Immunohistochemical study of osteopontin in oral squamous cell carcinoma allied to fractal dimension.

The aim of the study was to consider a possible correlation between the intensity of expression of osteopontin and grading established by the pathologist. Furthermore, a correlation was investigated between the increase of fractal dimension and osteopontin in order to use this marker as an early and reliable diagnostic tool for the degree of cell transformation in oral squamous carcinoma. Ten histologically healthy oral samples and sixty-four primary oral squamous cell carcinomas specimens were analysed by a single pathologist. Immunohistochemical analysis and Fulgen stain were performed in order to evaluate intensity of expression of osteopontin and fractal dimension. Data obtained were presented as mean and standard deviation and processed for the statistical analysis. Ostepontin expression revealed a statistical significance between groups (P less than 0.001). Fractal dimension in oral squamous cell carcinoma groups vs controls revealed statistically significant differences (P less than 0.001). The fractal dimension value and the osteopontin expression were compared, using two-dimensional scatter. The correlation was relevant in the G3 group. The results demonstrated a correlation between the growths of osteopontin expression and nuclear abnormality measured by fractal dimension. These results support the hypothesis that the level of osteopontin expression might be used as a marker for the evaluation of oral squamous cell carcinoma differentiation. Osteopontin and fractal dimension could support the histological grading to increase the predictability of the diagnosis, choices of treatment procedure and long-term prognosis.

Immunoexpression of angiogenesis and proliferation markers in patients treated with cyclosporin A.

AIM: In the present immunohistochemical study, the expression of vascular endothelial growth factor, nitric oxide synthase 1 and 3, and Ki-67 in the gingival tissues of renal transplant patients treated with cyclosporin A was assessed. Gingival overgrowth (GO) frequently occurs in transplant patients receiving immunosuppressive drugs such as cyclosporine and this gingival inflammation might play an important role in the pathogenesis of drug-induced GO. METHODS: Twenty-eight human gingival biopsies were taken from healthy patients with chronic periodontitis (N.=14 control group), and from renal transplant recipients treated with cyclosporin A (N.=14 test group). The retrieved specimens were immunohistochemically processed and stained for vascular endothelial growth factor, nitric oxide synthase 1 and 3, and Ki-67. RESULTS: The levels of vascular endothelial growth factor, nitric oxide synthase 1 and 3, and Ki-67 were found to be significantly different among groups (P>0.001), with patients treated with cyclosporin A showing higher levels of all the analyzed markers compared to control group. CONCLUSION: In summary, the data from this pilot study suggests that the investigated factors have a role in the inflammation processes associated to immunosuppressive therapy. However, further studies with a larger sample population need to be conducted for an exhaustive knowledge of the mechanisms leading to GO.

Management of compound odontoma in a 10-year-old girl preserving the associated impacted permanent tooth.

AIM: Compound odontoma has been reported to be the most common of all odontogenic neoplasms and tumor- like lesions. Only rarely the treatment of this lesion in association with an impacted tooth has been reported. CASE REPORT: A compound odontoma in a 10-year-old girl, associated with an impacted permanent incisor is described, focusing on the diagnosis and the importance of early treatment of this lesion. The patient underwent surgical excision of the lesion and it was decided to wait for the spontaneous eruption of the impacted tooth. After 6 months no eruption was observed and thus the orthodontic treatment was deemed necessary. At the one-year follow-up, the tooth was brought into the maxillary arch.

Morphometric and ultrastructural analysis of human granulosa cells after gonadotrophin-releasing hormone agonist or antagonist.

Morphological features of granulosa cells can reflect their functional status. The present study was aimed at comparing possible differences in the fine structure of human granulosa cells exposed to gonadotrophin-releasing hormone (GnRH) agonist or antagonist treatment during ovarian stimulation. Cells were obtained from follicular aspirates of 21 women treated with recombinant follicle-stimulating hormone (rFSH) plus either a GnRH agonist or a GnRH antagonist. Conventional light microscopy procedures and computerized image analysis systems were used to identify different cell type morphological patterns and to quantify different cells distribution. Two morphologically distinct granulosa cell populations, defined as large/pale and small/dark cells, were identified and a different distribution in the two groups of women under investigation was found: a significantly higher percentage in large/pale cells was detected in the agonist-treated women (P<0.05), whereas the percentage of small/dark cells was significantly higher in the antagonist-treated group (P<0.05). Ultrastructural observations showed the presence in both cell populations of typical hallmarks of steroidogenic cells, highlighting signs of functional activity in the large/pale cell population. Further investigations are needed to define the possible clinical significance of these morphological findings.

VEGF and MVD expression in sinus augmentation with autologous bone and several graft materials.

OBJECTIVE: The aim of this study was to assess vascular endothelial growth factor (VEGF) expression and microvessel density (MVD) in maxillary sinus augmentation with autogenous bone and different graft materials for evaluating their angiogenic potential. METHODS: Biopsies were harvested 10 months after sinus augmentation with a combination of autogenous bone and different graft materials: hydroxyapatite (HA, n = 6 patients), demineralized freeze-dried bone allograft (DFDBA, n = 5 patients), calcium phosphate (CP, n = 5 patients), Ricinus communis polymer (n = 5 patients) and control group--autogenous bone only (n = 13 patients). RESULTS: In all the samples, higher intensities of VEGF expression were prevalent in the newly formed bone, while lower intensities of VEGF expression were predominant in the areas of mature bone. The highest intensity of VEGF expression in the newly formed bone was expressed by HA (P < 0.001) and CP in relation to control (P < 0.01) groups. The lowest intensities of VEGF expression in newly formed bone were shown by DFDBA and polymer groups (P < 0.05). When comparing the different grafting materials, higher MVD were found in the newly formed bone around control, HA and CP (P < 0.001). CONCLUSION: Various graft materials could be successfully used for sinus floor augmentation; however, the interactions between bone formation and angiogenesis remain to be fully characterized.

Neuroglobin in aging carotid bodies.

Neuroglobin (Ngb) is a member of the vertebrate globin family expressed particularly in the brain and in the retina. Ngb is concentrated in the mitochondria-containing areas of neurons, and its distribution is correlated with oxygen consumption rates. Previously we have shown that Ngb is expressed in carotid body (CB) tissues. Considering that hypoxia and aging may be linked through a series of adaptive and protective mechanisms (e.g. reduction in mitochondrial numbers), we investigate the role of Ngb during aging and hypoxia. Two groups of six rats (age-matched 3 and 24 months old) were kept in room air as a control groups, the others two groups were kept in a Plexiglas chamber for 12 days in chronic hypoxia (10-12% inspired oxygen). The presence of Ngb in the CB tissue was detected by immunohistochemistry using a polyclonal antibody. Ngb immunoreactivity was significantly higher in CB tissues from young rats exposed to chronic intermittent hypoxia, whereas CB tissues from old rats did not show any significant increase in Ngb levels after hypoxia. Similar to hemoglobin, Ngb may act as a respiratory protein by reversibly binding gaseous ligands NO and O(2) and could act as a NO scavenger and participate in detoxification of Reactive Oxygen Species (ROS) generated under hypoxic conditions.

Peripheral giant cell granuloma associated with a dental implant. A case report.

Peripheral giant cell granuloma (PGCG) is a tumor-like pathologic condition arising on the buccal or lingual attached gingival or alveolar mucosa and the crest of the edentulous alveolar ridge, of uncertain etiology; it is probably a reactive lesion caused by chronic local irritants or trauma rather than a true neoplasm. PGCG is thought to originate from elements of the periodontal ligament or from the periosteum. Clinically, it appears as a sessile or broadly pedunculated, bluish to purple-red, fleshy or firm swelling with a frequently ulcerated surface. The occurrence of such a lesion may be significant for the prognosis of dental implants, and they can lead to integration failure. Treatment of choice is conservative surgical excision with total removal of the base of the lesion and with curettage of the underlying bone. However, some of these lesions recur. Clinically, it may be difficult to distinguidsh PGCG from pyogenic granuloma. Moreover, also peripheral odontogenic tumors have to be considered in the differential diagnosis of PGCG. The clinical appearance of all these lesions are similar and so the definitive diagnosis is only histological and a biopsy with micreoscopical examination is mandatory.

Vascular endothelial growth factor expression (VEGF) in salivary glands of diabetic rats.

This study is aimed at evaluating vascular endothelial growth factor (VEGF) expression in the salivary glands of diabetic rats. Our study was carried out on 1 group of male Wistar rats and 1 group of male Bio Breeding Wistar (BB/W) pre-diabetic rats. The first group (control) was composed of 11 rats (4 months old), and the other (test) of 11 rats (4 months old) which showed diabetes at day 85 of life. This kind of diabetes can be compared to human diabetes mellitus type 1. The submandibular glands were removed and processed for immunohistochemical analysis of VEGF. Diabetes increased salivary gland VEGF expression in the rats. The tissues analyzed (vascular endothelium, ductal endothelium, mucinous glands) always expressed VEGF, thus demonstrating that not only vascular endothelial cells, but also the other elements evaluated, have a role in the neoangiogenesis process. In both control and diabetic rats, the VEGF expression was constantly negative only in serous acini; thus demonstrating that serous acini are not involved in the neoangiogenetic processes. The vascular growth is a fundamental part of normal salivary gland development, therefore we speculate that strategies aimed at preservation or promotion of salivary gland VEGF expression may mitigate or attenuate diabetic-induced gland microvascular injury.

MIB-1, Bcl-2 and p53 in odontogenic myxomas of the jaws.

Odontogenic myxoma is a rare benign neoplasm occurring in the jaws. Microscopically, it is composed of spindle or stellate-shaped cells arranged in a mucinous matrix. In some cases (20%), odontogenic epithelial islands may be found. The Authors evaluated p53, MIB-1, and Bcl-2 expressed by the epithelial and stromal elements in 12 cases of odontogenic myxoma of the jaws. The cells of the odontogenic epithelium were positive for Bcl-2, p53 and MIB-1. The stromal cell showed a high positivity for MIB-1. Proliferation of both the epithelial and stromal components could be related to the growth of this odontogenic tumour.

Factors associated with apoptosis in symptomatic and asymptomatic carotid atherosclerotic plaques.

The aim of this study was to investigate the differences that are present between apoptosis in symptomatic (with symptoms of cerebral ischemic attack) and asymptomatic carotid atherosclerotic plaques. The apoptotic process in macrophages and smooth muscle cells was evaluated. Cellular markers and products of immune cells in symptomatic and asymptomatic atherosclerotic plaque and endoarterectomy specimen were analyzed by immunohistochemistry. No statistically significant differences were present regarding the mean SMC actin-positive area. Using double staining of alpha-smooth muscle actin and TUNEL techniques, the number of smooth muscle cells in apoptosis was statistically higher in symptomatic plaque as compared with asymptomatic plaque. Statistically significant differences (p=0.009) were also found in the CD45-positive cells in the inflammatory infiltrate. The CD68-positive macrophages showed statistically significant differences (p=0.0001). Similarly, the double staining with CD68 and TUNEL revealed that apoptotic macrophages were mainly present in asymptomatic plaques rather than symptomatic plaques. Statistically significant differences (p<0.001) were found in the Bcl-2 expression, with higher values in asymptomatic plaques. Our data showed that the increase of the inflammatory cells contributes to plaque instability and that death due to apoptosis of smooth muscle cells in symptomatic plaques could contribute to their destabilization and explains their tendency to fracture.

Vascular endothelial growth factor (VEGF) in human tooth germ center.

Many oncogenis and tumour suppressor genes found inside normal and pathological cells are fundamental for the processes of development, proliferation and tissue differentiation. The purpose of our study is to show the presence and a possible relationship of the VEGF protein during different phases of the development of human dental germ centers. After cephalometric investigation in 8 orthodontic patients with a mean age of 13 years, (4 females and 4 males), hyperdivergence of the third molars were extracted. The 40 surgical samples were tested with monoclonal human anti-VEGFs antibodies carrying out a semi-quantitative analysis to look for a positive reaction. Reaction for anti-VEGF antibodies was detected in normal embryological tissues and in microvessels near odontogenic cells. During different phases of embryologic development of the dental bud our search showed intracytoplasmatic positive immunoreactions both in the ameloblastic and odontoblastic cells. Additionally, a positive reaction was observed for the VEGF protein in the cells of the stellate reticulum and in those endothelial tissue surrounding the microvessels in all the samples examined.

Microvessel density (MVD) and vascular endothelial growth factor expression (VEGF) in human oral squamous cell carcinoma.

BACKGROUND: Carcinogenesis is thought to be dependent on neovascularization. Vascular endothelial growth factor (VEGF) is a glycoprotein that has the capability of increasing vascular proliferation and permeability. VEGF has been found to be expressed in several different types of tumours and it may contribute to the progression of malignant tumours. Increased microvessel density (MVD) has been described in oral squamous cell carcinoma (OSCC) and seems to be related to patient prognosis. MATERIALS AND METHODS: Fifty-two cases of OSCC were evaluated in the present study. Immunostaining for VEGF and Factor-VIII was performed. The MVD was evaluated in G1, G2 and G3 tumours. RESULTS: The differences between these 3 groups were statistically significant (p = 0.0331). MVD was also evaluated in lymph-node negative and lymph-node positive cases: the differences between these two groups were statistically significant (p < 0.0001). VEGF expression was evaluated in G1, G2 and G3 tumours. The differences between the 3 groups were not statistically significant (p = 0.289), even if an increasing trend in the VEGF positivity was evident from G1 to G3. The difference of VEGF expression between tumours with and without lymph node metastases was not significant (p = 0.196). No correlation was present between intensity of VEGF positivity and histological grading or lymph-node status and between VEGF and MVD. CONCLUSION: Our data showed that MVD was correlated with grading and lymph-node status, while no similar correlation was found for VEGF.

Ultrastructure of the testis in rats with blood hypertension induced by long-term lead exposure.

Male Sprague-Dawley rats received 60 micrograms/ml of lead (as acetate) in drinking water for 18 months. Their blood pressure and cardiac inotropism were increased. Lead was augmented in blood, kidney and brain, but not in testis. Examination by light microscopy of the organs did not reveal alterations. Ultrastructural examination of the testis with both transmission and scanning electron microscopy (EM) did not evidence modifications in the external part of the seminiferous tubules, in the spermatogenetic cells and in the connective tissue including the Leydig cells; only Sertoli cells presented increased size of lysosomes.

Immune cells in periapical granuloma: morphological and immunohistochemical characterization.

Samples of periapical granulomas obtained from 12 patients were examined using light and electron microscopes and monoclonal antibodies. Monocytes/macrophages, lymphocytes, and plasma cells were nearly always the most abundant cell populations. Ultrastructural analysis showed close contacts between macrophages and cells of the lymphoid lineage, with the lymphoid cells frequently demonstrating blastic features. Immunohistochemical staining with the anti-interleukin 2 receptor antibody showed that the concentration of labeled cells was quite low. The vast majority were lymphocytes, though some mast cells were also labeled. Mast cells were chiefly located in perivascular areas and interleukin 2 receptor-positive mast cells were frequently associated with lymphoid cells. mast cells could be part of a negative feedback mechanism in the immune response. By releasing histamine, they would block the immune response and by absorbing interleukin 2 they would remove it as an immune system stimulant.

Immunoreactivity for interleukin 1-beta and tumor necrosis factor-alpha and ultrastructural features of monocytes/macrophages in periapical granulomas.

Interleukin 1 (IL-1) and tumor necrosis factor (TNF) have recently been shown to be involved in bone resorption, and because macrophages constitute a significant part of human periapical granulomas, it is reasonable to suspect that they may secrete IL-1 and TNF. The purpose of our investigation was to detect and characterize IL-1 beta- and TNF-alpha-producing cells in human periapical granulomas. Fresh tissue samples obtained during surgery from 10 patients with previously untreated teeth and histologically established periapical granulomas were studied with light and electron microscopy and with immunohistochemical analysis with monoclonal antibodies against IL-1 beta and TNF-alpha. There were very few IL-1 beta + and TNF-alpha + cells present in periapical granulomas, and the positive cells had monocyte/macrophage morphology. The IL-1 beta + cells were located mainly in areas of active exudation and were surrounded by and/or were in close contact with lymphoid cells, whereas TNF-alpha + cells were scattered and in contact with or near other inflammatory cells at the periphery of active granulation tissue. This suggests that the IL-1 beta + cells may act in a paracrine manner to activate lymphoid cells. The ultrastructural findings showed that only some macrophages are adapted to extracellular secretion rather than phagocytosis. These modified macrophages could be the major producers of interleukins in tissues. Occasionally, they have plasmacytic or plasmacytoid features resembling the so-called "plasmacytoid monocytes". Only a minor fraction of the monocytes/macrophages (representing about 40% of the inflammatory cells) is in an active cytokine-producing state.(ABSTRACT TRUNCATED AT 250 WORDS)

Vascular endothelial growth factor and microvessel density around healthy and failing dental implants.

Inflammatory infiltrate may be important in the evolution of inflammatory processes involving peri-implant tissues. Angiogenesis is an important feature of inflammation and healing, but its role in the development and progression or in the healing of periodontal lesions has not been elucidated. Vascular endothelial growth factor (VEGF) is a potent inducer of endothelial cell proliferation. The aim of the present study was to conduct a comparative immunohistochemical evaluation of VEGF and microvessel density (MVD) in normal keratinized gingiva and in peri-implant soft tissues surrounding failing implants. Fifteen patients participated in this study. Ten biopsies were taken from healthy keratinized gingiva, and 10 were taken from peri-implant soft tissues surrounding failing non-submerged implants. In healthy sites, the endothelial lining cells of the vessels always tested positive for VEGF; also, VEGF intensity was high in most cases. Stromal cells were positive for VEGF in 70% to 90% of samples. The MVD was 60.250 +/- 5.123. In peri-implantitis samples, the cells of the inflammatory infiltrate were positive for VEGF in 80% to 100% of cases, and the VEGF intensity was low in all cases. The stromal cells were positive for VEGF in 90% to 100% of cases, and in most cases the intensity was low. The MVD was 101.800 +/- 11.256. The difference in MVD between healthy sites and peri-implantitis was statistically significant (P = .0158). Expression of VEGF was lower in peri-implantitis samples, and this difference was statistically significant (P = .0373). Because of its extensive presence, VEGF is probably a factor in both the maintenance of periodontal physiology and in the progression of peri-implant inflammatory disease.

Tissue response to an expanded polytetrafluoroethylene external valve support device: a histologic study in dogs.

Expanded polytetrafluoroethylene (ePTFE) external valve support devices (EVS) have been used successfully in patients to restore valve function in leg veins with incompetent valves when incompetence is due to dilatation of the vein walls or elongation of the valve leaflet edges. To assess tissue response to these devices, the authors implanted 12 of them in dogs, wrapping the devices around veins in the head and neck. The dogs recovered from the implantation procedure uneventfully, and the veins remained patent on color flow Doppler scanning. Gross and histologic evaluations of vein segments and attached EVS devices after sacrifice of the dogs 30 days postoperatively showed that the ePTFE devices did not affect vein patency or the cellular composition or architecture of vein walls. There were no adverse tissue reactions to the EVS and no thrombus formation in the veins to which the EVS had been applied. Tissue attachment to the EVS was apparent in all specimens. These histologic results support clinical experiences indicating that the ePTFE EVS device is safe to use in external valvuloplasty for the treatment of venous incompetence.

Shock waves in vascular diseases: an in-vitro study.

UNLABELLED: Three human aortic specimens were used for this in-vitro study on the effects of shock waves on the arterial wall. Specimen one was from a normal (for age) healthy aorta. The full abdominal length was used (including mesenteric and renal arteries and the aortoiliac bifurcation), divided into six pieces (3 cm). The pieces were placed and fixed into degassed water. Shock waves (SW) were focused onto the aortic wall by means of a B-mode ultrasound imager. An SW generator (Minilith SL1, Storz Medical AG, Kreuzlingen, Switzerland) was used for setting of energy flux density between 0.03 and 0.5 mJ/mm2. The six aortic pieces (excluding piece 1, placed in water and left untreated as control) were treated with SW at increasing energy levels. A second aortic specimen of a man with arteriosclerotic plaques was also used and the experiment repeated at energy levels 1, 5, and 8. Another specimen of normal thoracic aorta was exposed at energy levels 1 and 8 only. Energy levels delivered onto the aortic walls were selected from theoretically destructive levels to minimal levels known not to alter vascular tissues. High-resolution ultrasounds of the aortic segments were performed with a 10 MHz high-resolution, broad-band (ATL 3000, USA) probe in water before and after SW application to detect structural changes in the wall after SW. Histology was performed with a standard hematoxylin-eosin staining. RESULTS: The aortic pieces did not show macroscopic damages at visual examination, and at the ultrasound examination no visible changes were observed even at higher levels of SW energy. Also no effects were seen by histology. In conclusion, no damaging effects were observed, visually, by ultrasound, or by histology. At these energy levels SW appear to be safe and do not produce any damage to the aortic wall. Therefore, SW could be considered a safe, nondamaging procedure for potential treatment (ie, thrombolysis) in which vessel walls could be involved. Theoretically it is possible that functional changes could be observed in vivo including cell permeability modifications and other alterations (including changes in the potential of the cells in SW fields to modify themselves and to divide). At the energy levels described in this study SW could, theoretically be, safely used for vascular applications (ie, treating venous and arterial thrombi or in arterial plaques modification) without altering major, structural, arterial wall characteristics. Lesions or alterations that have a different density from the normal wall (thrombi or plaques) could be differently sensitive to the same dosage of SW. These differences in acoustic impedance characteristics could be used for potential treatments with SW without damaging the arterial wall.

Circadian pattern of post-surgical fatal pulmonary embolism.

BACKGROUND: The circadian distribution of fatal pulmonary thromboembolism in general surgical patients is unknown. PATIENTS AND METHODS: One hundred consecutive cases of pulmonary embolism, with reliable clinical notes and data, were studied (67 men and 33 women; mean age 71 years). Only post-surgical cases were considered in this analysis. Patients had undergone elective (78%) or emergency abdominal surgery (22%). Correct prophylaxis (according to the Windsor Consensus Statement) had been used in 12%. Cases were grouped according to the time of onset of signs and symptoms related to pulmonary embolism at one hour intervals. RESULTS: The maximum incidence of fatal pulmonary embolism was between 7.00 a. m. and 1.00 p. m. with the highest peaks at 9.00 and 11.00 a. m. 9% of deaths) (P < 0.02). When results from this study were compared to a previous study no significant difference was observed between the distribution profile of cases from general medical wards and surgical wards. CONCLUSION: It appears that in surgical patients there is a circadian pattern in pulmonary embolism as already documented in medical patients.

[Aortic valve replacement in a case of Takayasu's disease with polydistributed localization]. / Sostituzione valvolare aortica in un caso di malattia di Takayasu a localizzazione polidistrettuale.

We report a case of a 49 year-old woman suffering from severe aortic regurgitation due to Takayasu's disease involving the ascending and abdominal aorta and the brachial arteries. The patient was managed successfully with aortic valve replacement. Considering the Literature data, the pathogenesis of the aortic insufficiency and the main surgical and anaesthesiologic problems related to the Takayasu's disease are discussed.