Peri-implantitis increases the risk of medication-related osteonecrosis of the jaws associated with osseointegrated implants in rats treated with zoledronate.

This study evaluated the peri-implant tissues under normal conditions and under the influence of experimental peri-implantitis (EPI) in osseointegrated implants installed in the maxillae of rats treated with oncologic dosage of zoledronate. Twenty-eight senescent female rats underwent the extraction of the upper incisor and placement of a titanium dental implant (DI). After eight weeks was installated a transmucosal healing screw on DI. After nine weeks, the following groups were formed: VEH, ZOL, VEH-EPI and ZOL-EPI. From the 9th until the 19th, VEH and VEH-EPI groups received vehicle and ZOL and ZOL-EPI groups received zoledronate. At the 14th week, a cotton ligature was installed around the DI in VEH-EPI and ZOL-EPI groups to induce the EPI. At the 19th week, euthanasia was performed, and the maxillae were processed so that at the implanted sites were analyzed: histological aspects and the percentage of total bone tissue (PTBT) and non-vital bone tissue (PNVBT), along with TNFα, IL-1ß, VEGF, OCN and TRAP immunolabeling. ZOL group presented mild persistent peri-implant inflammation, higher PNVBT and TNFα and IL-1ß immunolabeling, but lower for VEGF, OCN and TRAP in comparison with VEH group. ZOL-EPI group exhibited exuberant peri-implant inflammation, higher PNVBT and TNFα and IL-1ß immunolabeling when compared with ZOL and VEH-EPI groups. Zoledronate disrupted peri-implant environment, causing mild persistent inflammation and increasing the quantity of non-vital bone tissue. Besides, associated with the EPI there were an exacerbated inflammation and even greater increase in the quantity of non-vital bone around the DI, which makes this condition a risk factor for medication-related osteonecrosis of the jaws.

Genomic Evaluation of Formed Biofilm on Dental Implants with Different Surface Treatments Associated with Zirconia or Titanium Abutments: An In Vitro Study.

PURPOSE: The aim of this study was to evaluate the formed biofilm on two types of implant surfaces (hydrophilic or hydrophobic) associated with titanium (Ti) or zirconia (Zn) abutments. MATERIALS AND METHODS: Samples were separated into four groups according to type of surface and abutment used (n = 10): (1) hydrophobic/Ti abutment, (2) hydrophilic/Ti abutment, (3) hydrophobic/Zn abutment, and (4) hydrophilic/Zn abutment. Implant-abutment assemblies were incubated with human saliva and supragingival biofilm. Samples of biofilm were evaluated by DNA Checkerboard hybridization, identifying up to 41 species. Scanning electron microscopy (SEM) images were obtained from the implants and abutments. RESULTS: The microbial count was higher for samples from groups with the hydrophilic/Ti abutment, followed by hydrophobic/Zn abutment, hydrophilic/Ti abutment, and hydrophobic/Zn abutment (P < .05). Hydrophilic surfaces and Zn abutments showed the highest counts of microorganisms. Individual bacterial counts were variable between groups; the hydrophilic/Zn abutment group had the highest microbial diversity, including T forsythia, P nigrescens, S oralis, S sanguinis, L casei, M orale, P aeruginosa, P endodontalis, S aureus, S gallolyticus, S mutans, S parasanguinis, S pneumoniae, and C albicans. The hydrophilic/Ti abutment group had the highest count of T forsythia and T denticola, microorganisms of Socransky red complex. The SEM images showed the bacterial colonization in both surfaces of the implant and abutment. CONCLUSION: Different surfaces of implants and abutments showed significant differences in the count and diversity of species. The hydrophilic/Zn abutment group presented the highest count and diversity of target species.

The use of fluorescein for labeling genomic probes in the checkerboard DNA-DNA hybridization method.

Molecular methods that permit the simultaneous detection and quantification of a large number of microbial species are currently employed in the evaluation of complex ecosystems. The checkerboard DNA-DNA hybridization technique enables the simultaneous identification of distinct bacterial species in a large number of dental samples. The original technique employed digoxigenin-labeled whole genomic DNA probes which were detected by chemiluminescence. In this study, we present an alternative protocol for labeling and detecting whole genomic DNA probes in the Checkerboard DNA-DNA hybridization method. Whole genomic DNA was extracted from five bacterial species and labeled with fluorescein. The fluorescein labeled whole genomic DNA probes were hybridized against whole genomic DNA or subgingival plaque samples in a checkerboard hybridization format, followed by chemiluminescent detection. Our results reveal that fluorescein is a viable and adequate alternative labeling reagent to be employed in the checkerboard DNA-DNA hybridization technique.

Immunohistochemistry and ultrastructural characteristics of nerve endings in the oral mucosa of rat.

The sensory nerve endings of the rat tongue, cheek and palate were studied using immunohistochemical staining and transmission electron microscopy analysis. The specimens were fixed in modified Karnovsky solution and embedded in Spurr resin. Substance P, calcitonin gene-related peptide (CGRP)- and protein gene product 9.5 (PGP b9.5)-containing nerve fibers in the rat tongue, cheek and palate were examined by electronic microscopical analysis and immunohistochemical localization. These fibers run very close to the basal lamina of the epithelium and extend into the filliform and fungiform papillae. Numerous plexiform fibers immunoreactive for substance P, CGRP and PGP 9.5 were found in the connective tissue of mucosa. Electron microscopic observations showed clearly immunostained nerve fibers, which are located very close to the basal lamina of epithelial cells. Some electron-dense granules may be observed in the axoplasms of both substance P and CGRP immunoreactive fibers. Several lamellar corpuscles into the subepithelial connective tissue papillae, Merkel corpuscles and numerous thin unmyelinated and myelinated axons were observed. The terminal axons revealed numerous mitochondria, neurofilaments, microtubules and clear vesicles in the base of axoplasmic protrusions. The lamellar cells showed caveolae and interlamelar spaces filled by amorphous substance. Between the lamellar cells and axoplasmic membrane, and in the adjacent lamellae region, desmosome-type junctions were observed. The quantitative and morphometric analysis showed nerve endings with an average area of 4.83 ± 3.4 µm(2) and 19.4 internal mitochondria in this site and the organized corpuscles with an average area of 79.24 ± 27.24 µm(2) and 24.23 internal mitochondria in this place. All the structures observed are involved in the transmission of pain and mechanoreceptors stimulus of these oral mucosae.

Trigeminal nitric oxide synthase expression correlates with new bone formation during distraction osteogenesis.

Nitric oxide synthase (NOS) has been reported to be involved with both bone healing and bone metabolism. The aim of this study was to test the null hypothesis that there is no correlation between new bone formation during mandibular distraction osteogenesis and NOS expression in the trigeminal ganglion of rats. Newly formed tissue during distraction osteogenesis and trigeminal NOS expression measured by the NADPH-diaphorase (NADPH-d) reaction were evaluated in 72 male Wistar rats by histomorphometric and histochemical methods. In animals submitted to 0.5 mm/day distraction osteogenesis, the percentage of bone tissue was higher in the basal area of the mandibles compared with the center and significantly increased through the experimental periods (P < 0.05). At the sixth postoperative week, the difference in bone formation between the continuous and acute distraction osteogenesis groups was the highest. Significant correlation between new bone formation by distraction osteogenesis and NADPH-d-reactive neurons was found, varying according to neuronal cell size (r = -0.6, P = 0.005, small cells strongly stained; r = 0.5, P = 0.018, large cells moderately stained). The results suggest that NOS may play a role in the bone healing process via neurogenic pathways, and the phenomenon seems to be neuronal cell morphotype-dependent. Further studies are now warranted to investigate the mechanistic link between the expression of trigeminal NOS and mandibular new bone formation by distraction osteogenesis.

Morphological and histochemical study of the masseter muscle after occlusal alteration

The aim of this study was to evaluate alterations in the masseter muscle of 30 male guinea-pigs submitted to occlusal alteration. The animals were divided into 2 equal groups, the control group (C) only submitted to surgical stress, and the occlusal altered group (T) submitted to teeth extraction. Each group was subdivided into 3 groups, with 5 animals, for the following studies: macroscopy and vessels distribution, light microscopy and histochemical analysis, with animals' perfusion 2 months after surgery. Data were submitted to statistical analysis. Macroscopically, an anteroposterior orientation of the muscular fibers was found and statistical difference between C and T groups in width (p<0.05). Microscopically, C and T groups showed polygonal muscular fibers with variable diameters, and on the left side of the T group these differences were more pronounced. Histochemically, in both groups, the prevalence of intermediate reactivity fibers and several high reactivity fibers spread out in the deeper area was observed, with no significant differences between superficial fibers on both sides of C and T groups. It was possible to conclude that the masseter muscle in guinea-pigs was sensible to functional chewing alteration.

Periimplantitis: checkerboard DNA-DNA hybridization for bacterial pathogen identification

A recent innovation in medical field is the use of DNA probes in microbiological diagnosis of the oral cavity. Thus, this study has the objective to present the mainly characteristics of Checkerboard DNA-DNA Hybridization method for bacterial pathogens identification related to periimplantitis, commonly disease found in the oral cavity, as wells as, to show the uses and applications of this technique.

Una innovación reciente en medicina es la utilización de sondas de DNA para diagnóstico microbiológico. Este trabajo tiene como objetivo, presentar las principales características del método Checkerboard DNA-DNA Hybridization para la identificación de bacterias patógenas associadas a periimplantite en la cavidad oral, mostrando las diferentes utilizaciones y aplicaciones de esta técnica.

Morphological alterations in the masseter muscle induced by unilateral extraction of the upper molars in guinea pigs (Cavia porcellus)

El objetivo del presente estudio fue analizar los aspectos morfológicos del sistema estomatognático y su relación con la mala oclusión. Con tal propósito, determinamos las alteraciones morfológicas que ocurren en el músculo masetero de cobayas, Cavia porcellus después de la inducción de mala oclusión a través de la extracción de los molares superiores del lado izquierdo. Los animales (n=20) fueron divididos en dos grupos (control y experimental), los cuales fueron sometidos a análisis biométricos, angioarquitecturales macroscópicos y al microscopio de luz. Los resultados obtenidos permitieron concluir que, después de 60 días de extracción dental unilateral hubo reducción en el ancho y en el grosor del músculo masetero del grupo experimental. Microscópicamente, se observaron fibras musculares de variados diámetros, de forma redondeada y poligonal, fibras musculares alteradas con un núcleo central, además de fibras en degeneración y macrófagos evidentes en la parte profunda del músculo.