Implications for Prognosis and Survival, Oral Cancer, Kalu U. E.Ogbureke, IntechOpen, 2012. https://doi.org/10.5772/30591.
- 5I. J. C. Schneider, M. E. Flores, D. A. Nickel, L. G. T. Martins, J. Traebert, Rev. Bras. Epidemiol. 2014, 17, 680.
- 6A. Trakroo, M. K. Sunil, A. Trivedi, R. Garg, A. Kulkarni, S. Arora, J. Int. Oral Health 2015, 7(3), 33.
- 7N. V. Babshet M, K. Nandimath, S. Pervatikar, J. Cytol. 2011, 28(4), 165.
- 8T. A. Winning, G. C. Townsend, Clin. Dermatol. 2000, 18(5), 499.
- 9C. A. Squier, M. J. Kremer, J. Natl. Cancer Inst. Monogr. 2001, 52242(29), 7.
10.1093/oxfordjournals.jncimonographs.a003443 Google Scholar
- 10V. Kumar, N. Fausto, A. Abbas, Robbins and Cotran Pathologic Basis of Disease, 7th ed., Elsevier, Amsterdam, 2005.
- 11C. Scully, J. V. Bagan, C. Hopper, Am. J. Dent. 2008, 21(4), 200.
- 12K. Papamarkakis, B. Bird, J. M. Schubert, M. Miljković, R. Wein, K. Bedrossian, N. Laver, M. Diem, Lab. Invest. 2010, 90(4), 589.
- 13V. G. Patel, K. S. PV, V. Kumar, J. Cytol. 2011, 28(2), 66.
- 14D. Wynford-Thomas, J. Pathol. 1999, 187(1), 100.
10.1002/(SICI)1096-9896(199901)187:1<100::AID-PATH236>3.0.CO;2-T CAS PubMed Web of Science® Google Scholar
- 15A. V. O.-P. Alejandra Isabel Orellana-Bustos, I. L. Espinoza-Santander, M. E. Franco-Martínez, N. Lobos-James-Freyre, Med. Oral 2004, 9(3), 197.
- 16N. K. Proia, G. M. Paszkiewicz, M. A. S. Nasca, G. E. Franke, J. L. Pauly, Cancer Epidemiol. Biomarkers Prev. 2006, 15, 1061.
- 17P. A. Wu, C. H. Loh, L. L. Hsieh, T. Y. Liu, C. J. Chen, S. H. Liou, Mutat. Res., Genet. Toxicol. Environ. Mutagen. 2004, 562(1–2), 27.
- 18M. R. Khairnar, U. Wadgave, S. M. Khairnar, J. Alcohol. Drug Depend. 2017, 5(3), 3.
10.4172/2329-6488.1000266 Google Scholar
- 19S. Regina, D. A. Reis, M. Gustavo, S. Andrade, M. Sadigursky, Development 2006, 20(2), 97.
- 20M. Diem, K. Papamarkakis, J. Schubert, B. Bird, M. J. Romeo, M. Miljković, Appl. Spectrosc. 2009, 63(11), 307.
- 21M. Diem, A. Mazur, K. Lenau, J. Schubert, B. Bird, M. Miljković, C. Krafft, J. Popp, J. Biophotonics 2013, 6(11–12), 855.
- 22M. Miljković, B. Bird, K. Lenau, A. I. Mazur, M. Diem, Analyst 2013, 138, 3975.
- 23A. Sahu, N. Shah, M. Mahimkar, M. Garud, S. Pagare, S. Nair, C. M. Krishna, in Proc. SPIE 8926, Photonic Therapeutics and Diagnostics X, 89262N, 2014.
- 24A. Sahu, P. Gera, V. Pai, A. Dubey, G. Tyagi, M. Waghmare, S. Pagare, M. Mahimkar, C. M. Krishna, J. Biomed. Opt. 2017, 22(11), 1.
- 25A. Sahu, S. Tawde, V. Pai, P. Gera, P. Chaturvedi, C. M. Krishna, Anal. Methods 2015, 7, 7548.
- 26I. Behl, G. Calado, O. Ibrahim, A. Malkin, S. Flint, H. J. Byrne, F. M. Lyng, Anal. Methods 2017, 9(6), 937.
- 27A. K. El-Naggar, J. K. C. Chan, J. R. Grandis, T. Takata, P. J. Slootweg Eds., WHO Classification of Head and Neck Tumours, 4th ed., IARC, France, 2017.
- 28H. Nawaz, F. Bonnier, A. D. Meade, F. M. Lyng, H. J. Byrne, Analyst 2011, 136, 2450.
- 29F. Bonnier, H. J. Byrne, Analyst 2012, 137(2), 322.
- 30Z. Farhane, F. Bonnier, A. Casey, A. Maguire, L. O'Neill, H. J. Byrne, Analyst 2015, 140, 5908.
- 31L. T. Kerr, B. M. Hennelly, Chemom. Intel. Lab. Syst. 2016, 158, 61.
- 32M. Urvoy, F. Autrusseau, in Proc. 2nd ACM Work. Inf. Hiding Multimed. Secur., IH&MMSec'14, 2014, pp. 49–60.
- 33R. G. Brereton, G. R. Lloyd, J. Chemometr. 2014, 28(4), 213.
- 34R. Gautam, S. Vanga, F. Ariese, S. Umapathy, EPJ Tech. Instrum. 2015, 2(1), 8.
- 35S. F. Sawyer, J. Man. Manip. Ther. 2009, 17(2), 27E.
10.1179/jmt.2009.17.2.27E Google Scholar
- 36D. C. Montgomery, G. C. Runger, Appl. Stat. Probab. Eng. 7th ed., Wiley, Hoboken, NJ, 2018.
- 37C. Krafft, L. Neudert, T. Simat, R. Salzer, Spectrochim. Acta, Part A 2005, 61, 1529.
- 38M. Gniadecka, H. C. Wulf, N. N. Mortensen, O. F. Nielsen, D. H. Christensen, J. Raman Spectrosc. 1997, 28(2–3), 125.
- 39Z. Movasaghi, S. Rehman, I. Rehman, Appl. Spectrosc. 2007, 42(5), 493.
- 40H. Byrne, G. Sockalingum, N. Stone, Raman Microscopy: Complement or Competitor. in Biomedical Applications of Synchrotron Infrared Microspectroscopy, RSC Analytical Spectroscopy Series. 2011, p. 105.
- 41N. Stone, C. Kendall, J. Smith, P. Crow, H. Barr, Faraday Discuss. 2004, 126(1), 141.
- 42J. W. Chan, D. S. Taylor, T. Zwerdling, S. M. Lane, K. Ihara, T. Huser, Biophys. J. 2006, 90(2), 648.
- 43S. Kumar, N. Vezhavendhan, S. Priya, Int. J. Clin. Dent. Sci. 2011, 2(1), 293.
- 44S. Beloribi-Djefaflia, S. Vasseur, F. Guillaumond, Oncogenesis 2016, 5(1), e189.
- 45G. Cascianelli, M. Villani, M. Tosti, F. Marini, E. Bartoccini, M. V. Magni, E. Albi, Mol. Biol. Cell 2008, 19, 5289.
- 46F. L. Cals, T. C. B. Schut, J. A. Hardillo, R. J. B. de Jong, S. Koljenović, Lab. Invest. 2015, 95, 1186.
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