Rigid confocal endoscopy for in vivo imaging of experimental oral squamous intra-epithelial lesions.

BACKGROUND A rigid confocal endoscope has been developed to assess the oral squamous epithelium of mice and to determine sensitivity, specificity, and accuracy of this new technology. METHODS This endoscope is connected to the commercially available Heidelberg Retina Tomograph (HRT). HRT is a device with a 670-nm diode laser designed to acquire topographical measurements of the optic nerve head. Real-time rigid confocal endoscopy is demonstrated by imaging the epithelial lesions of a mice model. Six-week-old male C57Bl/6 mice were randomly divided into a non-treated group (n = 10) and into a 4-nitroquinoline 1-oxide (4-NQO)-treated group (n = 50). In the 4-NQO-treated group, the mice obtained 4-nitroquinoline 1-oxide in the drinking water (100 microg/ml) to induce tumourigenesis in the mouse tongue. The 4-NQO-solution was diluted in the drinking water for mice. After an 8-16-week carcinogen treatment with 4-NQO (ad libitum), mouse tongues were dissected within 3 h after CO(2) overdose. After confocal microscopy of all lesions of the tongue, conventional histopathological investigation was performed. RESULTS The inter-rater reliability for the two observers of the confocal microscopic findings was found to be Kappa = 0.59 (P < 0.001). The penetration depth varied in the healthy tissue of the underside of the tongue throughout this study and was measured between 104 and 240 microm. In keratotic lesions, the penetration depths were diminished and varied between 80 and 140 microm. Strong keratinization inhibits the evaluation of the epithelium. For differentiation between low-grade and high-grade squamous intra-epithelial lesions, a sensitivity and specificity of 73% and 88% was reached. CONCLUSIONS The animal experiment with this non-invasive new technology indicates that this imaging technology facilitates the detection of pre-cancerous lesions of the underside of the oropharynx. Human studies on oropharyngeal and laryngeal lesions are needed to prove the applicability of this method in the field of otorhinolaryngology.

[1]  Thomas Dreyer,et al.  Autofluorescence Endoscopy in the Diagnosis of Early Laryngeal Cancer and Its Precursor Lesions , 2002, The Laryngoscope.

[2]  R. Webb,et al.  In vivo confocal scanning laser microscopy of human skin: melanin provides strong contrast. , 1995, The Journal of investigative dermatology.

[3]  S. Tickoo,et al.  Oral Cavity and Esophageal Carcinogenesis Modeled in Carcinogen-Treated Mice , 2004, Clinical Cancer Research.

[4]  Joel B Epstein,et al.  Interobserver reliability in the histopathologic diagnosis of oral pre-malignant and malignant lesions. , 2004, Journal of oral pathology & medicine : official publication of the International Association of Oral Pathologists and the American Academy of Oral Pathology.

[5]  D. Macdonald,et al.  Oral epithelial atypia in denture induced hyperplasia, lichen planus and squamous cell papilloma. , 1975, International journal of oral surgery.

[6]  Rebecca R. Richards-Kortum,et al.  Fiber-optic confocal reflectance microscope with miniature objective for in vivo imaging of human tissues , 2002, IEEE Transactions on Biomedical Engineering.

[7]  S. Warnakulasuriya Histological grading of oral epithelial dysplasia: revisited , 2001, The Journal of pathology.

[8]  Michele Follen,et al.  Near real-time confocal microscopy of amelanotic tissue: detection of dysplasia in ex vivo cervical tissue. , 2002, Academic radiology.

[9]  Andreas Wree,et al.  Laser Scanning Microscopy of the Human Larynx Mucosa: A Preliminary, Ex Vivo Study , 2006, The Laryngoscope.

[10]  H. Pau,et al.  [In vivo diagnosis of epithelial changes of the oropharynx using confocal microscopy]. , 2007, Laryngo- rhino- otologie.

[11]  Pierre Lane,et al.  Direct fluorescence visualization of clinically occult high‐risk oral premalignant disease using a simple hand‐held device , 2007, Head & neck.

[12]  M Rajadhyaksha,et al.  Noninvasive Imaging of Human Oral Mucosa in Vivo by Confocal Reflectance Microscopy , 1999, The Laryngoscope.

[13]  A. Leunig,et al.  Detection of Squamous Cell Carcinoma of the Oral Cavity by Imaging 5‐Aminolevulinic Acid‐Induced Protoporphyrin IX Fluorescence , 2000, The Laryngoscope.

[14]  R Richards-Kortum,et al.  Near real time in vivo fibre optic confocal microscopy: sub‐cellular structure resolved , 2002, Journal of microscopy.

[15]  A. Karabulut,et al.  Observer variability in the histologic assessment of oral premalignant lesions. , 1995, Journal of oral pathology & medicine : official publication of the International Association of Oral Pathologists and the American Academy of Oral Pathology.

[16]  Paul M. Speight,et al.  The pathology of oral cancer and precancer , 1996 .

[17]  Calum MacAulay,et al.  Fluorescence Visualization Detection of Field Alterations in Tumor Margins of Oral Cancer Patients , 2006, Clinical Cancer Research.

[18]  J. R. Landis,et al.  An application of hierarchical kappa-type statistics in the assessment of majority agreement among multiple observers. , 1977, Biometrics.

[19]  H Stepp,et al.  Fluorescence staining of oral cancer using a topical application of 5-aminolevulinic acid: fluorescence microscopic studies. , 2001, Journal of photochemistry and photobiology. B, Biology.

[20]  J. Nakhosteen,et al.  A Pilot Study of Autofluorescent Endoscopy for the in Vivo Detection of Laryngeal Cancer , 2000, The Laryngoscope.

[21]  E. Eisenberg,et al.  Dysplasia of oral mucosa: a unified approach to proper evaluation. , 1991, Modern pathology : an official journal of the United States and Canadian Academy of Pathology, Inc.

[22]  E. Eisenberg,et al.  Intraexaminer and interexaminer reliability in the diagnosis of oral epithelial dysplasia. , 1995, Oral surgery, oral medicine, oral pathology, oral radiology, and endodontics.

[23]  G G Koch,et al.  A general methodology for the analysis of experiments with repeated measurement of categorical data. , 1977, Biometrics.

[24]  J. Bouquot,et al.  Oral epithelial dysplasia classification systems: predictive value, utility, weaknesses and scope for improvement. , 2008, Journal of oral pathology & medicine : official publication of the International Association of Oral Pathologists and the American Academy of Oral Pathology.