Assessment of Tumor Angiogenesis by Expression of CD 105 in Ameloblastoma, Odontogenic Keratocyst and Central Giant Cell Lesion.

BACKGROUND: Angiogenesis is critical for tumor growth and reflects the aggressive behavior of invasive odontogenic lesions [like Ameloblastoma (AM), Odontogenic Keratocyst (OKC) and Central giant cell lesion (CGCL)]. Mean vascular density (MVD) shows the angiogenic potential and CD105 is an ideal endothelial biomarker due to its specificity to new blood vessels for MVD detection. The aim of the study was to compare the MVD (angiogenic potential) among AM, OKC and CGCL in comparison to Pyogenic Granuloma (PG) using CD105 biomarker. METHODS: Sixty-four primary cases of odontogenic invasive tumors (AM, OKC and CGCL) and PG, diagnosed clinically and histologically were included in the study, with 16 samples in each group. Tissue samples of peripheral AM, Peripheral GCL of jaws, malignant AM, and specimen with insufficient tissue were excluded. Tissue sections were embedded, processed and stained using Hematoxylin and Eosin (H and E). Immunohistochemistry was performed using antibodies against CD105, with positive brown cytoplasmic staining in the endothelial cells of neo-vasculature. Distinct countable, positively stained endothelial cell or clusters were evaluated under light microscope for identification of MVD. ANOVA and t-test were applied for statistical analysis of data. RESULTS: Highest MVD was displayed in CGCL (32.99±0.77) and the minimum was observed in OKC (7.21± 0.75) respectively. CGCL showed significantly higher MVD to AM, OKC and PG lesions (p <0.05). AM (8.07± 0.36) and Odontogenic Keratocyst (7.21± 0.75) showed comparable MVD, which was lower than PG (14.7± 0.96) and CGCL vascular density (p < 0.01) respectively. CONCLUSION: CGCL was most aggressive, with highest MVD among the investigated odontogenic lesions (OKC, AM and PG). The proliferative aggressive behavior of Odontogenic Keratocyst is comparable to AM due to comparable mean vascular density.
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Can Differences in Vascularity Serve as a Diagnostic Aid in Fibro-Osseous Lesions of the Jaws?

PURPOSE: Different lesions in the fibro-osseous group share microscopic features; thus, establishing a definitive diagnosis based on microscopic features alone can be a challenge. There is a need for additional microscopic tools to aid in differentiating these lesions. This study compared parameters related to vascularity among 3 lesions in the fibro-osseous group: fibrous dysplasia (FD), central ossifying fibroma (COF), and cemento-osseous dysplasia (COD). MATERIALS AND METHODS: This study was a cross-sectional analysis of biopsied lesions retrieved from 3 medical centers over a 14-year period. The primary predictor variables were the vascularity parameters (number, perimeter, and area). The outcome variables were diagnoses of FD, COF, and COD. Diagnosis was based on clinical, microscopic, and radiologic correlations. From each histopathologic slide, 5 representative fields were captured with a computerized digital camera. The number of blood vessels was counted, and the surface area and vascular perimeter were measured by tracing the perimeter of each vessel. Data were statistically analyzed using analysis of variance with logarithmic transformation and a Tukey adjustment. RESULTS: Sixty-six cases were included in the study (26 in FD group, 26 in COF group, and 14 in COD group). The mean number of vessels showed only a tendency to be larger in the FD group compared with the COF and COD groups (5.4 ± 2.6, 3.7 ± 2.3, and 3.6 ± 1.7, respectively), but the results did not reach the threshold for significance. The mean vascular perimeter was 1,385.8 ± 859.2 pixels in the FD group and 742.6 ± 661.8 in COF group after logarithmic transformation (P = .012). The perimeter in the COD group was smaller (941.1 ± 502) compared with that in the FD group, but the difference did not reach the threshold for significance. The mean area was 25,061 ± 24,875.6 in the FD group and 11,773.8 ± 21,734.4 in the COF group after logarithmic transformation (P = .004). The perimeter in the COD group was smaller (13,011.1 ± 8,338.3) compared with the FD group, but the difference did not reach the threshold for significance. CONCLUSION: The vascular content of the FD group was markedly higher than of the COF group. These vascular changes can aid in differentiating these lesions microscopically.

Intra-epithelially entrapped blood vessels in ameloblastoma.

BACKGROUND: The ameloblastoma is a benign but locally aggressive odontogenic neoplasm with a high recurrence rate. While significant progress has been made in our understanding regarding the role of tumoral vasculature relative to the diverse behavioral characteristics of this tumor, no attention has been paid to a distinct subset of blood vessels entrapped within its epithelial compartment. As vascular niches are known to influence tumoral growth, clarification of these vessels is important. The objectives of this study were to investigate the morphologic characteristics of intra-epithelially entrapped blood vessels (IEBVs) in ameloblastoma and to speculate on their relevance. MATERIALS AND METHOD: Here, we evaluated the frequency, microvessel density (MVD), morphology, and distribution pattern of IEBVs in 77 ameloblastoma of different subtypes based on their immunoreactivity for endothelial markers (CD34, CD31, CD105), vascular tight junction protein (claudin-5), pericyte [α-smooth muscle actin (α-sma)], and vascular basement membrane (collagen IV). RESULTS: IEBVs were heterogeneously detected in ameloblastoma. Their mean MVD (CD34 = 15.46 ± 7.25; CD31 = 15.8 ± 5.04; CD105 = 0.82 ± 0.51) showed no significant correlation with different subtypes, and between primary and recurrent tumors (P > 0.05). These microvessels may occur as single/clusters of capillary sprouts, or formed compressed branching/non-branching slits entrapped within the epithelial compartment, and in direct apposition with polyhedral/granular neoplastic epithelial cells. They expressed proteins for endothelial tight junctions (claudin-5-positive) and pericytes (α-sma-positive) but had deficient basement membrane (collagen IV weak to absent). Aberrant expression for CD34, CD31, and CD105 in tumor epithelium was variably observed. CONCLUSIONS: Although rare in occurrence, identification of IEBVs in ameloblastoma could potentially represent a new paradigm for vascular assessment of this neoplasm.

Expression of CD34 and maspin in ameloblastoma from a West African subpopulation.

Ameloblastoma is a locally invasive odontogenic tumor with a high recurrence rate. Its local invasiveness is aided by angiogenesis, which can be correctly estimated by CD34. On the other hand, maspin decreases the local invasive and metastatic capability of cancer cells and functions as an angiogenesis inhibitor. We aim to assess the association between maspin expression and microvessel density in ameloblastoma. Twenty-five formalin-fixed paraffin-embedded (FFPE) blocks of ameloblastoma cases were prepared for antibody processing to CD34 and maspin. Positive immunohistochemical staining was marked by brown cytoplasmic/membrane coloration for CD34 and by nuclear/cytoplasmic coloration for maspin. At the ×40 magnification, we counted blood vessels in two areas of dimension; 300 × 400 µm (area A) and 150 × 200 µm (area B) adjacent to the tumor region to assess relative dispersion of the vessels bordering the tumor. The overall approximate microvessel density (MVD) for area A = 11 (minimum 2, maximum 21) and that for area B = 5 (minimum 1, maximum 10). The MVD in the area A of plexiform ameloblastoma was similar to that of the unicystic, while the hemangiomatous variant had the highest MVD for area A. Maspin positivity was present only in the cytoplasm of ameloblast, stellate reticulum, and the fibrous connective tissue in varying proportions. There was no evidence of the anti-angiogenesis effect of maspin in ameloblastoma from this study. The significance of cytoplasmic localization of maspin in the ameloblasts and stellate reticulum cells needs further investigation.

Tumor-associated macrophages and angiogenesis: a statistical correlation that could reflect a critical relationship in ameloblastoma.

Neoplasm growth is determined not only by the tumor cells themselves, but also by the tumor microenvironment. Increased densities of macrophages and activation of angiogenesis have been identified as common events in the progression of several neoplasms. Ameloblastoma is one of the most frequent odontogenic tumors and an excellent model for the study of neoplasm progression due to the different clinical variants that it exhibits. Here, by immunohistochemical studies using antibodies against CD68 and CD34, we evaluated the density of macrophages and microvessels associated to 45 paraffin-embedded ameloblastomas. In solid/multicystic ameloblastoma (SMA), we observed significantly higher densities of both macrophages and microvessels than in unicystic (UA) and desmoplastic (DA) ameloblastomas. Likewise, higher densities of macrophages and microvessels were found in UA than in DA. Furthermore, a predominance of intratumoral and peritumoral macrophage infiltrates was seen in SMA, while in UA, both macrophages and microvessels were also detected in the wall of the cysts. In contrast, DA had scant macrophages and microvessels, mainly situated distant from tumoral cells. In addition, a high correlation between macrophage and microvessel densities was observed in the samples (r=0.9623). Our results suggest that these two tumor microenvironmental elements could have an important role during ameloblastoma progression.

Comparison of angiogenesis in keratocystic odontogenic tumours, dentigerous cysts and ameloblastomas.

OBJECTIVE: The aim of the present study was to evaluate and compare angiogenesis in keratocystic odontogenic tumours, dentigerous cysts (DCs) and ameloblasomas using monoclonal antibody against CD34. MATERIALS AND METHODS: Microvessel density was assessed in a total of 53 cases including 20 keratocystic odontogenic tumours, 13 DCs and 20 ameloblastomas (14 solid and six unicystic variants). Microvessel density was expressed as the mean number of microvessels per high-power-field. RESULTS: Statistically significant differences in mean microvessel density were observed between keratocystic odontogenic tumours, DCs and solid ameloblastomas (P < 0.001). Mean microvessel density was significantly higher in solid ameloblastomas compared with both keratocystic odontogenic tumours and DCs; and was also significantly higher in keratocystic odontogenic tumours than in DCs. CONCLUSION: Within the limitations of the present study, it can be suggested that angiogenesis may be one of the mechanisms possibly contributing to the different biological behaviours of keratocystic odontogenic tumours, DCs and solid ameloblastomas.

Expression and significance of hypoxemia-inducible factor-1alpha in osteosarcoma of the jaws.

OBJECTIVE: The objective of this study was to examine the expression of hypoxia-inducible factor-1alpha (HIF-1alpha) in osteosarcoma of the jaw (OSJ) and its relationships with tumoral angiogenesis and clinicopathologic characteristics. STUDY DESIGN: Streptavidin peroxidase immunohistochemical staining was used to detect the expression of HIF-1alpha and CD34 in paraffin-embedded samples from 25 patients. There were 14 male and 11 female patients, ages ranged from 3 to 63 years. RESULTS: HIF-1alpha was overexpressed in OSJ. HIF-1alpha expression was correlated with tumoral microvascular density, as well as with the size, pathologic grade, and clinical stage of OSJ. Expression of HIF-1alpha was also correlated with clinicopathologic characteristics, such as primary occurrence or recurrence of the tumor, but not with metastasis. CONCLUSION: HIF-1alpha was significantly correlated with the size, pathologic grade, clinical stage, and primary occurrence or recurrence of the OSJ. HIF-1alpha appeared to promote tumoral angiogenesis in OSJ and may be an important target in antitumoral therapy.

Immunohistochemical evaluation of giant cell tumors of the jaws using CD34 density analysis.

PURPOSE: To compare CD34 expression in both aggressive and nonaggressive giant cell lesions of the jaws and identify any associations between tumor vascular density and biologic behavior. MATERIALS AND METHODS: This was a retrospective study of subjects treated for giant cell lesions of the jaws at Massachusetts General Hospital from 1992 to 2006. The primary predictor variable was tumor classification (aggressive or nonaggressive); tumors were considered aggressive if they were greater than 5 cm in size, recurred after treatment, or exhibited 3 of the following: presence of root resorption, tooth displacement, or cortical bone thinning or perforation. Secondary predictor variables, recorded for each patient, were demographic, anatomic, and clinical measures. The outcome variable was the average CD34 staining density of histologic specimens quantified in 2 different areas. Descriptive and bivariate statistics were computed to identify predictors associated with vascular density. RESULTS: The study sample was composed of 32 subjects with a mean age of 24.4 +/- 19.77 years (range: 2-83); 23 subjects (71.8%) were female. Of the tumors included, 11 (34.4%) were located in the maxilla, 21 (65.6%) in the mandible. Twenty-six tumors (81.2%) were classified as aggressive; the remainder (18.8%) were nonaggressive. There were no statistically significant differences between subjects with aggressive versus nonaggressive tumors with regard to age, gender, or location. Subjects with aggressive tumors had a significantly higher CD34 staining density (P = .02). None of the secondary predictors was associated with vascular density. CONCLUSION: Vascular density of giant cell tumors of the jaws is significantly increased in aggressive tumors.

Expression and pathological relevance of inducible nitric oxide synthase in osteosarcoma of the jaws.

The aim of the present study was to examine the expression of inducible nitric oxide synthase (iNOS) in osteosarcoma of the jaw, and its relationship with tumour angiogenesis and clinicopathological characteristics. Streptavidin peroxidase immunohistochemical staining was used to detect the expression level of iNOS and CD34 in paraffin-embedded samples from 25 patients. Osteosarcoma of the jaw was associated with overexpression of iNOS, which correlated with tumour microvessel density (MVD). iNOS expression correlated with the size, pathological grade and clinical stage of the osteosarcoma, and also with clinicopathological characteristics such as primary occurrence or recurrence of tumours. There was no correlation with metastasis. iNOS may promote tumour angiogenesis in osteosarcoma of the jaw, and so may represent an important target in anti-tumour therapy.

Antiangiogenic therapy with interferon alpha for giant cell lesions of the jaws.

PURPOSE: Giant cell tumors are classified and treated based on their biologic behavior. We hypothesize that they are proliferative vascular lesions and would be expected to respond to antiangiogenic therapy. The purpose of this report is to present a treatment protocol consisting of enucleation, with preservation of vital structures, followed by subcutaneous interferon alpha. MATERIALS AND METHODS: Patients with a biopsy-confirmed giant cell lesion satisfying criteria for "aggressive giant cell tumor" were included. Instead of wide en bloc resection, lesions were enucleated and the patients started on interferon alpha-2 or beta (3,000,000 units/m(2)) 48 to 72 hours postoperatively. The subjects were followed by clinical examination and radiography, immediately after surgery and every 3 months until the bone cavity completely healed. Thereafter, follow-up was every 6 months. RESULTS: Eight patients (7 females), with a mean age of 18.7 +/- 11.1 years, have been enrolled. Six tumors were in the posterior mandible, and 2 were in the anterior maxilla. The mean size was 29.0 mm (range, 15 to 70 mm). All patients underwent enucleation. There were no postoperative complications, and all patients tolerated interferon. There was no evidence of tumor growth during treatment. Seven of 8 patients have completed interferon therapy, and there have been no recurrences during 1 to 6 years of follow-up. The other patient continues on treatment with no evidence of disease. CONCLUSION: Antiangiogenic therapy, in combination with curettage, is a promising strategy for treatment of aggressive giant cell tumors. Combined treatment results in a high rate of tumor control with decreased operative morbidity compared with conventional treatment.

[Angiomas of the maxillofacial area: a clinical study with MR and angio-MR]. / Angiomi del distretto maxillo-facciale: studio clinico con RM ed angio-RM.

Reporting 3 cases of hemangiomas of the maxillo-facial area the authors describe, proper use, of RBc scintigraphy, CT with contrast media, MR, angio-MR. Comparing these methods it will results that RBc scintigraphy is a very useful method in a screening phase providing data concerning kind of blood flow, morphology of lesions and also detecting unknown lesions by total body scanning. CT imaging with contrast media is a useful method in a presurgical phase providing the exact anatomical limits of the lesions especially when involving bone tissue. MR imaging provides precise anatomic limits specially for low flow lesions; it also provides a vascular map of the arterial afferent vessels in high flow vascular lesions, without any use of ionogenic radiations. For these reasons MR is advisable in pediatric age and allergic patients. Super-selective embolization, sclerosant therapy and surgery therapy are indicated for treatment of high flow and low flow hemangiomas.

Immunohistochemical and ultrastructural evidence of a modified microvasculature in the giant cell granuloma of the jaws.

A panel of five immunohistochemical markers, MB1, leukocyte common antigen, S-100 protein, smooth muscle specific actin, and factor VIII related antigen, were used to study 20 giant cell lesions. These included eight central giant cell granulomas, nine peripheral giant cell granulomas, and three giant cell tumors of bone. The multinucleated giant cells stained positively with MB1, the mononuclear round cells were positive to leukocyte common antigen and the spindle cells were unreactive to all the markers chosen in all the lesions. The most interesting finding was the staining pattern of the blood vessels to factor VIII related antigen in the giant cell granuloma. The blood vessels on the periphery of the lesions were strongly positive for this antibody. However, reaction product was not evident deeper in the lesion within the aggregations of giant cells. Two other endothelial cell markers, Ulex europaeus 1 lectin and QBend 10 were used to study 10 giant cell lesions and a similar pattern of staining was observed. Transmission electron microscopy was subsequently used to study the ultrastructure of the microvasculature of three peripheral giant cell granulomas, and the findings indicated that the reasons for the differential staining may lie in the differences in the structure of the microcirculation.

[Vascular anomalies of the maxillofacial region].

513 cases with vascular lesions of oral and maxillofacial regions were followed from 1986 to 1990 by history, physical examination and pathological check. The vascular anomalies which had been termed "hemangioma" in the past were diagnosed anew according to new biological classification of vascular lesions proposed by Mulliken and Glowacki. Hemangioma and vascular malformation were completely different in clinical behavior and endothelial cell characteristics. Hemangiomas are often not present at birth, but appear during the 1st month. Its clinical behavior is that a proliferative phase is followed by a slow involution. Vascular Malformations are always present at birth and never regress, therefore the treatment of hemangioma and vascular malformation is different.

Carotid ligation. A justifiable extension of ablative surgery for malignant tumours in the maxillo-facial region.

In 30 cases operated on by ligation of the internal carotid artery, at the Vienna clinic, the average survival time was only 120 days (minimum 2 days, maximum 613 days). However, the immediate surgical risk of the operation is smaller than expected. Long-term postoperative care of the patient appears to be necessary, for psychological reasons.