The clinical effectiveness of reflectance optical spectroscopy for the in vivo diagnosis of oral lesions.

Optical spectroscopy devices are being developed and tested for the screening and diagnosis of oral precancer and cancer lesions. This study reports a device that uses white light for detection of suspicious lesions and green-amber light at 545 nm that detect tissue vascularity on patients with several suspicious oral lesions. The clinical grading of vascularity was compared to the histological grading of the biopsied lesions using specific biomarkers. Such a device, in the hands of dentists and other health professionals, could greatly increase the number of oral cancerous lesions detected in early phase. The purpose of this study is to correlate the clinical grading of tissue vascularity in several oral suspicious lesions using the Identafi(®) system with the histological grading of the biopsied lesions using specific vascular markers. Twenty-one patients with various oral lesions were enrolled in the study. The lesions were visualized using Identafi(®) device with white light illumination, followed by visualization of tissue autofluorescence and tissue reflectance. Tissue biopsied was obtained from the all lesions and both histopathological and immunohistochemical studies using a vascular endothelial biomarker (CD34) were performed on these tissue samples. The clinical vascular grading using the green-amber light at 545 nm and the expression pattern and intensity of staining for CD34 in the different biopsies varied depending on lesions, grading ranged from 1 to 3. The increase in vascularity was observed in abnormal tissues when compared to normal mucosa, but this increase was not limited to carcinoma only as hyperkeratosis and other oral diseases, such as lichen planus, also showed increase in vascularity. Optical spectroscopy is a promising technology for the detection of oral mucosal abnormalities; however, further investigations with a larger population group is required to evaluate the usefulness of these devices in differentiating benign lesions from potentially malignant lesions.

Narrow-band imaging system with magnifying endoscopy for early oral cancer.

It is often difficult to detect early oral cancer due to the specificity of the oral mucosa structure. The aim of this study was to investigate the potential of narrow band imaging (NBI) as an effective and non-invasive diagnostic tool in early oral cancer and other oral diseases. A magnifying endoscopy system manufactured by Olympus Corporation was used. A total of 121 subjects were included in the study. Subepithelial capillary loops were identified and categorized according to the classification of Inoue, with healthy mucosa graded as Type I or II, and that showing evidence of cancer-induced morphological change as Type III or IV. Sensitivity and specificity for the identification of oral cancer were estimated at 92.3% and 88.2%, respectively. Examination under a microscope with H&E staining and immunostaining for CD34 revealed dilation and extension of the capillaries in epithelial dysplasia, in addition to thickening of the epithelial layer. The present results indicate that use of NBI in conjunction with conventional magnifying endoscopy has great potential as an effective and non-invasive diagnostic tool in the early detection of oral cancer.

Light sources used in evaluating oral leukoplakia: broadband white light versus narrowband imaging.

This study aimed to investigate the clinical efficacy of using broadband white light (BWL) to observe morphologic appearance, narrow-band imaging (NBI) to observe intraepithelial microvasculature, and both BWL and NBI for the detection of high-grade dysplasia and carcinoma in oral leukoplakia. Among 317 patients (274 males and 43 females; aged 52.4±10.7 years), the odds ratio (95% confidence interval) for detecting high-grade dysplasia and carcinomatous lesions based on morphologic appearances of BWL, and microvasculature patterns of NBI, were 39.12 (9.33-64.10), and 97.16 (38.19-247.21), respectively, which were significantly better than BWL (p<1×10(-15)). The sensitivity, specificity, positive and negative predictive values, and accuracy of use of traditional BWL classification, NBI classification, and combined BWL and NBI classification for detecting high-grade dysplasia and carcinomatous lesions were 96.30, 60.08, 33.12, 98.75, 66.25, 39.92, and 3.70%; 87.04, 93.54, 73.44, 97.23, and 92.43%; and 100.00, 60.08, 33.96, 100.00, and 66.88%, respectively. In conclusion, the diagnostic accuracy by NBI classification of oral leukoplakia based on the intraepithelial microvasculature patterns is significantly better than BWL indicating that NBI is a promising non-invasive tool in detecting high-grade dysplasia and carcinomatous lesions in oral leukoplakia.

Clinical appraisal of endoscopy with narrow-band imaging system in the evaluation and management of homogeneous oral leukoplakia.

PURPOSE: To evaluate the clinical usefulness of endoscopy with a narrow-band imaging (NBI) system for the evaluation and management of homogeneous oral leukoplakia. METHODS: The chart records, morphology of vascular architecture of NBI, and histopathology of patients with homogeneous leukoplakia were retrospectively reviewed and analyzed. RESULTS: A total of 160 patients, with an average age of 50.96 ± 10.25 years, were enrolled. In 35 cases of thin leukoplakia, only intraepithelium papillary capillary loop (IPCL) type I was shown by NBI, and only squamous hyperplasia was revealed pathologically. In 125 cases of thick leukoplakia, IPCL type I was found in 94, IPCL type II in 29, and IPCL type III in 2. The Kendall rank correlation between pathology and NBI images was significant (p < 0.0001). CONCLUSIONS: The dichotomous classification of homogeneous leukoplakia by NBI is meaningful, and endoscopy with the NBI system is a promising tool for the evaluation and management of homogeneous oral leukoplakia.

Role of angiogenesis in oral squamous cell carcinoma development and metastasis: an immunohistochemical study.

Although a few studies have shown that vascularity is increased from normal mucosa to dysplasia to carcinoma suggesting that disease progression in the oral mucosa is accompanied by angiogenesis. The role in lymph node metastasis in oral squamous cell carcinoma (OSCC) is equivocal. Role of angiogenesis in OSCC development and metastasis is evaluated in this study. This retrospective study of 50 samples consisted of 9 normal buccal mucosa, 22 leukoplakias, and 19 OSCC. Polyclonal antibodies to von-Willebrand factor were used to highlight the microvessels. Images were captured and morphometric image analysis was done for microvessel density (MVD), area, and perimeter. Highest, as well as mean values of these three parameters were compared. MVD and perimeter, but not area, are significantly different between normal mucosa and OSCC, and leukoplakia and OSCC. There were no differences between normal mucosa and leukoplakia. MVD, area, and perimeter were not significantly different between the OSCC with and without lymph node metastasis. The highest and mean values of MVD are significantly correlated. In the development of OSCC, angiogenic phenotypic change occurs in carcinomas rather than in the pre-cancerous stage, and quantification of angiogenesis in OSCC does not predict the risk of lymph node metastasis.

Increased subepithelial vascularization and VEGF expression reveal potentially malignant changes in human oral mucosa lesions.

OBJECTIVE: The objective of this study was to provide evidence that the magnitude of angiogenesis induced by oral mucosa epithelium with potentially malignant lesions is related to the degree of epithelial aggressiveness. METHODS AND RESULTS: We evaluated 96 biopsies that included: (1) leukoplakia with and without dysplasia, (2) nontumoral borders adjacent to squamous cell carcinomas with and without dysplasia, and (3) normal oral mucosa. Number, size, and localization of vessels labeled immunohistochemically for the antigen CD34 were assessed by image analysis using a software developed "ad hoc." All vascular sections and those localized immediately below the epithelium (sub-basal vessels) were separately evaluated in areas 30-µm deep. Vascular endothelial growth factor (VEGF) expression was labeled immunohistochemically and evaluated semiquantitatively against a standard. Leukoplakia and nontumoral borders adjacent to carcinomas exhibited an increase in VEGF expression and in subepithelial vascularization. This increase was significantly greater in leukoplakia with dysplastic changes than in leukoplakia without dysplasia. Conversely, no differences were observed between epithelia with and without dysplasia adjacent to carcinomas. CONCLUSIONS: Demonstration of expression of epithelial VEGF and sub-basal vascularization could be an additional aid for evaluation of the severity of potentially malignant lesions in oral mucosa routine biopsies.

Utility of real-time diagnosis using contact endoscopy for oral and lingual diseases.

OBJECTIVE: In this study, we prospectively investigated the diagnostic accuracy of CE findings in oral and lingual diseases. METHODS: Between January 2004 and December 2009, CE was used to examine 66 patients with oral and lingual diseases at Hyogo College of Medicine Hospital. Blood vessel networks and superficial cell layers in the mucosal epithelium of normal and lesion sites were observed after staining with 1% methylene blue. Endoscopic diagnoses (CE diagnosis) were compared with subsequent definitive diagnoses based on pathological findings. The sensitivity and specificity for CE diagnosis were calculated. RESULTS: On CE findings, SCC showed the characteristics of absent and tortuous blood vessels, heterogeneous distribution, and increased nucleus/cytoplasm (N/C) ratio. Leukoplakia showed no atypical cells, abundant cornified layers, or cytoplasm without nuclei. Lesions were pathologically classified into three groups: 46 squamous cell carcinomas (SCC), 10 leukoplakias, and 10 other benign lesions (n=66). In 4 patients with SCC, malignancy was underestimated by CE findings. The overall diagnostic rate of the CE was 93.9% (62/66 patients). The sensitivity and specificity of SCC were 0.913 (42/46 patients) and 1.0 (20/20 patients), respectively. CONCLUSION: The usefulness of contact endoscopy (CE) as an in vivo real-time diagnostic instrument that can deliver results prior to pathological confirmation was suggested.

Detecting early oral cancer: narrowband imaging system observation of the oral mucosa microvasculature.

The aim of this study was to analyze and describe the intrapapillary capillary loops (IPCL), which are a feature of early oral neoplastic lesions, using a narrowband imaging (NBI) system. Forty-one patients (26 men, 15 women; mean age, 52.34 years; range, 23-83 years) presenting with non-neoplastic or neoplastic lesions, and normal cases, were examined using the prototype Evis Lucera Spectrum (Olympus Co.). The images were analyzed and an IPCL classification was devised. All normal cases (n=10) had regularly distributed capillary loops of the same shape (type I). Non-neoplastic lesions (n=8) had mild changes of the capillary loops (types II and III) and neoplastic lesions (n=23) were irregularly distributed and had several loop shapes (types III and IV). The microvascular organization of non-neoplastic lesions was notably different from that of neoplastic lesions. A brownish area was found in five cases of early carcinoma. The narrowband imaging system is a potential approach for clinically analyzing microvascular organization and IPCL. It could be useful for diagnosing oral squamous cell carcinoma at an earlier stage and for determining the margin of resection.

[Content of protein lactoferrin in precancer laryngeal disease]. / Soderzhanie belka laktoferrina pri predrakovykh zabolevaniiakh gortani.

Lactoferrin was studied in 50 patients with laryngeal precancer: chronic hyperplastic laryngitis (n=28), leukoplakia (n=22). Lactoferrin was measured before the treatment, 10-12 days and 12 months after the treatment. Blood serum concentration of lactoferrin was significantly increased before the treatment. By the authors, it was due to inflammation in laryngeal tissues. Conservative treatment of patients with chronic hyperplastic laryngitis resulted in a considerable reduction in this protein concentration. Surgical removal of leukoplakias stimulated laryngeal inflammation and raised lactoferrin concentration in blood serum. One year after the examination, those patients who had neither exacerbation of chronic hyperplastic laryngitis no recurrent leukoplakia exhibited serum lactoferrin in concentrations close to the baseline but still higher than in healthy individuals. This is explained by chronicity of inflammation in laryngeal tissues. These findings can be used in diagnosis and prognosis of the disease course in patients with laryngeal precancer.

Increase of mast cells and tumor angiogenesis in oral squamous cell carcinoma.

BACKGROUND: Although mast cells (MCs) have been implicated in promoting angiogenesis in some malignant tumors, especially of the aerodigestive tract, little is known in oral squamous cell carcinoma (SCC). METHODS: A retrospective study was conducted to elaborate upon the correlation between MCs and tumor angiogenesis in 26 cases of oral SCC, six cases of oral pre-malignant dysplasia, 10 cases of oral hyperkeratosis, and six cases of normal oral mucosa by means of immunohistochemical technique. RESULTS: The MCs in all lesions and normal oral mucosa strongly expressed tryptase. The densities of MCs and microvessels appeared to increase with disease progression. The MC and microvascular counts were significantly higher in oral SCC than in hyperkeratosis and normal oral mucosa (P < 0.05). A significant correlation between MC and microvascular densities was observed in oral SCC (r = 0.5; P = 0.012). CONCLUSIONS: These findings suggest that MCs may upregulate tumor angiogenesis in oral SCC, perhaps via MC tryptase.

Expression of granulocyte colony-stimulating factor receptor and platelet-derived endothelial cell growth factor in oral and oropharyngeal precancerous lesions.

Epithelial hyperplasia and dysplasia have been diagnosed as precancerous lesions and have been discussed in relationship to carcinogenesis. We analyzed the immunohistochemical expression of granulocyte colony-stimulating factor receptor (G-CSFR) and platelet-derived endothelial cell growth factor (PD-ECGF) in oral and oropharynx; 33 samples of normal epithelium, 28 samples of hyperplasia, 16 samples of dysplasia and 58 samples of squamous cell carcinoma. Also, we examined mean vessel density (MVD) by using CD34 staining and proliferating cell nuclear antigen (PCNA) staining. Dysplasia and head and neck Squamous Cell Carcinoma (SCC) exhibited higher G-CSFR expression and MVD than normal or hyperplastic epithelium (p <0.01). In the PD-ECGF staining, significant differences were found between SCC and normal epithelium, hyperplasia and dysplasia (p<0.001). In dysplasia and hyperplasia, PD-ECGF expression was significantly correlated with PCNA expression (r=0.345, p=0.025), however it was not correlated with the MVD. G-CSFR expression was not correlated with either PCNA or MVD. These results suggest that G-CSFR and PD-ECGF might be concerned with different carcinogenesis pathways of the squamous cells in the oral region and that PD-ECGF may be concerned with epithelial proliferation rather than angiogenesis.

Doppler sonography of the deep lingual artery.

PURPOSE: To clarify the Doppler sonographic features of the lingual artery in normal subjects and to evaluate those of patients with cancer of the tongue. MATERIAL AND METHODS: Sixty-seven volunteers and 12 patients with cancer and/or leukoplakia of the tongue were examined with an intraoral sonographic probe. The visibility of the deep lingual artery was determined on transverse and anteroposterior images. On the transverse images, the vascular index, which was defined as the number of colored pixels, was measured on bilateral lingual arteries. Thereafter, the degree of symmetry was evaluated for normal subjects and patients. RESULTS: In normal subjects, between younger and older volunteers, there were no significant differences in visibility of the trunk but differences were found between the two groups for the dorsal branches. The vascular indices of the right and left sides were not different. The characteristic Doppler sonographic feature was vasculature in and around the tumors in the patients with cancer of the tongue. The symmetry indices of the cancer patients were significantly different from those of normal subjects. CONCLUSION: Doppler sonography should be an important procedure for evaluation of tongue neoplasms.

[Study of resisting leukoplakia canceration and angiopoiesis by herba erigerontis].

OBJECTIVE: To evaluate the effect of resisting canceration of herba erigerontis and that of angiopoiesis. METHODS: Carcinomatous conversions of golden hamster cheek pouches induced by Salley way and obstructed by herba erigerontis were studied. Ink perfusion, analysis of images, resin cast form of micro-vessel, alpha-SMA detection and histopathological examination were applied to observe their changing patterns. RESULTS: The rate of leukoplakia canceration of herba erigerontis group was one time lower than that of model group. The rate of normal cells was about four times more than that of model. alpha-SMA of herba erigerontis was higher than that of model (P < 0.001). There was no significant difference between angeion vessel area and density (P > 0.05). Nevertheless, the resin cast form of micro-vessel showed that its configuration and spacious arrangement was similar to control group. CONCLUSIONS: Herba erigerontis has the function of preventing leukoplakia canceration. It has no obvious effect on angio-hyperplasia and expansion in the course of leukoplakia canceration. However, it has the protecting function on angio-configuration and spacious arrangement and the complete of angeion vessel wall. Probably its mechanism is to promote blood circulation by removing blood stasis and benign angiopoiesis so as to resist canceration.

Leukoplakia and angiopoiesis resistance of Herba Erigerontis.

OBJECTIVE: To evaluate the effects of Herba Erigerontis on angiopoiesis and carcinogenesis; to seek an effective drug for prevention of leukoplakia progression to carcinoma and to stop the mechanism of angiopoiesis. METHODS: Carcinomatous conversions of golden hamster cheek pouches induced by Salley way and inhibited by Herba Erigerontis. Ink perfusion, analysis of images, resin cast form of micro-vessel, a-SMA detection and histopathological examination were applied to observe their changing patterns. RESULTS: The rate of leukoplakia conversion to tumor of the Herba Erigerontis group was half that of model group. The proportion of normal cells was approximately 4 times greater than that of model. The a-SMA levels of the Herba Erigerontis treated group was higher than that of model (P < 0.001). There was no significant difference between angeion vessel area and density (P > 0.05) in these 2 groups, which were elevated compared to the control. Nevertheless, the resin casts of the microvasculature in the Herba Erigerontis group showed that its configuration and spatial arrangement was similar to the control group. CONCLUSIONS: Herba Erigerontis has the function of inhibiting leukoplakia progression to tumor. This may be due to its ability to upregulate the expression of a-SMA. It has no obvious effect on angio-hyperplasia and expansion in the course of leukoplakia progression to tumor. However, it may preserve the angio-configuration and spatial arrangement and keep intact the angeion vessel wall. Its mechanism of promoting blood circulation may be through removing blood stasis and benign angiopoiesis so as to resist carcinogenesis.

Study of the vascular pattern in oral lichen planus.

Oral Lichen Planus is a relatively frequent disease. Its etiopathogenesis is still unknown and it can undergo malignant transformation during its evolution. Thus, data which could contribute to the knowledge of the biology of this disease are particularly significant. The present study involves a quantitative evaluation of the vascular pattern of oral lichen planus. A portion of biopsy specimens taken for histopathologic diagnosis was processed to mark vascular walls using the histoenzymic technique for ATPase activity demonstration. Stained Sections were then evaluated in a semi-automatic magnetic image analyser. The stereologic parameters studied, showed there is no vascular increase in lichen with regard to normal mucosae or leukoplakias, since the number of vascular walls did not show significant differences. Instead, a significant increase was observed in the vascular area. The association of these parameters, indicates that lichen is a more congestive lesion than the other two conditions studied. These findings indicate that the modifications of the vascular pattern could play a role in the etiopathogenesis of oral lichen planus and suggest that the observation of these changes could be a useful element in the histopathologic diagnosis.