Orale Zytologie

Oral cytology has aroused new interest caused by introduction of the cytobrush as a sampling device and the use of additional analytical methods. By brushing it is possible to reach deeper layers of the oral mucosa where squamous intraepithelial neoplasia (SIN) begins. The biological potential of the oral epithelial cells obtained can be evaluated by the following additional methods: computer-assisted image analysis (OralCDx^®), DNA cytometry, immunohistochemistry, monolayer cytology, and molecular biological analysis. All of those methods can increase sensitivity (up to 100%) and specificity (up to 100%) of oral brush biopsy. Nevertheless, there are reports that oral epithelial carcinomas were not identified. No comparative study exists allowing conclusions to be drawn about the value of the single methods. Immunocytochemistry with commercial antibodies against laminin-5 is generally available and methodologically easy. Oral brush biopsy as a non invasive diagnostic method can be useful for the early detection of oral mucosal lesions. Positive findings or progression of the lesion despite negative findings are indications to refer the patient to a specialized clinic where a surgical biopsy should be performed, followed by histopathological analysis. Histopathology remains the gold standard for the definitive diagnosis of oral malignant lesions.     

The utility of toluidine blue staining and brush cytology as adjuncts in clinical examination of suspicious oral mucosal lesions

The objective of this study was to investigate the utility of toluidine blue and brush cytology in patients with clinically detected oral mucosal lesions. Clinical examination of 35 patients was completed before toluidine blue application, oral brush cytology and scalpel biopsy. Lesions were photographed before and after stain application; followed by brush cytology. All findings were compared with histopathologic results. Severe dysplasia and carcinoma-in-situ were determined as ‘positive’; no dysplasia and mild to moderate dysplasia were defined as ‘negative’. The sensitivity, specificity, positive and negative predictive values of clinical examination and toluidine blue were the same: 0.923, 0.433, 0.414, and 0.929, respectively. Those of brush cytology were 0.923, 0.517, 0.462, and 0.938. The concordance of all methods was 30% for benign and 61% for malignant lesions. Adjuncts identified 92% of carcinoma-in-situ and squamous cell carcinoma as confirmed by histopathology, in contrast to clinical findings alone in which 62% of these lesions were identified (p = 0.046). In conclusion, adjunct diagnostic methods decreased the level of uncertainty for the diagnosis of oral malignancies and lichenoid dysplasias when applied as adjuncts to clinical examination.     

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提要：口腔癌是全身第6位常见肿瘤,近几十年5年存活率没有明显改善,严重危害患者的生命和生存质量。其中一个主要原因是由于缺乏行之有效的监测和筛查早期口腔癌的方法。本文旨在结合笔者多年从事临床病理的经验,对检测早期口腔癌的方法进行评述。除作为金标准的组织学检查外,细胞学检查、DNA定量分析、分子生物学检查将有望成为早期口腔癌检测及筛查的重要手段。     

Evaluation of nuclear morphometry and DNA ploidy status for detection of malignant and premalignant oral lesions: quantitative cytologic assessment and review of methods for cytomorphometric measurements.

PURPOSE: Detection of a precancerous or cancerous lesion when small is one of the most important factors to improve 5-year survival rates of oral cancer. Although surgical biopsy is the most definitive method for diagnosing oral lesions, it is impractical to routinely subject large numbers of patients to biopsy. Recently, cytomorphometric assessments improved by advanced computer-assisted image analysis systems have gained importance. This study was established to evaluate the efficacy of nuclear cytomorphometric analysis and DNA ploidy status for the detection of oral malignancies. Methods used for cytomorphometric analysis were also reviewed. PATIENTS AND METHODS: Oral mucosal smears (n = 44) were obtained from patients (n = 22) presenting with various oral lesions using a cytobrush immediately before biopsy. Cytomorphometric measurements and nuclear Feulgen DNA content analysis were carried out after the Feulgen staining procedure. Smears from the lesion site constituted the study group whereas contralateral healthy mucosal sites served as control. RESULTS: DNA ploidy analysis revealed 20 diploid (90.9%) and 2 aneuploid DNA patterns (9.1%) sampled from the lateral margin of the tongue and floor of the mouth. When only malignant lesions were considered, aneuploidy rate was 16.7% whereas a diploid pattern was indicated for 83.3% of the sample. With cytomorphometric measurements, a statistically significant difference was shown for nuclear perimeter, area, diameter equivalent to circle, minimum and maximum Feret, intensity, DNA content (c) and DNA index values. CONCLUSIONS: Cytomorphometric analysis via oral brush biopsy is a valuable adjunct to biopsy for identification of premalignant and early stage cancerous oral lesions as a rapid and minimally invasive procedure with high specificity and sensitivity rates, requiring no topical or local anesthetic.     

Cytologic and DNA-cytometric very early diagnosis of oral cancer

BACKGROUND: The aim of this study was to evaluate the diagnostic accuracy of exfoliative cytology (EC) and DNA-image cytometry applied to suspicious oral lesions compared with synchronous histology. METHODS: Brush- and scalpel biopsies were obtained from 98 patients with suspicious oral lesions. In cases, in which EC revealed malignant or suspicious cells, nuclear DNA-contents were measured using a TV image analysis system. RESULTS: Among 98 oral lesions both cytological and histological diagnosis showed no sign of malignancy or dysplasia in 75. In 23 cases cytology yielded tumor cell-positive (15), suspicious (four) or doubtful (four) results. DNA-cytometry showed aneuploidy in 19 of these. The comparison between cytological diagnosis combined with DNA-cytometry and biopsy-histology resulted in a sensitivity of 100% and a specificity of 97.4%. CONCLUSION: In conclusion, cytology with DNA-cytometry is a highly sensitive, specific and non-invasive method for the early diagnosis of oral epithelial neoplasia, showing excellent compliance among patients.     

Patients’ views on a proposed oral cancer screening technology

Brush cytology and telemedicine platforms are used with proven benefits in remote diagnosis of malignancies like cervical cancer. In low-resource settings access to the current standard of care, biopsy and histopathology grading of oral lesions, can be unfeasible. Risk-stratifying early malignant and oral potentially-malignant lesions can help instigate therapeutic treatment and improve prognosis. Therefore, a telecytology platform enabling remote connectivity to specialist centres may aid early detection of oral cancer. We outline head and neck (H&N) cancer patients’ views on an oral brush cytology investigation.     

Epstein-Barr virus in tobacco-induced oral cancers and oral lesions in patients from India

D'Costa J, Saranath D, Sanghvi V, Mehta AR: Epstein-Barr virus in tobacco-induced oral cancers and oral lesions in patients from India. J Oral Pathol Med 1998; 27: 78–82. © Munksgaard, 1998.
We examined 103 oral squamous cell carcinomas (OSCC), 100 oral lesions consisting primarily of leukoplakia (82 cases), and 76 clinically normal mucosa specimens from the contralateral site in the oral cavity of individuals with oral lesions, for the presence of Epstein-Barr virus (EBV). Polymerase chain reaction (PCR) was used to amplify a 239 bp fragment of the BamHIL region of the EBV genome, followed by Southern blot hybridization with EBV oligonucleotide probe to increase further the specificity and sensitivity of the assay system. Since EBV seropositivity is frequent in populations, we also examined the peripheral blood cells (PBC) from 141 patients (50 oral cancer patients, 91 patients with oral lesions) for the presence of EBV We detected EBV in 25 of 103 (25%) OSCC, 13 of 100 (13%) oral lesions, 3 of 76 (4%) clinically normal mucosa samples and 10 of 141 (7%) PBC. Our results indicate that EBV may contribute as one of the multiple factors in oral cancers, in a certain proportion of Indian patients.     

High-molecular tenascin-C as an indicator of atypical cells in oral brush biopsies

Tumour-invasion like wound healing is characterised by the formation of an extracellular matrix with a high tenascin-C content. The tenascin-C molecule undergoes alternative splicing. Analysis using antibody BC2 indicates that especially the high-molecular tenascin-C (hm tn-C) variants are typically tumour-associated, while distribution in normal tissue is restrictive. This study investigated whether hm tn-C is a suitable indicator of atypical cells with invasive potential in oral brush biopsies. One hundred fifty nine consecutive oral brush biopsies with histopathological diagnoses were analysed for the identification of atypical cells. A standardised haematoxylin and eosin staining plus standardised immunocytochemistry using the monoclonal anti-hm tn-C antibody was performed. The bound hm tn-C antibodies were detected with the streptavidine/alkaline phosphatase technique in the autostainer. Conventional cytology produced four false-positives when identifying atypical cells in brush biopsies of inflammatory/benign hyperproliferative mucosa (specificity 96%), while 10 in 52 carcinomas and three of eight recurrences were not identified (sensitivity 78%). Ten of these 13 non-identified tumours could be marked when adding the hm tn-C assay (increasing specificity to 99%). Combining the two assays also reduced the false-positive outcomes from four to one (increasing sensitivity to 95%). The positive and negative predictive values were 92 and 88% for conventional cytology vs 98 and 97% for the dual assay. (1) A 95%-sensitivity proves hm tn-C assisted conventional cytology to be a suitable means of identifying atypical cells in oral brush biopsies. (2) The positive (98%) and negative (97%) predictive values obtained approximate hm tn-C assisted conventional cytology to laminin-5 (100/97%). Lecture given at the 56th Congress of the German Society of Oral and Maxillofacial Surgery (DGMKG) in Dresden, Germany, June 7–10, 2006.     

Utility of chemiluminescence (ViziLite™) in the detection of oral potentially malignant disorders and benign keratoses

J Oral Pathol Med (2011) 40 : 541–544 Background: Oral potentially malignant disorders (OPMD) are known to precede the development of oral cancer. Detection of OPMD allows delivery of interventions that may reduce the evolution of these disorders to malignancy. Following oral examinations, the accuracy of detection of OPMD by chemiluminescence was evaluated using a commercially available detection kit – ViziLite. Data derived were compared in relation to conventional oral examination and surgical biopsy. Methods: A total of 126 patients, 70 men and 56 women (mean age 58.5 ± 11.9 years) attending Oral Medicine Clinics at King’s and Guy’s Hospitals, London, with oral white, red, and mixed white and red patches were enrolled. Sixty‐one patients were current smokers, 28 were ex‐smokers, while 92 were alcohol users. In a detailed investigation, these patients underwent ViziLite examination followed by surgical biopsy. Results: Based on the clinical diagnosis, 70 patients had oral leukoplakia/erythroplakia, 32 had oral lichen planus, nine had chronic hyperplastic candidiasis, and rest had frictional keratosis (13) or oral submucous fibrosis (2). Of 126 lesions, 95 (75.4%) showed aceto‐whitening. Most oral leukoplakias had enhanced visibility and sharpness of the lesion when viewed with the ViziLite system. Following biopsy, 44 had oral epithelial dysplasia (29 mild, eight moderate, and seven severe). The sensitivity (se) and specificity (sp) of chemiluminescence for the detection of a dysplastic lesion were 77.3% and 27.8%, respectively. Conclusion: While ViziLite has the ability to detect OPMD, it does not accurately delineate dysplastic lesions. The device can be used as a general oral mucosal examination system and may in particular improve the visualization of leukoplakias.     

Dental hygienists and dentists as providers of brush biopsies for oral mucosa screening

Background

Oral cancer is a severe and potentially fatal disease usually starting in the squamous epithelium lining the oral cavity. Together with oropharyngeal carcinoma, it is the fifth to sixth most common malignancy worldwide. To limit the increase in the global oral cancer incidence over the past two decades, the World Health Assembly adopted a resolution urging member states to integrate preventive measures such as engagement and training of dental personnel in screening, early diagnosis, and treatment into their national cancer control programs.

Aim

The aim of this study was to investigate if dental hygienists (DHs) and dentists (Ds) in general dental practice care can be entrusted to perform brush sampling of oral potentially malignant disorders (OPMDs), and to evaluate their level of comfort in performing brush biopsies.

Methods

Participants were five DHs and five Ds who received one day of theoretical and clinical training in oral pathology to identify OPMDs (leukoplakia [LP], erythroplakia [EP], and oral lichen planus [OLP]), and perform brush sampling for PAP cytology and high-risk human papillomavirus (hrHPV) analysis.

Results

Out of 222 collected samples, 215 were adequate for morphological assessment and hrHPV analysis. All the participants agreed that sample collection can be incorporated in DHs and Ds routine clinical duties, and most of them reported that sample collection and processing was easy/quite easy.

Conclusion

Dentists and DHs are capable of collecting satisfactory material for cytology and hrHPV analysis. All the participating DHs and Ds were of the opinion that brush sampling could be handled routinely by DHs and Ds in GDP.     

Diagnostic efficiency of differentiating small cancerous and precancerous lesions using mucosal brush smears of the oral cavity—a prospective and blinded study

The aim of this study was to evaluate the diagnostic accuracy of oral brush biopsy to identify early malignancy. One hundred and eighty-six brush biopsies of suspicious mucosal lesions were obtained, haematoxilin and eosin (H&E)-stained and compared with the histology of conventional excision biopsies of the same site performed concomitantly. The sensitivity for identifying squamous cell carcinoma (SCC) was 88.5%. High-risk lesions including squamous intraepithelial neoplasia (SIN II, SIN III) and SCC were identified with a sensitivity of 86.4%, using a pap-analogous classification, which is considered to be carcinomatous, as well as moderate and severe dysplastic cells positive. Depending on the cytopathologic definition for malignancy and the tumour size, the test accuracy varied: Extending the cytopathologic criteria for malignancy by defining all dysplastic or malignant cytopathologic findings as positive, the sensitivity was increased to 95.2% at the expense of the specificity, which was reduced from 94.9% to 82.3%. Separately analysing SCCs of less than 20 mm, the sensitivity was reduced by 9.5% to 78%. Although small malignant lesions seem to be less reliable by the conventional oral brush biopsy, it is a useful screening instrument for early diagnosis of suspicious, epithelial lesions and could therefore contribute to improved cancer prognosis.     

Cytologic and DNA-cytometric examination of oral lesions in lichen planus

BACKGROUND: The aim of the present study was to evaluate the diagnostic value of exfoliative cytology (EC) and DNA image cytometry applied to oral lesions of lichen planus (LP; n = 56), in order to detect or exclude malignant transformation. METHODS: Brush and excisional biopsies were obtained from 56 patients. In cases of oral LP in which brush biopsies were suspicious for tumor cells, nuclear DNA contents were measured, using a TV Image Analysis System. RESULTS: In 50 patients EC yielded tumor cell-negative, doubtful in four cases and suspicious results obtained in two cases. DNA image cytometry revealed DNA-aneuploidy only in the two suspicious cases. The comparison between cytologic/DNA-cytometric diagnosis and biopsy histology resulted in a total agreement (LP without dysplasia: 54 and squamous cell carcinoma in LP: two cases). CONCLUSIONS: In conclusion, cytology with DNA-cytometry is a highly sensitive, specific, and non-invasive method, which can be used for periodical follow up of oral LP lesions in order to early detect or exclude malignancy.     

用DNA倍体分析评估口腔黏膜疾病严重程度

目的:用DNA倍体分析法判断口腔黏膜病病变的严重程度。方法:用小刷刷取口腔黏膜病变处的上皮细胞,直接涂片制成1-2张玻片,经Feulgen染色,用全自动DNA图像分析仪测定细胞核内DNA含量。部分患者在可疑处活检做组织病理检查,将活检病例分成DNA倍体分析组和阴性组,比较两组病例病变的严重程度。结果:303例口腔黏膜病例中有口腔溃疡237例、口腔扁平苔藓17例、红斑34例、白斑25例。经DNA倍体分析其中267例为阴性,36例为阳性。将经活检组织病理检查97例患者分成DNA倍体分析阳性组和阴性组。在36例DNA倍体分析阳性病例中有11例组织像正常或炎性病变、6例轻度异常增生、9例中度异常增生、6例重度异常增生和4例浸润癌,中度异常增生以上改变19例,其阳性率为52.78%;61例DNA倍体分析阴性病例中发现有52例组织像正常或炎性病变、6例轻度异常增生、3例中度异常增生和1例浸润癌,中度异常增生以上改变3例,其阳性率为4.91%。经方差分析,两组之间有显著性差别(x2=30.98,P<0.005)。结论:测定口腔黏膜病变处细胞DNA含量是一种无创伤且能判别出病变严重程度的方法。     

Exfoliative cytology: a helpful tool for the diagnosis of paracoccidioidomycosis

OBJECTIVE: To describe the main cytological findings associated with smears collected from oral lesions of paracoccidioidomycosis and to appraise the use of cytology as a diagnostic tool for the disease. STUDY DESIGN: Cytological smears and biopsies were collected from 40 lesions with a clinical suspicion of paracoccidioidomycosis. Evaluation of the sensitivity, specificity, positive and negative predictive values, accuracy and the positive likeness ratio of the oral smear when compared with the histological diagnosis, was performed. The latter is considered the 'gold standard' for comparison. RESULTS: The main morphological findings were the rounded-shaped, birefringent and multiple-budded fungi, Langhans' giant cells and epithelioid cells. The following associative measures were found: sensitivity, 67.9%; specificity, 91.7%; positive predictive value, 95.0%; negative predictive value, 55.0%; accuracy, 75.0%; and positive likeness ratio, 8.14. CONCLUSION: The cytological findings of paracoccidioidomycosis are characteristic and cytology is accurate in the diagnosis of the disease. Positive patients should be treated. Negative patients should be submitted to biopsy to confirm or to dismiss the diagnosis of this mycosis.     

Computer-assisted analysis of the oral brush biopsy

Computer-assisted analysis of the oral brush biopsy is a recently introduced tool that determines the significance of an oral lesion. The oral brush biopsy is minimally invasive, requires no anesthesia, and definitively distinguishes benign from precancerous and cancerous lesions. Oral brush biopsy specimens are analyzed with the aid of a highly specialized neural network-based computer system specifically designed to detect oral epithelial precancerous and cancerous cells.     

Improving detection of precancerous and cancerous oral lesions. Computer-assisted analysis of the oral brush biopsy. U.S. Collaborative OralCDx Study Group

BACKGROUND: A study group composed of researchers from across the United States undertook a study to evaluate the sensitivity and specificity of OralCDx (OralScan Laboratories Inc.), a computer-assisted method of analysis of the oral brush biopsy, in the detection of precancerous and cancerous lesions of the oral mucosa. METHODS: The study group conducted a multicenter double-blind study comparing results of OralCDx analysis with those of scalpel biopsy of suspicious oral lesions, as well as using OralCDx on oral lesions that appeared benign clinically. RESULTS: In 945 patients, OralCDx independently detected every case of histologically confirmed oral dysplasia and carcinoma (sensitivity = 100 percent, false-negative rate = 0 percent). Every OralCDx "positive" result was subsequently confirmed by histology as dysplasia or carcinoma. The specificity for the OralCDx "positive" result was 100 percent, while the specificity for the OralCDx "atypical" results was 92.9 percent. In 4.5 percent of clinically benign-appearing lesions that would not have received additional testing or attention other than clinical follow-up, OralCDx uncovered dysplasia or carcinoma (statistical sensitivity > 96 percent, P < .05, n = 131; statistical specificity for the OralCDx "positive" result > 97 percent and for the "atypical" result > 90 percent, P < .05, n = 196). CONCLUSIONS: The authors propose that this multicenter trial demonstrates that OralCDx is a highly accurate method of detecting oral precancerous and cancerous lesions. OralCDx can aid in confirming the nature of apparently benign oral lesions and, more significantly, revealing those that are precancerous and cancerous when they are not clinically suspected of being so. All OralCDx "atypical" and "positive" results should be referred for scalpel biopsy and histology to completely characterize the lesion. CLINICAL IMPLICATIONS: Given the difficulty in clinically differentiating premalignant and malignant lesions from benign lesions with a similar appearance, OralCDx appears to determine the significance of an oral lesion definitively and detect innocuous-appearing oral cancers at early, curable stages.     

甲苯胺蓝活体染色在口腔癌早期诊断中的临床价值

对８４例早期口腔病损患者用１％甲苯胺蓝染色，观察着色情况，与病理结果对照。结果显示甲苯胺蓝对口腔癌诊断的敏感性９７．３％，特异性为４１．２％，假阳性率４２．２％，假阴性率５．０％；白斑癌变及上皮异常增生性白斑的染色阳性率明显高于上皮单纯增生性白斑；４３例溃疡病变都呈阳性染色，炎性溃疡染色，周界与恶性溃疡无显著性差异。     

Image processing analysis of oral cancer,oral potentially malignant disorders,and other oral diseases using optical instruments

Oral cancer screening is important for early detection and early treatment, which help improve survival rates. Biopsy is invasive and painful, while fluorescence visualization using optical instruments is non-invasive, convenient, and provides results in real time, and examinations can be repeated. The purpose of this study was to determine the usefulness of optical instruments in oral screening. A total of 314 patients who were examined using optical instruments at Tokyo Dental College between 2014 and 2018 were enrolled in this study. Fluorescence visualization images were analyzed using subjective and objective evaluations. Subjective evaluation for detecting oral cancer offered 98.0% sensitivity and 43.2% specificity. Regarding the objective evaluations for detecting oral cancer, sensitivity and specificity were 61.9% and 62.7% for mean luminance, 90.3% and 55.7% for luminance ratio, 56.5% and 67.7% for standard deviation of luminance, and 72.5% and 85.4% for coefficient of variation of luminance. Fluorescence visualization with subjective and objective evaluation using optical instruments is useful for oral cancer screening.     

光动力治疗口腔黏膜癌及癌前病变的临床进展

口腔癌是发生在口腔的恶性肿瘤之总称,严重影响人类生命和生活,发病率高,致残率和病死率高.许多口腔癌由癌前病变发展而来,最常见的是口腔黏膜白斑、扁平苔藓等.目前针对口腔癌一线治疗方案分为:手术切除、放射线治疗、化学治疗以及联合治疗.特别是高龄口腔癌症患者,传统治疗尚有一定的局限性和毒副作用.针对癌前病变常用药物和手术治疗...     

Cytomorphometric Analysis for Evaluation of Cell Diameter,Nuclear Diameter and Micronuclei for Detection of Oral Premalignant and Malignant Lesions

Purpose: Detection of a precancerous or cancerous lesion when small is one of the most important factors to improve 5-year survival rates of oral cancer. Although surgical biopsy is the Gold Standard for diagnosing oral lesions, it is impractical to routinely subject large numbers of patients to biopsy. This study had been undertaken to evaluate quantitatively the cellular changes in exfoliated cells from different premalignant and malignant lesions in terms of cell diameter (CD), nuclear diameter (ND) and nucleocytoplasmic ratio and micronucleus frequency. Patients and Methods: Oral mucosal smears were obtained from patients presenting with precancer and cancer using a cytobrush immediately before biopsy. The study group consisted of Group I: lesions with no dysplasia, Group It: lesions with mild and moderate dysplasia, Group Ul: lesions with severe dysplasia and carcinoma in situ, Group IV: lesions with invasive carcinoma. Results: Comparison of study group and controls showed a highly significant decrease in mean cellular diameter, increase in the nuclear diameter and a larger nucleo cytoplasmic ratio of exfoliated cells. (p <0.0001) Also micronuclei (MN) frequencies were found higher in SCC patients and in precancer than in control subjects. Conclusions: Cytomorphometric analysis via oral brush biopsy is a valuable adjunct to biopsy & CD, ND and micronuclei evaluation serves as important diagnostic markers.