Expression of Secreted Aspartyl Proteinases in an Experimental Model of Candida albicans-Associated Denture Stomatitis.

PURPOSE: Candida albicans is known to produce secreted aspartyl proteinases (SAPs) to aid adhesion, invasion, and host tissue destruction. SAPs may contribute to denture stomatitis (DS) pathogenesis. The aim of this study was to develop an in vivo experimental model for Candida-associated DS that allows the analysis of SAP2, SAP5, and SAP9 expression by C. albicans from biofilm induced on the denture surface. MATERIALS AND METHODS: Thirty-five male Wistar rats were divided into three groups: control, denture, and denture/Candida group. The last two groups remained with dentures for 2, 4, and 6 days, with or without induced biofilm. SAP expression was concomitant with leukocyte counts as well as clinical and histological changes shown by animal palate. RESULTS: The signs observed at 4 days in the denture/Candida group were clinically closer to the Candida-associated DS, showing a significant increase of neutrophils and decrease of lymphocytes in peripheral blood, presence of inflammation signs on the palate similar to DS Newton type I, and fungal invasion in the epithelial layer. Accordingly, the denture/Candida group at 4 days presented the highest relative expression of all SAPs studied. CONCLUSION: The results showed a coincidence between SAP expression and clinical, microscopic, and blood data. Finally, the molecular findings were consistent with the virulence capacities of C. albicans from biofilm formed on the denture resin, which possibly allowed epithelial invasion by the fungus.

Le Fort I osteotomy for the removal of a rare unicystic ameloblastoma lesion in the maxillary sinus.

The unicystic ameloblastoma (UA) presents the clinical and radiographic characteristics of a maxillary cyst, making early diagnosis difficult. A 30-year-old man had an extensive, asymptomatic lesion in the right maxillary sinus. Radiographic examinations demonstrated a retained tooth in association with a lesion. Histopathologic examination revealed the presence of UA with intraluminal and mural infiltration and a follicular pattern. Le Fort I access was chosen for enucleation of the lesion and curettage of the site, which were followed by cryotherapy. The treatment provided adequate intraoperative visibility, enabled the preservation of the surrounding bone, and eliminated postoperative complications. Follow-up over 5 years demonstrated no recurrence.

Preliminary Histological Evaluation of the Application of Ozone in the First Days of Orthodontic Force Induction in Animal Model.

OBJECTIVES: The aim of the study was to histologically evaluate the effect of ozone therapy on orthodontic force induction in an animal model. MATERIALS AND METHODS: Twenty-four Wistar rats were divided into three groups (n = 8). A NiTi coil spring was installed from the maxillary first molar to the maxillary central incisor. G1 was control and G2/G3 received 1 mL of ozonated gas at concentrations of 10 and 60 µg/mL, in the buccal mucosa above the first molar roots. The animals were euthanized 3 and 5 days after the procedure. Histological sections were obtained, longitudinally of the first molar' long axis, in the mesiodistal direction. The number of osteoclasts, osteoblasts, blood vessels, polymorphonuclear and mononuclear cells, formation of osteoid tissue and hyaline areas, and root resorption were evaluated with light microscope, in tension and pressure sides. Intergroup comparisons were performed with Kruskal-Wallis, Dunn, and Chi-square tests. RESULTS: At 3-days pressure side, a greater number of osteoclasts was observed in ozone groups and greater number of blood vessels and polymorphonuclear cells were observed in G2. On the tension side, there was a significantly greater number of blood vessels, osteoblasts, and mononuclear cells in G2. At 5-days pressure side, there was a significantly greater number of osteoclasts in G2, blood vessels and osteoblasts in the ozone groups, and lesser number of polymorphonuclear cells in G3. CONCLUSION: Ozone therapy increased the number of osteoclasts on the pressure side and osteoblasts on tension side, in 10 µg/mL concentration, demonstrating histological parameters favorable to bone remodeling. The 60 µg/mL ozone concentration accelerated the periodontal ligament reorganization process.

Study of the effectiveness of propolis extract as a storage medium for avulsed teeth.

The purpose of the present study was to evaluate the efficacy of propolis extract in maintaining the viability of human periodontal ligament (PDL) cells, and to radiographically analyze tooth replantation and the adjacent periodontium in dogs after storage in this extract. Human PDL cells were incubated with the experimental media propolis, milk, saliva, Hank's balanced salt solution (HBSS), and Dulbecco's modified Eagles medium (DMEM, positive controls), and distilled water (negative control). Cell viability was determined 0, 1, 3, 6, 12, and 24 h later by colorimetric MTT assay. Thirty incisors from dogs were divided into two storage time blocks (1 and 3 h) and were maintained in the experimental media. HBSS served as a positive control, and dry teeth (on gauze) as a negative control. The replanted teeth were radiographed once per month for 6 months. The radiographic images were standardized by the shortening/lengthening factor, and were both qualitatively and quantitatively analyzed. The in vitro results showed that the efficacy of propolis in maintaining functional viability of PDL cells was similar to that of milk. Propolis and milk were significantly better than controls from the 6-h time period. The in vivo results showed that teeth maintained in propolis medium exhibited replacement resorption with significant reduction in tooth length, similar to teeth maintained in saliva and dried teeth. This resorption was less intense with the 3-h storage time than the 1-h storage time. Conditions close to normal were found in teeth maintained in milk, similar to the HBSS control. Therefore, although propolis was effective in maintaining the viability of human PDL cells, resorption of the tooth replantation in dogs occurred under these experimental conditions.

Candida albicans-Cell Interactions Activate Innate Immune Defense in Human Palate Epithelial Primary Cells via Nitric Oxide (NO) and ß-Defensin 2 (hBD-2).

The presence of Candida albicans in the biofilm underlying the dental prosthesis is related to denture stomatitis (DS), an inflammatory reaction of the oral mucosa. The oral epithelium, a component of the innate immune response, has the ability to react to fungal invasion. In this study, we evaluated the in vitro effect of viable C. albicans on the apoptosis, nitric oxide (NO) production, and ß-defensin 2 (hBD-2) expression and production of human palate epithelial cells (HPECs). We further determined whether or not these effects were correlated with fungal invasion of epithelial cells. Interaction between HPEC primary culture and C. albicans was obtained through either direct or indirect cell-cell contact with a supernatant from a hyphal fungus. We found that the hyphae supernatants were sufficient to induce slight HPEC apoptosis, which occurred prior to the activation of the specific mechanisms of epithelial defense. The epithelial defense responses were found to occur via NO and antimicrobial peptide hBD-2 production only during direct contact between C. albicans and HPECs and coincided with the fungus's intraepithelial invasion. However, although the hBD-2 levels remained constant in the HPEC supernatants over time, the NO release and hBD-2 gene expression were reduced at a later time (10 h), indicating that the epithelial defense capacity against the fungal invasion was not maintained in later phases. This aspect of the immune response was associated with increased epithelial invasion and apoptosis maintenance.

Ewing's Sarcoma Family Tumors in the Jaws: Case Report, Immunohistochemical Analysis and Literature Review.

Due to the low incidence of the Ewing's Sarcoma (ES) family tumors, the available epidemiology is likely to be unreliable, and at present, there are no standard diagnostic or clinical guidelines outlining their management. This report describes a case of peripheral primitive neuroectodermal tumor (ES/pPNET) which initially mimicked cystic lesions, and describes a comparison between ES and ES/pPNET in the jaws by the World Health Organization classification. This review addressed 63 cases published in the English literature between 1950 and 2016. The majority of cases were ES. Both ES and ES/pPNET mimicked other benign entities such as traumatic, cystic and inflammatory lesions. The patients who died of their disease had a history of metastatic tumors, and primary tumor located in the mandible and maxilla for ES and ES/pPNET, respectively. The differentiation of the ES family tumors from other small blue-cell tumors may be difficult and requires familiarity with histological and immunohistochemical features.

Ameloblastic carcinoma, primary type: case report, immunohistochemical analysis and literature review.

Ameloblastic carcinoma (AC) is a rare malignant odontogenic neoplasm with scarce reported cases in the literature and it can be confused with benign ameloblastoma (AM). This study reports a case of AC, and presents a literature review of AC classified into primary type (ACPt) or secondary type (ACSt) by the World Health Organization (WHO). The review addressed 31 cases published in the English literature between the years 2005 and 2011. The majority of cases were ACSt. The mandible was the most common site of occurrence for both AC types. All patients who died of their disease had ACSt. Tumors with plexiform pattern, hyperchromatism, mitosis and necrosis were associated with a higher ratio of histories of recurrence and death by disease, as well as the tumors with clear cells, especially in the ACSt. ACSt appeared to correlate with recurrence and mortality. The histological features may have different prognostic importance depending on the AC type.

Psammomatoid juvenile ossifying fibroma: an analysis of 2 cases affecting the mandible with review of the literature.

Juvenile ossifying fibroma (JOF) is a rare fibro-osseous neoplasm, defined as a variant of the ossifying fibroma that arises within the craniofacial bones. Two subgroups, juvenile psammomatoid ossifying fibroma (PsJOF) and juvenile trabecular ossifying fibroma, have been delineated by their histology. PsJOF occurs predominantly in the sinonasal and orbital bones. This work reports on 2 cases of extensive PsJOF in the body of the right mandible as well as reviews the literature regarding the radiographic and histologic features, treatment, and prognosis of PsJOF of the jaws.

Phytomedicines for Candida-associated denture stomatitis.

Phytomedicines are becoming more popular all over the world. Denture stomatitis (DS) presents as an inflammatory reaction in denture-bearing patients, under maxillary prosthesis with Candida albicans being the principal etiological agent. Many different methods of treatment for DS have been observed. The effects of the main medicinal plants claimed to be useful as antifungal agents in the treatment of DS are reviewed. Herbal remedies that have shown potential promise are mentioned, although much research is still required.

Removal efficiency of propolis paste dressing from the root canal.

OBJECTIVES: The aim of this study was to investigate through scanning electron microscopy (SEM) the cleaning of root canal walls after the use of experimental propolis or calcium hydroxide root canal dressings. MATERIAL AND METHODS: Twenty single-rooted teeth were used. After conventional cleaning and shaping procedures and removal of the smear layer with 17% EDTA, the teeth were divided into four groups according to the medication used (N=5): Group I (control) - No drug, Group II - Calcium hydroxide dressing, Group III - Propolis paste A70D and Group IV - Propolis paste D70D. The medications were introduced into the root canals and maintained for 7 days, then removed with a K-file and 5 mL of 1% sodium hypochlorite irrigation. Finally, the canals were flushed with 2 mL of 17% EDTA for 3 min. For SEM analysis, the roots were cleaved and microphotographs from the middle third of the root canal were taken at 750x. The cleaning of the root canal walls was determined by the number of open dentinal tubules as verified with the software Image Tool 3.1. The statistical analysis was performed by ANOVA and Tukey's test (p<0.05). RESULTS: The results showed no statistically significant difference between the calcium hydroxide and propolis groups. CONCLUSIONS: The experimental propolis pastes presented acceptable physical characteristics to be used as intracanal medicaments.

Manutenção da viabilidade das células mononucleares de sangue periférico humano em extratos e formulações de própolis / Maintenance of human peripheral blood mononuclear cell viability in propolis extracts and formulations

Neste estudo comparou- se a viabilidade das células mononucleares de sangue periférico (PBMCs) humano quando mantidas em diferentes extratos e formulações de própolis. As PBMCs (106 cels mL-1), provenientes de doadores saudáveis (n=5), foram estocadas a 20ºC nas diferentes soluções de própolis, assim como em solução salina balanceada de Hank's (HBSS), utilizada como controle do experimento. A viabilidade celular foi determinada pelo método de exclusão com azul de Tripan. Quando incubadas por 1h, apenas dois extratos, denominados A70D e D70D, apresentaram desempenho satisfatório na manutenção da viabilidade celular, semelhante (p > 0,05) ao controle HBSS e diferindo estatisticamente (p < 0,05) das demais formulações. A70D e D70D foram então testados em cinco diluições em propilenoglicol, ao longo de 24h, com análise nos tempos 0, 30 min., 1, 3, 6, 10 e 24h. As frações mais concentradas apresentaram pior desempenho (p < 0,05) em relação aos seus extratos originais que mantiveram viabilidade próxima a 80% ao longo de 24h. A redução da viabilidade proporcional ao aumento da concentração das frações foi observada. Os resultados sugerem que soluções de própolis, em concentrações adequadas, podem ser utilizadas nos estudos futuros sobre alternativas aos meios de conservação de dentes avulsionados rotineiramente utilizados na prática odontológica.

In this study a comparison was made of human mononuclear cell (PBMCs) viability, when cells were kept in different propolis extracts and formulations. PBMCs (106 cell mL-1), obtained from healthy donors (n=5), were incubated at 20ºC in the different propolis solutions, as well as in Hank's balanced salt solution (HBSS), used as experimental control. The cell viability was analyzed by Trypan blue exclusion assay. When incubated for 1h, only two extracts, denominated A70D and D70D, showed appropriate results for maintaining cell viability. A70D and D70D showed better viability (p < 0.05) than other formulations and no difference (p > 0.05) from the HBSS control. A70D and D70D were tested in five dilutions in propylene glycol, over 24h, with analysis at 0, 30 minutes, 1, 3, 6, 10 and 24h. The most concentrated fractions showed the worst performance (p < 0.05) in comparison with their original extracts, which remained close to 80% viability over 24h. Reduction in viability proportional to increase in concentration of formulations was observed. The results suggest that propolis solutions in appropriate concentrations may be used in future studies on alternatives to mediums routinely used in dental practice for storing avulsed teeth.

Removal efficiency of propolis paste dressing from the root canal

OBJETIVES: The aim of this study was to investigate through scanning electron microscopy (SEM) the cleaning of root canal walls after the use of experimental propolis or calcium hydroxide root canal dressings. MATERIAL AND METHODS: Twenty single-rooted teeth were used. After conventional cleaning and shaping procedures and removal of the smear layer with 17 percent EDTA, the teeth were divided into four groups according to the medication used (N=5): Group I (control) - No drug, Group II - Calcium hydroxide dressing, Group III - Propolis paste A70D and Group IV - Propolis paste D70D. The medications were introduced into the root canals and maintained for 7 days, then removed with a K-file and 5 mL of 1 percent sodium hypochlorite irrigation. Finally, the canals were flushed with 2 mL of 17 percent EDTA for 3 min. For SEM analysis, the roots were cleaved and microphotographs from the middle third of the root canal were taken at 750x. The cleaning of the root canal walls was determined by the number of open dentinal tubules as verified with the software Image Tool 3.1. The statistical analysis was performed by ANOVA and Tukey's test (p<0.05). RESULTS: The results showed no statistically significant difference between the calcium hydroxide and propolis groups. CONCLUSIONS: The experimental propolis pastes presented acceptable physical characteristics to be used as intracanal medicaments.

Resposta imune inata na estomatite protética: interação in vitro entre células epiteliais de palato humano e Candida albicans / Innate imune response in denture stomatitis: in vitro interaction between human palatal epithelial cells and Candida albicans

A presença de Candida albicans nos biofilmes microbianos da superfície interna das próteses totais superiores está relacionada com uma doença inflamatória no palato, a estomatite protética. Constituinte da defesa inata do hospedeiro, o epitélio bucal, por sua vez, tem a capacidade de reconhecer e reagir aos fatores fúngicos a fim de evitar a invasão pelo microrganismo. O objetivo deste trabalho foi avaliar in vitro o efeito direto e indireto de C. albicans viável sobre as células epiteliais de palato humano (CEPH) ao longo do tempo. Objetivamos correlacionar os eventos de agressão, apoptose e invasão das CEPH provocados pelo fungo, com as respostas de defesa epitelial mediante produção de óxido nítrico (NO) e expressão gênica do peptídeo antimicrobiano β-defensina 2 (hBD-2). Material e Métodos: As CEPH foram obtidas, parte pelo método explante e parte pelo método enzimático, e mantidas em co-cultivo sobre uma camada de sustentação feederlayer (fibroblastos gengivais humanos mitoticamente inativados). Após desafios das CEPH com C. albicans ATCC 90028 por contato direto fungo-epitélio (D.D.) e indireto pelo sobrenadante da cultura do fungo hifal (D.I.), proporções de desafio de 0,01/1; 0,025/1 e 0,1/1 levedura/queratinócito (FUN/EPI) e tempos experimentais de 3, 6 e 10 h foram determinados; via ensaios de viabilidade celular por imunofluorescência (LIVE/DEAD), e análise qualitativa da invasão celular pelo fungo por meio do método colorimétrico com laranja de acridina. A apoptose epitelial foi determinada pela marcação nuclear fluorescente com Hoechtst 33258. A produção de óxido nítrico (NO) e a expressão de RNAm de hBD-2 foram avaliados por reação colorimétrica de Griess e RT-qPCR, respectivamente. Os resultados foram expressos como média ± desvio padrão e submetidos aos testes estatísticos ANOVA Fatorial, Teste de Contraste; ou Teste de Mann-Whitney (p<0,05). Resultados: Em 3 h, foi detectado aumento da apoptose das células epiteliais em relação ao...

The presence of the fungus Candida albicans in the microbial biofilm underlying maxillary prosthesis is related to an inflammatory reaction of the palatal mucosa, the denture stomatitis. As a component of the host innate defense, the oral epithelium has the ability to recognize and react to fungal factors in order to prevent the microrganism invasion. The aim of this study was to in vitro evaluate the direct and indirect effect of viable C. albicans on the human palatal epithelial cells (HPEC) over time. The aggressive events, such as apoptosis and HPEC invasion by the fungus, were correlated with epithelial defense responses through the nitric oxide (NO) production and antimicrobial peptides β-defensin (hBD-2) mRNA expression. Methods: The HPEC were obtained by explant and enzymatic methods, and were maintained in co-culture on a feeder-layer support (mitotically inactivated human gingival fibroblasts). After the HPEC challenges with C. albicans ATCC 90028 by direct contact fungus-epithelium (D.D.) and indirect contact by supernatant from hyphal fungus (D.I.), defiance ratios of 0.01/1, 0.025/1 and 0.1/1 yeast/keratinocyte (FUN/EPI) and experimental times of 3, 6 and 10 h were determined. These conditions were standardized by cell viability immunofluorescence assay (LIVE/DEAD), and cell invasion qualitative analysis (colorimetric method with acridine orange). The apoptotic cells were determined by fluorescent nuclear staining with Hoechtst 33258. The nitric oxide (NO) production and hBD-2 gene expression were evaluated by Griess colorimetric reaction and RT-qPCR, respectively. The results were expressed as mean ± standard deviation and were analyzed using the factorial ANOVA, Contrast Test; or Mann-Whitney Test (p<0,05). Results: At 3 h, the apoptotic epithelial cells under 0.1/1 FUN/EPI increased compared to epithelium unchallenged (p<0,05) that remained over time with increasing concentration and independent of D.D. and D.I. The onset...