Diverse effect of BMP-2 homodimer on mesenchymal progenitors of different origin.

Bone morphogenetic protein-2 (BMP-2), is a potential factor to enhance osseointegration of dental implants. However, the appropriate cellular system to investigate the osteogenic effect of BMP-2 in vitro in a standardized manner still needs to be defined. The aim of this study was to examine the effect of BMP-2 on the cell proliferation and osteogenic differentiation of human osteogenic progenitors of various origins: dental pulp stem cells (DPSC), human osteosarcoma cell line (Saos-2) and human embryonic palatal mesenchymal cell line (HEPM). For induction of osteogenic differentiation, cell culture medium was supplemented with BMP-2 homodimer alone or in combination with conventionally used differentiation inducing agents. Differentiation was monitored for 6-18 days. To assess differentiation, proliferation rate, alkaline phosphatase activity, calcium deposition and the expression level of osteogenic differentiation marker genes (Runx2, BMP-2) were measured. BMP-2 inhibited cell proliferation in a concentration and time-dependent manner. In a concentration which caused maximal cell proliferation, BMP-2 did not induce osteogenic differentiation in any of the tested systems. However, it had a synergistic effect with the osteoinductive medium in both DPSC and Saos-2, but not in HEPM cells. We also found that the differentiation process was faster in Saos-2 than in DPSCs. Osteogenic differentiation could not be induced in the osteoblast progenitor HEPM cells. Our data suggest that in a concentration that inhibits proliferation the differentiation inducing effect of BMP-2 is evident only in the presence of permissive osteoinductive components. ß-glycerophosphate, was identified interacting with BMP-2 in a synergistic manner.

Fitoterápico Equisetum giganteum e estomatite protética: estudo da ação antimicrobiana, antiaderente e anti-inflamatória contra Candida albicans, e potencial citotóxico sobre células epiteliais do palato humano / Phytotherapeutic Equisetum giganteum and denture stomatitis: study of antimicrobial, antiadherent and anti-inflammatory action against Candida albicans, and cytotoxic potential in human palatal epithelial cells

A presença de Candida albicans nos biofilmes microbianos aderidos na superfície interna das próteses removíveis, principalmente totais superiores, está relacionada com uma doença inflamatória no palato, conhecida como estomatite protética (EP). Assim, torna-se fundamental a realização de novos estudos sobre alternativas terapêuticas, direcionados à prótese e não somente à mucosa, que sejam simultaneamente antimicrobianas, anti-inflamatórias, não tóxicas para os tecidos bucais e que produzam menos danos à prótese que os métodos convencionais. Os fitoterápicos podem representar uma destas alternativas. Objetivos: O objetivo deste trabalho foi estudar a ação fitoterápica do Equisetum giganteum, nas concentrações de 50, 25, 16, 8 e 4 mg/mL, sobre C. albicans e descartar sua ação citotóxica sobre o palato humano bem como sobre monócitos humanos. Material e Métodos: Após coleta, obtenção e identificação de compostos por espectrometria de massas do extrato hidroalcoólico de E. giganteum, sua atividade antimicrobiana foi determinada pela concentração inibitória mínima em meio líquido, contra as cepas clínicas Candida albicans SC 5314 e Escherichia coli O:124, e a cepa padrão Staphylococcus aureus ATCC 6538. Propriedades antiaderentes do extrato, sobre biofilmes de C. albicans induzidos sobre corpos de prova de resina acrílica, foram determinadas por imunofluorescência (LIVE/DEAD) e pela análise em microscópio de varredura confocal a laser. A atividade anti-inflamatória do fitoterápico foi averiguada através da análise da produção de espécies reativas de oxigênio (ROS) por monócitos humanos estimulados por C. albicans e LPS, por meio da marcação fluorescente utilizando o reagente Cell Rox Deep Red^®. Avaliação de citotoxicidade foi realizada in vitro com células epiteliais de palato humano e monócitos humanos, por meio do ensaio colorimétrico MTT. Os resultados foram expressos como média ± desvio padrão...

The presence of Candida albicans in the microbial biofilms adhered to the internal surface of the removable denture, mainly the full upper ones, is related to an inflammatory palate disease known as denture stomatitis (DS). Thus, it is essential that new studies are done about therapeutic alternatives directed to the dentures, not only to the buccal mucosa, and which are, at the same time, antimicrobial, antiinflammatory, non-poisoning to the buccal tissues and that they produce less harm to the denture than the current methods. The phytotherapeutic (herbal) remedies may represent a good alternative. Objectives: The aim of this paper is to study the phytotherapeutic action of Equisetum giganteum in the concentrations of 50, 25, 16, 8 and 4 mg/mL on C.albicans and discard the cytotoxic action on the human palate, as well as on human monocytes. Material and Methods: After collecting, obtaining and identifying the compounds by means of mass spectrometry of the hydroalcoholic extract of E. giganteum, its antimicrobial activity was determined by the inhibitory minimum concentration in liquid media, against clinic strains of Candida albicans SC 5314 and Escherichia coli O:124, and standard Staphylococcus aureus ATCC 6538 strains. The antiadherent, properties of the extract on biofilms of C. albicans over acrylic resin proof specimens were determined by immunofluorescence test (LIVE/DEAD) and by the analysis in a Confocal Laser Microscope Scanning. The anti-inflammatory activity of the phytotherapeutic (herbal) remedy was assessed through the analysis of the production of reactive oxygen species (ROS) to human monocytes stimulated by C. albicans and LPS, through fluorescent lighting using the reagent Cell Rox Deep Red®. The evaluation of cytotoxicity was done in vitro with epithelial cells of human palate and human monocytes, through colorimetric MTT assay. The results were expressed in means ± standard deviation and submitted to statistics Kruskal-Wallis Test...

Effect of local antimicrobial agents on excisional palatal wound healing: a clinical and histomorphometric study in rats.

AIM: To assess the effect of topically applied antimicrobial agents on palatal excisional wound in rats. MATERIALS AND METHODS: Excisional wounds, 5 mm in diameter, were made in the centre of the palate of 125 Wistar male rats. In four experimental groups, chlorhexidine digluconate (CHX) 0.12% solution, 1% CHX gel, phenolic compounds solution (Listerine), amine/stannous fluoride solution (Meridol) and saline solution as a control group were applied daily for 1 min. The wound area was measured photographically and the epithelialization rate was determined histologically at 3, 7, 14 and 21 days post-surgery. RESULTS: The mean wound area and mean distance between the epithelial margins decreased significantly with time (p<0.001) in experimental and control groups, with the greatest wound area reduction and rate of epithelialization on day 14. A significantly superior rate of wound epithelialization (p=0.03) was presented following use of 1% CHX gel and Listerine and a comparatively inferior one when the Meridol solution was applied. CONCLUSIONS: Each tested antimicrobial agent when applied on an excisional wound with epithelial and connective tissue deficiency did not have a negative effect on the rate of wound closure. The best results were achieved with 1%CHX gel and Listerine.

[Effectiveness and mechanism of action of a new plant complex drug in nonspecific inflammatory processes in the bronchopulmonary system]. / Effektivnost' i mekhanizm deistviia novogo kompleksnogo sredstva rastitel'nogo proiskhozhdeniia pri nespetsificheskikh vospalitel'nykh protsessakh v bronkholegochnoi sisteme.

The elixir Bronchofit is an aqueous-alcoholic extract from 7 kinds of plants. It is a balanced compound, rich in biologically active substances including essential oils and flavonoids, possessing a wide spectrum of pharmacological activity. On the model of frog's palate the elixir enhanced the transport function of the ciliated epithelium. On the model "karragenin edema" bronchofit showed a marked anti-inflammatory activity. A noticeable therapeutic effect was registered in rats with induced bronchopulmonary inflammation. Bronchial secretion contained reduced content of glycosaminoglycans while bronchial lavage did of histamine. Also, bronchofit demonstrated a prominent antioxidative activity, an antibacterial action in relation to Str. pyogenes and Bac. cereus. By a total complex of the activities bronchofit is a promising medicine against bronchopulmonary inflammation.

Effect of hyperthermia on rat fetus palate epithelium / Efeito da hipertemia no epitélio palatino fetal do rato

Maternal hyperthermia during gestation induces delayed cellular growth and differentation in fetal rat palate epithelium, with increased nuclear, cytoplasmic and cellular volumes, increased epithelial thickness, decreased keratin and cellular numerical density

Epithelial bridging of the primary palate: I. Characterization of sub-cultured epithelial cells.

Primary palatogenesis involves an intricate array of events. Cell migration, proliferation, differentiation, programmed death, and fusion occur. Prior to fusion, the morphology of the epithelium undergoes marked changes. Epithelial projections form and extend across the fusion site attaching by filopodia to the opposite prominence. By appearance, the epithelium plays a critical role in facial development. In order to monitor epithelial activities, a study was done to isolate and characterize epithelial cells derived from the primary palate. The primary palate was microdissected from day 13 Sprague-Dawley rat embryos, and the epithelium and mesenchyme were separated by enzymatic digestion with a 3% trypsin-pancreatin solution (3:1). All explants were cultured in Dulbecco's modified Eagle's medium (DMEM) and Ham's F-12 medium (1:1) supplemented with 10% fetal calf serum (FCS), 20 ng/ml epidermal growth factor (EGF), and antibiotics. Explant cells were gathered by trypsin harvesting and sub-cultured. These sub-cultured cells were further characterized. Transmission and scanning electron microscopy showed that the cells retained many morphological features observed in vivo. In passaged cells, type IV collagen, laminin, and cytokeratins were visualized by immunocytochemistry. Gel electrophoresis analysis of the water-insoluble extracts demonstrated major bands of proteins of 50 kD and 44 kD that were synthesized by the epithelial cells but not by the mesenchymal cells. These cytokeratin types are suggestive of a simple undifferentiated embryonic epithelium. The effect of all-trans retinoic acid (RA) on cell number and [3H]-proline incorporation was assessed. At [10(-4)M] and [10(-6)M] retinoic acid resulted in significant inhibition in cell proliferation and amount of proline incorporated, with the greater inhibition occurring in the mesenchymal cells. In the concentrations studied, retinoic acid has an inhibitory effect on the two differently derived cell types. This study established that sub-cultured epithelial cells maintain their phenotype and can be used to study fusion processes. Part 2 will demonstrate how the morphology of the epithelial cells can be modified to produce the changes that are observed during fusion of the primary palate.