Oral Exfoliative Cytology Research Articles

Advances in Oral Exfoliative Cytology: From Cancer Diagnosis to Systemic Disease Detection.

Oral exfoliative cytology has emerged as a valuable tool in the early detection of oral cancer and other systemic diseases. This review comprehensively examines the current applications and recent advancements in oral exfoliative cytology techniques. We analyzed published literature from the past decade, focusing on methodological improvements, diagnostic accuracy, and emerging applications. Key findings include: (1) Enhanced cell collection and preparation methods have significantly improved sample quality and diagnostic reliability. (2) Integration of molecular markers and DNA analysis with traditional cytomorphological assessment has increased diagnostic sensitivity and specificity for oral cancer detection. (3) Novel applications in systemic disease detection, including diabetes and iron overload disorders, demonstrate the expanding utility of this technique. (4) Computer-assisted analysis and deep learning algorithms show promise in improving diagnostic accuracy and efficiency. Despite these advancements, challenges remain in standardization and widespread clinical implementation. This review provides a critical evaluation of oral exfoliative cytology's current status and future potential in oral and systemic disease diagnosis.

Training high-performance deep learning classifier for diagnosis in oral cytology using diverse annotations

The uncertainty of true labels in medical images hinders diagnosis owing to the variability across professionals when applying deep learning models. We used deep learning to obtain an optimal convolutional neural network (CNN) by adequately annotating data for oral exfoliative cytology considering labels from multiple oral pathologists. Six whole-slide images were processed using QuPath for segmenting them into tiles. The images were labeled by three oral pathologists, resulting in 14,535 images with the corresponding pathologists’ annotations. Data from three pathologists who provided the same diagnosis were labeled as ground truth (GT) and used for testing. We investigated six models trained using the annotations of (1) pathologist A, (2) pathologist B, (3) pathologist C, (4) GT, (5) majority voting, and (6) a probabilistic model. We divided the test by cross-validation per slide dataset and examined the classification performance of the CNN with a ResNet50 baseline. Statistical evaluation was performed repeatedly and independently using every slide 10 times as test data. For the area under the curve, three cases showed the highest values (0.861, 0.955, and 0.991) for the probabilistic model. Regarding accuracy, two cases showed the highest values (0.988 and 0.967). For the models using the pathologists and GT annotations, many slides showed very low accuracy and large variations across tests. Hence, the classifier trained with probabilistic labels provided the optimal CNN for oral exfoliative cytology considering diagnoses from multiple pathologists. These results may lead to trusted medical artificial intelligence solutions that reflect diverse diagnoses of various professionals.

Cytopathological Analysis of Oral Buccal Mucosa of Dental College Students

In this study Cytopathological observations were recorded, including inflammation, microbial colonies, micronuclei, keratinisation, overlapping and hemorrhage of cells which are obtained from the buccal mucosa of the dental students to assess the fluctuation of the cellular characteristics among relatively normal subjects. This prospective study included 50 dental students with no detectable oral alterations and are submitted to brush cytology. The smears were fixed with 95% ethyl alcohol and stained with hematoxylin and eosin stain. The stained section were observed. Chi-square tests were applied for non-parametric variables. A p-value < 0.05 was considered statistically significant. The results showed close proximity among subjects in the matter of cytopathological observation and measures; no significant influence of sex was found on cytopathological observation, and vice versa. Oral exfoliative cytology combined with cytopathological analysis can be helpful for the study of normal individuals in various investigations of oral and systemic diseases.

The awareness and attitudes of oral disease specialists, pathologists, and maxillofacial surgeons in oral exfoliative cytology in Kerman, Iran

Background: The present work examines the awareness and attitudes of specialists in oral diseases, pathology, and maxillofacial surgery in 2022 regarding oral exfoliative cytology. Methods: This study was an analytical and cross-sectional research. The statistical population of the present study included Iranian oral disease specialists, pathologists, and maxillofacial surgeons. A researcher-made questionnaire was given to the specialists by a senior (final-year) student. The results were analyzed using the chi-square test and t-test using SPSS 18 software. The significance level in data analysis was P<0.05. Results: A total of 192 questionnaires were completed in the study; the study revealed that only 18 participants used the exfoliative cytology technique. Moreover, 62% of people had a positive attitude towards the application and performance of cytology. There was a significant relationship between positive attitude, field of study, graduation year, and age. The mean awareness scores in men and women were 32.38±4.21 and 34.42±3.89, respectively. The participants had a high awareness and positive attitude towards this technique. Additionally, the mean awareness scores of oral specialists, surgeons, and pathologists were 33.12±4.23, 33.65±5.12, and 33.45±5.34, respectively. Conclusion: The study revealed that only 9.4 percent of participants used the exfoliative cytology technique. However, they were highly aware of and had a positive attitude to this technique.

Automated Analysis of Nuclear Parameters in Oral Exfoliative Cytology Using Machine Learning.

As oral cancer remains a major worldwide health concern, sophisticated diagnostic tools are needed to aid in early diagnosis. Non-invasive methods like exfoliative cytology, albeit with the help of artificial intelligence (AI), have drawn additional interest. The study aimed to harness the power of machine learning algorithms for the automated analysis of nuclear parameters in oral exfoliative cytology. Further, the analysis of two different AI systems, namely convoluted neural networks (CNN) and support vector machine (SVM), were compared for accuracy. A comparative diagnostic study was performed in two groups of patients (n=60). The control group without evidence of lesions (n=30) and the other group with clinically suspicious oral malignancy (n=30) were evaluated. All patients underwent cytological smears using an exfoliative cytology brush, followed by routine Hematoxylin and Eosin staining. Image preprocessing, data splitting, machine learning, model development, feature extraction, and model evaluation were done. An independent t-test was run on each nuclear characteristic, and Pearson's correlation coefficient test was performed with Statistical Package for the Social Sciences(SPSS) software (IBM SPSS Statistics for Windows, Version 28.0. IBM Corp, Armonk, NY, USA). The study found substantial variations between the study and control groups in nuclear size (p<0.05), nuclear shape (p<0.01), and chromatin distribution (p<0.001). The Pearson correlation coefficient of SVM was 0.6472, and CNN was 0.7790, showing that SVM had more accuracy. The availability of multidimensional datasets, combined with breakthroughs in high-performance computers and new deep-learning architectures, has resulted in an explosion of AI use in numerous areas of oncology research. The discerned diagnostic accuracy exhibited by the SVM and CNN models suggests prospective improvements in early detection rates, potentially improving patient outcomes and enhancing healthcare practices.

Nuclear Changes in Controlled and Uncontrolled Type-2 Diabetic Patients In Buccal Exfoliative Cytology Smears Using Pap And Giemsa Staining – A Comparative Study

Background: Diabetes is a third leading cause of mortality and morbidity in the world. Oral exfoliative cytology may be a more appropriate adjunctive diagnostic tool in conditions like diabetes mellitus, where the invasive techniques lose viability. Exfoliative cytology helps to detect cellular aging, cell atrophy, alterations in nucleus and cytoplasm, binucleation and occasional karyorrhexis in type-2diabetic patients which proves to be a valuable tool for early diagnosis of diabetes. The purpose of this research was to develop a new method for an early diagnosis of diabetes mellitus by exfoliative cytology using PAP stain and GIEMSA stains which is a nuclear specific stain. The aim of the study was to compare the exfoliative cytology of oral mucosa in controlled and uncontrolled type-2 diabetic patients using PAP stain and GIEMSA stain detect the nuclear and cytoplasmic changes. Materials And Methods: The smears were obtained from buccal mucosa of the mouth in total of 25 individuals out of which 10 are controlled (&lt;120 mg/dl) and 10 are uncontrolled (&gt;130 md/dl) diabetic patients and 5 normal individuals. These smear samples was obtained from the clinical lab of Saveetha Dental College. Results: The nuclear changes include nuclear bud, binucleate/micronucleate, karyolysis, increased nuclear cytoplasmic ratio &amp; hyperchromatism. The binucleated cells were found to have higher differences between controlled &amp; uncontrolled whereas karyorrhexis showed no significant changes between controlled &amp; uncontrolled diabetes. Conclusion: This is a preliminary study conducted to analyse the potential interaction between the nuclear changes and the amount of genetic damage caused in diabetes. However, our result clearly demonstrated that uncontrolled diabetic patients have significantly more genetic damage (in terms of MN frequency) compared to controlled diabetic patients. This indicates than MN may be a useful constituent in a panel of biomarkers for the risk of diabetes.

Study the cytologic changes In Oral Mucosa of Normal and Diabetic patient using H&amp;E, PAP, PAS and PAS-Diastase Stain

Introduction: Diabetes mellitus is the fifth most common chronic condition and the sixth most frequent cause of death among elderly. An estimated 100 million people are affected by diabetes mellitus world-wide. It represents an extreme disturbance in glucose metabolism with severe hyperglycemia and insulin deficiency Objective: The objective of this research was to study the effect of diastase enzyme on the glycogen content of the epithelial cells of normal and diabetic patients in oral exfoliative cytology. Materials and Methods: Ten control subjects and ten diabetic patients (study group) were taken, four oral smears for both control and study group from the buccal mucosa were taken and stained with hematoxylin and eosin stain, Papanicolaou (PAP) stain, periodic acid Schiff (PAS) stain and PAS-Diastase (PAS-D) stain. Results: Glycogen is stained magenta with PAS and was absent on PAS(D) stained slide because of glycogen digestion by diastase enzyme. Such detection of glucose can help in the diagnosis of diabetes. Conclusion: The results of our study showed that exfoliative cytology of the oral cavity, when stained with PAS-D, can be used as an effective screening and diagnostic tool for DM patients.

Utility of Perl's Prussian Blue Stain in Exfoliated Buccal Cells of Thalassemia and Sickle Cell Anemia Patients and Their Correlation With Serum Ferritin Levels.

Introduction Iron is essential for all living beings. Excess iron, on the other hand, is dangerous because it causes the creation of free radicals. As a result, iron absorption is carefully managed to maintain a balance between absorption and iron loss in the body. Due to the lack of particular excretory channels for iron in humans, iron excess in the tissues is common. It can be caused by a number of factors, including increased iron absorption, as seen in hemochromatosis, or frequent parenteral iron treatment, as seen in thalassemia and sickle cell anemia patients (a transfusional overload). Aim The study aims to demonstrate Perl's Prussian blue stain to identify iron overload at a preliminary stage and correlate with serum ferritin levels in patients withthalassemia and sickle cell anemia who frequently receive blood transfusions. Materials and methods The present study comprised 62 confirmed cases of thalassemia and sickle cell anemia patients undergoing repeated blood transfusions of a minimum of 15/more, along with 62 clinically healthy individuals between December 2016 and November 2018. The patients with thalassemia and sickle cell anemia were confirmed by hemoglobin electrophoresis (Bio-Rad D-10, Bio-Rad Laboratories, Inc, California, United States). The buccal smears were obtained from these patients along with the controls, and these slides were stained by Perl's Prussian blue stain and were examined under a light microscope. Results Sixty-two cases and 62 controls were considered in the current investigation. Forty-seven of the 62 people had thalassemia, and 15 had sickle cell anemia. Thirty-nine out of the 47 patients with thalassemia and six of the 15 individuals with sickle cell anemia had positive results for Perl's Prussian blue stain. All patients had elevated blood ferritin levels, with varying ranges associated with positive results for Perl's Prussian blue stain. Conclusion The objective of this study was to demonstrate the utility of oral exfoliative cytology in thalassemia and sickle cell anemia patients who often receive blood transfusions as a screening and diagnostic tool. The exfoliative cytology methods' acceptability and simplicity, along with their correlation with serum ferritin levels and Perl's Prussian blue reaction, make this noninvasive procedure an excellent screening and diagnostic tool for all patients who receive repeated blood transfusions.

Micronuclei detection in oral cytologic smear: does it add diagnostic value?

BackgroundScreening and early diagnosis of oral squamous cell carcinoma (OSCC) are directly associated with increased survival rate and improved prognosis. Noninvasive diagnostic tools have been implemented in the early detection as toluidine blue staining, optical imaging, and oral cytology. This study aimed to assess and compare the presence of micronuclei (MN) in oral exfoliative cytology of healthy controls, subjects exposed to high-risk factors for oral cancer, subjects with oral potentially malignant lesions (OPMLs), and those with malignant oral lesions.Subjects and methodsA total number of 92 subjects were divided into 46 healthy controls with no oral mucosal lesions (23 with no evidence of cancer risk factors and 23 with cancer risk factors), 23 with OPMLs and 23 with oral malignant lesions. All the 92 participants were subjected to cytological sampling for detection of MN. The final diagnosis of the oral lesions was confirmed by the histopathological picture and compared to the cytological results.ResultsThe results showed that the diagnostic accuracy of MN was higher in OPMLs group (95.2%). The sensitivity of MN test in malignant group was much lower (52.2%); however, all the cytological criteria of malignancy were markedly detected as compared to the OPMLs group.ConclusionsConventional oral cytology supported by MN is highly beneficial as adjunctive tool in the screening for early detection of dysplastic oral lesions.

Comparative Analysis of Cytomorphology in Adolescent and Geriatric Patients: A Cross-Sectional Study.

Background Exfoliation of the surface cells of the normal epithelium occurs as a result of physiological turnover.Epithelial cells are constantly renewed, and they are exfoliated or shed as they migrate from the basal layer to the uppermost layer of the epithelium. Oral exfoliative cytology involves the collection and microscopic evaluation of these shed cells or scraped epithelial cells, quantitatively and qualitatively. The objective of the present study was to analyze and compare the cytomorphometric features like cellular area, nuclear area, and nuclear-cytoplasmic ratio from buccal mucosal smears of adolescent and geriatric patients. This study highlights the changes in cell morphology in different age groups (adolescent and pediatric), which could be attributed to hormonal, habitual, and aging factors. Material and methods Buccal smear sample was collected from a total of 60 individuals belonging to the age groups of 11-19 years and above 60 years. The smears were stained with H&E and PAP (Papanicolaou) stain. Photomicrographs were taken in 40x, and measurements were calculated using ImageJ software (National Institutes of Health, Bethesda, Maryland, United States). Cellular size, nuclear size, and nucleo-cytoplasmic ratio were analyzed and compared between the two age groups using independent t-tests using IBM SPSS Statistics for Windows, Version 22.0 (Released 2013; IBM Corp., Armonk, New York, United States). Results A significant difference was observed in the cellular area and nuclear area between the two age groups with a p-value of 0.00. No significant findings were present in the nucleo-cytoplasmic area of the two age groups. Conclusion Cytomorphometric analysis has shown that there were variations in the cytoplasmic and nuclear areas among different age groups.

Cytological Detection of Iron Overload and Cytomorphometric Changes in buccal Mucosa of Βeta Thalassemia Major Patients

Aim: In this study, the phenomenon of oral exfoliative cytology is explored and buccal squamous cells are examined for intracytoplasmic, Prussian blue stained, iron containing granules. In addition, these buccal squamous cells are subjected to morphometric measurements of their nuclear and cytoplasmic diameter along with their ratios. Methods: It was a Descriptive, Cross sectional study including 120patients of Thalassemia Major, ranging from 4 -18 years with repeated blood transfusions and a serum ferritin level of 1000ng/ml at minimum. Scrapings were obtained from buccal mucosa for cytological examination and stained with Prussian Blue Stain. Results: The study included 45 female and 75 male patients with a mean age 11.04(± 3.81) years. 56out of 120 patients were positive for Prussian blue and thus showed 46.6% positivity. The mean cytoplasmic diameter was 50.90 (± 0.64) microns and mean nuclear diameter was 9.31(±0.53) microns. Mean Nuclear to cytoplasmic (NC) ratio was 0.182(± 0.01) and ranged from 0.165 and 0.211 microns. Conclusion: The positivity of Prussian blue in our study is clearly less than most of the quoted figures while alterations in morphometric features of buccal squamous cells are quite consistent with reported data. In our study, we observed iron containing granules not only in the cytoplasm but also on surface of cells like a clue cell dispersed outside the cell, individually or in discrete clusters and as faint bluish hue in background. Keywords: Iron overload, Cytomorphometry, buccal smear, Prussian blue, Beta Thalassemia

Uncommon case of cytopathological features of sialadenoma papilliferum.

Sialadenoma papilliferum, a benign and rare salivary gland neoplasm, accounts for 0.4%-1.2% of all salivary gland tumors and occurs primarily in minor salivary glands of the oral cavity. Here, we report a case of sialadenoma papilliferum and its associated cytological findings. A papillary tumor was incidentally detected on the palate of an 86-year-old Japanese man. Conventional oral exfoliative cytology was performed; the cytology smear exhibited epithelial clusters composed of atypical epithelial cells with a high nuclear/cytoplasm ratio and arranged in sheet or small papillary-like projections. Cytoplasmic vacuoles were also observed in the papillae. It was difficult to make a definitive diagnosis due to the presence of uncommon cytological features. The excisional biopsy specimen revealed histological features characteristic of sialadenoma papilliferum. Mutational analysis detected BRAFV600E mutation, which confirmed the diagnosis of sialadenoma papilliferum. To the best of our knowledge, no prior cytomorphological evaluations of sialadenoma papilliferum have been reported in detail. Oral exfoliative cytology specimens from salivary gland tumors can demonstrate uncommon cytomorphological features. A differential diagnosis of sialadenoma papilliferum can be based on the observation of mildly atypical epithelial cells that form small papillary-like structures.

Detection and Comparision of Cytopathological Changes in Smokers and Non-Smokers: A Comparative Study

Background: Oral cancer is one of the sixth most common cancers in the world. Oral exfoliative cytology is one of the popular screening tool for oral cancer. Use of tobacco in any form are documented as the most common cause as initiators for dysplastic changes in oral mucosa. The purpose of the study was to detect the cytological changes in buccal mucosa, tongue and palate among non-smokers smokers. Material Methods: Smears sample were collected according to site (buccal mucosa, tongue palate) from 100 subjects among smokers non-smokers. Smears were then stained using Papanicolaou staining technique. Results: Among the smokers and non-smokers the results were statistically significant. Conclusion: Recent advances in the clinical visualization and detection of the oral mucosa have made the viability of cytological procedures more specific and sensitive. Contact endoscopy and use of autofluorescence devices are the forerunners in this group. The fluorescence characteristics of tissues depend upon their biochemical composition and histomorphological architecture, both of which undergo a change during malignant transformation. These changes are detectable as an alteration in the fluorescence spectral profile of the tissues21. Due to low feasibilities of such devices the benchmark of diagnosis will be microscopic tissue examination. Hence cytological smears will always be highly specific, sensitive, easy to use and reproducible procedures in routine screening of population for potentially and malignant conditions of the oral cavity.

Diagnostic and Prognostic Implications of the Oral Exfoliative Cytology in Oral precancer And Cancer Detection

Diagnostic and Prognostic Implications of the Oral Exfoliative Cytology in Oral precancer And Cancer Detection

Exfoliative Cytology and Genetic Analysis for a Non-Invasive Approach to the Diagnosis of White Sponge Nevus: Case Series

White Sponge Nevus (WSN) is a rare benign disorder associated with mutations in genes coding for cytokeratin 4 (KRT4) and 13 (KRT13) characterized by dyskeratotic hyperplasia of mucous membranes. This study was aimed at examining different approaches (cytology, pathology and genetic analysis) to WSN diagnosis. A series of four patients with asymptomatic white diffuse oral lesions were evaluated and, before performing an incisional biopsy for pathology, an oral brush Thin Prep was collected for exfoliative liquid-based cytology (LBC). DNA for genetic analysis was also obtained from patients and both their parents, using buccal swabs. Pathology and cytology showed similar results, leading to the same diagnosis of hyperkeratotic epithelium with acanthosis and spongiosis, without atypia, demonstrating the efficiency of LBC for the differential diagnosis. Sequencing analysis revealed at least 6 rare variants in the KRT4 and KRT13 genes in each patient, contributed in part by both unaffected parents. Thin Prep for oral exfoliative cytology and genetic analysis are sufficient for an accurate diagnosis of WSN. The combination of cytological and genetic analyses could substitute the histologic exam, providing a non-invasive alternative for incisional biopsy.

Cytomorphometric Analysis of Oral Buccal Mucosa of Dental Colleges' Students in Sulaimani City.

This study evaluates the cytomorphometric measures of cells obtained from the buccal mucosa of dental students to assess the fluctuation of the cellular characteristics among relatively normal subjects, with any potential correlations with demographic information, different habits, and hormonal disturbance. This prospective study included 100 dental students with no detectable oral alterations submitted to brush cytology. The smears were fixed with 95% ethyl alcohol and stained with hematoxylin and eosin stain. The stained section was observed under an image analyzer for cytomorphometric analysis. Cytopathological observations were recorded, including inflammation, microbial colonies, micronuclei, keratinization, overlapping, and hemorrhage. Chi-square tests were applied for non-parametric variables. One-way analysis of variance (ANOVA) was used to compare the cytometric parameters to habits and hormonal disturbances. A p-value &lt; 0.05 was considered statistically significant. The results showed close proximity among subjects in the matter of cytomorphometric measures; no significant influence of sex, smoking, alcohol drinking habits, and menstruation was found on cytomorphometric diameters or cytopathological observation, and vice versa, while polycystic ovary syndrome impacted nuclear and nuclear-cytoplasmic ratio (p = 0.003, p = 0.02), respectively. Oral exfoliative cytology combined with cytomorphometric analysis for the studied normal individuals can be helpful in various investigations of oral and systemic diseases.

Absence of Synergistic Effect of Toluidine Blue and Cytomorphometry in Discriminating Dysplasia in Oral Exfoliative Cytology

Numerous tools and techniques such as vital stain, autofluorescence, exfoliative cytology, histomorphometry, etc. were explored in the detection of dysplasia, but have variable sensitivity and specificity. The present study evaluated the effect of toluidine blue in increasing the accuracy of histomorphometry analysis in detection of the dysplasia in OPMDs on exfoliative cytology. An experimental, observational, cross-sectional research was performed on patients attending dental hospitals and in

Periodic acid–Schiff staining in oral exfoliative cytology of diabetic patients: The odyssey for noninvasive screening – A systematic review and meta-analysis

Periodic acid–Schiff staining in oral exfoliative cytology of diabetic patients: The odyssey for noninvasive screening – A systematic review and meta-analysis

Effectiveness of Rapid, Economic, Acetic Acid, Papanicolaou Stain (REAP) Over Conventional Papanicolaou Stain in Oral Exfoliative Cytology

Effectiveness of Rapid, Economic, Acetic Acid, Papanicolaou Stain (REAP) Over Conventional Papanicolaou Stain in Oral Exfoliative Cytology

Effective deep learning for oral exfoliative cytology classification

The use of sharpness aware minimization (SAM) as an optimizer that achieves high performance for convolutional neural networks (CNNs) is attracting attention in various fields of deep learning. We used deep learning to perform classification diagnosis in oral exfoliative cytology and to analyze performance, using SAM as an optimization algorithm to improve classification accuracy. The whole image of the oral exfoliation cytology slide was cut into tiles and labeled by an oral pathologist. CNN was VGG16, and stochastic gradient descent (SGD) and SAM were used as optimizers. Each was analyzed with and without a learning rate scheduler in 300 epochs. The performance metrics used were accuracy, precision, recall, specificity, F1 score, AUC, and statistical and effect size. All optimizers performed better with the rate scheduler. In particular, the SAM effect size had high accuracy (11.2) and AUC (11.0). SAM had the best performance of all models with a learning rate scheduler. (AUC = 0.9328) SAM tended to suppress overfitting compared to SGD. In oral exfoliation cytology classification, CNNs using SAM rate scheduler showed the highest classification performance. These results suggest that SAM can play an important role in primary screening of the oral cytological diagnostic environment.