The diagnostic value of Superb Microvascular Imaging in identifying benign tumors of parotid gland.

BACKGROUND: We compared the ultrasound features, superb microvascular imaging (SMI) and micro vessel density (MVD) of pleomorphic adenoma (PA), Warthin's tumor (WT) and basal cell adenoma (BCA) to explore the clinic value of SMI in differential diagnosis of benign tumors of parotid gland. METHODS: The vascular distributions and grade by color doppler flow imaging (CDFI) and SMI, as well as vascular index (VI) of 249 parotid gland masses from 217 patients were analyzed. RESULTS: The internal echogenicity of BCA are more homogeneous in comparing with WT and PA(P < 0.05). By SMI, the vascular distribution and vascular grade in PA were mainly peripheral (33.1%) and avascular (25.7%), Grade 1 (27.8%) and Grade 0 (25.7%). WT were mainly central (31.3%) and mixed distribution (34.9%), in Grade 3 (37.3%) and Grade 2 (36.2%). BCA was mainly peripheral (33.3%) and mixed distribution (33.3%), in Grade 2 (33.3%) and Grade 3 (33.3%). The overall detection rate of SMI for vascular Grade 2 and 3 was significantly higher than that of CDFI (P < 0.05). Both VI and MVD were lowest in PA, highest in WT (P < 0.001). The VI by SMI was correlated with MVD (P < 0.001). The correlation index between vascular distribution and grade by SMI and MVD were significantly higher than CDFI. CONCLUSION: SMI can provide low-velocity blood flow information, which is helpful for the differential diagnosis of common benign tumors of parotid gland, and is expected to be more widely used.

Vascular Pattern Analysis on Microvascular Sonography for Differentiation of Pleomorphic Adenomas and Warthin Tumors of Salivary Glands.

OBJECTIVES: Pleomorphic adenomas and Warthin tumors are the most common salivary gland tumors. It is important to differentiate between them because at least a partial parotidectomy is necessary for pleomorphic adenomas, whereas enucleation is sufficient for Warthin tumors. This study aimed to evaluate the usefulness of vascular pattern analysis using microvascular sonography to differentiate between the tumors. METHODS: Sixty-two patients with pathologically proven pleomorphic adenomas (n = 38) and Warthin tumors (n = 24) were included. For all tumors, grayscale, power Doppler, and microvascular sonographic examinations were performed. Differences in vascular patterns (vascular distribution and internal vascularity) on power Doppler and microvascular sonography as well as grayscale sonographic features (size, shape, border, echogenicity, heterogeneity, and cystic change) between pleomorphic adenomas and Warthin tumors were evaluated. A comparison of diagnostic performances of grayscale sonography with power Doppler sonography and grayscale sonography with microvascular sonography was performed. The level of interobserver agreement between 2 reviewers in diagnosing tumors was evaluated. RESULTS: No grayscale sonographic features showed a significant difference between the tumors. Vascular distributions and internal vascularity on power Doppler sonography (P = .01 and .002) and microvascular sonography (both P < .001) were all significantly different. The diagnostic accuracy of grayscale sonography with microvascular sonography (79.0%) was higher than that of grayscale sonography with power Doppler sonography (72.6%). This difference was significant according to the McNemar test (P = .004). Interobserver agreement was excellent in diagnosing tumors on both grayscale sonography with power Doppler sonography (κ = 0.83) and grayscale sonography with microvascular sonography (κ = 0.94). CONCLUSIONS: Vascular pattern analysis using microvascular sonography with other sonographic features is helpful for differentiating between pleomorphic adenomas and Warthin tumors.

Immunoexpression of GLUT-1 and angiogenic index in pleomorphic adenomas, adenoid cystic carcinomas, and mucoepidermoid carcinomas of the salivary glands.

This study aimed to evaluate and compare the immunoexpression of glucose transporter-1 (GLUT-1) and angiogenic index between pleomorphic adenomas (PAs), adenoid cystic carcinomas (ACCs), and mucoepidermoid carcinomas (MECs) of the salivary glands, and establish associations with the respective subtype/histological grade. Twenty PAs, 20 ACCs, and 10 MECs were submitted to morphological and immunohistochemical analysis. GLUT-1 expression was semi-quantitatively evaluated and angiogenic index was assessed by microvessel counts using anti-CD34 antibody. Higher GLUT-1 immunoexpression was observed in the MECs compared to PAs and ACCs (p = 0.022). Mean number of microvessels was 66.5 in MECs, 40.4 in PAs, and 21.2 in ACCs (p < 0.001). GLUT-1 expression and angiogenic index showed no significant correlation in the tumors studied. Results suggest that differences in biological behavior of the studied tumors are related to GLUT-1. Benign and malignant salivary gland tumors differ in the angiogenic index; however, angiogenesis may be independent of the tumor cell's metabolic demand.

Density of mast cells and microvessels in minor salivary gland tumors.

The aim of this study was to investigate the density of mast cells and microvessels in minor salivary gland tumors. Forty-one cases of minor salivary gland tumors (pleomorphic adenoma, n = 10; adenoid cystic carcinoma, n = 11; mucoepidermoid carcinoma, n = 10; and polymorphous low-grade adenocarcinoma) were investigated using immunohistochemistry for mast cell tryptase and von-Willebrand factor. Density of mast cells was higher in mucoepidermoid carcinoma; however, no differences in the number of these cells were observed between the different types of tumors (p > 0.05). The number of mast cells was higher in periparenchymal areas in all tumors, but the difference was not significant (p > 0.05). Mucoepidermoid carcinoma showed the largest number of periparenchymal mast cells, whereas pleomorphic adenomas showed the smallest number of intraparenchymal mast cells (p > 0.05). The highest microvessel density was observed in mucoepidermoid carcinomas, being this difference statistically significant when mucoepidermoid carcinoma was compared to pleomorphic adenoma (p = 0.0034) and polymorphous low-grade adenocarcinoma (p = 0.004). Microvessel density was significantly higher in adenoid cystic carcinoma when compared to pleomorphic adenoma (p = 0.0406) and polymorphous low-grade adenocarcinoma (p = 0.0123). Comparison of mast cells and microvessel densities showed no significant difference between tumors. A quantitative difference in mast cells and microvessels was observed, particularly in mucoepidermoid carcinoma, a finding supporting the aggressive behavior of malignant salivary gland tumors without myoepithelial differentiation. Further studies are needed to determine the role of mast cells in angiogenesis, as well as in the development and biological behavior of these tumors.

Extracellular matrix metalloproteinase inducer expression in salivary gland tumors: a correlation with microvessel density.

The purpose of this study was to investigate the expression pattern of extracellular matrix metalloproteinase inducer (EMMPRIN) as well as the correlation between EMMPRIN and microvessel density (MVD) in salivary gland tumors. Extracellular matrix metalloproteinase inducer expression and MVD were examined immunohistochemically on paraffin-embedded tissue specimens from 95 patients with salivary gland tumors, who underwent surgical resection from 1998 to 2006. Reverse transcription-polymerase chain reaction was used to monitor EMMPRIN mRNA expression in frozen samples. Extracellular matrix metalloproteinase inducer expression in mucoepidermoid carcinomas and adenoid cystic carcinomas was significantly higher than in normal salivary gland tissues and pleomorphic adenomas (P < 0.05). The MVD of mucoepidermoid carcinomas and adenoid cystic carcinomas was significantly higher compared with pleomorphic adenomas (P < 0.05). The MVD of the EMMPRIN-positive expression group was significantly higher than the MVD of the EMMPRIN-negative expression group (P < 0.05). Extracellular matrix metalloproteinase inducer mRNA expression in malignant salivary gland tumors was higher than that in pleomorphic adenomas (P < 0.05). This study suggests that EMMPRIN expression is an important feature of malignant salivary gland tumors and can be used as a biologic marker to characterize salivary gland tumors. Extracellular matrix metalloproteinase inducer is also a positive angiogenic factor in salivary gland tumors.

[Cervical giant tumor in a 56-year-old female patient]. / Zervikaler Riesentumor bei einer 56-jährigen Patientin.

We report the case of a 56-year-old female patient with a giant tumor on the left side of the neck, which appeared to be localised outside the large salivary glands. Due to the extensive vascularisation seen on MRI, digital subtraction angiography was performed. To reduce intraoperative bleeding, the vasculature was embolized. The tumor was resected together with the submandibular gland, since a connection between gland and tumor could not be excluded intraoperatively. Histology showed a pleomorphic adenoma with an intact capsule and no indication of malignancy. Pleomorphic adenomas are typical tumors of the salivary glands; however, they can also occur outside the gland. The tumor and its pseudocapsule must be fully resected to prevent recurrence.

Angiogenic and lymphangiogenic microvessel density in recurrent pleomorphic adenoma.

BACKGROUND: Recurrent pleomorphic adenoma (RPA) is an uncommon and challenging disease. The aim of this study was to determine if there is a difference between RPA and the pleomorphic adenoma (PA) without recurrence related to tumor blood and lymphatic vascularization. Moreover, we compared the microvessel density (MVD) between cell-rich areas (predominance of epithelial cells) and cell-poor areas (predominance of myxoid and chondroid areas) of the stroma of PA and RPA. In addition, immunohistochemical staining for the Ki-67 antigen was conducted simultaneously to evaluate cell proliferation in PA and RPA. METHODS: A total of 19 cases of PA and 24 cases of RPA, blood, and lymphatic vessels were analyzed by immunohistochemical technique using the antibodies CD34, CD105, D2-40, and Ki-67. RESULTS: Comparing no recurrent with recurrent tumor, no significant difference was found in terms of lymphatic vessel density, MVD, and proliferation index. When MVD and proliferation index were compared with different areas in cellular composition (cell-rich and cell-poor areas), there was a significant difference in PA, as well as in RPA. CONCLUSION: This study shows that although RPA presents more aggressive clinical behavior than PA, there is no difference between tumor blood and lymphatic vascularization, suggesting that there is no correlation between vascularity and risk of recurrence. Furthermore, vascularized stroma in PA, as well as RPA, depends on the proportion of the cellular composition.

Angiogenic switch during tumor progression of carcinoma ex-pleomorphic adenoma.

We analyzed the tumor vascularization in carcinomas ex-pleomorphic adenoma (CXPA) to investigate the angiogenic switch during the malignant transformation of pleomorphic adenoma (PA) to carcinoma and during tumor progression. In eight cases of early CXPA (intracapsular and minimally invasive tumors), eight of advanced CXPA (widely invasive tumors), and ten of PA without malignant transformation, tumor vascularization was assessed in histological samples by measuring total microvascular area (TVA) and microvessel density (MVD) using CD34 and CD105 antibodies. MVD for CD105 increased significantly during tumor progression, whereas this was not the case for CD34 MVD. Comparing widely invasive CXPA with and without myoepithelial differentiation, CXPA with myoepithelial differentiation showed a significantly lower number of CD105 positive vessels but revealed higher TVA values. In these tumors, the neoplastic cells usually formed larger hypovascularized aggregates that were often surrounded by large-sized vessels. In conclusion, the antibody CD105 reveals an angiogenic switch during the progression from adenoma to carcinoma in salivary glands. The degree of angiogenesis and the total vascular area have distinctive patterns in CXPA with and without myoepithelial differentiation. Low angiogenesis associated with high TVA value is more characteristic of CXPA with myoepithelial differentiation.

Evidence of lymphangiogenesis in Warthin's tumor of the parotid gland.

The details of the pathogenesis of cystadenolymphoma (Warthin's tumor) of the parotid gland are still unclear. Neovascularization is considered to be a pivotal factor for solid tumor progression and biological behavior of the tumor. Using double-labeling immunohistochemistry for LYVE-1 and CD34 (specific markers for lymphatic and vascular endothelial cells, respectively) this study analyzes lymphatic vessel density (LVD) and blood vessel density (BVD) in 10 Warthin's tumors and 10 pleomorphic adenomas of the parotid gland as well as in 5 normal parotid glands and 5 normal parotid lymph nodes. There was no significant difference in the intratumoral LVD and BVD among pleomorphic adenoma and normal parotid gland tissue. In contrast, the intratumoral LVD and BVD were significantly higher in Warthin's tumor than pleomorphic adenoma, normal parotid gland and parotid lymph node (P<0.0001 versus P<0.004). The increase in lymphatic vessels in Warthin's tumor suggests that epithelial tumor cells might promote lymphangiogenesis in this kind of lesions.

Vascular endothelial growth factor in salivary pleomorphic adenomas: one of the reasons for their poorly vascularized stroma.

To better understand the poorly vascularized background of the stroma of pleomorphic adenomas, we attempted to determine the expression of molecules related to blood vessels and hypoxic conditions in pleomorphic adenoma. Surgical specimens and tumor cells in primary culture of salivary pleomorphic adenomas were used for immunohistochemistry for CD31, vascular endothelial growth factor (VEGF) and its receptors Flk-1 and Flt-1, as well as for hypoxia markers, such as hypoxia-inducible factor-1alpha (HIF-1alpha) and lactate dehydrogenase-1 (LDH). At the same time, alternative splicing modes of the VEGF gene and expression levels of the HIF-1alpha gene were analyzed in surgical specimens by means of reverse-transcription polymerase chain reaction (RT-PCR) and direct sequencing of the PCR products. In addition to co-immunolocalization with CD31+ vascular endothelial cells, VEGF and its receptors were demonstrated in normal duct epithelial and myoepithelial cells as well as in tumor cells in ductal structures and in myxochondroid stromata. Immunolocalizations for HIF-1alpha and LDH were confirmed in the VEGF-positive area. Immunofluorescence signals for VEGF and others were confirmed in pleomorphic adenoma cells in culture. RT-PCR results showed that there were at least four splicing modes of the VEGF gene, among which VEGF(121) was most enhanced, and higher HIF-1alpha levels in pleomorphic adenomas. The results suggest that pleomorphic adenoma cells produce VEGF in several functional forms for their own proliferation or differentiation, and that the VEGF expression is controlled by hypoxic circumstances of poorly vascularized pleomorphic adenomas.

Spontaneous infarction of a pleomorphic adenoma in parotid gland: diagnostic problems and review.

Although infarction of parotid gland pleomorphic adenoma (PA) following fine-needle aspiration (FNA) has been well-documented, spontaneous infarction of PA has remained as an uncommon entity in the literature. To our knowledge, we report the second case of spontaneous infarction occurring in a parotid gland PA. A 44-yr-old man presented with a 2-yr history of slowly enlarging right parotid mass, which had become painful 1 mo before performing FNA biopsy. Smears revealed abundant necrotic debris, atypical squamous cells, and small cells with dark nuclei suggestive of a carcinoma. Histologic examination of the tissue fragments demonstrated degenerated clusters of cells and chondromyxoid matrix. The parotidectomy specimen had features consistent with those of an infarcted PA. Although appears to be rare, spontaneous infarction of PA should be considered in the differential diagnosis, since necrosis may mimic carcinoma and cause misinterpretation of necrosis as an indication of malignancy, in an otherwise benign salivary gland neoplasm.

Abnormal venous structures in salivary gland tumors: vasculature characteristics of pleomorphic adenoma.

To clarify the diagnostic significance of abnormal venous structures present in salivary gland tumors, we examined 21 pleomorphic adenomas, 14 Warthin tumors, 1 oncocytic adenoma, 3 myoepitheliomas, 7 basal cell adenomas, 5 mucoepidermoid carcinomas, and 6 adenoid cystic carcinomas. Verhoeffvan Gieson staining was carried out and the morphology of the veins within the tumors was observed microscopically. Branching veins, thickened intima of the veins, discontinuous elastic membrane and multilayered elastic membrane were seen in 71.4%, 76.2%, 47.6% and 85.7% of pleomorphic adenomas, respectively, and were abundant and easily found in most cases. The abnormal venous structures were also found in other salivary gland tumors examined, but they were few in number and lacked variety. Elastic fibers extending radially into the surrounding stroma were seen in 66.7% of pleomorphic adenomas, and were not seen in other salivary gland tumors. Our results showed that a variety of abnormal venous structures are more abundant and more easily found in pleomorphic adenoma compared with other salivary gland tumors, and, in particular, that perivascular radiating elastic fibers are characteristic of pleomorphic adenoma. We emphasize that the presence of perivascular radiating elastic fibers may be helpful in diagnosing pleomorphic adenoma in small biopsy specimens.

Salivary gland tumours: is colour Doppler imaging of added value in their preoperative assessment?

Using a Toshiba SSA-270A Colour Duplex Scanner, 15 patients with suspected parotid and submandibular gland tumours had preoperative duplex scanning done. Peak systolic doppler shifts (fmax) were recorded and their vascularity was also subjectively scored. There were no arteries identified within the substance of contralateral normal glands. All recordings were analysed retrospectively and an independent histological assessment of tumours was made. Pleomorphic adenomas (n = 9) had a median peak systolic frequency (fmax) of 0.8 kHz, while adenolymphomas (n = 4) had a fmax of 1.65 and the two malignant tumours had fmax of 4.5 (P = 0.007 Kruskal-Wallis test). There appears to be a strong correlation between the fmax and the subjective vascular score (Corr 0.92). Colour duplex scanning is a non-invasive procedure which may be of help in the preoperative assessment of salivary gland tumours.

Primary pleomorphic adenoma in posterior cranial fossa.

A 35-year-old woman had an intradural tumor in the posterior fossa adjacent to the posterior wall of the left pyramidal bone, which was totally removed and histologically diagnosed as a pleomorphic adenoma. Follow-up examination for 2 years showed no recurrence of the tumor. There was no primary lesion in any other gland of the body, and therefore there is no alternative but to conclude a "migration" of some gland cells. The pathogenesis of this tumor remains unclassified.

Dynamic scintigraphy with thallium-201 chloride (Tl-201) for the diagnosis of tumors of the head and neck.

OBJECTIVE: The purpose of this study was to differentiate between malignant tumors and benign tumors of the head and neck by means of thallium-201 chloride scintigraphy. STUDY DESIGN: Thallium-201 chloride (Tl-201) scintigraphy and Ga-67 citrate (Ga-67) scintigraphy were performed in 10 patients with a benign tumor and in 90 patients with a malignant tumor. The blood flow index and the tumor retention index were obtained by means of Tl-201 scintigraphy. The blood flow index = the count of tumor/the count of control in the early dynamic scan from 30 to 120 seconds after injection, and the tumor retention index = (the count of tumor/the count of control in the delayed dynamic scan 2.5 hours after injection)/(the count of tumor/the count of control in the early dynamic scan from 4 to 5 minutes after injection). RESULTS: The early and delayed scans with Tl-201 demonstrated a very high correlation with the blood flow index. The tumor retention index had a tendency to increase in malignant tumors, especially in the poorly differentiated tumors and the severely invasive tumors. Likewise, the tumor retention index decreased in benign tumors. These results proved to be statistically significant, and in turn Tl-201 scintigraphy was deemed superior to Ga-67 scintigraphy. Tl-201 scintigraphy was also useful in detecting small-size tumors. CONCLUSION: The tumor retention index of Tl-201 was useful for the differentiation of malignant tumors of the head and neck from benign tumors of the same areas.

Infarction after fine needle aspiration biopsy of pleomorphic adenoma of the parotid gland.

BACKGROUND: Fine needle aspiration (FNA) is a well-established and safe method for the rapid diagnosis of pleomorphic adenoma. A few clinically important complications, including bleeding, infection and inflammatory reactions, result from FNA. In a small number of cases FNA has been followed by varying degrees of necrosis in some organs. In the literature there are a few reports associated with necrosis in a pleomorphic adenoma of the parotid gland following FNA. CASE: A 27-year-old female had a two-year history of a right parotid mass. FNA revealed pleomorphic adenoma. A histologic diagnosis of pleomorphic adenoma of the right parotid with infarction was made. CONCLUSION: Necrosis associated with infarction may cause diagnostic problems. It is not a sufficient sign of malignant transformation.

Spontaneous infarction of a parotid gland pleomorphic adenoma. Report of a case with cytologic and radiographic overlap with a primary salivary gland malignancy.

Pleomorphic adenoma is the most common neoplasm of the parotid gland, generally presenting as a slowly enlarging, firm, well-circumscribed, painless nodule. Occasional cases have presented after a short period of rapid growth or have been associated with pain. The vast majority of these tumors are solid, but rare examples have been associated with cystic degeneration or hemorrhage. Spontaneous and tumor-associated infarction of the parotid has been reported, but these examples have been limited to infarctions of Warthin's tumors and postoperative infarctions of salivary glands. We present the case of a 48-year-old male with a one-year history of a painful, enlarging, left parotid mass associated with paresthesia of the tongue. Computed tomographic examination of the parotid demonstrated a left superficial lobe mass with a rim of enhancement and low attenuation center. Fine needle aspiration yielded necrotic debris and atypical squamous elements that were thought to be compatible with carcinoma. A superficial parotidectomy with intraoperative frozen section revealed a pleomorphic adenoma with extensive central necrosis. To our knowledge, this represents the first reported case of an infarcted pleomorphic adenoma and illustrates the potential for misinterpretation of these cytologic and radiologic findings as indicative of malignancy.

Vascular endothelial growth factor (VEGF) expression and microvascular density in salivary gland tumours.

This study investigates whether salivary tumours with different morphology and evolution also differ in terms of neovascularization and VEGF expression and the prognostic value of the results. Surgical specimens from 45 patients - 8 pleomorphic adenomas (PA), 7 Warthin tumours (WT), 5 basal cell adenomas (BA), 6 carcinomas ex-pleomorphic adenoma (CEPA), 6 mucoepidermoid carcinomas (MEC), 5 acinic cell carcinomas (AC), 4 adenoid cystic carcinomas (ACC) and 4 adenocarcinomas not otherwise specified (ADK NOS) - were immunostained. In malignant salivary tumours, the following mean microvascular density (MVD) values were recorded (± SD = Standard Deviation): 27.61 (SD ± 2.27) in cases with CEPA, 27.08 (DS ± 7.81) in AC and 32.93 (SD ± 7.76) in ADK NOS, with lower values for MEC 24.31(SD ± 2.88) and for ACC 22.13 (SD ± 5.44). For benign tumours, an MVD of 35.71 (SD ± 2.09) was recorded in WT and lower average values in PA (MVD = 14.84; SD ± 4.86) and in BA (MVD = 23.96; SD ± 9.13). MVD did not correlate with the investigated clinicopathological parameters. The VEGF expression is significantly more important (p = 0.001) in malignant salivary tumours as compared with benign ones. The VEGF expression and the microvascularization in salivary gland tumours are important elements to be considered when formulating a diagnosis and assessing case evolutions in patients with such tumours.

Expression of Ki67 and CD105 as proliferation and angiogenesis markers in salivary gland tumors.

OBJECTIVE: To investigate the association between CD105 and tumor cell proliferation in salivary gland tumors. METHODS: In this study, 59 samples of salivary tumors from Khalili Hospital archive, including 20 cases of pleomorphic adenoma (PA), 20 cases of mucoepidermoid carcinoma (MEC) and 19 cases of adenoid cystic carcinoma, as well as 10 cases of normal salivary gland tissue, were reviewed by immunohistochemistry (IHC) for CD105 and Ki67 staining. RESULTS: CD105 positive vessels were absent in normal salivary gland tissue in the vicinity of tumors (51.6% of all tumors were positive). There was a statistically significant difference in frequency of CD105 staining between PA and malignant tumors and between four groups of different lesions (p<0.000) being highest in MEC. Intratumoral microvessel density was also elevated in malignant neoplasms (2.61 ± 3.1) as compared to PA (0.46 ± 0.6). Normal salivary glands did not express Ki67. There was a statistically significant difference in frequency and percentage of Ki67 immunoreactivity in malignant neoplasms (86.5% and 10.7 ± 10.8 respectively) compared to PA (50% and 0.78 ± 0.2) and among the four groups values were highest in MEC (p<0.000). CONCLUSION: n this study, it was observed a higher rate of angiogenesis and cellular proliferation was noted in malignant tumors compared to benign tumors, but no correlation was observed between these two markers.