Oral Rinse as a Simpler Approach to Exfoliative Cytology: A Comparative Study

Abstract

IntrOductIOnBiopsy is the diagnostic test of choice for oral potentially malignant disorders and malignant lesions.But since scalpel biopsy is an invasive procedure associated with potential morbidity,several adjunctive screening aids are used to assist clinicians with the detection of early cancerous changes [1]. Although oral exfoliative cytology is a simple non-invasive technique, the traditional exfoliative cytology methods show low sensitivity (i.e. a high proportion of false negatives), inadequate sampling, procedural errors, and the need for subjective interpretation of the findings [2]. Routinely shed oral epithelial cells can be detected in saliva and oral rinses, making cytologic and molecular analysis of this fluid attractive for oral cancer screening. In the present study epithelial cells collected via oral rinse method are subjected to routine cytologic analysis along with the conventional exfoliative cytology smear. The advantages of this method are increased patient comfort and the possibility of oral rinse collection by anyone, including the patient himself, without the requirement of health personnel or armamentarium, especially in resource challenged areas. MAterIAl And MethOdsHundred and five subjects from the Department of Oral Medicine, A.B. Shetty Memorial Institute of Dental Sciences, Deralkatte, India with normal oral mucosa were selected for the study. The study protocol was approved by the Committee on Ethics of the Nitte University, Mangalore, Karnataka, India. Patients were informed with regard to the research objectives, methods, possible benefits and potential risks, and a written consent was obtained from all participants.Careful examination of the oral cavity was followed by exfoliative cytology (oral rinse based technique followed by wooden spatula based cytology).Techniques employed: Oral rinse technique was used to collect oral cells. Patient was asked to swish his/her mouth with water and expectorate. Then, the clinically normal buccal mucosa was rubbed on firmly by the patient themselves using their tongue for 30 seconds. While swishing phosphate buffered saline, pH -7.2 was used and patient was asked to expectorate into a sterile container. Once the sample was obtained, it was labelled and centrifuged at 1,000 rpm for 5 minutes. Supernatant fluid was discarded and smears were prepared from the cell plug. For exfoliative cytology, scrapings were obtained from clinically normal buccal mucosa using a standard moistened wooden spatula. Scrapings were smeared on labelled glass slides. All the slides thus prepared were immediately fixed in absolute alcohol and consequently stained with Papanicoloau stain. All slides were assessed by 2 trained cytopathologists, who were not aware of the type of the technique from which the material was collected. For comparative analysis of both techniques three parameters were used: a) cell yield b) cellular dispersion and d) cellular clarity. Both the observers were asked to grade each parameter into Good /Average / Poor in a data sheet comprisin...

Evaluation of the Cytomorphometric Changes Observed in the Oral Mucosal Cells Exposed to Computed Tomography

“…Since then, exfoliative cytology has gained a widespread use, not only confined to gynecologic endocrinology but also to other various departments including that of oral cancer detection and in early diagnosis of Acquired Immunodeficiency Syndrome (AIDS). [5][6][7][8] Many techniques have been employed for collecting the exfoliated cells of the oral mucosa like oral rinse technique, scraping the oral mucosa by means of a steel spatula, wooden tongue depressor, and oral CDX brushes 9 . In the present study, wooden tongue depressor was used as it is easy to use, disposable, leads to good amount of exfoliated sample collection and even distribution of obtained sample onto a glass slide.…”

Improvization of conventional cytology by centrifuged liquid-based cytology in oral exfoliative cytology specimen

“…[ 7 ] It is well-established that only a small percentage of the harvested epithelial cells obtained during conventional smear are transferred to the glass slide and there is always a possibility of potential source for false negative smears. [ 8 ] It is shown that a maximum of only 20% of the cells collected on a variety of collection devices can be mechanically transferred to the flat surface of a glass slide. [ 9 ]…”

“…In a study conducted by Shaila et al ., slides prepared by the conventional wooden spatula method in normal oral mucosa were disregarded due to either excessive clumping or scarcity of cells. [ 8 ] Ogden et al . found less cell yield and cell dispersion,[ 10 ] whereas Ahmed et al .…”

Prevalence of human papillomavirus in oral rinse samples from healthy individuals in northern Thailand

“…Currently, there is no universally agreed‐on method of oral sampling for the detection of HPV DNA. The most common method is using an oral rinse swirl or gargle to obtain an oral rinse sample, which is easy and non‐invasive suitable for oral screening . DNA can be extracted, and polymerase chain reaction (PCR) amplified after storage of mouthwash solution at room temperature for periods of up to 30 days .…”

“…The most common method is using an oral rinse swirl or gargle to obtain an oral rinse sample, which is easy and noninvasive suitable for oral screening. 1,[20][21][22] DNA can be extracted, and polymerase chain reaction (PCR) amplified after storage of mouthwash solution at room temperature for periods of up to 30 days. 23 However, the prevalence of HPV varies according to sample size, sampling methods 24 (probability or non-probability sampling methods), analysis, specimens (such as fresh, frozen, formalin-fixed paraffin-embedded samples with tissue/cell or plasma/ serum), and methodology (such as PCR and nested PCR using GP5+/6+ primer, PGMY09/11 primer MY09/11 with GP5+/6+ primer; Southern blotting assay; real-time PCR, DNA/RNA In situ hybridization; hybrid capture II; antibodies against early/late HPV protein; and DNA/RNA microarray).In this study, we investigated the prevalence of HPV infection in 594 oral rinse samples from healthy northern Thai population using PCR (GP5+/6+ primers) 25 and HPV genotyping by sequencing.Demographic data and behavioral factors for these subjects were collected and evaluated for association with HPV.…”