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.

Microcystin-LR at sublethal concentrations induce rapid morphology of liver and muscle tissues in the fish species Astyanax altiparanae (Lambari).

The process of eutrophication and consequent proliferation of cyanobacteria in rivers and lakes leads to increasing numbers of harmful algal blooms and higher concentration of toxic metabolites in freshwater bodies. Microcystin is a toxic metabolite produced by cyanobacteria that is frequently detected and can pose health risks to important freshwater species including fish. The aim of the present study was to evaluate the effects of microcystin-LR on the morphology of Astyanax altiparanae's liver and muscle. One hundred (n = 100) Astyanax altiparanae were divided into 5 groups (n = 20) with 24 h and 96 h of microcystin exposition at two doses of 0.5 and 1.0 µg/L. Differences were observed in the microcystin treatment with respect to histopathological analyses including cytoplastic degradation, displacement, and increase in nuclei volume and area of hepatocytes. Hyperemia and dilation of blood capillaries were seen in the liver. There were also observable changes in the size of muscle fibers and muscle inflammation. Our results demonstrate that microcystins can impact the integrity of both tissues even at sublethal concentrations. Low doses of microcystins are therefore sufficient to intoxicate fish livers and muscle tissues.

Development and femoral growth of rabbits of the New Zealand line / Desarrollo y crecimiento femoral de conejas de la línea new zealand

RESUMEN En este proyecto se investigan los cambios que acontecen en el desarrollo y crecimiento de conejos hembras de la línea New Zealand (CoNZ) en sus huesos femorales. Los animales fueron mantenidas en jaulas individuales desde las dos semanas de edad, con comida y agua ad libitum y se sacrificaron en tiempos mensualmente consecutivos: 1, 2, 3, 4, 5, y 6 meses. Tras la obtención de las piezas femorales, y a partir de estudios imagenológicos se determinaron los ángulos del cuello femoral (Af), la longitud total (L), la densidad mineral ósea total, del centro óseo y de la metáfisis femoral (DMOt, DMOco y DMOmf respectivamente), analizándose las variaciones intergrupales por el test Wilcoxon, y corrección de Bonferroni. Se realizaron estudios histológicos de los cortes descalcificados de las piezas femorales. Los análisis sobre los Af mostraron un incremento significativo durante el primer mes mientras que L se estabilizó a partir del 4to mes. Los valores de DMOt mostraron un plateau a partir del cuarto mes, si bien las DMOco y DMOmf ya a partir del tercer mes no mostraron incrementos significativos. Histológicamente se observó para el cuarto mes ausencia de las diferentes zonas características del cartílago de crecimiento metafisiario, con presencia únicamente de un pequeño remanente de células condrales. Desde el quinto mes se observa ausencia total de cartílago, con presencia únicamente de tejido osteoide (TO). La interpretación integrada de los resultados nos permite afirmar, que a partir del cuarto mes de desarrollo, el fémur de CoNZ adquiere características compatibles con un periodo de adultez.

ABSTRACT In this project we investigated the changes of femoral development and growth of female New Zealand rabbits (NZr). Animals were maintained in individual cages since they were two weeks old with food and water ad libitum, and were sacrificed monthly consecutively: 1, 2, 3, 4, 5 and 6. Radiological studies were made with femoral pieces to determine femoral neck angle (fnA), total length (L), total bone mineral density (tBMD), bone center mineral density (bcBMD) and femoral metaphysis bone mineral density (fmBMD). We analyzed intergroup variations with Wilcoxon test and Bonferroni correction. We also performed histologic studies with femoral pieces. The fnA analyzes showed a significant increase in the first month while L stabilized since the fourth month. tBMD showed a plateu since the fourth month, even though bcBMD and fmBMD did not show any significant changes since the third month. In histology it was observed the absence of all typical growth cartilage zones since the fourth month, with the only presence of small remaining cartilage cells. In the fifth month we observed complete absence of cartilage, and presence of osteoid tissue only. The integrated interpretation of the results allows us to affirm that since the fourth month of development the femur of NZr acquires characteristics compatible with the adulthood.

Recombinant Proteins-Based Strategies in Bone Tissue Engineering.

The increase in fracture rates and/or problems associated with missing bones due to accidents or various pathologies generates socio-health problems with a very high impact. Tissue engineering aims to offer some kind of strategy to promote the repair of damaged tissue or its restoration as close as possible to the original tissue. Among the alternatives proposed by this specialty, the development of scaffolds obtained from recombinant proteins is of special importance. Furthermore, science and technology have advanced to obtain recombinant chimera's proteins. This review aims to offer a synthetic description of the latest and most outstanding advances made with these types of scaffolds, particularly emphasizing the main recombinant proteins that can be used to construct scaffolds in their own right, i.e., not only to impregnate them, but also to make scaffolds from their complex structure, with the purpose of being considered in bone regenerative medicine in the near future.

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.

Matrices de tercera generación en la ingeniería de tejidos ósea / Third-generation matrices in bone tissue engineering

Existen numerosas patologías que generan situaciones invalidantes debido a problemas asociados a nivel de defectos óseos. Esto genera, en muchas oportunidades, cuestiones sanitarias de alto impacto. La ingeniería de tejidos óseos pretende generar propuestas novedosas para reparar pérdidas o fracturas óseas, promoviendo regenerar el tejido mediante el implante de matrices biodegradables que puedan actuar como estructuras para la adhesión celular, favoreciendo el crecimiento y la diferenciación hasta formar hueso de novo. El incremento notable de los conocimientos en las áreas biotecnológicas, de síntesis química, así como de biomedicina, permiten el desarrollo de numerosos tipos de matrices de tercera generación, biodegradables y no tóxicas, con características que proponen sean consideradas en la regeneración tisular ósea. Este trabajo intenta resumir los tipos de matrices que mayor impacto han tenido hasta el momento en la medicina regenerativa ósea, mostrando los casos más relevantes de resultados experimentales y clínicos, y propone algunas perspectivas que se deberían considerar para poder aplicarlas a la práctica clínica. Esta es un área que invita a los investigadores a posicionarse en un pensamiento complejo desde el punto de vista científico-filosófico. (AU)

There are several pathologies that generate disability due to complications associated with bone defects. This often generates high impact health troubles. Bone tissue engineering aims to generate novel means to repair bone loss or bone fractures, promoting tissue regeneration through the implantation biodegradables scaffolds, which can act as structures for cell adhesion, that promts cell growth and differentiation for the novo bone formation. The remarkable for the novo bone formation in biotechnology, chemical synthesis, and biomedical knowledge allows the development of numerous types of third generation scaffolds, applied to promote bone tissue regeneration. This brief report aims to review the scaffolds that have had more impact in bone regenerative medicine so far, describing the most relevant experimental and clinical results. This is an area that invites researchers to situate themselves in a complex thought of scientific-philosophical point of view. (AU)

Morphometric, quantitative, and three-dimensional analysis of the heart muscle fibers of old rats: transmission electron microscopy and high-resolution scanning electron microscopy methods.

This research investigated the morphological, morphometric, and ultrastructural cardiomyocyte characteristics of male Wistar rats at 18 months of age. The animals were euthanized using an overdose of anesthesia (ketamine and xylazine, 150/10 mg/kg) and perfused transcardially, after which samples were collected for light microscopy, transmission electron microscopy, and high-resolution scanning electron microscopy. The results showed that cardiomyocyte arrangement was disposed parallel between the mitochondria and the A-, I-, and H-bands and their M- and Z-lines from the sarcomere. The sarcomere junction areas had intercalated disks, a specific structure of heart muscle. The ultrastructural analysis revealed several mitochondria of various sizes and shapes intermingled between the blood capillaries and their endothelial cells; some red cells inside vessels are noted. The muscle cell sarcolemma could be observed associated with the described structures. The cardiomyocytes of old rats presented an average sarcomere length of 2.071 ± 0.09 µm, a mitochondrial volume density (Vv) of 0.3383, a mitochondrial average area of 0.537 ± 0.278 µm(2), a mitochondrial average length of 1.024 ± 0.352 µm, an average mitochondrial cristae thickness of 0.038 ± 0.09 µm and a ratio of mitochondrial greater length/lesser length of 1.929 ± 0.965. Of the observed mitochondrial shapes, 23.4% were rounded, 45.3% were elongated, and 31.1% had irregular profiles. In this study, we analyzed the morphology and morphometry of cardiomyocytes in old rats, focusing on mitochondria. These data are important for researchers who focus the changes in cardiac tissue, especially changes owing to pathologies and drug administration that may or may not be correlated with aging.

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.

Osteoconductive capacity of hydroxyapatite implanted into the skull of diabetics.

Diabetes mellitus can cause various diseases, including loss of bone mineral density as a characteristic manifestation of osteoporosis. In this condition, bone is more vulnerable to pathologic fractures that can be treated by implantation of biomaterial grafts. The aim of this study was to evaluate the osteogenic capacity of hydroxyapatite implanted into bone defects in the skull of nonobese diabetic mice. Fifteen nonobese diabetic mice were divided into 3 groups: control (nondiabetic), spontaneously diabetic, and spontaneously diabetic receiving insulin replacement applied subcutaneously into the dorsum. Defects were created experimentally in the skull with a surgical bur and filled with hydroxyapatite granules. The animals were killed 4 weeks after surgery, and samples were obtained for analysis. Quantitative methods were used for measurement of the new bone formation. Data were analyzed by analysis of variance followed by the Tukey test (P < 0.05). Radiographic results showed good radiopacity of the hydroxyapatite; however, radiolucent spots were seen between the hydroxyapatite granules in the diabetic groups, indicating infiltration of connective tissue. Microscopic results showed projections of newly formed bone from the margin of bone defect toward the implant. The quantity of newly formed bone was significantly higher (P < 0.05) than that observed in the diabetic groups. The recipient area of diabetic groups contained a larger amount of connective tissue as demonstrated by radiographic analyses. In conclusion, the osteogenesis guided by the properties of hydroxyapatite may even occur in bone suffering from the effects of diabetes, but the quantity of newly formed bone is lower, and the process is slower.

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.

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.

Microwave Fixation in Rat Fetuses Tissues: Histological and Immunohistochemical Analysis / Fijación de tejidos de Fetos de Ratas por Microondas: Análisis Histológico e Inmunohistoquímico

The aim of this study was to show the microwaves action in fixation of rat fetuses, dermal and cartilaginous tissues, using histological and immunohistochemistry methods for analysis. It was possible to conclude in this study using the rat as experimental model that the two methods for antibody retrieval, presented an excellent ways for the use of Ki67 antibody in the immunohistochemical analysis.

El objetivo de este estudio fue evaluar la acción de las microondas en la fijación de los tejidos dérmico y cartilaginoso de fetos de ratas, usando para el análisis métodos histológico e inmunohistoquímico. Fue posible concluir en este estudio usando la rata como modelo experimental, que los dos métodos empleados para la recuperación antigénica representan excelentes medios para el uso del anticuerpo Ki67, en el análisis inmunohistoquímico.

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.

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.(AU)

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.