Diagnosis of differentiated dysplasia as a variant of oral epithelial dysplasia.

OBJECTIVES: The World Health Organization's definition of oral epithelial dysplasia includes differentiated dysplasia, which is defined by purely architectural abnormalities of oral mucosa without cytological changes. We analysed differentiated dysplasia's frequency, progression risk and correlation with oral brush cytology. MATERIALS AND METHODS: Cytoarchitectural criteria and expression patterns of keratin 13/17 and ki67 were studied in oral biopsies clinically diagnosed with leukoplakia. Biopsies were assessed for dysplasia and its grade. Available brush cytology findings were obtained from clinical records. RESULTS: We included 159 biopsies from 112 patients (33% differentiated dysplasia; 27% keratosis without dysplasia; oral epithelial dysplasia with atypia of mild, moderate and severe degree including invasive cancers in 9%, 8% and 7%, respectively). Keratin 13 loss and keratin 17 gain were higher in differentiated-dysplasia cases (p < 0.0001), which had the highest hypergranulosis frequency. Keratin 17 expression was associated with higher malignant-transformation rates (p = 0.0028). The transformation rate and time were comparable between dysplasia with atypia and differentiated-dysplasia cases, which had higher progression rates and shorter time periods than keratosis cases without dysplasia (p = 0.08). Cytology prior to differentiated dysplasia all indicated normal oral mucosa. CONCLUSIONS: Keratin 17 but not oral brush cytology can help identify patients with differentiated dysplasia with higher risk for malignant transformation.

Retrospective evaluation of the oral brush biopsy in daily dental routine - an effective way of early cancer detection.

OBJECTIVES: Oral brush biopsies are a well researched index for early detection of oral cancer in specialised centers. But the performance of the exfoliative biopsy is not yet researched in daily dental routine. METHODS: Private dentists and private oral surgeons in Germany took brush biopsies out of 814 suspicious lesions from 670 patients using the Orcellex brush while regular dental appointments. The analyses of the biopsies were performed by the Cytological Laboratory of Bonn (CLB) using liquid-based cytology. RESULTS: The final results were 74 oral squamous cell carcinomas and one verrucous carcinoma, histological proven, 232 cases of leukoplakia, 242 cases of lichen planus, 17 cases of erythroplakia, 259 cases of benign inflammatory, traumatic or hyperplastic oral lesions. The sensitivity for the detection of cancer cells using brush biopsy archived 100%, the specificity for the detection of non-neoplastic cells was 86.5%. The positive predictive value was 43.1%, the negative predicative value was at 100%. CONCLUSION: The oral brush biopsy seems to be a sufficient tool for early cancer detection in private dental offices. CLINICAL RELEVANCE: Generally, practicing dentists do not see various oral squamous cell carcinomas in their careers, so the experience in identifying oral squamous cell carcinomas as such is very low. The brush biopsy might help them in cases of doubt to prevent tumors from expansive growth.

Comparison between two cell collecting methods for liquid-based brush biopsies: a consecutive and retrospective study.

BACKGROUND: This study compares two different cell collectors, the Orcellex Brush (rigid brush) and the Cytobrush GT (nylon brush), using liquid-based cytology. A comparison of their obtainment procedures was also considered. The aim was to determine the diagnostic accuracy for detection of malignancy in oral brush biopsies. PICO-Statement: In this consecutive and retrospective study we had as population of interests, patients with oral lesions, the intervention was the brush biopsy with two different cell collectors and the control was healthy oral mucosa. The outcome of the study was to compare both cell collectors. METHODS: From 2009 to 2018, 2018 patients with oral lesions were studied using the nylon brush (666 cases) and rigid brush (1352 cases). In the first cohort five smears per patient were taken with the nylon brush, while each patient received one smear with the rigid brush in the second cohort. These were further processed in a liquid-based procedure. Cytological evaluations were categorised into 'negative', which were considered as negative, whereas 'doubtful', 'suspicious' and 'positive' cytological results were overall considered as positive for malignancy in comparison to the final histological diagnoses. Additionally, the clinical expenditure for each collector was estimated. RESULTS: 2018 clinically and histologically proven diagnoses were established, including 181 cases of squamous cell carcinomas, 524 lichen, 454 leukoplakias, 34 erythroplakias and 825 other benign lesions. The sensitivity and specificity of the nylon brush was 93.8% (95% CI 91.6-95.5%) and 94.2% (95% CI 91.8-95.5%) respectively, whereas it was 95.6% (95% CI 94.4-96.6%) and 84.9% (95% CI 83.8-87.5%) for the rigid brush. The temporal advantage using the plastic brushes was 4× higher in comparison to the nylon brush. The risk suffering from a malignant oral lesion when the result of the brushes was positive, suspicious, or doubtful was significantly high for both tests (nylon brush OR: 246.3; rigid brush OR: 121.5). CONCLUSIONS: Both systems have a similar sensitivity, although only the rigid brush achieved a satisfactory specificity. Additional methods, such as DNA image cytometry, should also be considered to improve the specificity. Furthermore, the rigid brush proved to be more effective at taking a sufficient number of cells, whilst also being quicker and presenting less stress for the patient.

Assessment of the cancerization risk for oral potentially malignant disorders by clinical risk model combined with autofluorescence and brush biopsy with DNA-image cytometry.

PURPOSE: To explore the feasibility of assessing the cancerization risk of oral potentially malignant disorders (OPMD) through a clinical risk model combined with autofluorescence and brush biopsy with DNA-image cytometry. METHODS: We collected the baseline clinical data of 269 patients; then, performed autofluorescence, brush biopsy with DNA-image cytometry and histopathological examination. Then, we obtained the significant factors by univariate logistic analysis, constructed the clinical risk model by multiple logistic regression and selected the optimal cutoff value according to the maximum Youden index. Finally, we calculated the sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) of the clinical risk score ≥ cutoff value, autofluorescence and brush biopsy with DNA-image cytometry, and plotted the receiver-operating characteristic (ROC) curves and decision curve analysis (DCA). RESULTS: The clinical risk model is represented by the formula: 1 × gender + 1.6 × age group + 1 × lesion site + 1.4 × local stimulus + 1.5 × drink. The area under the curve (AUC) was 0.83, and the optimal cutoff score was 3. The AUC indicated that the clinical risk score ≥ 3 (0.74) and autofluorescence (0.77) had a certain diagnostic values, while brush biopsy with DNA-image cytometry (0.92) displayed a good value. Besides, the DCA showed that all three tests had clinical significance. CONCLUSIONS: The cancerization risk of patients can be assessed by the clinical risk model combined with sequence application of autofluorescence and brush biopsy with DNA-image cytometry, to decide whether histopathological examination or other intervention measures should be selected.

Liquid-based oral brush cytology in the diagnosis of oral leukoplakia using a modified Bethesda Cytology system.

OBJECTIVES: This study aimed to develop diagnostic criteria to identify oral squamous cell carcinoma (OSCC) and oral potentially malignant disorders (OPMDs) using oral liquid-based brush cytology (OLBC), and to compare its accuracy with the gold standard of surgical biopsy and histopathological diagnosis of oral leukoplakia. METHODS: A total number of 134 samples were collected. All patients underwent Orcellex® brush biopsy with liquid-based cytology immediately prior to diagnostic surgical biopsy. A preliminary study was first performed utilizing samples from 4 distinct lesion groups (20 samples) to revise the 2014 Bethesda Cytology system for use with OLBC specimens. RESULTS: Five diagnostic groups of OLBC for the diagnosis of OSCC and OPMDs with relevant cytopathological features were established. From the 114 samples in the test group, 101 were included. The other 13 were excluded due to inadequate cellularity. The test showed sensitivity, specificity, positive predictive value and negative predictive value of 75%, 76%, 76% and 75%, respectively, and an accuracy of 75%. The use of the oral brush sampling technique was well accepted by multiple clinicians; however, local anaesthetic was suggested to be useful prior to performing the brush biopsy. CONCLUSIONS: Oral liquid-based brush cytology using the Orcellex® brush and ThinPrep® system is a simple and minimally invasive procedure for adequate intraepithelial sampling and can be used as an adjunct for the early detection of oral cancer. The modified Bethesda system established useful means for OLBC assessment that can be utilized in future studies to increase the standardization of oral cytology assessment.

The prognostic value of GLUT-1 staining in the detection of malignant transformation in oral mucosa.

BACKGROUND: Head and neck squamous cell carcinoma (HNSCC) ranks as the sixth most common tumor entity worldwide. Unfortunately, the multimodal treatment consisting of surgery, radiation, and chemotherapy does not show the desired efficacy. The intent of this study was to evaluate the sensitivity and specificity of an oral brush biopsy in combination with glucose transporter (GLUT)-1 staining in identifying premalignant and malignant lesions. METHODS: A total of 72 patients were included in the study, divided into four diagnostic subgroups (24 healthy, 15 carcinoma, 18 leukoplakia, 15 oral lichen planus). Oral brush biopsies were taken and analyzed for GLUT-1 expression by immunocytologic staining. Incisional biopsy served as the gold standard. RESULTS: Twelve (80 %) of the 15 carcinomas, nine (50 %) of the 18 leukoplakia, nine (60 %) of the 15 oral lichen planus, and none of the healthy specimens stained positive for GLUT-1. This resulted in a sensitivity rate of 80 % and a specificity rate of 68.42 %. Diagnostic accuracy was 70.83 % based on the correct diagnoses in 51 of 72 patients. CONCLUSION: An oral brush biopsy can easily be performed throughout the entire oral cavity, is noninvasive, and shows high sensitivity and specificity rates with conventional cytology or computer-assisted analysis. CLINICAL RELEVANCE: The significance of GLUT-1-specific staining with an oral brush biopsy is more limited than expected but could be used as an additional tool in detecting malignant transformation in the oral cavity.

Liquid-based versus conventional cytology of oral brush biopsies: a split-sample pilot study.

OBJECTIVE: The aim of this prospective split sample study was to evaluate the applicability of liquid-based cytology (LBC) of oral brush biopsies for detection of oral cancer. METHODS: Two different preparation techniques were investigated: the conventional transfer procedure to glass slides and the LBC preparation method. The obtainments of epithelial cells were performed five times with a nylon brush and transferred onto five glass slides. Additionally, the brushes, which were normally discarded, were stored in a fixative solution. Conventional slides and respective thin layers from a total of 113 cases were reviewed with both techniques. RESULTS: Thin layers showed excellent morphology on a clear background, which allowed an accurate diagnosis. In contrast, the conventional glass slides showed significantly more blood contamination and cell overlapping. The sensitivity of conventional cytological diagnosis was 96.3%, the specificity archived 90.6%, the positive predictive value was 96.3% and the negative predictive value scored 90.6%. The sensitivity of the cytological diagnosis using thin layers archived 97.5%, the specificity was 68.8%, the positive predictive value revealed 88.76% and negative predictive value was 91.7%. CONCLUSION: Our findings indicate that in oral cytology, LBC may replace other types of wet-fixed preparations using the full amount of collected cells, resulting in enhanced specimen quality archiving comparable values of diagnostic accuracy. CLINICAL RELEVANCE: LBC facilitates the cell collection due to simpler handling and less transfer errors by dentists and may improve the overall diagnostic accuracy of oral brush biopsies in future.

Adjunctive screening devices for oral lesions: their use by Canadian Dental Hygienists and the need for knowledge translation.

Screening for oral cancer and other mucosal conditions is a knowledge-to-action objective that should be easy: there is supportive evidence, it is fast and non-invasive, and the oral cavity is easy to visualize. However, over 60% of oral cancers are diagnosed late, when treatment is complex and prognosis poor. Adjunctive screening devices (ASDs), e.g. toluidine blue (TB), fluorescence visualization (FV), chemiluminescence (CL) and brush biopsies, were designed to assess risk of oral lesions or aid in identification and localization of oral premalignant and malignant lesions. Little is known on how clinicians feel about using ASDs. OBJECTIVES: To evaluate use and level of comfort in using ASDs for oral cancer screening among dental hygienists. METHODS: Online email survey of a stratified random sample of nearly 3000 dental hygienists from four Canadian provinces. RESULTS: A total of 369 hygienists responded about ASDs. Ninety-three (25%) had used an ASD. Use was associated with six or more continuing education (CE) courses per year (P = 0.030), having a CE course in oral pathology within the last 3 years (P = 0.003) and having a screening protocol (P = 0.008). The most commonly used ASD is FV, which was the tool hygienists felt most comfortable using. Few used brush biopsies. Older graduates were more comfortable using TB (P = 0.014) and CL (0.033). CONCLUSION: Current evidence and education through CE appears to bolster knowledge translation efforts for hygienists to become more comfortable in the use of ASDs. ASDs with minimal supporting evidence and not specifically targeted to hygienists, such as the brush biopsies, are not well utilized.

Programmable bio-nanochip-based cytologic testing of oral potentially malignant disorders in Fanconi anemia.

Fanconi anemia (FA) is caused by mutations of DNA repair genes. The risk of oral squamous cell carcinoma (OSCC) among FA patients is 800-folds higher than in the general population. Early detection of OSCC, preferably at it precursor stage, is critical in FA patients to improve their survival. In an ongoing clinical trial, we are evaluating the effectiveness of the programmable bio-nanochip (p-BNC)-based oral cytology test in diagnosing oral potentially malignant disorders (OPMD) in non-FA patients. We used this test to compare cytomorphometric and molecular biomarkers in OSCC cell lines derived from FA and non-FA patients to brush biopsy samples of a FA patient with OPMD and normal mucosa of healthy volunteers. Our data showed that expression patterns of molecular biomarkers were not notably different between sporadic and FA-OSCC cell lines. The p-BNC assay revealed significant differences in cytometric parameters and biomarker MCM2 expression between cytobrush samples of the FA patient and cytobrush samples of normal oral mucosa obtained from healthy volunteers. Microscopic examination of the FA patient's OPMD confirmed the presence of dysplasia. Our pilot data suggests that the p-BNC brush biopsy test recognized dysplastic oral epithelial cells in a brush biopsy sample of a FA patient.

The impact of GSTM1/GSTT1 polymorphism for the risk of oral cancer.

OBJECTIVES: Since development of oral squamous cell cancer (OSCC) is triggered by various noxa, different variants of the antioxidant glutathione S-transferases (GSTs) can counteract toxic compounds (e.g., tobacco smoke). Because different polymorphisms of GST are known to have an increased sensitivity to carcinogenic agents, the aim of this study was to analyze whether GSTM1 or GSTT1 polymorphisms increase the risk for the development of OSCC. MATERIALS AND METHODS: GSTM1 and GSTT1 polymorphism was examined in healthy volunteers (n = 93) and in patients with OSCC (n = 100) by PCR after brush biopsy of oral mucosa. Odds ratio (OR) was calculated to evaluate the risk of oral cancer development. RESULTS: GSTM1 and GSTT1 deletion was found in 57% (53/93) and 18% (17/93), respectively, in healthy patients, while the OSCC group showed 57% (57/100) for GSTM1 deletion and 22% (22/100) with a deletion of GSTT1. Odds ratio for GSTM1 polymorphism was 1.00 and for GSTT1 1.26. Comparing smokers and nonsmokers with GSTM1 deletion polymorphism, OR was 4.35, while smokers without GSTM1 deletion showed an OR of 1.45. Adapting these data to the smoking habits of the general population in Germany, the OR was 9.25 for smokers with a GSTM1 deletion and OR 6.68 for smokers without a GSTM1 deletion. In smokers with GSTT1 deletion polymorphism, OR was 1.6 (adapted to the smoking habits of the general population: OR 6.16) and 3.16 (OR 8.56) in smokers without deletion in GSTT1 gene. CONCLUSIONS: Analysis of GST-M1 polymorphism in smokers could help to identify patients with a higher risk for the development of oral cancer. CLINICAL RELEVANCE: Early detection of OSCC due to a close meshed monitoring program for patients with GST-M1 polymorphism could help to improve the patient outcome. For polymorphism investigations, the oral brush biopsy is a sufficient method to gain DNA material.

An alternative method in the diagnosis of intrabiliary lesions: Percutaneous endobiliary brush biopsy.

Background and Aim: Biliary strictures can occur as a result of various benign or malignant processes. The aim of this study is to evaluate the effectiveness and reliability of percutaneous endobiliary brush biopsy in the diagnosis of intrabiliary lesions. Materials and Methods: This retrospective, single-center study was conducted between January 2022 and April 2023, involving a total of 16 patients. Of the patients, 10 were male (62.5%), and 6 were female (37.5%). The average age of the patients was 68.1±8. All patients underwent the procedure using an endobiliary biopsy brush under ultrasound and fluoroscopic guidance. Results: Technical success was achieved in all patients (100%). Cell detection was not observed in biopsy samples from 2 patients (12.5%), resulting in a diagnostic success rate of 87.5%. Access was made to the right biliary system in 14 patients (87.5%) and to the left biliary system in 2 patients (12.5%). Biopsy locations included the common bile duct in 12 patients (75%), hepatic hilum in 2 patients (12.5%), and bilioenteric anastomosis line in 2 patients (12.5%). The mean fluoroscopy time was 16.2±7.1 minutes. The average radiation dose was 660±370 mSv. Pathological diagnosis revealed malignancy in 8 patients (50%) and benign findings in 6 patients (37.5%). Liver abscess requiring drainage developed in 2 patients (12.5%). Conclusion: Percutaneous endobiliary brush biopsy performed under imaging guidance is an effective and reliable method for the diagnosis of biliary lesions.

Evaluation of Usage of Different Diagnostic Aids for Oral Cancer by Oral and Maxillofacial Surgeons: An Original Research.

Objective: The purpose of this study was to assess how oral and maxillofacial surgeons used various diagnostic tools for oral cancer. Materials and Methods: A cross-sectional methodology was used, and a standardized questionnaire was given to oral and maxillofacial surgeons randomly chosen sample. The questionnaire gathered information on demographics and the use of diagnostic tools. Data analysis methods included Chi-square testing and descriptive statistics. Results: The study included 200 oral and maxillofacial surgeons in total. The most often used diagnostic tool (95%) was visual inspection, followed by toluidine blue staining (48%) and brush biopsy (32%). Less frequently used were newer methods like optical coherence tomography (12.5%) and autofluorescence imaging (15%). No significant correlations between demographic factors and patterns of use of diagnostic tools were found by Chi-square tests. Conclusion: The results show that oral and maxillofacial surgeons frequently use brush biopsy, toluidine blue staining, and ocular evaluation. However, there is a need for more widespread adoption of cutting-edge technologies. By removing obstacles and offering training opportunities, one can increase the use of diagnostic tools, improving patient outcomes and the diagnosis of oral cancer.

Assessment of brush biopsy findings and salivary LDH levels in oral mucosal lesions of tobacco users.

Introduction: The oral brush cytology is an alternative method developed to improve the efficacy of conventional cytology in oral potentially malignant disorder (OPMD), and salivary lactate dehydrogenase (LDH) which is a cytoplasmic enzyme has been widely used as a marker for diagnosing various diseases. The purpose of the study is to evaluate the brush biopsy findings and salivary LDH levels for the early diagnosis of premalignant and malignant lesions of the oral cavity. Materials and Methods: Patients with deleterious habits including tobacco-related lesions such as leukoplakia, tobacco pouch keratosis, and oral cancer were included in the study. For each patient, saliva sample was collected, brush biopsy was done and smears were prepared. Collected saliva samples were analysed for salivary LDH levels and prepared smears were analysed for dysplastic changes and statistical analysis was performed. Results: Out of 80 samples, 30 were leukoplakia, 45 were tobacco pouch keratosis and 5 were oral cancer, and 13 samples showed positive dysplastic changes, 26 samples showed atypical dysplastic changes and 41 samples showed no signs of dysplastic changes and concluded as negative. On comparing the results of brush biopsy findings and salivary LDH levels, the mean salivary LDH value for positive dysplasia was elevated and the P value was statistically significant (P value: 0.00). Conclusion: Brush biopsy showed good potential in detecting premalignant lesions and salivary LDH levels showed a marked increase which can be used as a diagnostic biomarker and serve as a potent diagnostic aid for early detection of malignancy.

Culture expansion of primary human nasal epithelial cells (NEC) isolated with a nasal scraping spoon.

OBJECTIVE: To obtain high-purity nasal epithelial cells (NEC) while avoiding the irritation experienced by patients during nasal biopsies. METHODS: This prospective, observational study enrolled patients undergoing surgical treatment for nasal septum deviation. After general anaesthesia, a novel nasal scraping spoon was used to collect epithelial cells from the mid-part of the inferior turbinate. The cells were evenly plated on six-well plates coated with rat tail collagen. The morphology and growth of the cells were observed at different time-points using an inverted phase-contrast microscope. Immunofluorescent staining of cytokeratin 18 was used to identify NEC. Ki67 staining was used to check cell viability. RESULTS: This study collected samples from 19 patients during a short procedure. No postoperative complications were observed. Cell samples ranging from 8.31 × 105 to 2.04 × 106 cells/sample were obtained. The culture model was suitable for primary NEC culture as demonstrated by the faster proliferation (5-7 days). There was no fungal or bacterial contamination. Immunofluorescent staining confirmed the presence and proliferative activity of NEC in the cultures. CONCLUSION: A novel nasal scraping spoon provided an easy sampling method, avoided nasal injuries and psychological barriers to sampling and sufficient viable NEC to establish primary cultures.

Assessment of Ploidy Status in Oral Potentially Malignant Disorders - A Systematic Review.

Malignant and potentially malignant epithelial lesions are often associated with various abnormalities such as epithelial dysplasia, abnormal DNA content, loss of heterozygosity, and chromosomal number aberrations. Screening and early detection of such abnormalities facilitates proper care and also helps to prevent further progression of potentially malignant lesions to malignancy. In such way, the presence of DNA aneuploidy in oral potentially malignant disorders (OPMDs) may serve as an indicator for the malignant transforming potential. Various assessment methods have been proposed to find the DNA ploidy status of cells. This current systematic review is mainly designed to assess the importance of ploidy status in OPMD while measuring the feasibility of using this biomarker for evaluating the hazard of malignant transformation. As an upshot of this systematic review, we can conclude that use of DNA ploidy status can serve as an independent bio-marker for predicting the malignant transformation of lesions. Furthermore, as a future scope the use of DNA ploidy analysis in normal mucosa of smokers will help to assess the malignancy risk and this technique might also help to predict the genetic predisposition of patients with malignancy.

Analysis of laryngeal brush biopsy-based cytology results in patients of the 4th Military Teaching Hospital and Polyclinic in Wroclaw in years 2019-2020.

<b>Introduction:</b> Cytological examination of exfoliated epithelial cells of the uterine cervix, oral cavity, or rectum has been successfully used in the diagnostics of pathological conditions of these organs for many years. In these cases, the test material is collected from the available regions. </br></br> <b>Aim:</b> The aim of the study consisted in the analysis of cytological smears of laryngeal epithelial cells from patients hospitalized at the Department of Otolaryngology, Head and Neck Surgery of the 4th Military Teaching Hospital and Polyclinic in Wroclaw in years 2019-2020. The analysis was aimed at demonstrating whether representative laryngeal epithelial material could be obtained from brush biopsies. </br></br> <b>Material and methods:</b> The study was carried out in 92 subjects aged between 26 and 85 years, including 34 women (37.0%), from whom material for cytological examination had been collected from the larynx in the course of microsurgical procedures carried out using the Kleinsasser laryngeal instrument set in 2019-2020. </br></br> <b>Results: </b>Analysis was performed on 90 out of 92 cell smears (97.8%). Two smears were not qualified for analysis due to ille-gibility. The smears were assessed using a proprietary scale consisting in a modification of the Bethesda system. Abnormal results of cytological examinations were obtained in a majority of cases. HSILs with invasive features were the most common abnormal results of cytological examinations. </br></br> <b>Conclusions:</b> Laryngeal epithelial cells can be successfully evaluated by means of cytological examination. Abnormal presen-tation of cytological smear is frequently hypercellular, with inflammatory cells being observed less frequently. No statistically significant relationship was observed between the results of the cytological examination and the overall quality of the smear, number of cells, number of erythrocytes, or the severity of inflammation.

Biobanked tracheal basal cells retain the capacity to differentiate.

Objective: While airway epithelial biorepositories have established roles in the study of bronchial progenitor stem (basal) cells, the utility of a bank of tracheal basal cells from pediatric patients, who have or are suspected of having an airway disease, has not been established. In vitro study of these cells can enhance options for tracheal restoration, graft design, and disease modeling. Development of a functional epithelium in these settings is a key measure. The aim of this study was the creation a tracheal basal cell biorepository and assessment of recovered cells. Methods: Pediatric patients undergoing bronchoscopy were identified and endotracheal brush (N = 29) biopsies were collected. Cells were cultured using the modified conditional reprogramming culture (mCRC) method. Samples producing colonies by day 14 were passaged and cryopreserved. To explore differentiation potential, cells were thawed and differentiated using the air-liquid interface (ALI) method. Results: No adverse events were associated with biopsy collection. Of 29 brush biopsies, 16 (55%) were successfully cultured to passage 1/cryopreserved. Samples with higher initial cell yields were more likely to achieve this benchmark. Ten unique donors were then thawed for analysis of differentiation. The average age was 2.2 ± 2.2 years with five donors (50%) having laryngotracheal pathology. Nine donors (90%) demonstrated differentiation capacity at 21 days of culture, as indicated by detection of ciliated cells (ACT+) and mucous cells (MUC5B+). Conclusion: Pediatric tracheal basal cells can be successfully collected and cryopreserved. Recovered cells retain the ability to differentiate into epithelial cell types in vitro. Level of Evidence: Level 3.

Oral brush biopsy using liquid-based cytology is a reliable tool for oral cancer screening: A cost-utility analysis: Oral brush biopsy for oral cancer screening.

BACKGROUND: This study aimed to assess the diagnostic utility and associated cost of oral liquid-based brush cytology (OLBC) in the diagnosis of oral cancer and oral potentially malignant disorders (OPMDs). METHODS: A total of 284 patients with oral mucosal lesions were included. OLBC samples were collected from all patients immediately before undergoing surgical biopsies. A liquid-based cytology slide was prepared from each OLBC sample for cytological evaluation using the modified 2014 Bethesda cytology system. The results and the cost were compared with the histopathological outcomes. RESULTS: The level of agreement between the two approaches was very good (weighted kappa = 0.824). The accuracy of OLBC in differentiating between the different diagnostic groups was 91.69%, whereas the associated sensitivity and specificity were 79.23% and 94.81%, respectively. The estimated cost of each OLBC sample was at least 26% less than the cost of a single biopsy and more than 42% less in cases of multiple biopsied lesions. CONCLUSIONS: The proposed modifications of the Bethesda system can be adopted as a standardized system for oral cytological assessment. Our findings support OLBC as a reliable adjunct to surgical biopsy in the diagnosis of OPMDs. This tool has potential for oral cancer-finding and surveillance programs.

Evaluation of Static DNA Ploidy Analysis Using Conventional Brush Biopsy-Based Cytology Samples as an Adjuvant Diagnostic Tool for the Detection of a Malignant Transformation in Potentially Oral Malignant Diseases: A Prospective Study.

BACKGROUND: The accuracy of DNA image cytometry as an investigation method for potentially malignant disorders of the oral cavity is currently still a subject of controversy, due to inconsistently applied definitions of DNA aneuploidy, small cohorts and different application techniques of the method. The aim of this study was to examine the accuracy of the method as a supplementary diagnostic tool in addition to the cytological examination using internationally consented definitions for DNA aneuploidy. METHODS: A total of 602 samples from 467 patients with various oral lesions were included in this prospective study. Brush biopsies from each patient were first cytologically examined and categorized by a pathologist, second evaluated using DNA image cytometry, and finally compared to either histological biopsy result or clinical outcome. RESULTS: Using the standard definition of DNA aneuploidy, we achieved a sensitivity of 93.5%, a positive predictive value for the detection of malignant cells of 98.0%, and an area under the curve of 0.96 of DNA ploidy analysis for the detection of severe oral epithelial dysplasia, carcinoma in situ or oral squamous cell carcinoma. Importantly, using logistic regression and a two-step model, we were able to describe the increased association between DNA-ICM and the detection of malignant cells (OR = 201.6) as a secondary predictor in addition to cytology (OR = 11.90). CONCLUSION: In summary, this study has shown that DNA ploidy analysis based on conventional specimens of oral brush biopsies is a highly sensitive, non-invasive, patient-friendly method that should be considered as an additional diagnostic tool for detecting malignant changes in the oral cavity.