Serum-based diagnostic prediction of oral submucous fibrosis using FTIR spectrometry Vertika Rai a,1 , Rashmi Mukherjee b,1 , Aurobinda Routray c , Ananta Kumar Ghosh d , Seema Roy e , Barnali Paul Ghosh e , Puspendu Bikash Mandal f , Surajit Bose g , Chandan Chakraborty a, ⁎ a School of Medical Science & Technology, Indian Institute of Technology, Kharagpur, India b Advanced Technology Development Centre, Indian Institute of Technology, Kharagpur, India c Dept. of Electrical Engineering, Indian Institute of Technology, Kharagpur, India d Dept. of Biotechnology, Indian Institute of Technology, Kharagpur, India e B C Roy Technology Hospital, Indian Institute of Technology, Kharagpur, India f Dept. of ENT, Midnapur Medical College and Hospital, West Midnapur, India g Awadh Dental College and Hospital, Tata Jamshedpur, India abstract article info Article history: Received 9 December 2016 Received in revised form 4 August 2017 Accepted 7 August 2017 Available online 08 August 2017 Oral submucous fibrosis (OSF) is found to have the highest malignant potentiality among all other pre-cancerous lesions. However, its detection prior to tissue biopsy can be challenging in clinics. Moreover, biopsy examination is invasive and painful. Hence, there is an urgent need of new technology that facilitates accurate diagnostic pre- diction of OSF prior to biopsy. Here, we used FTIR spectroscopy coupled with chemometric techniques to distin- guish the serum metabolic signatures of OSF patients (n = 30) and healthy controls (n = 30). Serum biochemical analyses have been performed to further support the FTIR findings. Absorbance intensities of 45 infrared wavenumbers differed significantly between OSF and normal serum FTIR spectra representing alterations in car- bohydrates, proteins, lipids and nucleic acids. Nineteen prominent significant wavenumbers (P ≤ 0.001) at 1020, 1025, 1035, 1039, 1045, 1078, 1055, 1100, 1117, 1122, 1151, 1169, 1243, 1313, 1398, 1453, 1544, 1650 and 1725 cm -1 provided excellent segregation of OSF spectra from normal using multivariate statistical techniques. These findings provided essential information on the metabolic features of blood serum of OSF patients and established that FTIR spectroscopy coupled with chemometric analysis can be potentially useful in the rapid and accurate preoperative screening/diagnosis of OSF. © 2017 Elsevier B.V. All rights reserved. Keywords: Oral submucous fibrosis (OSF) Metabolite profiling Multivariate analysis 1. Introduction Oral cancer (OC), a worldwide health problem has a global annual incidence of around 300,000 new cases. World Health Organisation (WHO) reported death rate of 45% OC patients of all stages within five years from diagnosis. Oral squamous cell carcinoma (OSCC) patients who receive early treatment show better prognosis with N 80% 5-year survival rates [1]. It is now well established that OC is preceded by pre- malignant lesions such as oral submucous fibrosis (OSF), leukoplakia (OLK), erythroplakia and lichen planus (OLP) [2]. OSF is a chronic de- ceptive disease which affects oral cavity including the pharynx [3]. It is generally characterized by idiopathic scleroderma of mouth, idiopathic palatal fibrosis, sclerosing stomatitis and juxta-epithelial fibrosis. The clinical signature of OSF is submucosal fibrosis affecting oral cavity, pro- gressive trismus due to rigid lips, cheeks, pharynx and upper third of the esophagus leading to dysphagia [4]. OSF has a tendency of high malig- nant transformation all cases of epithelial dysplasia at the time of biopsy. Malignant transformation of OSF to OSCC ranges from 3% to 19% according to epidemiologic studies. Generally, diagnosis of OC patients and precancerous lesions is carried out by conventional oral examination. Poor survival among OC patients results due to advanced extent malignancy at the time of diagnosis [5] (N 60% of OC cases). Al- though, histological biopsy is the gold standard method for diagnosis of pre-/malignant oral lesions; however, lack of expertise, infrastruc- ture, and negative psychological effects hinders the process. Moreover, it becomes difficult to demarcate the appropriate site for biopsy when the lesions are large [6]. Histopathological study remains a diagnostic technique with limited sensitivity, which depends heavily on the sub- jective interpretation of the examining pathologist. As such, it leads to inter and intra -observer variations along with high false positive and false negative rates [7]. Consequently, there has been a far-reaching shift from histopathological to molecular approaches for disease diagnosis since alterations first occur at the molecular level followed by clinical changes, which can be visible under the microscope. This Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 189 (2018) 322–329 ⁎ Corresponding author. E-mail address: chandanc@smst.iitkgp.ernet.in (C. Chakraborty). 1 Both the authors have contributed equally. http://dx.doi.org/10.1016/j.saa.2017.08.018 1386-1425/© 2017 Elsevier B.V. All rights reserved. Contents lists available at ScienceDirect Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy journal homepage: www.elsevier.com/locate/saa