Microstructural changes in the irradiated and osteoradionecrotic bone: a SEM study.

Radiation exposure is a major health concern due to bone involvement including mandible, causing deleterious effects on bone metabolism, and healing with an increasing risk of infection and osteoradionecrosis. This study aims to investigate the radiotherapy-induced microstructural changes in the human mandible by scanning electron microscopy (SEM). Mandibular cortical bone biopsies were obtained from control, irradiated, and patients with osteoradionecrosis (ORN). Bone samples were prepared for light microscopy and SEM. The SEM images were analyzed for the number of osteons, number of Haversian canal (HC), diameter of osteon (D.O), the diameter of HC (D.HC), osteonal wall thickness (O.W.Th), number of osteocytes, and number of osteocytic dendrites. The number of osteons, D.O, D.HC, O.W.Th, the number of osteocytes, and osteocytic dendrites were significantly decreased in both irradiated and ORN compared to controls (p < .05). The number of HCs decreased in irradiated and ORN bone compared to the control group. However, this was statistically not significant. The deleterious effect of radiation continues gradually altering the bone quality, structure, cellularity, and vascularity in the long term (>5 years mean radiation biopsy interval). The underlying microscopic damage in bone increases its susceptibility and contributes further to radiation-induced bone changes or even ORN.

Effects of irradiation in the mandibular bone loaded with dental implants. An experimental study with a canine model.

Radiation therapy may compromise the quality of bone around dental implants, and its ability to regenerate, remodel, and revascularize. This study aimed to describe the irradiation effect on the bone microstructure of the mandible using dental implants in a canine model. Five beagle dogs were exposed to 40 Gy fractionated radiation. In total, 20 dental implants were inserted, two in the irradiated and two in the non-irradiated side. The mandible bone blocks were subjected to 3D micro-computed tomography (µCT) imaging, later evaluated histomorphometrically by light microscopy and scanning electron microscopy. Alterations in irradiated bone were observed under µCT imaging showing an increased anisotropy, porosity, and pore volume. Bone surface-to-bone volume decreased. The bone to implant contact index was significantly reduced in the irradiated bone (75.6% ± 5.8%) as compared to the non-irradiated bone (85.1% ± 6.8%). In the irradiated mandible, osteocytes with their filopodial processes, the bone beneath the periosteum, and subperiosteal veins showed structural differences but were not significant, whereas the diameter of Haversian canals were smaller statistical significant as compared to the control side. The study highlights that radiation dosage of fractioned 40 Gy causes alterations in the alveolar bone microstructure with compatible osseointegration and clinically stable dental implants.

Localization of transmembrane mucin MUC1 on the apical surface of oral mucosal cells.

The purpose of the present study was to demonstrate the localization of transmembrane mucin MUC1 on the outer layer of oral mucosal cells and the involvement of apical cell surface microplicae (MPL) in bioadhesion of MUC1. Tissue samples of six healthy subjects were obtained. First, the presence of MUC1 was examined with an immunohistochemical method using a monoclonal MUC1 antigen called HFMG1. Second, the localization of MUC1 was examined with immuno-scanning electron microscopy. Immunohistochemically, high intense staining for MUC1 (antigen HFMG1) was detected in the epithelial superficial layers. In the superficial layer, intense MUC1 expression was seen predominantly on the apical cell surface. On the apical epithelial cells, MUC1 was associated predominantly with MPL towards the oral cavity. The novelty of the results of the present study is that MPL serves a harbor of MUC1 in superficial epithelial cells towards the oral cavity. It is speculated that the transmembrane MUC1 is one component of the "oral mucosal barrier complex" representing a signaling pathway between saliva and mucosal cells.Abbreviations: MUC1: mucin1; MAM: membrane-anchored mucin; OMBC: oral mucosal barrier complex; LM: light microscopy; TEM: transmission electron microscopy; SEM: scanning electron microscopy; iSEM: immuno-scanning electron microscopy; MPL: microplicae.

Alveolar bone remodeling after tooth extraction in irradiated mandible: An experimental study with canine model.

OBJECTIVES: The aim of the present study is to investigate the morphological and cellular changes in dental extraction socket that has been irradiated after the tooth extraction and to describe morphological characteristics of the osteocytes and osteocyte-lacunar-canalicular network (LCN) by scanning electron microscopy (SEM). MATERIAL AND METHODS: Five beagle dogs aged 1-2 years were used in this study. One side of each mandible was irradiated in two sessions and the other side of mandible (non-irradiated) served as a control. The mandible bone blocks were processed by bulk staining en bloc in basic fuchsin and the specimens were embedded routinely in polymethyl methacrylate resin without preliminary decalcification. All blocks were subjected to micro-CT imaging, after that the specimens were prepared for light microscopy and SEM. RESULTS: Alterations in bone macrostructure are minimal in irradiated bone, but the changes in LCN are clear. In the area of the tooth extraction socket, the connections of osteocytes to the vessels and to neighboring osteocytes were not observed both in irradiated and nonirradiated bone. However, osteoclasts were located in the bone surface entering inside to the bone between osteons. In the lamellar bone of lateral sides, a decrease in canalicular connections between osteocytes and periosteum was found in irradiated bone as compared to the non-irradiated side. CONCLUSIONS: The novelty of the present study is that radiation disrupts osteocytes and their dendrites.

Acid-etching technique of non-decalcified bone samples for visualizing osteocyte-lacuno-canalicular network using scanning electron microscope.

The aim of this study was to define the acid-etching technique for bone samples embedded in polymethyl metacrylate (PMMA) in order to visualize the osteocyte lacuno-canalicular network (LCN) for scanning electron microscopy (SEM). Human jaw bone tissue samples (N = 18) were collected from the study population consisting of patients having received dental implant surgery. After collection, the bone samples were fixed in 70% ethanol and non-decalcified samples embedded routinely into polymethyl metacrylate (PMMA). The PMMA embedded specimens were acid-etched in either 9 or 37% phosphoric acid (PA) and prepared for SEM for further analysis. PMMA embedded bone specimens acid-etched by 9% PA concentration accomplishes the most informative and favorable visualization of the LCN to be observed by SEM. Etching of PMMA embedded specimens is recommendable to start with 30 s or 40 s etching duration in order to find the proper etching duration for the samples examined. Visualizing osteocytes and LCN provides a tool to study bone structure that reflects changes in bone metabolism and diseases related to bone tissue. By proper etching protocol of non-decalcified and using scanning electron microscope it is possible to visualize the morphology of osteocytes and the network supporting vitality of bone tissue.

Radiation-induced changes in the microstructure of epithelial cells of the oral mucosa: A comparative light and electron microscopic study.

OBJECTIVES: The microplicae is a typical structure of the epithelial cell surface of the oral mucosa. The cell surface is potentially of great significance, as it provides the underlying basis for the protective function of the salivary pellicle. The aim of this study was to investigate whether radiation therapy affects the surface morphology of the superficial cells of the human oral mucosa in patients who have received radiotherapy for oral cancer. MATERIAL AND METHODS: Oral mucosal tissue samples from 91 patients were collected during dental implant surgery or ablative surgery. Study group 1 consisted of 28 patients who underwent dental implant surgery after radiotherapy. Group 2 consisted of five patients who developed osteoradionecrosis. Group 3 consisted of eight oral cancer patients without radiotherapy. Group 4 consisted of 50 clinically healthy subjects as controls. The samples were studied with scanning electron microscopy and compared with both light and transmission electron micrographs. RESULTS: Radiation therapy (RT) induces breakage and destruction in the microplicae morphology and declines the density of the microplicae surface structures. In some of the irradiated cells, the microplicae were completely vanished, especially in patients who developed osteoradionecrosis. In non-irradiated tissue, the microplicae of the superficial epithelial cells were intact in all cases. CONCLUSION: Scanning electron microscopy, in contrast to light microscopy, appears to be a useful tool to reveal the condition of superficial oral mucosal cells. In respect of the possible pathogenesis of osteoradionecrosis, the radiation-induced damage of the microplicae and its influence on the mucosal salivary pellicle is discussed.

Surface morphology of superficial cells in irradiated oral mucosa: an experimental study in beagle dog.

OBJECTIVES: The aim of the present study is to investigate if radiation induces changes in the superficial cells of the oral mucosa and secondly to describe morphological characteristics of the cell surface structure by scanning electron microscopy (SEM). MATERIALS AND METHODS: Ten beagle dogs aged 1-2 years were used in this study. One side of each mandible was irradiated in two sessions, each lasting 1 week. The total dosage was 40 Gy (Group A; 5 dogs) and 50 Gy (Group B; 5 dogs), in five fractions of 4 Gy. The other side of mandible (non-irradiated) served as a control. The specimen was harvested with a scalpel from the alveolar mucosa of the irradiated area 1 year after irradiation and studied with SEM. RESULTS: In the control side, the surface structure of the cell contains straight parallel or branched microplicae (MPL), which were equally spaced over the cell surfaces. Discontinuous and short MPL were typical cell structure of irradiated mucosa. In 50 Gy group, the surface structure of epithelial cell was pitted and the cell boundaries were thick. CONCLUSIONS: The novelty of the present study is that radiation disrupts superficial cells of the oral mucosa. The role of the MPL structure of the superficial cells in mucositis development is discussed.

Microstructure of the superficial epithelial cells of the human oral mucosa.

BACKGROUND: The apical cell membrane of the oral mucosa adjacent to the saliva interface is thrown into membrane folds, termed microplicae (MPL) with variation in morphology. The present study classifies morphological changes undergone by MPL into qualitative and quantitative categories. MATERIAL AND METHODS: Oral mucosal specimens were obtained from 32 healthy patients. Half of each specimen was prepared routinely for light microscopy, and the other part for scanning and transmission electron microscopy. Different measurements of cell structure were presented: the density of MPL, the width and height of MPL, the width of furrows between two adjacent MPL and the distance of the centre of MPL. Morphometric measurements were obtained using a semiautomatic ImageJ analysis software (W Rasband, National Institutes of Health, Bethesda, MD). RESULTS: Parallel and branching MPL was common observation in the area of lining mucosa and in the tongue between the filiform papillae. The density of MPL was higher in the cells of the buccal mucosa than in the cells of the tongue, 43.69 + 11.43% and 31.68 + 10.32%, respectively. The difference was significant (p < 0.001). The width of MPL was 0.16 µm in cells of the buccal mucosa and 0.12 µm in cells of the tongue. CONCLUSIONS: Our findings support the idea that MPL structure is a determining factor for the functionality of the oral epithelium since the values of the MPL were kept relatively stable. The role of MPL structure of the oral mucosal cells is discussed.

Microstructure of oral epithelial cells as an underlying basis for salivary mucosal pellicle.

BACKGROUND: Salivary mucosal pellicle forms the structural basis of the local innate immune defense mechanism of the oral mucosa. At the surface of the oral mucosa, the apical cell membrane adjacent to the saliva interface contains short membrane folds, termed microplicae (MPL). This MPL structure of oral epithelial cells and its function as a basis to the salivary mucosal pellicle is unclear. In this preliminary study, we describe the ultrastructural morphology of cell membrane of superficial cells of the oral mucosa and study the membrane-associated mucins (MAMs), MUC1 and MUC4, with immunohistological methods. MATERIALS AND METHODS: Oral mucosal specimens were obtained from six healthy patients. Half of each specimen was prepared routinely for light microscopy, and the other part for scanning and transmission electron microscopy. The presence of MUC1 and MUC4 were studied by immunohistochemical methods in oral mucosal specimens. RESULTS: Morphologically, the cell membrane of MPL is partly discontinuous and membrane-associated molecules extrude from the cell membrane. MUC1 expression was detected in the superficial part of the buccal epithelium, while MUC4 had no expression in the oral squamous epithelium. CONCLUSIONS: The novel of this study is that the membrane-tethered molecules seem to occur onto the cell membrane of the superficial epithelial cells of the oral mucosa. Furthermore, the stratified squamous epithelium of the buccal mucosa produces MUC1 for the surface-saliva pellicle interface. The interaction between MPL structure, MUC1 mucin, and salivary mucosal pellicle is discussed.

Irradiation affects the structural, cellular and molecular components of jawbones.

PURPOSE: Emerging evidence shows that changes in the bone and its microenvironment following radiotherapy are associated with either an inhibition or a state of low bone formation. Ionizing radiation is damaging to the jawbone as it increases the complication rate due to the development of hypovascular, hypocellular, and hypoxic tissue. This review summarizes and correlates the current knowledge on the effects of irradiation on the bone with an emphasis on jawbone, as these have been a less extensively studied area. CONCLUSIONS: The stringent regulation of bone formation and bone resorption can be influenced by radiation, causing detrimental effects at structural, cellular, vascular, and molecular levels. It is also associated with a high risk of damage to surrounding healthy tissues and an increased risk of fracture. Technological advances and research on animal models as well as a few human bone tissue studies have provided novel insights into the ways in which bone can be affected by high, low and sublethal dose of radiation. The influence of radiation on bone metabolism, cellular properties, vascularity, collagen, and other factors like inflammation, reactive oxygen species are discussed.

Effect of smoking on MUC1 expression in oral epithelial dysplasia, oral cancer, and irradiated oral epithelium.

OBJECTIVES: The aim of this study was to assess the MUC1 expression in the oral epithelium of normal, oral epithelial dysplasia (OED), oral squamous cell carcinoma (OSCC), and irradiated oral epithelium (IROE) and its association with smoking habits in non-smokers and smokers. DESIGN: Oral mucosal biopsies from controls, OED, OSCC, and IROE groups were obtained and categorized based on the smoking history as non-smokers, smoker I (25 pack-years), and smoker II (>25 pack-years). Immunohistochemical staining of MUC1 using human milk fat globule 1 (HMFG 1) antibody was performed, and the MUC1 score was calculated. The relation between MUC1 expression and clinicopathological findings was examined. RESULTS: MUC1 staining of superficial oral epithelial cells with mild MUC1 score was detected in all control samples. The MUC1 staining extended from superficial to basal cell layer of oral epithelium with the increase in MUC1 score from moderate to strong in OED, OSCC, and IROE, and the difference was significant (p < 0.004, p < 0.002 and p < 0.004, respectively) compared to controls. A positive association between smoking and MUC1 score was observed within groups (p < 0.05). CONCLUSION: The depolarization of MUC1 protein expression is associated with smoking habits in OED and OSCC. In the IROE, the radiation causes subcellular and molecular changes, observed as altered MUC1 expression and accelerated by smoking, furthermore, complicating the oral mucosal adaptation and progress to radiation-induced lesions as a delayed effect.

Irradiation Alters the Expression of MUC1, CD44 and Hyaluronan in Oral Mucosal Epithelium.

Purpose: It is well established that cancer cells exploit aberrant synthesis of mucin 1 (MUC1) and hyaluronan (HA) synthesis along with HA's physiological cell surface receptor CD44. However, their role in irradiated oral tissue has not been reported previously. We, therefore, aimed to study MUC1, CD44 and HA immunohistochemically in irradiated oral mucosa and their role in the long-term effects after radiotherapy. Materials and Methods: Oral mucosal biopsies were obtained from healthy subjects as controls and from patients after radiotherapy for head and neck cancer (irradiated group) during dental implant surgery.The presence of MUC1, CD44, and HA in oral mucosa was studied by immunohistochemical methods. The differences in the localization and intensity in the oral epithelium between control and irradiated tissue were analyzed. Results: The staining intensity of MUC1 was confined to the superficial epithelial layer, whereas HA and CD44 were found in the cell membranes in the epithelial basal and intermediate layers of control specimens. In irradiated epithelium, MUC1 staining was distributed throughout all the layers of the oral epithelium, with significant staining in the basal and intermediate layers. Accordingly, HA and CD44 staining extended to involve the superficial cells of the irradiated epithelium. The staining pattern of MUC1 and CD44 showed significant changes in irradiated samples. Conclusions: Our results showed that the staining intensities of MUC1, CD44, and HA were significantly elevated in irradiated tissue compared to controls. MUC1, CD44, and HA are important markers and take part in long-term changes in the oral mucosa after radiotherapy.

Irradiation Induced Biochemical Changes in Human Mandibular Bone: A Raman Spectroscopic Study.

Understanding the biochemical changes in irradiated human mandible after radiotherapy of cancer patients is critical for oral rehabilitation. The underlying mechanism for radiation-associated changes in the bone at the molecular level could lead to implant failure and osteoradionecrosis. The study aimed to assess the chemical composition and bone quality in irradiated human mandibular bone using Raman spectroscopy. A total of 33 bone biopsies from 16 control and 17 irradiated patients were included to quantify different biochemical parameters from the Raman spectra. The differences in bone mineral and matrix band intensities between control and irradiated groups were analyzed using unpaired Student's t-test with statistical significance at p < 0.05. Findings suggest that the intensity of the phosphate band is significantly decreased and the carbonate band is significantly increased in the irradiated group. Further, the mineral crystallinity and carbonate to phosphate ratio are increased. The mineral to matrix ratio is decreased in the irradiated group. Principal component analysis (PCA) based on the local radiation dose and biopsy time interval of irradiated samples did not show any specific classification between irradiation sub-groups. Irradiation disrupted the interaction and bonding between the organic matrix and hydroxyapatite minerals affecting the bone biochemical properties. However, the normal clinical appearance of irradiated bone would have been accompanied by underlying biochemical and microscopical changes which might result in radiation-induced delayed complications.

Salivary Metabolomics for Diagnosis and Monitoring Diseases: Challenges and Possibilities.

Saliva is a useful biological fluid and a valuable source of biological information. Saliva contains many of the same components that can be found in blood or serum, but the components of interest tend to be at a lower concentration in saliva, and their analysis demands more sensitive techniques. Metabolomics is starting to emerge as a viable method for assessing the salivary metabolites which are generated by the biochemical processes in elucidating the pathways underlying different oral and systemic diseases. In oral diseases, salivary metabolomics has concentrated on periodontitis and oral cancer. Salivary metabolites of systemic diseases have been investigated mostly in the early diagnosis of different cancer, but also neurodegenerative diseases. This mini-review article aims to highlight the challenges and possibilities of salivary metabolomics from a clinical viewpoint. Furthermore, applications of the salivary metabolic profile in diagnosis and prognosis, monitoring the treatment success, and planning of personalized treatment of oral and systemic diseases are discussed.

Low-Dose Doxycycline Treatment Normalizes Levels of Some Salivary Metabolites Associated with Oral Microbiota in Patients with Primary Sjögren's Syndrome.

Saliva is a complex oral fluid, and plays a major role in oral health. Primary Sjögren's syndrome (pSS), as an autoimmune disease that typically causes hyposalivation. In the present study, salivary metabolites were studied from stimulated saliva samples (n = 15) of female patients with pSS in a group treated with low-dose doxycycline (LDD), saliva samples (n = 10) of non-treated female patients with pSS, and saliva samples (n = 14) of healthy age-matched females as controls. Saliva samples were analyzed with liquid chromatography mass spectrometry (LC-MS) based on the non-targeted metabolomics method. The saliva metabolite profile differed between pSS patients and the healthy control (HC). In the pSS patients, the LDD treatment normalized saliva levels of several metabolites, including tyrosine glutamine dipeptide, phenylalanine isoleucine dipeptide, valine leucine dipeptide, phenylalanine, pantothenic acid (vitamin B5), urocanic acid, and salivary lipid cholesteryl palmitic acid (CE 16:0), to levels seen in the saliva samples of the HC. In conclusion, the data showed that pSS is associated with an altered saliva metabolite profile compared to the HC and that the LLD treatment normalized levels of several metabolites associated with dysbiosis of oral microbiota in pSS patients. The role of the saliva metabolome in pSS pathology needs to be further studied to clarify if saliva metabolite levels can be used to predict or monitor the progress and treatment of pSS.

Variability of salivary metabolite levels in patients with Sjögren's syndrome.

PURPOSE: To investigate inter- and intra-individual variation in the levels and outputs (concentration multiplied by salivary flow rate) of salivary metabolites in patients with primary Sjögren's syndrome (pSS). METHODS: A total of 56 samples of stimulated saliva were collected from 14 female pSS patients during four laboratory visits within 20 weeks and analyzed using proton nuclear magnetic resonance spectroscopy. Single saliva samples from each of 15 controls were also analyzed. RESULTS: Among 21 quantified metabolites, choline was significantly elevated in the pSS patients at each time point (P ≤ 0.015), taurine at the last three time points (P ≤ 0.013), alanine at the last two time points (P ≤ 0.007) and glycine at the last time point (P = 0.005). Inter-individual variation in metabolite concentrations was generally larger among the patients than among the controls, and significantly large variations were observed for glycine (P ≤ 0.007, all time points), choline (P ≤ 0.033, three last time points) and alanine (P = 0.028, baseline). Metabolite output analysis showed that choline had the lowest intra-patient variation. CONCLUSION: In spite of considerable intra- and inter-individual variation, levels and outputs of specific metabolites in patients with pSS differ from those in controls, and may be potentially applicable as new biological markers for monitoring of the response to treatment.

Biochemical Changes in Irradiated Oral Mucosa: A FTIR Spectroscopic Study.

Radiation exposure during the course of treatment in head and neck cancer (HNC) patients can induce both structural and biochemical anomalies. The present study is focused on utilizing infrared imaging for the identification of the minor biochemical alterations in the oral mucosa. Chemical maps generated using glycoprotein band indicates its differential distribution along the superficial layer. Spectra extracted from this layer suggests changes in overall nucleic acid and protein content in response to the therapeutic irradiation. Discrimination among control and irradiated groups have been achieved using principal component analysis. Findings of this preliminary study further support prospective utilization of Fourier Transform InfraRed (FTIR) imaging as a non-destructive, label-free tool for objective assessment of the oral mucosa in patient groups with or without radiation therapy.

Potential role of nuclear magnetic resonance spectroscopy to identify salivary metabolite alterations in patients with head and neck cancer.

The analysis of the salivary metabolomic profile may offer an early phase approach to assess the changes associated with a wide range of diseases including head and neck cancer. The aim of the present study was to investigate the potential of nuclear magnetic resonance (NMR) spectroscopy for detecting the salivary metabolic changes associated with head and neck squamous cell carcinoma (HNSCC). Unstimulated whole-mouth saliva samples collected from HNSCC patients (primary tumour was located either in the larynx or in the oral cavity) and healthy controls were analysed by 1H-NMR spectroscopy. Reliably identified salivary metabolites were quantified and the determined concentration values were compared group-wise using a Mann-Whitney U-test. Multivariate discrimination function analysis (DFA) was conducted to identify such a combination of metabolites, when considered together, that gives maximum discrimination between the groups. HNSCC patients exhibited significantly increased concentrations of 1,2-propanediol (P=0.032) and fucose (P=0.003), while proline levels were significantly decreased (P=0.043). In the DFA model, the most powerful discrimination was achieved when fucose, glycine, methanol and proline were considered as combined biomarkers, resulting in a correct classification rate of 92.1%, sensitivity of 87.5% and specificity of 93.3%. To conclude, NMR spectrometric analysis was revealed to be a feasible approach to study the metabolome of saliva that is sensitive to metabolic changes in HNSCC and straightforward to collect in a non-invasive manner. Salivary fucose was of particular interest and therefore, controlled longitudinal studies are required to assess its clinical relevance as a diagnostic biomarker in HNSCC.

Oral mucosal epithelial cells express the membrane anchored mucin MUC1.

OBJECTIVE: The presence of a stable salivary pellicle (SP) is essential to provide a wet surface for the oral mucosal epithelia. The oral mucosa is covered by the SP which is suggested to be a mixed film of both salivary and epithelial components. Our aim was to analyse the presence of membrane-anchored mucin MUC1 in the oral mucosal epithelia. DESING: The presence of MUC1 was studied by immunohistochemical and immunoelectron microscopical methods in 19 buccal mucosal specimens. The localization and intensity of the epithelial expression were analyzed. RESULTS: Strong staining of MUC1 was found in the epithelial cells of intermediate and superficial layers. Some basal cells were shown faint expression. In the intermediate and superficial layers, the MUC1 expression was seen mainly on the upper cell surface. Furthermore, the expression of MUC1 was noted in the cytoplasm near the nucleus and in the rough granules. By electron microscopy, extracellular domain of membrane-anchored molecules extruded about 15-30nm above the cell surface in the apical cells of the oral epithelium. Immunoelectron microscopic examination shows that MUC1 is mainly localized in the plasma membrane of epithelial cells and also in small vesicles (75-100nm) just below the plasma membrane. CONCLUSION: The membrane-anchored MUC1 is expressed in the superficial layer of the oral mucosal epithelium, especially on the upper surface of epithelial cells. MUCI may be the anchoring protein of the salivary pellicle stabilization.

Fourier Transform Infrared Spectroscopy and Photoacoustic Spectroscopy for Saliva Analysis.

Saliva provides a valuable tool for assessing oral and systemic diseases, but concentrations of salivary components are very small, calling the need for precise analysis methods. In this work, Fourier transform infrared (FT-IR) spectroscopy using transmission and photoacoustic (PA) modes were compared for quantitative analysis of saliva. The performance of these techniques was compared with a calibration series. The linearity of spectrum output was verified by using albumin-thiocyanate (SCN(-)) solution at different SCN(-) concentrations. Saliva samples used as a comparison were obtained from healthy subjects. Saliva droplets of 15 µL were applied on the silicon sample substrate, 6 drops for each specimen, and dried at 37 ℃ overnight. The measurements were carried out using an FT-IR spectrometer in conjunction with an accessory unit for PA measurements. The findings with both transmission and PA modes mirror each other. The major bands presented were 1500-1750 cm(-1) for proteins and 1050-1200 cm(-1) for carbohydrates. In addition, the distinct spectral band at 2050 cm(-1) derives from SCN(-) anions, which is converted by salivary peroxidases to hypothiocyanate (OSCN(-)). The correlation between the spectroscopic data with SCN(-) concentration (r > 0.990 for transmission and r = 0.967 for PA mode) was found to be significant (P < 0.01), thus promising to be utilized in future applications.