Keratocystic odontogenic tumors and Carnoy's solution: results and complications assessment

Oral Diseases (2012) 18, 548557 Objective: Keratocystic odontogenic tumors (KOTs) can be treated with Carnoys solution, although this treatment modality is not free from complications. It is important to verify the incidence of complications after the use of Carnoys solution and compare these with the literature. Materials and methods: This study verified the effects of a complementary treatment for KOTs and assessed the incidence of such complications as recurrence, infection, sequestrum formation, mandibular fracture, dehiscence, and neuropathy. Results: Twenty-two KOTs treated with Carnoys solution combined with peripheral ostectomy were included, and the follow-up period varied from 12 to 78 months with a mean of 42.9 months. Complications included recurrence (4.5%), dehiscence (22.7%), infection (4.5%), and paresthesia (18.2%). No difference was found among lesions associated (9.1%) or not (0%) with nevoid basal cell carcinoma syndrome (P > 0.05). Dehiscence was influenced by marsupialization (P < 0.05), and paresthesia was observed exclusively in cases of mandibular canal fenestration (P < 0.01). Conclusions: Complementary treatment with Carnoys solution and peripheral ostectomy appear to provide efficient treatment for KOTs. Complications originating from the use of the solution are less frequent and less serious than complications associated with cryotherapy. Neuropathy seems to be related to direct contact between the solution and the epineurium.

Bone repair investigation using rhBMP-2 and angiogenic protein extracted from latex

Background: The aim of this work was to study the new bone tissue formation after bone morphogenetic protein type 2 (rhBMP-2) and P-1 application, using 5 and 10 mu g of each, combined to a material carrier, in critical bone defects. Methods: It was used 70 Wistar rats (male, similar to 250 g) that were divided in 10 groups with seven animals on each. Groups are the following: critical bone defect only, pure monoolein gel, 5 mu g of pure P-1, 5 mu g of pure rhBMP-2, 5 mu g of P-1/monoolein gel, 5 mu g of rhBMP-2/monoolein gel, 10 mu g of pure P-1, 10 mu g of pure rhBMP-2, 10 mu g of P-1/monoolein gel, 10 mu g of rhBMP-2/monoolein gel. Animals were sacrificed after 4 weeks of the surgical procedure and the bone samples were submitted to histological, histomorphometrical, and immunohistochemical evaluations. Results: Animals treated with pure P-1 protein, in both situations with 5 mu g and 10 mu g, had no significant difference (P > 0.05) for new bone formation; other groups treated with 10 mu g were statistically significant (P < 0.05) among themselves and when compared with groups in which it was inserted the monoolein gel or critical bone defect only (P < 0.05). In the group involving the 10 mu g rhBMP-2/monoolein gel association, it was observed an extensive bone formation, even when compared with the same treatment without the gel carrier. Conclusion: Using this experimental animal model, more new bone tissue was found when it was inserted the rhBMP-2, especially when this protein was combined to the vehicle, and this process seems to be dose dependent. Microsc. Res. Tech., 2011.(c) 2011 Wiley Periodicals, Inc.

Immunolocalization of podoplanin in benign odontogenic tumours with and without ectomesenchyme

Objectives: To investigate podoplanin expression in epithelial odontogenic tumours with and without ectomesenchyme and verify the association between its immunoexpression and proliferative activity in keratocystic odontogenic tumours (KCOTS) and orthokeratinized odontogenic cysts (OOCs). Design: Eight ameloblastomas, nine adenomatoid odontogenic tumours, twenty KCOTS, five OOC, one calcifying epithelial odontogenic tumour, two ameloblastic fibromas, four ameloblastic fibro-odontomas and five calcifying cystic odontogenic tumours were immunohistochemically analysed with anti-podoplanin antibody. For KCOTS and OOC, the cell proliferation index was determined with Ki-67 immunostaining and compared by Spearman correlation coefficient. Results: Podoplanin was expressed in the peripheral odontogenic epithelium of most tumours. Ectomesenchyme was negative, except for odontoblasts. KCOTS exhibited positive podoplanin expression while in OOC it was absent/weak. There was statistically significant correlation ( p = 0.006) between podoplanin expression and cellular proliferation index of KCOTS and OOC. Conclusion: Podoplanin seems to be related to the proliferative activity of KCOTS and may have a role in the process of local invasion of odontogenic tumours with and without ectomesenchyme

Distintas manifestações do tumor odontogênico cístico calcificante

O tumor odontogênico cístico calcificante normalmente se manifesta como uma massa indolor, de crescimento lento, que acomete tanto a maxila como a mandíbula, tendo predileção pelo segmento anterior (área dos incisivos e caninos). Geralmente, ocorre em adultos jovens, na terceira a quarta década de vida, sem preferência por gênero. Imagens de tomografia computadorizada revelaram características importantes não detectadas na radiografia panorâmica, tais como fenestração, calcificação e estruturas dentiformes. A característica microscópica típica dessa lesão é a presença de células epiteliais aberrantes anucleadas, em quantidades variáveis, denominadas "células fantasmas". Também se pode encontrar dentina displásica e, ocasionalmente, os cistos estão associados a tecido dentário duro, semelhante ao odontoma. O tratamento do tumor odontogênico cístico calcificante incluiu simples enucleação e curetagem. O objetivo deste trabalho foi mostrar duas manifestações diferentes do tumor odontogênico cístico calcificante, em que a tomografia computadorizada, associada às características clínicas, foi ferramenta importante para o diagnóstico, o planejamento cirúrgico e o acompanhamento dos pacientes.

E-cadherin in canine mast cell tumors: Decreased expression and altered subcellular localization in Grade 3 tumors

Mast cell tumors (MCTs) are the most frequent round cell tumors in dogs and comprise approximately 21% of all canine cutaneous tumors. MCTs are highly invasive and metastatic corresponding to the histological grade. E-cadherin is an adhesion molecule expressed in epithelial cells and although it is an epithelial cellular marker, studies have shown expression of E-cadherin in canine round cell tumors. To better characterize the expression pattern of E-cadherin in several different histological grades of MCTs in dogs, the expression and localization of the adhesion molecule was investigated using immunohistochemistry. For this purpose, 18 cutaneous MCTs were classified into three histological grades, 1, 2 or 3. Clinical history and follow-up data were available for all of the dogs. Cytoplasmic and nuclear expressions of E-cadherin in all three types of tumors were verified by immunostaining using two different antibodies. There was decreased E-cadherin expression in the more aggressive MCTs (Grade 3), suggesting an association between E-cadherin and tumor aggressiveness. Additionally, the loss of E-cadherin expression in either the cytoplasm or nucleus in more aggressive and undifferentiated tumor types confirmed the importance of cellular adhesion in tumor behavior. (C) 2012 Published by Elsevier Ltd.

RGMa and RGMb expression pattern during chicken development suggest unexpected roles for these repulsive guidance molecules in notochord formation, somitogenesis, and myogenesis

Background: Repulsive guidance molecules (RGM) are high-affinity ligands for the Netrin receptor Neogenin, and they are crucial for nervous system development including neural tube closure; neuronal and neural crest cell differentiation and axon guidance. Recent studies implicated RGM molecules in bone morphogenetic protein signaling, which regulates a variety of developmental processes. Moreover, a role for RGMc in iron metabolism has been established. This suggests that RGM molecules may play important roles in non-neural tissues. Results: To explore which tissues and processed may be regulated by RGM molecules, we systematically investigated the expression of RGMa and RGMb, the only RGM molecules currently known for avians, in the chicken embryo. Conclusions: Our study suggests so far unknown roles of RGM molecules in notochord, somite and skeletal muscle development. Developmental Dynamics, 2012. (C) 2012 Wiley Periodicals, Inc.

Single-cell expression analysis of BMP15 and GDF9 in mature oocytes and BMPR2 in cumulus cells of women with polycystic ovary syndrome undergoing controlled ovarian hyperstimulation

To detect expression of bone morphogenetic protein 15 (BMP15) and growth differentiation factor 9 (GDF9) in oocytes, and their receptor type 2 receptor for BMPs (BMPR2) in cumulus cells in women with polycystic ovary syndrome (PCOS) undergoing in vitro fertilization (IVF), and determine if BMPR2, BMP15, and GDF9 expression correlate with hyperandrogenism in FF of PCOS patients. Prospective case-control study. Eighteen MII-oocytes and their respective cumulus cells were obtained from 18 patients with PCOS, and 48 MII-oocytes and cumulus cells (CCs) from 35 controls, both subjected to controlled ovarian hyperstimulation (COH), and follicular fluid (FF) was collected from small (10-14 mm) and large (> 18 mm) follicles. RNeasy Micro Kit (Qiagen(A (R))) was used for RNA extraction and gene expression was quantified in each oocyte individually and in microdissected cumulus cells from cumulus-oocyte complexes retrieved from preovulatory follicles using qRT-PCR. Chemiluminescence and RIA assays were used for hormone assays. BMP15 and GDF9 expression per oocyte was higher among women with PCOS than the control group. A positive correlation was found between BMPR2 transcripts and hyperandrogenism in FF of PCOS patients. Progesterone values in FF were lower in the PCOS group. We inferred that BMP15 and GDF9 transcript levels increase in mature PCOS oocytes after COH, and might inhibit the progesterone secretion by follicular cells in PCOS follicles, preventing premature luteinization in cumulus cells. BMPR2 expression in PCOS cumulus cells might be regulated by androgens.

Tomographic, Histological, and Immunohistochemical Evidences on the Use of N-Butyl-2-Cyanoacrilate for Onlay Graft Fixation in Rabbits

Background: The bone tissue responses to Cyanoacrylate have been described in the literature, but none used N-butyl-2-cyanoacrilate (NB-Cn) for bone graft fixation. Purpose: The aims of the study were: (a) to analyze the bone grafts volume maintenance fixed either with NB-Cn or titanium screw; (b) to assess the incorporation of onlay grafts on perforated recipient bed; and (c) the differences of expression level of tartrate-resistant acid phosphatase (TRAP) protein involved in bone resorption. Materials and Methods: Eighteen New Zealand White rabbits were submitted to calvaria onlay grafting on both sides of the mandible. On one side, the graft was fixed with NB-Cn, while on the other hand the bone graft was secured with an osteosynthesis screw. The computed tomography (CT) was performed just after surgery and at animals sacrifice, after 1 (n = 9) and 6 weeks (n = 9), in order to estimate the bone grafts volume along the experiments. Histological sections of the grafted areas were prepared to evaluate the healing of bone grafts and to assess the expression of TRAP protein. Results: The CT scan showed better volume maintenance of bone grafts fixed with NB-Cn (p = 0.05) compared with those fixed with screws, in both experimental times (analysis of variance). The immunohistochemical evaluation showed that the TRAP expression in a 6-week period was significantly higher compared with the 1-week period, without showing significant difference between the groups (Wilcoxon and MannWhitney). Histological analysis revealed that the NB-Cn caused periosteum damage, but provided bone graft stabilization and incorporation similar to the control group. Conclusion: The perforation provided by screw insertion into the graft during fixation may have triggered early revascularization and remodeling to render increased volume loss compared with the experimental group. These results indicate that the NB-Cn possesses equivalent properties to titanium screw to be used as bone fixation material in osteosynthesis.

Influence of low-level laser associated with osteogenic proteins recombinant human BMP-2 and Hevea brasiliensis on bone repair in Wistar rats

This study analyzed the newly formed bone tissue after application of recombinant human BMP-2 (rhBMP-2) and P-1 (extracted from Hevea brasiliensis) proteins, 2 weeks after the creation of a critical bone defect in male Wistar rats treated or not with a low-intensity laser (GaAlAs 780 nm, 60 mW of power, and energy density dose of 30 J/cm2). The animals were divided into two major groups: (1) bone defect plus low-intensity laser treatment and (2) bone defect without laser irradiation. The following subgroups were also analyzed: (a) 5 mu g of pure rhBMP-2; (b) 5 mu g of pure P-1 fraction; (c) 5 mu g of rhBMP-2/monoolein gel; (d) 5 mu g of P-1 fraction/monoolein gel; (e) pure monoolein gel. Comparisons of the groups receiving laser treatment with those that did not receive laser irradiation show differences in the areas of new bone tissue. The group treated with 5 mu g of rhBMP-2 and laser irradiation was not significantly different (P >0.05) than the nonirradiated group that received the same treatment. The irradiated, rhBMP-2/monoolein gel treatment group showed a lower area of bone formation than the nonirradiated, rhBMP-2/gel monoolein treatment group (P < 0.001). The area of new bone tissue in the other nonirradiated and irradiated groups was not significantly different (P > 0.05). Furthermore, the group that received the 5 mu g of rhBMP-2 application showed the greatest bone formation. We conclude that the laser treatment did not interfere with the area of new bone tissue growth and that the greatest stimulus for bone formation involved application of the rhBMP-2 protein. Microsc. Res. Tech. 2011. (c) 2011 Wiley Periodicals, Inc.

Biological evaluation of the bone healing process after application of two potentially osteogenic proteins: an animal experimental model

doi: 10.1111/j.1741-2358.2011.00526.x Biological evaluation of the bone healing process after application of two potentially osteogenic proteins: an animal experimental model Objective: The aim of this work was to analyse qualitatively and quantitatively the newly formed bone after insertion of rhBMP-2 and protein extracted from Hevea brasiliensis (P-1), associated or not with a carrier in critical bone defects created in Wistar rat calvarial bone, using histological and histomorphometrical analyses. Materials and methods: Eighty-four male Wistar rats were used, divided into two groups, according to the period of time until the sacrifice (2 and 6 weeks). Each one of these groups was subdivided into six groups with seven animals each, according to the treatments: (1) 5 mu g of pure rhBMP-2, (2) 5 mu g of rhBMP-2/monoolein gel, (3) pure monoolein gel, (4) 5 mu g of pure P-1, (5) 5 mu g of P-1/monoolein gel and (6) critical bone defect controls. The animals were euthanised and the calvarial bone tissue removed for histological and histomorphometrical analyses. Result and conclusion: The results showed an improvement in the bone healing process using the rhBMP-2 protein, associated or not with a material carrier in relation to the other groups, and this process demonstrated to be time dependent.

Immunohistochemistry Analysis of Proteins Related with Apoptosis as Prognostic Factor in Epidermoid Carcinoma of Penis

The aim was to analyze the protein expression of apoptotic genes caspase-3, caspase-8 and bcl-2 with the immunohistochemistry technique, correlating with tumor grade (I, II and III) and with the patient survival in order to understand the basic mechanism of tumoral transformation. The immunohistochemistry reactions on 50 samples of squamous cell carcinoma were carried out with the avidin-biotin immunoperoxidase method and antigen recovery. The analyses were made using the graduation method "in crosses" (0 to 4 crosses - no stain to more than 75% of positives cells) and in categories (low, intermediate, high) of the cytoplasm immunoreactivity of the epidermoid penile carcinoma cells. It was observed a statistically significant difference when the expression of caspase-3 were compared with the grades land II of the tumor (p=0.0010) and when comparing the patient survival with the grades I and II of the tumor (p=0.0212). The protein bcl-2 was more expressed than caspase-3 and caspase-8 proteins, suggesting that the apoptotic rate in this carcinoma is low. The higher expression of the anti-apoptotic protein bcl-2 suggests a higher preservation of the tumoral cells.

Ameloblastoma demographic, clinical and treatment study - analysis of 40 cases

Dental lesions represent about 1% of oral cavity tumors being ameloblastoma the most common one. It is a tumor of epithelial origin that mainly affects the jaw, and less commonly the maxilla. Its clinical presentation is that of an asymptomatic slow-growing tumor. Despite being a benign tumor, it has an invasive behavior with a high rate of recurrence if not treated properly. Objective: To describe the cases of ameloblastoma in a reference department. Methods: Retrospective analysis of 40 cases. The variables analyzed were: age, gender, ethnicity, tumor location, type of treatment, complications and recurrence. Results: The most affected gender was male - 21 cases (52.5%); with a predominance of Caucasians - 24 cases (60%). The mean age was 35.45 years; the most common location was in.the jaw - 37 cases (92.5%). Facial asymmetry was the most frequent complaint. Of the 40 cases, 33 were submitted to surgery. Of those submitted to surgery, 24 (72.72%) underwent segmental resection, with recurrence in 4 (12.12%) cases. Conclusion: Ameloblastoma may relapse when treatment is not performed with broad surgical resection of the lesion with wide safety margins.

Changes in the expression of proteins associated with aerobic glycolysis and cell migration are involved in tumorigenic ability of two glioma cell lines

Background: The most frequent and malignant brain cancer is glioblastoma multiforme (GBM). In gliomas, tumor progression and poor prognosis are associated with the tumorigenic ability of the cells. U87MG cells (wild-type p53) are known to be tumorigenic in nude mice, but T98G cells (mutant p53) are not tumorigenic. We investigated the proteomic profiling of these two cell lines in order to gain new insights into the mechanisms that may be involved in tumorigenesis. Results: We found 24 differentially expressed proteins between T98G and U87MG cells. Gene Ontology supports the notion that over-representation of differentially expressed proteins is involved in glycolysis, cell migration and stress oxidative response. Among those associated with the glycolysis pathway, TPIS and LDHB are up-regulated in U87MG cells. Measurement of glucose consumption and lactate production suggests that glycolysis is more effective in U87MG cells. On the other hand, G6PD expression was 3-fold higher in T98G cells and this may indicate a shift to the pentose-phosphate pathway. Moreover, GRP78 expression was also three-fold higher in T98G than in U87MG cells. Under thapsigargin treatment both cell lines showed increased GRP78 expression and the effect of this agent was inversely correlated to cell migration. Quantitative RT-PCR and immunohistochemistry of GRP78 in patient samples indicated a higher level of expression of GRP78 in grade IV tumors compared to grade I and non-neoplastic tissues, respectively. Conclusions: Taken together, these results suggest an important role of proteins involved in key functions such as glycolysis and cell migration that may explain the difference in tumorigenic ability between these two glioma cell lines and that may be extrapolated to the differential aggressiveness of glioma tumors.

Dental pulp stem cells express proteins involved in the local invasiveness of odontogenic myxoma

Little is known about the histogenesis of the odontogenic myxoma (OM). Dental pulp stem cells could be candidate precursors of OM because both OM and the dental pulp share the same embryological origin: the dental papilla. For the purpose of comparing OM and stem cells, this study analyzed the expression of two proteins related to OM invasiveness (MMP-2 and hyaluronic acid) in human immature dental pulp stern cells (hIDPSCs). Three lineages of hIDPSCs from deciduous and permanent teeth were used in this study. Immunofluorescence revealed positive reactions for MMP-2 and hyaluronic acid (HA) in all hIDPSCs. MMP-2 appeared as dots throughout the cytoplasm, whereas HA appeared either as diffuse and irregular dots or as short fibrils throughout the cytoplasm and outside the cell bodies. The gene expression profile of each cell lineage was evaluated using RT-PCR analysis, and HA was expressed more intensively than MMP-2. HA expression was similar among the three hIDPSCs lineages, whereas MMP-2 expression was higher in DL-1 than in the other cell lines. The expression of proteins related to OM invasiveness in hIDPSCs could indicate that OM originates from dental pulp stem cells.

Implant Osseointegration in Circumferential Bone Defects Treated with Latex-Derived Proteins or Autogenous Bone in Dog's Mandible

Background: In sites with diminished bone volume, the osseointegration of dental implants can be compromised. Innovative biomaterials have been developed to aid successful osseointegration outcomes. Purpose: The aim of this study was to evaluate the osteogenic potential of angiogenic latex proteins for improved bone formation and osseointegration of dental implants. Materials and Methods: Ten dogs were submitted to bilateral circumferential defects (5.0 x 6.3 mm) in the mandible. Dental implant (3.3 x 10.0 mm, TiUnite MK3 (TM), Nobel Biocare AB, Goteborg, Sweden) was installed in the center of the defects. The gap was filled either with coagulum (Cg), autogenous bone graft (BG), or latex angiogenic proteins pool (LPP). Five animals were sacrificed after 4 weeks and 12 weeks, respectively. Implant stability was evaluated using resonance frequency analysis (Osstell Mentor T, Osstell AB, Goteborg, Sweden), and bone formation was analyzed by histological and histometric analysis. Results: LPP showed bone regeneration similar to BG and Cg at 4 weeks and 12 weeks, respectively (p >= 3.05). Bone formation, osseointegration, and implant stability improved significantly from 4 to 12 weeks (p <= 2.05). Conclusion: Based on methodological limitations of this study, Cg alone delivers higher bone formation in the defect as compared with BG at 12 weeks; compared with Cg and BG, the treatment with LPP exhibits no advantage in terms of osteogenic potential in this experimental model, although overall osseointegration was not affected by the treatments employed in this study.