Digital Planning for Immediate Implants in Anterior Esthetic Area: Immediate Result and Follow-Up after 3 Years of Clinical Outcome-Case Report.

In this case report, we demonstrate how the correct positioning of implants, associated with optimal gingival conditioning, and the correct choice of biomaterial can yield very predictable and fantastic aesthetic results. OBJECTIVE: We aimed to use dental implants to rehabilitate the area of elements #11 and #21 in a satisfactory surgical and prosthetic manner, using guided surgery, connective tissue, nano-biomaterials, and a porcelain prosthesis. CASE REPORT: A 32-year-old male patient presented with bone loss of elements #11 and #21, which was proven radiographically and clinically. Thus, oral rehabilitation with the use of dental implants was required. It was decided to proceed via digital planning with the DSD program (Digital smile design) and with the software Exoplan, (Smart Dent-Germany) whenever it was possible to plan immediate provisional and accurate dental implant positioning through reverse diagnostics (Software Exoplan, Smart Dent-German). The dental elements were extracted atraumatically; then, a guide was established, the implants were positioned, the prosthetic components were placed, the conjunctive tissue was removed from the palate and redirected to the vestibular wall of the implants, the nano-graft (Blue Bone®) was conditioned in the gaps between the vestibular wall and the implants, and, finally, the cemented provision was installed. RESULTS: After a 5-month accompaniment, an excellent remodeling of the tissues had been achieved by the implants; consequently, the final prosthetic stage could begin, which also achieved a remarkable aesthetic result. CONCLUSIONS: This report demonstrates that the correct planning of dental implants, which is associated with appropriate soft tissue and bone manipulation, allows for the achievement of admirable clinical results.

Cytokines and inflammatory mediators: Markers involved in interstitial damage to the pancreas in two dengue fever cases associated with acute pancreatitis.

Dengue viral (DENV) infections can lead to acute pancreatitis and associated tissue damage. This study examined the pancreas from two fatal cases of DENV for histopathological changes as well as for the detection of cytokines, and other inflammatory mediators. Tissue sections were prepared for examination by ultrastructural and histopathological techniques. Sections from the pancreas of non-infected individuals were prepared in parallel as a control. The presence of viral replication in macrophages was detected by co-staining for the proteins NS3 and CD68 by immunofluorescence. Immunohistochemistry was used to detect cells that expressed cytokines and inflammatory mediators to characterize the inflammatory response. Edema, acinar necrosis and fibrosis areas associated with a mononuclear infiltrate were found in infected tissues. The major site of virus replication appeared to be macrophages based on their exclusive presentation of the viral protein NS3. Pancreatic tissues from the infected individuals also displayed increased levels of high mobility group box-1, caspase-3, gelatinase B and tumor necrosis factor alpha compared to controls. The presence of virus replicating macrophages in the pancreas was associated with multiple changes in tissue structure that included elevated levels of cytokines and inflammatory markers that may differentiate acute pancreatitis due to DENV infections from other causes.

Ultrastructural and Physicochemical Characterization of a Non-Crosslinked Type 1 Bovine Derived Collagen Membrane.

In this work, in vitro testing was used to study the properties of non-crosslinked type 1 bovine derived collagen membranes used in bone regeneration surgery. Collagen membranes were prepared, their surface roughness was quantified by interferometry, their morphology was observed by scanning electron microscopy (SEM) and transmission electron microscopy (TEM), their wettability was measured by the contact angle technique, their mechanical properties were investigated by tensile testing, their phase transformation temperatures were measured by Differential Scanning Calorimetry (DSC), and their biocompatibility was evaluated by immunological testing. The calorimetry tests showed that the membrane is formed only by type 1 collagen. The SEM observations showed that the morphology consists of layers of highly organized collagen fibers and patterns of striated fibrils typical of type 1 collagen. The small contact angle showed that the membrane is hydrophilic, with the possibility of rapid absorption of body fluids. The tensile tests showed that the membrane has enough elasticity, ductility, and mechanical strength for use in tissue regeneration. With the immunostaining technique, it was possible to confirm the membrane biocompatibility.

Repair of Critical Size Bone Defects Using Synthetic Hydroxyapatite or Xenograft with or without the Bone Marrow Mononuclear Fraction: A Histomorphometric and Immunohistochemical Study in Rat Calvaria.

Bone defects are a challenging clinical situation, and the development of hydroxyapatite-based biomaterials is a prolific research field that, in addition, can be joined by stem cells and growth factors in order to deal with the problem. This study compares the use of synthetic hydroxyapatite and xenograft, used pure or enriched with bone marrow mononuclear fraction for the regeneration of critical size bone defects in rat calvaria through histomorphometric (Masson's staining) and immunohistochemical (anti-VEGF, anti-osteopontin) analysis. Forty young adult male rats were divided into five groups (n = 8). Animals were submitted to critical size bone defects (Ø = 8 mm) in the temporoparietal region. In the control group, there was no biomaterial placement in the critical bone defects; in group 1, it was filled with synthetic hydroxyapatite; in group 2, it was filled with xenograft; in group 3, it was filled with synthetic hydroxyapatite, enriched with bone marrow mononuclear fraction (BMMF), and in group 4 it was filled with xenograft, enriched with BMMF. After eight weeks, all groups were euthanized, and histological section images were captured and analyzed. Data analysis showed that in groups 1, 2, 3 and 4 (received biomaterials and biomaterials plus BMMF), a significant enhancement in new bone matrix formation was observed in relation to the control group. However, BMMF-enriched groups did not differ from hydroxyapatite-based biomaterials-only groups. Therefore, in this experimental model, BMMF did not enhance hydroxyapatite-based biomaterials' potential to induce bone matrix and related mediators.

Resveratrol promotes liver regeneration in drug-induced liver disease in mice.

Studies suggest that the bioactive polyphenolic compound resveratrol (RESV, trans-isomer), found naturally in certain foods such as red grapes and peanuts, may be able to ameliorate liver damage. However, the effects and efficacy of long-term treatment with RESV remain unclear. Here, we used an acetaminophen (APAP; 400 mg/kg/d for 15 days) overdose model to induce liver damage in C56BL/6 mice. Three days after the intoxication was stopped, we observed biochemical, histological and ultrastructural alterations in the livers of these mice. The APAP-treated animals were then given RESV (10 mg/kg/d) for 60 days. Blood and tissue were analyzed at days 7, 30 and 60. Our data show that long-term RESV treatment (60 days) ameliorates the liver injury caused by APAP intoxication, restoring histological features, ultrastructural organization and serum biochemical parameters (albumin, alanine aminotransferase). Ck18- and F4/80-positive cells (indicators of hepatocyte recovery) were reestablished and the number of α-SMA positive cells was normalized after long-term RESV treatment. Additionally, downregulation of the drug transporter BCRP was observed. Electron microscopy revealed that treatment with RESV was effective in restoring the shape and size of hepatic microvilli and normalizing both the number and viability of mitochondria. Taken together, these results indicate that long-term treatment with RESV is effective in alleviating liver injury caused by APAP administration.

Pancreatic steatosis in adult rats induced by nicotine exposure during breastfeeding.

PURPOSE: Maternal nicotine exposure negatively impacts offspring's health and metabolism, leading to obesity and insulin resistance. Here we investigated the pancreatic islet function, glycemic homeostasis, and insulin signaling in adult rat offspring that were nicotine-exposed during breastfeeding. METHODS: For this, lactating Wistar rat dams were divided into two groups: Nicotine (implanted with osmotic minipumps containing 6 mg/Kg, NIC) and Control (saline, CON). Solutions were released from postnatal (PN) day 2-16. At PN110 and PN170, 10 offspring per litter/sex/group were submitted to the oral glucose tolerance test (OGTT). PN180 offspring were killed and glycemia, insulinemia, adiponectinemia, pancreas morphology as well as pancreatic islet protein expression (related to insulin secretion) and skeletal muscle (related to insulin action) were evaluated. Males and females were compared to their respective controls. RESULTS: Adult NIC offspring of both sexes showed glucose intolerance in the OGTT. Despite normoglycemia, NIC males showed hyperinsulinemia while females, although normoinsulinemic, had hyperglycemia. Both sexes showed increased IRI, reduced adiponectin/visceral fat mass ratio and higher ectopic deposition of lipids in the pancreatic tissue adipocytes. In pancreatic islets, NIC males showed lower PDX-1 expression while females had higher PDX-1 and GLUT2 expressions plus lower α2 adrenergic receptor. In the muscle, NIC offspring of both sexes showed reduction of GLUT4 expression; NIC males also had lower insulin receptor and pAKT expressions. CONCLUSIONS: Thus, glycemic homeostasis and peripheral insulin signaling in adult offspring of both sexes are affected by nicotine exposure through the milk, increasing the risk for type 2 diabetes development.

Histomorphometric, Immunohistochemical, Ultrastructural Characterization of a Nano-Hydroxyapatite/Beta-Tricalcium Phosphate Composite and a Bone Xenograft in Sub-Critical Size Bone Defect in Rat Calvaria.

Nowadays, we can observe a worldwide trend towards the development of synthetic biomaterials. Several studies have been conducted to better understand the cellular mechanisms involved in the processes of inflammation and bone healing related to living tissues. The aim of this study was to evaluate tissue behaviors of two different types of biomaterials: synthetic nano-hydroxyapatite/beta-tricalcium phosphate composite and bone xenograft in sub-critical bone defects in rat calvaria. Twenty-four rats underwent experimental surgery in which two 3 mm defects in each cavity were tested. Rats were divided into two groups: Group 1 used xenogen hydroxyapatite (Bio Oss™); Group 2 used synthetic nano-hydroxyapatite/beta-tricalcium phosphate (Blue Bone™). Sixty days after surgery, calvaria bone defects were filled with biomaterial, animals were euthanized, and tissues were stained with Masson's trichrome and periodic acid-Schiff (PAS) techniques, immune-labeled with anti-TNF-α and anti-MMP-9, and electron microscopy analyses were also performed. Histomorphometric analysis indicated a greater presence of protein matrix in Group 2, in addition to higher levels of TNF-α and MMP-9. Ultrastructural analysis showed that biomaterial fibroblasts were associated with the tissue regeneration stage. Paired statistical data indicated that Blue Bone™ can improve bone formation/remodeling when compared to biomaterials of xenogenous origin.

Açaí (Euterpe oleracea Mart.) seed extract improves aerobic exercise performance in rats.

The purpose of this study was to examine whether the supplementation with an açai (Euterpe oleracea Mart.) seed extract (ASE) would affect the aerobic exercise performance in rats and correlate with the vascular function, muscle oxidative stress and mitochondrial biogenesis. Male Wistar rats were divided into five groups: Sedentary, Sedentary with chronic supplementation of ASE, Training, Training with chronic (200 mg/Kg/day intragastric gavage for 5 weeks) or acute (30 min before the maximal treadmill stress test (MST) supplementation with ASE. The exercise training was performed on a treadmill (30 min/day; 5 days/week) for 4 weeks. The chronic supplementation with ASE increased the exercise time (58%) and the running distance (129%) in relation to the MST, while the Training group increased 40% and 78% and the Training with acute ASE group increased 30% and 63%, respectively. The training-induced increase of ACh vasodilation was not changed by ASE, but the norepinephrine-induced vasoconstriction was reduced by chronic and acute supplementation with ASE. The increased levels of malondialdehyde in soleus muscle homogenates from the Training group was reduced only by chronic supplementation with ASE. The muscle antioxidant defense, NO2 levels, and expression of the mitochondrial biogenesis-related proteins (PGC1α, SIRT-1, p-AMPK/AMPK, Nrf-2) were not different between Training and Sedentary groups, but all these parameters were increased in the Training with Chronic ASE compared with the Sedentary groups. In conclusion, chronic supplementation with ASE improves aerobic physical performance by increasing the vascular function, reducing the oxidative stress, and up-regulating the mitochondrial biogenesis key proteins.

Commentary on mitochondrial stereology in transmission electron microscopy / Comentario sobre estereología mitocondrial en microscopía electrónica de transmisión

Mitochondria (m) are responsible for the energy availability of cells, and their analysis is indicated for example, in studies related to metabolism and oxidative stress. The direct measurement of mitochondria (morphometry) is biased because of the section obliquity and position relative to the mitochondria length (non-equatorial cut). Therefore, stereology is an appropriate technique to evaluate mitochondria. However, before beginning the study, it is necessary to consider the premises to obtain random and uniform samples to be analyzed stereology. Mitochondria must have the chance to appear in all the possibilities of cut and orientation in the micrographs. The number of micrographs to be analyzed will depend on the distribution and occupation of mitochondria in the cell. After this is resolved, a proposal is the estimation of the following stereological data: volume density (Vv), surface density (Sv), and mean cross-sectional area (A). Overlapping a known test area at each micrograph, the density by area of mitochondria is estimated (NAT). Vv [m] can easily be estimated by point-counting (Vv = Pp/PT; Pp are the points hitting the structure, PT are the number of points of the test system). Sv is estimated overlaying a test-line (LT) on the micrographs and counting the intersections of the lines (I) with the outer membrane (om), inner membrane (im), and crests (c), thus, Sv [om], Sv [im], Sv [c] (Sv = 2I / LT). A [m] is obtained as the ratio: A = Vv / 2NAT.

Las mitocondrias (m) son responsables de la disponibilidad de energía de las células, y su análisis está indicado, por ejemplo, en estudios relacionados con el metabolismo y el estrés oxidativo. La medición directa de las mitocondrias (morfometría) está sesgada debido a la oblicuidad de la sección y la posición relativa a la longitud de las mitocondrias (corte no ecuatorial). Por lo tanto, la estereología es una técnica apropiada para evaluar las mitocondrias. Sin embargo, antes de comenzar el estudio, es necesario considerar las premisas para obtener muestras aleatorias y uniformes para analizar estereológicamente. Es esencial que las mitocondrias tengan la posibilidad de aparecer en todas las posibilidades de corte y orientación en las micrografías. El número de micrografías que se analizarán dependerá de la distribución y ocupación de las mitocondrias en la célula. Una vez resuelto esto, una propuesta es la estimación de los siguientes datos estereológicos: densidad de volumen (Vv), densidad de superficie (Sv) y área de sección transversal media (A). Superponiendo un área de prueba conocida en cada micrografía, se estima la densidad por área de mitocondrias (NAT). Vv [m] se puede estimar fácilmente contando puntos (Vv = Pp / PT; Pp son los puntos que llegan a la estructura, PT son el número de puntos del sistema de prueba). Sv se estima superponiendo una línea de prueba (LT) en las micrografías y contando las intersecciones de las líneas (I) con la membrana externa (om), la membrana interna (im) y las crestas (c), por lo tanto, Sv [om], Sv [im], Sv [c] (Sv = 2I / LT). A [m] se obtiene como la relación: A = Vv / 2NAT.

Nanosized hydroxyapatite and ß-tricalcium phosphate composite: Physico-chemical, cytotoxicity, morphological properties and in vivo trial.

The objective of this work was to characterize the properties of a synthetic biomaterial composite with nanoparticles size (Blue Bone). This biomaterial is a composite recommended for dental and orthopedic grafting surgery, for guided bone regeneration, including maxillary sinus lift, fresh alveolus filling, and treatment of furcation lesions. The nano biomaterials surface area is from 30% to 50% higher than those with micro dimensions. Another advantage is that the alloplastic biomaterial has homogeneous properties due to the complete manufacturing control. The analyzed biomaterial composite was characterized by XRD, cytochemistry, scanning electron microscopy, porosimetry and in vivo experiments (animals). The results showed that the analyzed biomaterial composite has 78.76% hydroxyapatite [Ca5(PO4)3(OH)] with monoclinic structure, 21.03% ß-tricalcium phosphate [ß -Ca3(PO4)2] with trigonal structure and 0.19% of CaO with cubic structure, nanoparticles with homogeneous shapes, and nanoporosity. The in vivo experiments showed that the composite has null cytotoxicity, and the site of insertion biomaterials has a high level of vascularization and bone formation. The conclusion is that the synthetic biomaterial with Blue Bone designation presents characteristics suitable for use in grafting surgery applications.

Research proposal: inflammation and oxidative stress in coronary artery bypass surgery graft: comparison between diabetic and non-diabetic patients.

BACKGROUND: Diabetes mellitus patients (DM) have more severe progression of atherosclerotic disease than non-diabetic (NDM) individuals. In situ inflammation and oxidative stress are key points in the pathophysiology of atherosclerosis, a concept largely based on animal model research. There are few studies comparing inflammation and oxidative stress parameters in medium-sized arteries between DM and NDM patients. A fragment of the internal mammary artery used in coronary artery bypass grafting (CABG) will be employed for this purpose OBJECTIVE: To assess the expression of inflammatory markers tumor necrosis factor-α, transforming growth factor-ß1, nuclear factor kappa B, the enzymes superoxide dismutase, and catalase in the vascular wall of the arterial graft used in CABG, comparing DM and NDM patients RESULTS: The present study will add information to the vascular degenerative processes occurring in diabetic patients.

Radiotherapy-Induced Skin Reactions Induce Fibrosis Mediated by TGF-ß1 Cytokine.

PURPOSE: This study aimed to investigate radiation-induced lesions on the skin in an experimental animal model. Methods and Materials: Cutaneous wounds were induced in Wistar rats by 4 MeV energy electron beam irradiation, using a dose rate of 240 cGy/min, for 3 different doses (10 Gy, 40 Gy, and 60 Gy). The skin was observed 5, 10, and 25 days (D) after ionizing radiation exposition. RESULTS: Infiltrate inflammatory process was observed in D5 and D10, for the 40 Gy and 60 Gy groups, and a progressive increase of transforming growth factor ß1 is associated with this process. It could also be noted a mischaracterization of collagen fibers at the high-dose groups. CONCLUSION: It was observed that the lesions caused by ionizing radiation in rats were very similar to radiodermatitis in patients under radiotherapy treatment. ADVANCES IN KNOWLEDGE: This study is important to develop strategies to prevent radiation-induced skin reactions.

Fat tissue morphology of long-term sex steroid deficiency and estrogen treatment in female rats.

After long-term estradiol deficiency, female rats displayed body mass gain accompanied by an increase in the size of adipocytes, an increase in hyperglycemia, and a decrease in insulinemia. The effects were reversed by daily estradiol treatment. Adiposity was suggested by the increased vascular endothelial growth factor expression in castrated rats, whereas the proliferative effect of estradiol was suggested by the increased fibronectin expression in treated rats.

Expressions of vascular endothelial growth factor and nitric oxide synthase III in the thyroid gland of ovariectomized rats are upregulated by estrogen and selective estrogen receptor modulators.

BACKGROUND: Estrogen promotes the growth of thyroid cells. Therefore, we analyzed the influence of estrogen and selective estrogen receptor modulators (SERMs) on the expression of vascular endothelial growth factor (VEGF) and nitric oxide synthase III (NOS III) in the thyroid gland of ovariectomized (Ovx) rats. METHODS: Wistar rats were divided into five groups, and bilateral ovariectomies were performed, except on the Sham-operated controls (Sham). Rats were grouped as follows: Sham; Ovx; and Ovx rats treated with daily subcutaneous injections of estradiol benzoate 3.5 microg/kg, tamoxifen 2.5 mg/kg, or raloxifene 2.5 mg/kg for 50 consecutive days. Control animals received vehicle (propyleneglycol), and at the end of the treatment, rats were sacrificed. The thyroid glands were excised, weighed, and processed for analysis of the expression of VEGF or NOS III by immunohistochemistry. The mean vascular areas were evaluated by immunodetection of alpha-smooth muscle actin. RESULTS: Thyroid weight and mean vascular area were lower in Ovx as compared with Sham, Ovx + estradiol benzoate, Ovx + Tam, or Ovx + Ral (p < 0.01). VEGF (p < 0.01) and NOS III expressions (p < 0.05) were significantly lower in the Ovx group, as compared with Sham, Ovx + estradiol benzoate, Ovx + Tam, and Ovx + Ral. Immunoreactivity for both VEGF and NOS III was mainly detected in the cytoplasm of the follicular epithelial cells. CONCLUSIONS: Our data suggest that estrogen and SERMs regulate the thyroid gland vascularization and that tamoxifen and raloxifene behave like estrogen does. Estrogen and SERMs upregulate VEGF and NOS III in such a way as to reverse the effects detected on the thyroid microvasculature of the Ovx rats.

Giardia lamblia: a report of drug effects under cell differentiation.

The Giardia lamblia life cycle is characterized by two phases during which two major cell differentiation processes take place: encystation and excystation. During encystation, the trophozoites transform into cysts, the resistance form. Once ingested by a susceptible host, the cysts are stimulated to excyst in the stomach, and the excysted trophozoites adhere to the epithelium of the upper small intestine. Our work analyses the effects of four benzimidazole derivatives during Giardia differentiation into cysts and evaluates the excystation efficiency of water resistant cysts. Albendazole (AB) showed the most significant results by inhibiting encystation about 30% and a decreasing rate of excystation efficiency. The ultrastructural organization of the cyst adhesive disk was notably affected by AB treatment. Although other benzimidazoles showed some effect on encystation, they were not able to inhibit the excystation process. It is known that the benzimidazoles affect the cytoskeleton of many organisms but how it interferes in Giardia differentiation processes is our main focus. The importance of studying Giardia's differentiation under drug action is reinforced by the following arguments: (1) Cysts eliminated by hosts undergoing treatment could still be potentially infective; (2) once the host has been treated, it would be desirable that the shedding of cysts into the environment is avoided; (3) the prevention of Giardia dissemination is a question of extreme importance mainly in underdeveloped countries, where poor sanitary conditions are related to high rates of giardiasis. This report concerns the importance of keeping the environment free from infective cysts and on Giardia's drug resistance and differentiating abilities.

Raloxifene effects on thyroid gland morphology in ovariectomized rats.

We aimed to analyze the effects of raloxifene and estrogen on thyroid gland morphology of ovariectomized rats. Raloxifene treatment led to effects similar to those of estrogen on thyroid glands from ovariectomized rats, so that both were able to normalize the changes detected after ovariectomy.

DNA typing from vaginal smear slides in suspected rape cases.

In an investigation of suspected rape, proof of sexual assault with penetration is required. In view of this, detailed descriptions of the genitalia, the thighs and pubic region are made within the forensic medical service. In addition, vaginal swabs are taken from the rape victim and some of the biological material collected is then transferred to glass slides. In this report, we describe two rape cases solved using DNA typing from cells recovered from vaginal smear slides. In 1999, two young women informed the Rio de Janeiro Police Department that they had been victims of sexual assaults. A suspect was arrested and the victims identified him as the offender. The suspect maintained that he was innocent. In order to elucidate these crimes, vaginal smear slides were sent to the DNA Diagnostic Laboratory for DNA analysis three months after the crimes, as unique forensic evidence. To get enough epithelial and sperm cells to perform DNA analysis, we used protocols modified from the previously standard protocols used for DNA extraction from biological material fixed on glass slides. The quantity of cells was sufficient to perform human DNA typing using nine short tandem repeat (STR) loci. It was 3.3 billion times more probable that it was the examined suspect who had left sperm cells in the victims, rather than any other individual in the population of Rio de Janeiro.