Behavior of Human Osteoblast Cells Cultured on Titanium Discs in Relation to Surface Roughness and Presence of Melatonin.

The aim of this work was to observe the behavior of osteoblast cells cultured in vitro on titanium discs in relation to disc surface roughness and the addition of melatonin to the culture medium. MG63 osteoblast cells were cultivated on 120 Grade 5 Ti divided into three groups: Group E, treated with dual acid etch; Group EP, treated with dual acid etch and calcium phosphate; and Group M, machined. Surface roughness was examined under a laser scanning confocal microscope (CLSM) and scanning electron microscopy (SEM). The proliferation and morphology of cells were determined under fluorescence microscopy and SEM. Messenger ribonucleic acid (mRNA) of different genes related to osteoblastic differentiation was quantified by means of real-time quantitative polymerase chain reaction (RT-PCR) assay. The greatest surface roughness was found in Group EP (Ra 0.354 µm), followed by Group E (Ra 0.266 µm), and Group M (Ra 0.131 µm), with statistically significant differences between the groups (p < 0.001). In the presence of melatonin a trend to a higher cell proliferation was observed in all groups although significant differences were only found in Group M (p = 0.0079). Among the genes studied, a significant increase in phosphate-regulating neutral endopeptidase, X-linked (PHEX) expression was observed in cells cultured on EP discs. The addition of melatonin increased osteoblast cell proliferation and differentiation, and may favor the osseointegration of dental implants.

Cladosporium cladosporioides and Cladosporium pseudocladosporioides as potential new fungal antagonists of Puccinia horiana Henn., the causal agent of chrysanthemum white rust.

Puccinia horiana Hennings, the causal agent of chrysanthemum white rust, is a worldwide quarantine organism and one of the most important fungal pathogens of Chrysanthemum × morifolium cultivars, which are used for cut flowers and as potted plants in commercial production regions of the world. It was previously reported to be controlled by Lecanicillium lecanii, Cladosporium sphaerospermum, C. uredinicola and Aphanocladium album, due to their antagonistic and hyperparasitic effects. We report novel antagonist species on Puccinia horiana. Fungi isolated from rust pustules in a commercial greenhouse from Villa Guerrero, México, were identified as Cladosporium cladosporioides and Cladosporium pseudocladosporioides based upon molecular analysis and morphological characters. The antagonism of C. cladosporioides and C. pseudocladosporioides on chrysanthemum white rust was studied using light and electron microscopy in vitro at the host/parasite interface. Cladosporium cladosporioides and C. pseudocladosporioides grew towards the white rust teliospores and colonized the sporogenous cells, but no direct penetration of teliospores was observed; however, the structure and cytoplasm of teliospores were altered. The two Cladosporium spp. were able to grow on media containing laminarin, but not when chitin was used as the sole carbon source; these results suggest that they are able to produce glucanases. Results from the study indicate that both Cladosporium species had potential as biological control agents of chrysanthemum white rust.

In vitro preliminary study of osteoblast response to surface roughness of titanium discs and topical application of melatonin

OBJECTIVES: To observe human osteoblast behavior cultured in vitro on titanium discs (Ti) in relation to surface roughness and melatonin application. Study DESIGN: Human osteoblasts (MG-63) were cultured on 60 Ti6Al4V discs divided into three groups: GroupI: discs treated with dual acid etching; Group II dual acid etching and blasting with calcium phosphate particles; Group III (control) machined discs. Surface roughness and topography of the discs were examined with scanning electron microscope (SEM) and confocal laser scanning electron microscope( CLSM).Osteoblast adhesion, proliferation and cell morphology were determined by means of fluorescence microscopy with Image-Pro Plus software and SEM. RESULTS: Group II presented the roughest discs, while the least rough were Group III. Cell adhesion was greatest in Group II. The addition of melatonin improved cell proliferation. CONCLUSIONS: 1. Surface treatments (dual acid etching, calcium phosphate impaction) increase surface roughness in comparison with machined titanium.2. Greater surface roughness tends to favor cell adhesion after 24-hour cell culture.3. The addition of melatonin tends to favor osteoblast proliferation

In vitro preliminary study of osteoblast response to surface roughness of titanium discs and topical application of melatonin.

OBJECTIVES: To observe human osteoblast behavior cultured in vitro on titanium discs (Ti) in relation to surface roughness and melatonin application. STUDY DESIGN: Human osteoblasts (MG-63) were cultured on 60 Ti6Al4V discs divided into three groups: Group I: discs treated with dual acid etching; Group II dual acid etching and blasting with calcium phosphate particles; Group III (control) machined discs. Surface roughness and topography of the discs were examined with scanning electron microscope (SEM) and confocal laser scanning electron microscope( CLSM). Osteoblast adhesion, proliferation and cell morphology were determined by means of fluorescence microscopy with Image-Pro Plus software and SEM. RESULTS: Group II presented the roughest discs, while the least rough were Group III. Cell adhesion was greatest in Group II. The addition of melatonin improved cell proliferation. CONCLUSIONS: 1. Surface treatments (dual acid etching, calcium phosphate impaction) increase surface roughness in comparison with machined titanium. 2. Greater surface roughness tends to favor cell adhesion after 24-hour cell culture. 3. The addition of melatonin tends to favor osteoblast proliferation.

Análisis de la variación intraespecífica de Conidiobolus coronatus usando RAPD y secuencias ITS / Intraspecific variation analysis of Conidiobolus coronatus using RAPD and ITS sequencing

Introducción El hongo Conidiobolus coronatus (C. coronatus) tiene una extensa distribución de hábitats y organismos; se encuentra en forma saprófita, ataca insectos y mamíferos, incluido el humano, aunque de este los reportes son pocos y están restringidos a zonas tropicales. El objetivo de este trabajo fue determinar si existe o no variación genética entre aislamientos de C. coronatus provenientes de lesiones en humanos y otras fuentes. Métodos Once aislamientos de C. coronatus obtenidos de suelo, insectos y humanos se analizaron con las técnicas de polimorfismo derivado de la amplificación aleatoria de ADN (RAPD) y secuenciación del espaciador interno transcrito (ITS1-5.8S rADN-ITS2); para el análisis de las regiones ITS se usaron los métodos de máxima parsimonia, neighbour-joining (NJ) y mínima evolución. Resultados El análisis de los polimorfismos de C. coronatus con las técnicas RAPD e ITS evidenció altos niveles de variación intraespecífica en los aislamientos evaluados. Los aislamientos obtenidos de lesiones en humanos presentan la mayor divergencia genética respecto de los obtenidos de otras fuentes; la mayor distancia genética entre grupos de aislamientos se da entre los provenientes de humanos y los tomados del insecto Lycoriella ingenua. Conclusiones Este trabajo es el primero en evaluar y demostrar que existe variación intraespecífica a nivel molecular de C. coronatus que, a su vez, está relacionada con la fuente de la que se obtuvieron (AU)

Introduction The fungus Conidiobolus coronatus (C. coronatus) has an extensive distribution of habitats and hosts. It is found saprophytically, and attacks insects and mammals, including humans. Although there are few reports on humans, and they are restricted to tropical areas. The aim of this work was to determine whether genetic variation exists between C. coronatus isolates coming from human lesions and other sources. Methods A total of 11 C. coronatus isolates obtained from soil, insects and humans were analyzed with the random amplification of polymorphic ADN (RAPD) and internal transcribed spacer (ITS) techniques; the maximum parsimony, neighbour-joining and minimum evolution methods were used for the ITS analysis. Results The analysis of the C. coronatus polymorphisms showed high intra-species variation levels between the evaluated isolates. The isolates coming from human lesions showed the greatest genetic divergence compared with the remaining isolates. The greatest genetic distance between isolate groups was found between those coming from humans and those taken from the insect Lycoriella ingenua. Conclusions This is the very first work evaluating and demonstrating that within species variation exists at molecular level in C. coronatus, and is related to the source where the isolates were taken from (AU)

[Intraspecific variation analysis of Conidiobolus coronatus using RAPD and ITS sequencing]. / Análisis de la variación intraespecífica de Conidiobolus coronatus usando RAPD y secuencias ITS.

INTRODUCTION: The fungus Conidiobolus coronatus (C. coronatus) has an extensive distribution of habitats and hosts. It is found saprophytically, and attacks insects and mammals, including humans. Although there are few reports on humans, and they are restricted to tropical areas. The aim of this work was to determine whether genetic variation exists between C. coronatus isolates coming from human lesions and other sources. METHODS: A total of 11C. coronatus isolates obtained from soil, insects and humans were analyzed with the random amplification of polymorphic ADN (RAPD) and internal transcribed spacer (ITS) techniques; the maximum parsimony, neighbour-joining and minimum evolution methods were used for the ITS analysis. RESULTS: The analysis of the C. coronatus polymorphisms showed high intra-species variation levels between the evaluated isolates. The isolates coming from human lesions showed the greatest genetic divergence compared with the remaining isolates. The greatest genetic distance between isolate groups was found between those coming from humans and those taken from the insect Lycoriella ingenua. CONCLUSIONS: This is the very first work evaluating and demonstrating that within species variation exists at molecular level in C. coronatus, and is related to the source where the isolates were taken from.