Computer assisted analysis of the microvasculature in metastasized and nonmetastasized squamous cell carcinomas of the tongue

Hannen, E.J.M.; Laak, Jeroen van der; Manni, Johannes J.; Freihofer, Hans Peter M.; Slootweg, Piet J.; Koole, R.; Wilde, P.C.M. de

doi:10.1002/hed.10100

Cited by 26 publications

(18 citation statements)

References 33 publications

(46 reference statements)

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“…Moreover, the whole procedure could be automated through specialized software, using a PC connected to a light microscope. 35 Some limitations of our study should be discussed. First, we estimated the total number of vessels (blood and lymphatics) using CD34.…”

Section: Discussionmentioning

confidence: 94%

Comparison of the diagnostic accuracy of lymphatic endothelium markers: Bayesian approach

2005

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Tumor lymphatic density is evaluated by means of specific lymphatic endothelium markers, and is a potential predictor of clinically meaningful outcomes. There are many claims on the postulated superiority of some of these markers to identify lymphatics, always in the absence of quantitative data. We therefore compared the diagnostic accuracy of the antibody against podoplanin and the commercially available D2-40, employing Bayesian statistics to account for the absence of a gold standard. We used the pan-endothelial marker CD34 to identify 23 542 distinct blood and lymphatic vessels in sections from 30 formalin-fixed, paraffin-embedded archival tissue blocks of head and neck squamous cell carcinoma specimens. We stained two adjacent sections with podoplanin and D2-40 and identified the continuum of each stained vessel in the sections with a comprehensive method. Overall, 1864 vessels were stained with both markers, 119 only with podoplanin and 391 only with D2-40. Significantly more vessels with intraluminal red blood cells were stained with D2-40 compared to podoplanin (McNemar's Po0.0001). Both antibodies had extremely high specificity (99.7% (95% credible interval (CrI): 99.5-99.9%) and 98.8% (95% CrI: 98.3-99.5%) for podoplanin and D2-40, respectively) and very high sensitivity (92.6% (95% CrI: 86.1-97.9%) and 97.3% (95% CrI: 94.9-99.2%) for podoplanin and D2-40, respectively). Inferences were qualitatively similar when we took into account in the analyses the possibility that the two tests (antibodies) may be correlated. We calculated that 96.3% (95% CrI: 94.2-98.6%) of the vessels stained with podoplanin and 88.9% (95% CrI: 83.9-95.7%) of the vessels stained with D2-40 were truly lymphatics. These numbers were in agreement with the observed number of stained vessels without intraluminal red blood cells. Our results suggest that both antibodies are excellent lymphatic endothelium markers and that there may be little reason to prefer either of them in most settings.

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Section: Discussionmentioning

confidence: 94%

Comparison of the diagnostic accuracy of lymphatic endothelium markers: Bayesian approach

2005

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“…MVD was assessed by hot spot method using computerized image morphometric analysis as described in previous studies [13][14]. Briefly, in the carcinoma cases, areas representative of the invasive component of the cancer were selected from sections stained with hematoxylin and eosin ( Figure 1A).…”

Section: Tissue Samplingmentioning

confidence: 99%

“…It was done as described previously [14] using: Motic binocular microscope, computer system, Motic images plus software, version 2.0 (Motic China Group Co., Ltd, Shenzhen, China), "Image Pro ® Express", version 4.0 (Media Cybernetics L.P., Bethesda, USA).…”

Section: Computerized Image Analysismentioning

confidence: 99%

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

et al. 2011

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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.

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“…The obtained results were tabulated and compared. [6] Areas representative of the invasive component of the cancer were selected from sections stained with hematoxylin and eosin. Microvessels were counted and assessed for morphometric parameters (area and perimeter) using "Image Pro" software.…”

Section: Methodsmentioning

confidence: 99%

Comparison of Vascular Hot Spots Method with Randomly Chosen Microscopic Fields Method in the Morphometric Assessment of Neovascularization in OSCC

Kushwaha¹

2017

AIMDR

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Background: Despite all efforts made to develop predictive biomarkers for antiangiogenic therapies, no unambiguous markers have been identified so far. The most widely used method of quantifying neovascularity is immunohistochemistry, by which microvessel density is determined using endothelial markers, such as CD34. There are several methods to morphometrically assess neovascularization in OSCC. Aims and Objectives: To compare vascular hot spots method with randomly chosen microscopic fields method in the morphometric assessment of neovascularization in OSCC. Methods: Our study included 30 paraffin embedded tissue blocks of diagnosed cases of OSCC and 10 controls. Tissue sections of 4µ thickness were taken and immunostaining by anti CD34 monoclonal antibody was performed to demonstrate endothelial cells. Endothelial areas and Microvascular density was assessed by hot spot method and randomly chosen microscopic fields method using computerized image morphometric analysis and. The obtained results were tabulated and compared. Results: A statistically significant difference was found between normal mucosa and carcinoma (P<0.001). Endothelial areas values measured by the hot spot method were superior than the values obtained by the random field method. Conclusion: Among both the methods, hot spots method was found to be superior than random chosen microscopic fields method.

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Computer assisted analysis of the microvasculature in metastasized and nonmetastasized squamous cell carcinomas of the tongue

Cited by 26 publications

References 33 publications

Comparison of the diagnostic accuracy of lymphatic endothelium markers: Bayesian approach

Comparison of the diagnostic accuracy of lymphatic endothelium markers: Bayesian approach

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

Comparison of Vascular Hot Spots Method with Randomly Chosen Microscopic Fields Method in the Morphometric Assessment of Neovascularization in OSCC

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