Oral Brush Biopsy Research Articles

Abstract A036: The genetic landscape of head and neck cancer using brush biopsy

Abstract Oral potentially malignant disorders (OPMDs) are characterized by the presence of dysplasia and DNA copy number aberrations (CNAs) and subsequently progress to oral squamous cell carcinoma (OSCC). OSCC has a 5-year survival rate of only about 50% and varies greatly depending on site and stage. The combination of next-generation sequencing and advanced computational data analysis approaches has revolutionized our understanding of the genomic underpinnings of cancer development and progression. This can reveal individual targetable alterations, mutational load, complex mutation signatures, and tumor-specific antigens, which might inform the utilization of targeted therapies and early diagnosis. Oral brush biopsy represents an attractive non-invasive advantage in the characterization of premalignant and malignant oral lesions. Still, a sufficient number of abnormal cells is mandatory to diagnose. The current study tested the possibility of recreating the genomic landscape of oral cancer using brush biopsy. Materials and methods: As a pilot study to address this question, we analyzed matched tissue and brush biopsy of a patient with synchronous OPMD and OSCC. Brush and tissue biopsy was taken from normal epithelium, OPMD, and OSCC. We scrutinized the evolution and heterogeneity of these three brush/tissue biopsy pairs. The techniques used include whole exome sequencing (WES), mutational signature analysis to evaluate smoking and other aberrant signatures, copy number analysis (using ASCAT), and subclonal architecture reconstruction (using DPclust). Results: To analyze intra-tumoral heterogeneity (ITH), ASCAT and DPclust were used to call copy number aberrations (CNA) and cluster somatic SNVs based on cancer cell fraction (CCF), respectively. We observed a large number of shared SNVs between the OPMD and OSCC lesions suggesting a common evolutionary origin. After adjusting the ploidy and purity based on DPClust results, ASCAT profiles revealed differences in aberrant tumor cell fraction, gains and loss of heterozygosity, and copy number neutral events between the samples. It showed almost similar profiles between the respective lesions' brush and tissue samples, suggesting that brush biopsy can recapitulate the genomic landscape. The OPMD and OSCC lesions showed loss of heterozygosity in chromosome 9 and chromosome 17, which play a potential role in tumorigenesis with loss of function of P53. We inferred the subclonal architecture of each sample, illuminating the clonal relationships between samples and distinct lineages of precursor clones developing independently. Conclusion: Collectively, these results emphasize the possibility of using brush biopsy to reconstruct the genomic characterization of OPMD and OSCC show prognostic potential for diagnosis and intervention strategies, and accelerate progress in precision cancer care. Citation Format: Evit John, Tom Lesluyes, Xiao Zhao, Peter Van Loo. The genetic landscape of head and neck cancer using brush biopsy [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Translating Cancer Evolution and Data Science: The Next Frontier; 2023 Dec 3-6; Boston, Massachusetts. Philadelphia (PA): AACR; Cancer Res 2024;84(3 Suppl_2):Abstract nr A036.

Abstract PO-015: The genetic landscape of head and neck cancer using brush biopsy

Abstract Oral potentially malignant disorders (OPMDs) are characterized by the presence of dysplasia and DNA copy number aberrations (CNAs) and subsequently progress to oral squamous cell carcinoma (OSCC). OSCC has a 5-year survival rate of only about 50% and varies greatly depending on site and stage. The combination of next-generation sequencing and advanced computational data analysis approaches has revolutionized our understanding of the genomic underpinnings of cancer development and progression. This can reveal individual targetable alterations, mutational load, complex mutation signatures, and tumor-specific antigens, which might inform the utilization of targeted therapies and early diagnosis. Oral brush biopsy represents an attractive non-invasive advantage in the characterization of premalignant and malignant oral lesions. Still, a sufficient number of abnormal cells is mandatory to diagnose. The current study tested the possibility of recreating the genomic landscape of oral cancer using brush biopsy. Materials and methods: As a pilot study to address this question, we analyzed matched tissue and brush biopsy of a patient with synchronous OPMD and OSCC. Brush and tissue biopsy was taken from normal epithelium, OPMD, and OSCC. We scrutinized the evolution and heterogeneity of these three brush/tissue biopsy pairs. The techniques used include whole exome sequencing (WES), mutational signature analysis to evaluate smoking and other aberrant signatures, copy number analysis (using ASCAT), and subclonal architecture reconstruction (using DPclust). Results: To analyze intra-tumoral heterogeneity (ITH), ASCAT and DPclust were used to call copy number aberrations (CNA) and cluster somatic SNVs based on cancer cell fraction (CCF), respectively. We observed a large number of shared SNVs between the OPMD and OSCC lesions suggesting a common evolutionary origin. After adjusting the ploidy and purity based on DPClust results, ASCAT profiles revealed differences in aberrant tumor cell fraction, gains and loss of heterozygosity, and copy number neutral events between the samples. It showed almost similar profiles between the respective lesions' brush and tissue samples, suggesting that brush biopsy can recapitulate the genomic landscape. The OPMD and OSCC lesions showed loss of heterozygosity in chromosome 9 and chromosome 17, which play a potential role in tumorigenesis with loss of function of P53. We inferred the subclonal architecture of each sample, illuminating the clonal relationships between samples and distinct lineages of precursor clones developing independently. Conclusion: Collectively, these results emphasize the possibility of using brush biopsy to reconstruct the genomic characterization of OPMD and OSCC and show prognostic potential for diagnosis and intervention strategies and accelerate progress in precision cancer care. Citation Format: Evit John, Tom Lesluyes, Toby Baker, Xiao Zhao, Peter Van Loo. The genetic landscape of head and neck cancer using brush biopsy [abstract]. In: Proceedings of the AACR-AHNS Head and Neck Cancer Conference: Innovating through Basic, Clinical, and Translational Research; 2023 Jul 7-8; Montreal, QC, Canada. Philadelphia (PA): AACR; Clin Cancer Res 2023;29(18_Suppl):Abstract nr PO-015.

THE USE OF CDX BRUSH BIOPSY IN ORAL CANCER DETECTION

Oral squamous cell carcinoma is the most common group of cancers that affect the head and neck region. Detection in its early stages is a key factor to improve the survival rate and oral health professionals must play a leading role in this endeavor. Various diagnostic aids have been developed as an adjunct to the routine oral cancer screening, one of which is the oral brush biopsy (known as the OralCDx® Brush Test). The technique is non-invasive and relatively simple, allowing a painless and easy way to evaluate a suspicious lesion. The purpose of this article is to review the OralCDx test including its indications, advantages, disadvantages and applications in dentistry

Conspectus on nanodiagnostics as an incipient platform for detection of oral potentially malignant disorders and oral squamous cell carcinoma.

Conspectus on nanodiagnostics as an incipient platform for detection of oral potentially malignant disorders and oral squamous cell carcinoma.

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

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. 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. 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). 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.

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

ObjectivesOral 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.MethodsPrivate 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.ResultsThe 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%.ConclusionThe 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.

A New Multi-Color FISH Assay for Brush Biopsy-Based Detection of Chromosomal Aneuploidy in Oral (Pre)Cancer in Patients with Fanconi Anemia.

Simple SummaryPatients with Fanconi Anemia (FA) are at high risk of developing oral squamous cell carcinoma with poor prognosis. Oral brush biopsy–based cytology represents an attractive non-invasive advantage in the characterization of (pre) malignant oral lesions, but a sufficient number of abnormal cells is mandatory to make the diagnosis. The aim of this 2-phases retrospective study is to introduce a new multi-color FISH assay including a 9p21 FISH assay to determine the oral lesions that require treatment, with just a few cells. We demonstrate a good accuracy of the multicolor FISH assay if applied on oral brush biopsy-based specimens from FA patients, using different algorithms to determine chromosomal aneuploidy. Nevertheless, some false positive results were observed and the detection of a genetically altered field in the oral cavity as a possible reason for what may hamper the application of multi-color FISH is discussed.Background: Fanconi anemia (FA) is a rare inherited DNA instability disorder with a remarkably elevated risk of oral squamous cell carcinoma. These cancers can be detected with oral brush biopsy-based cytology even at early stages. This study aims to determine the diagnostic accuracy of a new multi-color fluorescent in situ hybridization (FISH) assay consisting of probes for CCND1, TERC, MYC and centromere of chromosome 6, as well as a 9p21 FISH assay consisting of probes for CDKN2A and centromere of chromosome 9 for the detection of oral (pre) malignant lesions in FA. Methods: (I) Cutoffs for the dichotomization of positive or negative multi-color FISH results are determined and (II) retrospectively validated by using archived oral brush biopsy specimens from individuals with Fanconi anemia. In addition, the specimens for cutoff determination were re-hybridized with the 9p21 FISH assay. Results: A cutoff of six or more chromosomal aneuploid cells for a positive FISH result was determined in the cutoff study on 160 biopsy specimens. The validating of this cutoff on 152 specimens showed at best a sensitivity of 87% and a specificity of 82.9%. Conclusion: Multi-color FISH is a sufficient tool to detect chromosomal aneuploidy in oral (pre) malignant lesions of individuals with Fanconi anemia. However, some false positive results may hamper the application as an adjuvant method to oral brush biopsy-based cytology in an oral cancer surveillance program.

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

BackgroundThis 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).MethodsA 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.ResultsThe 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.ConclusionsThe 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.

Immunoexpression of oral brush biopsy enhances the accuracy of diagnosis for oral lichen planus and lichenoid lesions.

BackgroundThis study assessed the efficacy of using oral liquid‐based brush cytology (OLBC) coupled with immunostained cytology‐derived cell‐blocks, quantified using machine‐learning, in the diagnosis of oral lichen planus (OLP).MethodsEighty‐two patients diagnosed clinically with either OLP or oral lichenoid lesion (OLL) were included. OLBC samples were obtained from all patients before undergoing surgical biopsy. Liquid‐based cytology slides and cell‐blocks were prepared and assessed by cytomorphology and immunocytochemistry for four antibodies (Ki‐67, BAX, NF‐κB‐p65, and AMACR). For comparison purposes, a sub‐group of 31 matched surgical biopsy samples were selected randomly and assessed by immunohistochemistry. Patients were categorized according to their definitive diagnoses into OLP, OLL, and clinically lichenoid, but histopathologically dysplastic lesions (OEDL). Machine‐learning was utilized to provide automated quantification of positively stained protein expression.ResultsCytomorphological assessment was associated with an accuracy of 77.27% in the distinction between OLP/OLL and OEDL. A strong concordance of 92.5% (κ = 0.84) of immunostaining patterns was evident between cell‐blocks and tissue sections using machine‐learning. A diagnostic index using a Ki‐67‐based model was 100% accurate in detecting lichenoid cases with epithelial dysplasia. A BAX‐based model demonstrated an accuracy of 92.16%. The accuracy of cytomorphological assessment was greatly improved when it was combined with BAX immunoreactivity (95%).ConclusionsCell‐blocks prepared from OLBC are reliable and minimally‐invasive alternatives to surgical biopsies to diagnose OLLs with epithelial dysplasia when combined with Ki‐67 immunostaining. Machine‐learning has a promising role in the automated quantification of immunostained protein expression.

A standalone approach to utilize telomere length measurement as a surveillance tool in oral leukoplakia.

Oral squamous cell carcinoma (OSCC) is often preceded by a white patch on a surface of the mouth, called oral leukoplakia (OL). As accelerated telomere length (TL) shortening in dividing epithelial cells may lead to oncogenic transformation, telomere length measurement could serve as a predictive biomarker in OL. However, due to high variability and lack of a universal reference, there has been a limited translational application. Here, we describe an approach of evaluating TL using paired peripheral blood mononuclear cells (PBMC) as an internal reference and demonstrate its translational relevance. Oral brush biopsy and paired venous blood were collected from 50 male OL patients and 44 male healthy controls (HC). Relative TL was measured by quantitative PCR. TL of each OL or healthy sample was normalized to the paired PBMC sample (TL ratio). In OL patients, the mean TL ratio was significantly smaller not only in the patch but also in distal normal oral tissue, relative to healthy controls without a high‐risk oral habit. Dysplasia was frequently associated with a subgroup that showed a normal TL ratio at the patch but significantly smaller TL ratio at a paired normal distal site. Our data suggest that evaluation of TL attrition using a paired PBMC sample eliminates the requirement of external reference DNA, makes data universally comparable and provides a useful marker to define high‐risk OL groups for follow‐up programs. Larger studies will further validate the approach and its broader application in other premalignant conditions.

Efficacy of oral brush biopsy in the early detection of oral cancer and oral potentially malignant disorders - a systematic review

Introduction: Oral cancer is one of the most prevalent cancers in South East Asian countries due to increased consumption of tobacco products and alcohol and ranks sixth among the malignancies worldwide. Several screening modalities such as toluidine blue, Lugol's iodine staining, exfoliative cytology, and biopsy are being used to detect suspicious oral potentially malignant disorders (PMDs) at an early stage. Oral brush biopsy is a painless and less invasive technique that uses variations of a small brush to collect cells from all three layers of epithelium. The technique is non-invasive and promises to be an adjunct to the gold-standard histopathological examination and diagnosis of oral PMD. Aim: The aim of this systematic review was to compile studies pertaining to diagnostic utility of oral brush biopsy with or without computer-assisted cytological analysis in the screening of innocuous oral PMDs for early detection of cancerous changes. Materials and Methods: Computerized literature search was performed to select eligible articles from the following databases: PubMed (MEDLINE), GOOGLE SCHOLAR, and SCIENCE DIRECT using specific keywords. The search was limited to articles published as full text in English, which were screened by two reviewers for eligibility. Results and Discussion: Six studies satisfied our inclusion criteria and found oral brush biopsy as a moderately reliable screening tool for early detection of dysplastic changes in oral PMDs. For oral brush biopsy, sensitivity and specificity obtained were 97.7% and 84.5%, respectively, in the detection of high-grade dysplasia. Conclusion: Brush cytology using a cytobrush is a reliable adjunct to histopathology in detecting oral premalignant and malignant oral lesions. Specificity and sensitivity of the technique requires further investigative clinical studies.

Comparison of transepithelial cytology and histopathology in the diagnosis of potentially malignant and malignant lesions of the oral mucosa

Background/Aim: The objective of the study was to compare cytological diagnoses using transepithelial cytology (oral brush biopsy) with histopathological diagnoses obtained by incisional biopsy in patients with benign lesions, oral potentially malignant disorders and oral squamous cell carcinoma. Material and Methods: The study included 57 patients. Brush biopsy was performed after local anaesthesia administration using the cervical brush. It was immediately followed by an incisional biopsy. Modified Bethesda System was used for cytological analysis and correlated with histopathological diagnoses according to intraepithelial neoplasia. Results: Good agreement was shown between cytological and histopathological diagnosis (kappa = 0.791). The sensitivity of the study was 92.85%, specificity 100%, positive predictive value (PPV) was 100% and negative predictive value (NPV) 93.54%. Conclusions: Oral brush biopsy, which allows the collection of epithelial cells of all layers can provide fast, precise and efficient cytological results which are in good agreement with the 'gold standard' - incisional biopsy followed by histopathology.

Detection of cytogenetic changes and chromosomal aneuploidy with fluorescent in situ hybridization in cytological specimens of oral cancers in Fanconi anemia-Proof of concept.

ObjectivesFanconi anemia (FA) is a rare inherited DNA instability disorder with a remarkably elevated risk of neoplasia compared with the general population, mainly leukemia and squamous cell carcinoma (SCC). Two thirds of the SCCs arise in the oral cavity and are typically preceded by visible lesions. These lesions can be classified with brush biopsy‐based cytological methods regarding their risk of a malignant transformation. As a proof of concept, this study aims to investigate genetic changes and chromosomal aneuploidy using fluorescent in situ hybridization (FISH) on oral squamous cells derived from FA affected individuals.Material and MethodsFive FA oral SCC (OSCC) tumor cell lines, one FA OSCC cervical lymph node metastasis as well as tumor‐negative and atypical smears from oral brush biopsies were analyzed with FISH probes covering 5p15.2, MYC, EGFR, TERC, 9q34.1, CCND1, 9p21 and centromeres of chromosomes 3, 6, 7, 9, 11, and 17.ResultsOSCC specimens showed gains of all analyzed chromosomal regions. Chromosomal aneuploidy was observed in five of the six OSCC specimens in two multicolor FISH assays with panels of four probes each. Five out of six OSCC specimens displayed a relative deletion of 9p21. Applied on atypical brush biopsy‐based smears, chromosomal aneuploidy was detected in malignant lesions but not in the smear derived from a severe parodontitis.ConclusionsAs proof of concept, FISH was able to detect genetic changes and chromosomal aneuploidy discriminating oral cancer from noncancerous lesions in individuals with FA. This supports its application on oral brush biopsy‐based cytology.

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

BackgroundThis 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.MethodsFrom 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.Results2018 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).ConclusionsBoth 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.

Recent Advances in Cytology-A Review

In olden days, specificity and sensitivity of oral cytology is poor. Utilizing traditional oral cytology for theconclusive diagnosis of malignancyand premalignant lesions. But with the recent cytological techniquespaves cytology to conclude a final diagnosis. This reestablished interest is somewhat because of thepresentation of a specific brush that gathers a full-thickness epithelial sample and not simply sloughed cells,just as examination of that sample with computer support and analysis. There is an assortment of adjunctivestrategies have been acquainted with possibly upgrade the finding of the cytologic examples including DNAexamination, immunocytochemistry, molecular analysis, and liquid based preparations. An expansion insensitivity (>96%) and specificity (>90%) of the oral brush biopsy with computer support, diagnosis hasbeen accounted for detection of malignant and premalignant lesions. Brush cytology is significant to forestallmisdiagnosing suspicious oral lesions.

Don't rush, first brush: A comparative study between Modified Brush Biopsy (MBB) and Liquid-Based Cytology (LBC)

Introduction: Oral cancer accounts for 2–4% of all malignant tumors worldwide with the percentage going much higher in the Indian subcontinent. Many of these lesions are in the advanced stage at the time of diagnosis. Research is always on for newer techniques to improve early detection and diagnosis of oral malignancy. Oral brush biopsy is one such technique. Aims and Objectives: The aim of the study was to evaluate and compare the clinical usefulness of two modified oral brush biopsy techniques in the early detection of oral cancer. Study Design: Two modified oral brush biopsy techniques, one performed with a baby brush spreading the cells directly on slide and then fixed and another with a specially prepared brush submerged in a liquid fixer, were performed in 24 patients having oral submucous fibrosis clinically. These were followed by punch biopsy (as a gold standard). All specimens were analyzed for cytology and histopathology manually. Results and Conclusion: Modified brush biopsy (MBB) and liquid-based cytology (LBC) both can be potential methods for early detection of oral cancer in different conditions.

Dental Hygienists and Dentists as Providers of Oral Mucosa Screening and Brush Biopsies

Dental Hygienists and Dentists as Providers of Oral Mucosa Screening and Brush Biopsies

CDK4, CDK6, cyclin D1 and Notch1 immunocytochemical expression of oral brush liquid-based cytology for the diagnosis of oral leukoplakia and oral cancer.

This study aimed to investigate the feasibility of using oral liquid-based brush cytology (OLBC) coupled with immunocytochemistry as a minimally invasive approach to stratify the cancer risk in patients with oral leukoplakia. Fifty-five patients diagnosed with either oral leukoplakia (OLK) or oral squamous cell carcinoma (OSCC) were recruited. All patients underwent oral brush biopsy followed by surgical biopsy. 275 liquid-based cytology preparations were made. Pap-stained OLBC slides were assessed using the modified 2014 Bethesda Cytology system. The expression of CDK4, CDK6, cyclin D1, and Notch 1 was immunocytochemically analysed and compared against the histopathological diagnosis. A combined index score of OLBC grading and protein expression was calculated. A significant association was found between the definitive histopathological diagnosis and the cytological interpretation (P=0.0005). The index scores of CDK4, CDK6, and cyclin D1 were significantly associated with the development of disease from non-dysplastic epithelium to OSCC. No significant association was observed between the Notch 1 index score and disease stage. The diagnostic accuracy of OLBC showed the highest values of sensitivity, specificity, positive predictive value, negative predictive value, and accuracy: 84.6%, 70.4%, 73.3%, 82.6%, and 78.8%, respectively, compared with the cumulative protein index, CDK4/6 index, and the combined OLBC grading and CDK4/6 index. This study has also demonstrated the efficacy of the use of OLBC in the detection of OED and OSCC, and showed that the use of CDK4, CDK6, cyclin D1, and Notch 1 immunocytochemistry failed to improve the diagnostic accuracy of OLBC suggesting they are not useful in the early detection of OSCC.

Effective early detection of oral cancer using a simple and inexpensive point of care device in oral rinses

ABSTRACTIntroduction: Head and neck cancer remains a challenging disease that is increasing in incidence with the majority of patients diagnosed at an advanced stage where 5-year survival is approximately 50%. Current approaches including oral-brush biopsies, fluorescence-based technologies, and salivary molecular profiling have demonstrated some success; however, cost, ease of use, and accuracy remain limiting factors.Areas covered: This is a profile of a novel, easy to use oral rinse point-of-care (POC) test to aid in the diagnosis of oral and oropharyngeal cancer. Background science related to the challenge of oral and oropharyngeal cancer and natural history of diagnostic aids for this disease are provided. Results of studies performed for validation of a POC and laboratory test are also discussed.Expert commentary: The POC test has been validated through a case : control clinical study and a prospective European trial, using version 1.0 (v1.0), which have demonstrated consistent performance including a > 90% negative predictive value, with a sensitivity of 80%. The assay was designed to identify malignant lesions in the oral cavity and oropharynx by improving upon standard clinical assessment.

Clinical comparison of liquid-based and conventional cytology of oral brush biopsies: a randomized controlled trial

BackgroundExfoliative cytology performed on oral brush samples can help dentists to decide, whether a given oral lesion is (pre-) malignant. The use of non-invasive brush biopsies as an auxiliary tool in the diagnosis of oral mucosal lesions has gained renewed interest since improvements in cytological techniques such as the development of adjuvant diagnostic tools and liquid-based cell preparation techniques.MethodsThe aim of this study was to compare the quality of two different preparation techniques (cell collectors): the conventional transfer procedure to glass slides and the so-called liquid-based cytology preparation method. Cell smears were collected from 10 orally healthy individuals (mean age: 24 years) from the palatine mucosa at two different times (baseline and 4 weeks later). Slides of both techniques were stained by Giemsa (n = 40) and May-Gruenwald Giemsa (n = 40). The statistical analysis was performed with Excel.ResultsOn specimen analysis, the liquid-based cytology showed statistically significant improvement compared to conventional glass sides (p < 0.001). Thin layers, which were performed by liquid-based cytology showed significantly better results in the parameters (p < 0.001): uniform distribution, cellular overlapping, cellular disformation, mucus, microbial colonies and debris. The conventional glass slides approach showed more cell overlapping and contamination with extraneous material than thin layers, which were performed by Orcellex® Brush cell collectors.ConclusionsBoth techniques are diagnostically reliable. The liquid-based method showed an overall improvement on sample preservation, specimen adequacy, visualization of cell morphology and reproducibility.Liquid-based cytology simplifies cell collection due to easier handling and less transfer errors by dentists.