Quantitative physiology and immunohistochemistry of oral lesions

Abstract

Angiogenesis and hypoxia are reported to correlate with tumor aggressiveness. In this study, we investigated the potential of optically measured total hemoglobin concentration (THC) and blood oxygen saturation (StO 2 ) as a quantitative measure of angiogenesis and hypoxia in oral lesions with an immunohistochemical comparison. 12 normal subjects and 40 oral patients (22 oral squamous cell carcinoma (SCC), 18 benign/premalignant lesions including 11 verrucous hyperplasia (VH) and 7 hyperkeratosis/parakeratosis (HK)) were studied. The results showed that the THC measurement was consistent with vascular endothelial growth factor (VEGF) and microvessel staining in the stromal area, but StO 2 was not associated with HIF-1α. We observed inflammation induced neovascular formation in the stromal area of VH and HK that were likely attributed to higher-than-control THC and StO 2 and resulted in no difference in optical measurements between all lesions. However, we found that in majority of SCC, the ratio of THC and StO 2 levels between lesions and the surrounding tissues provide potential distinguishing characteristics from VH, which are not visually differentiable from SCC, with a sensitivity, specificity, and accuracy of 91%, 68%, and 76%, respectively.

Current relevance of hypoxia in head and neck cancer

“…Chronic inflammation associated with poor oral hygiene and Lichen Planus (an autoimmune disease) and oral cancer has been shown in more than 1% of afflicted patients [62]. In early premalignant lesions, inflammation-induced neovascularisation has been observed in the stromal component of verrucous hyperplasia and hyperkeratosis [67]. The composition of cytokine/chemokines, liberated from monocytes at a site of inflammation and which are important in the development of a chronic state of disease, can broadly be divided into pro-inflammatory and anti-inflammatory cytokines.…”

Lesion oxygenation associates with clinical outcomes in premalignant and early stage head and neck tumors treated on a phase 1 trial of photodynamic therapy

“…Interpatient heterogeneities in S t O 2 of oral lesions have also been measured by others. Lee et al [37] report ranges of ~300 mM in tHB and up to 40% in S t O 2 among patients with squamous cell carcinoma, verrucous hyperplasia, or hyperkeratosis/parakeratosis. These ranges are similar to those in the present study.…”

“…We detect baseline S t O 2 to be ~40%. Some have reported average values in the range of 81-93% [37–39] or ~65-75% [40]. These differences can potentially be explained by the depth of tissue that was probed.…”

“…These differences can potentially be explained by the depth of tissue that was probed. Our instrument probed to tissue depths of ~1.7 mm, but less than half of this distance was probed by the above-mentioned studies [37]. We have previously examined the effect of probing depth, finding that superficial measurements detected higher values of S t O 2 in oral lesions [10].…”

Semaphorin 3E Inhibits House Dust Mite–Induced Angiogenesis in a Mouse Model of Allergic Asthma

“…Immunofluorescence staining of vWF and CD31 was analyzed quantitatively by counting the number of blood vessels positively stained for vWF and CD31 in five randomly chosen high-power fields of four tissue sections from each mouse. 28 The result was reported as an average of the number of red blood vessels in each group. 28 Moreover, the result of VEGF, VEGFR1, and VEGFR2 staining was analyzed quantitatively by calculating the signal mean intensity of 20 randomly chosen high-power fields from each group using ImageJ software version 1.48v (NIH, Bethesda, MD; https://imagej.nih.gov/ij).…”