Lymphangiogenesis and angiogenesis in oral cavity and lower lip squamous cell carcinoma / Linfangiogênese e angiogênese em carcinomas de células escamosas de lábio inferior e da cavidade oral

ABSTRACT INTRODUCTION: Tumors of the lip and oral cavity differ in various aspects; therefore a clarification of the distinctions among these sites may help to better understand the biologic behavior of neoplasms occurring in these locations. OBJECTIVE: Considering that angiogenesis and lymphangiogenesis are two major elements that can influence various aspects of tumor biology, we aimed to compare these factors between squamous cell carcinoma of the lower lip and oral cavity. METHODS: A total of 84 primary squamous cell carcinomas including 45 oral and 39 lower lip tumors were selected and immunohistochemically stained with monoclonal antibody against D2-40 and CD105. Mean microvessel density was assessed in tumoral tissue, while lymphatic vessel density was calculated in both neoplastic tissue and invasion front. Data were statistically analyzed using t-test and p-values of <0.05 were considered significant. RESULTS: We found a mean microvessel density ± standard deviation of 31.94 ± 18.9 in oral cavity and 27.54 ± 20.8 in lower lip squamous cell carcinomas, with no significant difference (p = 0.32). Mean lymphatic vessel density ± standard deviation was 13.05 ± 8.2 and 16.57 ± 10.79 in of oral cavity and lower lip neoplastic tissue, respectively. The corresponding values were 9.94 ± 5.59 and 12.50 ± 7.8 in the invasive front. Significant differences were not observed in either of the lymphatic vessel density variables between the two sites. CONCLUSION: According to our results, it seems that the search for additional factors other than those related to the vasculature should continue, to help clarify the differences in biologic behavior between lower lip and oral cavity squamous cell carcinomas.

Resumo Introdução: Os tumores de lábio e da cavidade oral diferem em vários aspectos; portanto, o conhecimento das diferenças entre eles pode ajudar na melhor compreensão do comportamento biológico das neoplasias que ocorrem nesses locais. Objetivo: Considerando que a angiogênese e a linfangiogênese são dois elementos importantes que podem influenciar diversos aspectos da biologia dos tumores, objetivamos comparar esses fatores entre o carcinoma de células escamosas (CCE) de lábio inferior e da cavidade oral. Método: No total, foram selecionados 84 CCEs primários (45 tumores da cavidade oral e 39 tumores de lábio). Esses tumores foram corados por processo imunohistoquímico com anticorpo monoclonal anti-D2-40 e CD105. Avaliamos a densidade média de microvasos (DMV) no tecido tumoral, enquanto que a densidade vascular linfática (DVL) foi calculada tanto no tecido neoplásico como no front de invasão. Os dados foram estatisticamente analisados com o uso do teste t e valores de p < 0,05 foram considerados significantes. Resultados: Chegamos a uma média para DMV ± DP de 31,94 ± 18,9 para CCEs na cavidade oral e de 27,54 ± 20,8 no lábio inferior, sem diferença significante (p = 0,32). As médias para DVL ± DP foram de 13,05 ± 8,2 e 16,57 ± 10,79 no tecido neoplásico da cavidade oral e lábio inferior, respectivamente. Os valores correspondentes foram 9,94 ± 5,59 e 12,50 ± 7,8 no front invasivo. Não foram observadas diferenças significantes nas duas variáveis DVL entre os dois locais. Conclusão: De acordo com os nossos resultados, a pesquisa por fatores adicionais, além daqueles relacionados à vasculatura, deve ter continuidade, para auxiliar no esclarecimento das diferenças do comportamento biológico entre CCEs no lábio inferior e na cavidade oral.

Lymphangiogenesis and angiogenesis in oral cavity and lower lip squamous cell carcinoma.

INTRODUCTION: Tumors of the lip and oral cavity differ in various aspects; therefore a clarification of the distinctions among these sites may help to better understand the biologic behavior of neoplasms occurring in these locations. OBJECTIVE: Considering that angiogenesis and lymphangiogenesis are two major elements that can influence various aspects of tumor biology, we aimed to compare these factors between squamous cell carcinoma of the lower lip and oral cavity. METHODS: A total of 84 primary squamous cell carcinomas including 45 oral and 39 lower lip tumors were selected and immunohistochemically stained with monoclonal antibody against D2-40 and CD105. Mean microvessel density was assessed in tumoral tissue, while lymphatic vessel density was calculated in both neoplastic tissue and invasion front. Data were statistically analyzed using t-test and p-values of <0.05 were considered significant. RESULTS: We found a mean microvessel density±standard deviation of 31.94±18.9 in oral cavity and 27.54±20.8 in lower lip squamous cell carcinomas, with no significant difference (p=0.32). Mean lymphatic vessel density±standard deviation was 13.05±8.2 and 16.57±10.79 in of oral cavity and lower lip neoplastic tissue, respectively. The corresponding values were 9.94±5.59 and 12.50±7.8 in the invasive front. Significant differences were not observed in either of the lymphatic vessel density variables between the two sites. CONCLUSION: According to our results, it seems that the search for additional factors other than those related to the vasculature should continue, to help clarify the differences in biologic behavior between lower lip and oral cavity squamous cell carcinomas.

VEGF, CD105 and α-SMA immunoexpression study in lips squamous cell carcinomas and associated dysplastic lesions.

This study analyzes the microvascular density (MVD) for CD105+ and α-SMA+ vessels and VEGF immunoexpression for 35 oral squamous cell carcinomas and for the associated dysplastic lesions of the lips. CD105+ MVD was superior in the advancing edge compared to the intratumoral area, no matter the analyzed clinico-pathological parameters (gender, age, differentiation degree, tumor stage) (p<0.05), MVD being significantly higher in poorly differentiated carcinomas (p<0.05). α-SMA+ MVD was also superior in the advancing edge compared to the intratumoral area (p<0.05), MVD values being significantly higher in well and moderately differentiated carcinomas (p<0.05). CD105+ MVD and α-SMA+ MVD were significantly lower compared to the analyzed tumor area (p<0.05), in the dysplastic lesions. VEGF score showed significantly higher values in well to moderately differentiated carcinoma and in the tumor area versus dysplastic associated lesions (p<0.05). CD105+ MVD and VEGF are markers able to characterize the angiogenic phenotype of carcinomas and of the dysplastic lesions of the lips, while α-SMA+ MVD quantification is useful in assessing the vascular maturity degree.

A comparative study of microvessel density in squamous cell carcinoma of the oral cavity and lip.

OBJECTIVES: The objective of this study was to comparatively evaluate the density of lymphatic vessels (LVD) and neoformed microvessels (NMVD) in squamous cell carcinoma of the oral cavity (OCSCC) and lip (LSCC). Association between LVD/NMVD and vascular endothelial growth factor (VEGF)-A/-C was also assessed. STUDY DESIGN: OCSCC and LSCC were compared with regard to immunoexpression of LVD, NMVD, and vascular endothelial growth factor-A (VEGF)-A/-C. Association between VEGF-A/-C with vascularity was also assessed. Statistical analyses were performed using t test, Pearson χ(2), and Mann-Whitney tests. Statistical significance was accepted at P less than .05. RESULTS: The NMVD and VEGF-C expressions were significantly higher in OCSCC compared with LSCC. NMVD was associated with VEGF-C in OCSCC, but not in LSCC. CONCLUSIONS: Differences in NMVD and VEGF-C were found between OCSCC and LSCC. Positive association between VEGF-C and NMVD was observed in OCSCC, but not in LSCC, which may be one of the contributing factors that account for the distinctive clinical-biological behavior of these lesions.

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.

Density and migration of mast cells in lip squamous cell carcinoma and actinic cheilitis.

Mast cells (MCs) display a diversity of roles that may contribute to the stromal microenvironment alterations during tumor progression. The aim of this study was to investigate MC populations expressing tryptase and c-kit in lip squamous cell carcinoma (lip SCC) (n=37), actinic cheilitis (AC) (n=15) and normal lip mucosa (control) (n=6), as well as their relationship with microscopic parameters (collagen degeneration, elastin changes, angiogenesis and proliferative index). Tryptase, c-kit, CD31 and Ki-67 expressions were analyzed by means of immunohistochemistry and collagen and elastic fibers were visualized with Picrosirus and Verhoeff's stain, respectively. The numbers of tryptase+ MC were significantly higher in lip SCC when compared with control (P=0.01), while a similar density of these cells was observed in AC and lip SCC (P=0.09). The density of c-kit+ MC was similar in all groups examined (P=0.65). MC migration (c-kit+/Tryptase+ relationship) was 69% in lip SCC, 60% in AC and 100% in control. The number of CD31+ blood vessels was significantly higher in the lip SCC when compared with control and AC (P<0.01). The increase of MCs and angiogenesis in lip SCC may reflect an important modification in the tumor microenvironment during squamous photo-carcinogenesis.

Carcinoma de células escamosas de labio inferior: asociación entre grado de angiogénesis, graduación histológica y frente de invasión tumoral / Lower lip squamous cell carcinoma: relationship between angiogenesis grade, histological grading and invasion tumoral front

El carcinoma de células escamosas de la cavidad oral presenta una alta prevalencia en nuestro país, siendo el labio inferior el sitio más comúnmente afectado. El objetivo de este estudio fue determinar el grado de angiogénesis en el carcinoma de células escamosas de labio inferior, y su asociación con el grado de diferenciación según la Clasificación Internacional de Tumores y el Frente de Invasión Tumoral según el sistema de medición de Bryne, en pacientes con carcinoma de células escamosas de labio inferior, diagnosticados en los hospitales de Talca y Curicó, entre los años 1995 y 2005.

Squamous cell carcinoma of the oral cavity present a high prevalence in our country, become the lower lip the most common site affected. The aim of this study was determinated the angiogenesis grade in the lower lip squamous cell carcinoma and their relationship whit the histological grading according to the International Tumours Classification and the Invasion Tumoral Front according to the Bryne's system measuring in patients with lower lip squamous cell carcinoma diagnosed in the Talca´s and Curico´s Hospital between 1995 and 2005 years.

Simultaneous demonstration of mast cells and blood vessels by the combined method CD34--alcian blue-safranin in lip tumors.

The aim of the study was to evaluate the mast cell-blood vessel relationship using double staining CD34/AAS. Sections from 14 cases with lip tumors have been stained with Hematoxylin-Eosin. On additional sections from each case, we highlighted blood vessels by immunohistochemistry for CD34 antigen using the method LSAB2-HRP/DAB, followed by alcian blue-safranin stain for mast cells. We quantified the density, distribution and the mast cell types as well as the correlation with the number of blood vessels. All cases have been positive for both staining. We observed a significant correlation between the number of vessels and the mast cells (p = 0.003). In one case, we observed the mast cells stained with safranin (red), the vascular density being less than the mast cells density. Our results confirmed the data from the literature with respect to the large number of mast cells observed in the malignant tumors. The increased vascular density together with the mast cell density suggests a correlation between these two elements in the tumor angiogenesis, possibly though the VEGF secretion. The CD34/AAS stain is a quick and simple method and it allows an optimal correlation between the number of mast cells and blood vessels on the same section. The type of mast cells correlated with microvessel density is a powerful argument towards the involvement of the mast cells in the tumor angiogenesis of the malignances of the lips.

Immunohistochemical expression of vascular endothelial growth factor correlates with positive surgical margins and recurrence in T1 and T2 squamous cell carcinoma (SCC) of the lower lip.

Tumor angiogenesis is associated with tumor progression and aggressiveness in a number of malignancies, and vascular endothelial growth factor (VEGF) is considered as a leading candidate in this process. The aim of the present study was to investigate the role of VEGF immunohistochemical expression and tumor angiogenesis in predicting local recurrence in surgically treated lip SCC. We performed a retrospective analysis of 50 patients with lip SCC in order to investigate whether VEGF immunohistochemical expression and tumor angiogenesis correlate with clinicopathologic parameters and outcome. Tumor angiogenesis was estimated by determining microvessel density (MVD) with the use of CD34 antibody. Our results showed that VEGF was strongly correlated with tumor invasion towards the surgical margin. There was also a significant association of high VEGF expression with a higher incidence of local recurrence (p < 0.001). We suggest that VEGF expression may be used as an index to distinguish patients with higher risk of relapse.

T1 and T2 lip cancer: a superselective method of facial arterial infusion therapy--preliminary experience.

PURPOSE: To formulate and evaluate a facial arterial infusion chemotherapy for squamous cell lip carcinoma. MATERIALS AND METHODS: The study included six patients (age range, 46-84 years) with squamous cell carcinoma of the lower lip. There were two T1 tumors, three T2 tumors, and one T1-compatible postoperative recurrent tumor. A 4-F, double-lumen balloon catheter was inserted into the external carotid artery through the superficial temporal artery and placed for selective infusion into the tumor-feeding facial artery. Patients received a combination of mitomycin C (4.4 mg/m2 per body surface area) on day 1 and 3.2 mg/m2 of peplomycin sulfate on days 1-7 (22.4 mg/m2 per week), or, when peplomycin sulfate was contraindicated, 16 mg/m2 of cisplatin only on days 1-5 (80 mg/m2 per week). Two to three cycles of chemotherapy were given until tumor disappearance was histologically confirmed. RESULTS: Complete tumor disappearance was achieved in all cases. One patient had a self-limiting asthma attack during peplomycin sulfate treatment, and another had transient partial hair loss. No disfigurement, recurrence, or late complications were observed at a mean follow-up of 5.0 years (range, 2.3-11.2 years). CONCLUSION: The described facial arterial infusion chemotherapy appears to be a safe and curative treatment for T1 and T2 squamous cell lip carcinomas.

Angiogenesis in invasive squamous cell carcinoma of the lip: tumor vascularity is not an indicator of metastatic risk.

BACKGROUND: Recent studies have shown that tumor growth beyond a certain size requires angiogenesis. Microvessel density has, moreover, correlated with metastatic risk in some tumors. Invasive squamous carcinoma (SCC) can develop in the epithelium of the lip and metastasize even when relatively small. This study investigates neovascularization and its relationship to metastatic risk in this tumor. METHODS: All 41 primary SCCs of the lip diagnosed at our institution from 1960 to 1991 were immunostained for factor VIII. Microvessel grade (Mv) from 1+ to 4+ and the average number of vessel profiles (TMvD) in the highest density 200 x (0.785 mm2) and 400 x (0.196 mm2) microscopic fields were determined. TMvDs were compared with those of adjacent non-tumor tissue (NTMvD). Normalized counts (TMvDns) were calculated as TMvD/NTMvD. TMvDs and TMvDns of metastasizing (N = 10) and non-metastasizing (N = 31) tumors were compared (student t-test). RESULTS: In all SCCs TMvDs exceeded NTMvDs (50 vs. 35 at 200 x, P = 0.0014, and 26 vs. 14 at 400 x, P < 0.0001). Metastasizing and non-metastasizing tumors did not, however, differ in Mv, TMvD, or TMvDn. CONCLUSIONS: Angiogenesis develops, but is not quantitatively related to metastatic risk, in primary invasive squamous cell carcinoma of the lip.

Multinucleate cell angiohistiocytoma of the upper lip.

Multinucleate cell angiohistiocytoma is a rare benign fibrohistiocytic vascular tumor with distinctive histologic features. It appears clinically as small, slow-growing, grouped reddish-brown to purple papules and nodules that may be mistaken for other vascular proliferative or reactive conditions. Multinucleate cell angiohistiocytoma tends to occur more frequently in women, to favor the extremities, and to appear as multiple lesions. We report a case of solitary multinucleate cell angiohistiocytoma that appeared as a nodule on the upper lip of a man but that otherwise demonstrated characteristic histologic and immunohistochemical features. Greater awareness of this uncommon benign lesion will facilitate diagnosis and appropriate management.

Prognostic factors in squamous cell carcinoma of the lower lip.

One hundred eighty-seven squamous cell carcinomas of the lower lip were examined microscopically to identify parameters that might predict metastasis and patient outcome. Excision specimens of 157 nonmetastasizing carcinomas (group I) were compared with specimens from 30 tumors that had metastasized (group II). The following features were recorded: architectural pattern; microscopic thickness (mm); cytologic grade; presence of muscle, nerve, or vessel invasion; inflammatory response; and mitotic rate. The mean thickness was 2.5 mm for group I tumors and 7.5 mm for group II tumors. Seventy-six per cent of the group I tumors were 3 mm thick or less, whereas only one group II lesion (3 per cent) was less than 3 mm thick. Five per cent of the group I neoplasms, compared with 77 per cent of the group II tumors, were at least 6 mm thick. Perineural invasion was seen in 5 per cent of the group I and 41 per cent of the group II lesions. Three per cent of the group I carcinomas had a dispersed pattern, compared with 57 per cent of those in group II. One group I lesion (0.6 per cent) and 37 per cent of the group II tumors were grade 4. Each of these differences was statistically significant (P less than 0.0001). For all lesions studied, metastases had occurred in 60 per cent with perineural invasion, 74 per cent measuring 6 mm or more, 77 per cent with a dispersed pattern, and 92 per cent that were grade 4. These important prognostic variables were best evaluated in the deeper portions of the lesions.

Complications of embolization treatment for problem cavernous hemangiomas.

Radiologists trained in selective angiography can now deliver sclerosing or occluding materials to the arterial supply of vascular tumors. Such skills require special training and there are limitations to the use of these materials. Specific obliteration can be carried out if this technique is quality controlled. Materials being used for embolization are continually being studied and are changing as new materials are developed. Radiologists can assist surgeons in managing some of these very difficult problems. A large series of patients with various vascular lesions treated at the Oregon Health Sciences University was reviewed. The complications of embolization used for cavernous hemangiomas have tempered our enthusiasm for it as an adjunct form of therapy. We do feel that this method of treatment remains useful in selected cases. The occlusion achieved caused a decrease in both symptoms and size of lesions treated. The decrease in size excluded the need for surgery in 3 cases, aided resection in 1, but caused significant complicating slough in 1. Five cases are presented to demonstrate problems of tissue necrosis encountered.

Angiography in the evaluation of cryosurgery efficiency in the treatment of a haemangioma of the face. A case report.

The authors present the use of carotid angiography for the diagnosis of maxillofacial cavernous haemangiomata. Radiological investigation of vessels permits the identification of the exact site and range of pathological change but also the evaluation of the mechanism of tissue healing following cryogenic treatment.