Connective tissue graft as a biological barrier for guided tissue regeneration in intrabony defects: a histological study in dogs

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Expressão imuno-histoquímica de KI-67, COX-2, MMP-9 e P53 nos tumores testiculares caninos

Pós-graduação em Ciência Animal - FMVA

Focal invasiveness in complete histological analyses of a large acral lentiginous melanoma

Background: Acral lentiginous melanoma is a melanoma with poor prognosis which is frequently diagnosed at an advanced stage. Since the thickness of tumour is one of the main prognostic factors, this case can exemplify how important complete histological analyses looking for focal invasiveness can be.Case report: A 77 year-old woman with a black spot with slow progressive growth on the left plantar region. She sought medical attention due to the expansion onto the dorsal surface of toes. The lesion had irregular borders and had spread to half the plantar surface. Histopathology confirmed the clinical suspicion of acral lentiginous melanoma Clark level IV and 2.6 mm Breslow thickness. The surgical specimen was entirely processed for histological evaluation, requiring 53 slides. Tumor dermal invasion was detected in only three out of 53 glass slides as the invasiveness was not identified by clinical, dermatoscopy or macroscopy exams.Conclusion: Sectioning through the entire lesion is considered very important to determinate the appropriate stage of the disease and the correct treatment and patient follow-up.

Classification of histological images based on the stationary wavelet transform

Non-Hodgkin lymphomas are of many distinct types, and different classification systems make it difficult to diagnose them correctly. Many of these systems classify lymphomas only based on what they look like under a microscope. In 2008 the World Health Organisation (WHO) introduced the most recent system, which also considers the chromosome features of the lymphoma cells and the presence of certain proteins on their surface. The WHO system is the one that we apply in this work. Herewith we present an automatic method to classify histological images of three types of non-Hodgkin lymphoma. Our method is based on the Stationary Wavelet Transform (SWT), and it consists of three steps: 1) extracting sub-bands from the histological image through SWT, 2) applying Analysis of Variance (ANOVA) to clean noise and select the most relevant information, 3) classifying it by the Support Vector Machine (SVM) algorithm. The kernel types Linear, RBF and Polynomial were evaluated with our method applied to 210 images of lymphoma from the National Institute on Aging. We concluded that the following combination led to the most relevant results: detail sub-band, ANOVA and SVM with Linear and RBF kernels.

Cleaning capacity promoted by motor-driven or manual instrumentation using ProTaper Universal system: Histological analysis

The aim of this study was to assess the cleaning capacity of the Protaper system using motor-driven or manual instrumentation. Materials and Methods: Ten mandibular molars were randomly separated into 2 groups (n = 5) according to the type of instrumentation performed, as follows: Group 1 - instrumentation with rotary nickel-titanium (Ni-Ti) files using ProTaper Universal System (Dentsply/Maillefer); and, Group 2 - instrumentation with Ni-Ti hand files using ProTaper Universal (Dentsply-Maillefer). Afterwards, the teeth were sectioned transversely and submitted to histotechnical processing to obtain histological sections for microscopic evaluation. The images were analyzed by the Corel Photo-Paint X5 program (Corel Corporation) using an integration grid superimposed on the image. Results: Statistical analysis (U-Mann-Whitney - P < 0.05) demonstrated that G1 presented higher cleaning capacity when compared to G2. Conclusions: The rotary technique presented better cleaning results in the apical third of the root canal system when compared to the manual technique.

Cutaneous healing after topical application of nebacetin and gingilone to infected wounds. Clinical and histological study in rats

It was performed a study to examine the cutaneous cicatrization of infected wound of skin after daily topic application of Nebacetin and Gingilone. Following clinical observations the animals were sacrificed after 4, 10 and 17 post-operative days. The pieces were treated, analyzed, in an optical microscope. The topic application of Nebacetin showed to be more efficient in the epithelial reparation comparatively to the Gingilone and control group.

Alpha 1-adrenoceptor subtypes mediating the norepinephrine-induced contractile response in the rat vas deferens

The effects of chlorethylclonidine (CEC), 5-methyl-urapidil (5-MU), ryanodine and prolonged exposition to norepinephrine (NE) on the concentration-response curves (CRC) to this agonist on the bisected rat vas deferens (RVD) were investigated. 2. CEC did not affect the 50% effective concentration (EC50) of NE in either the prostatic (PP) or the epididymal (EP) portions of the RVD. 3. 5-MU did not alter the EC50 of NE in the PP but caused a significant and concentration-dependent rightward shift of the CRC to NE in the EP. 4. Ryanodine caused a shift to the right of the CRC to NE in the PP associated to a decrease in maximal response, but did not affect the CRC to NE in the EP. 5. Incubation of the EP with NE for 6 hr elicited a significant decrease in the maximal response with no changes in the EC50. Similar treatment of the PP was associated with a significant shift to the right of the CRC to NE without modifications in the maximal response. 6. These results suggest that in the RVD, NE interacts with two different alpha 1-adrenoceptors subtypes which are disposed in a selective manner along the RVD: the alpha 1(a) subtype in the EP, and non-alpha 1b-non-alpha 1a adrenoceptor subtype mainly located at the PP.

Effects of castration and of testosterone replacement on alpha(1)-adrenoceptor subtypes in the rat vas deferens

The contractions of the rat vas deferens in response to noradrenaline are mediated through alpha(1A)-adrenoceptors. We observed participation of alpha(1B)-adrenoceptors in these contractions after castration. We now investigated the time course of this plasticity and the effects of testosterone by determining the actions of competitive antagonists on noradrenaline-induced contractions after 7, 14, 21 and 30 days of castration. BMY 7378 (8-[2-[4-(2-methoxyphenyl)-1-piperazinyl]ethyl]-8-azaspiro[4.5]decane-7,9-dione dihydrochloride) antagonised noradrenaline-induced contractions in control and castrated rats with low pA(2) values (approximately = 6.8). In control vas deferens, WB 4101 (2-(2,6-dimethoxyphenoxyethyl)aminomethyl-1,4-benzodioxane hydrochloride) had a slope in the Schild plot no different from 1.0, while slopes lower than 1.0 (approximately 0.6) were observed for vas deferens from castrated rats. Chloroethylclonidine was ineffective in the control vas while it inhibited noradrenaline-induced contractions in vasa from castrated rats and converted the complex antagonism by WB 4101 into simple competitive antagonism. Treatment of castrated rats with testosterone prevented the effects of castration. The results suggest that alpha(1B)-adrenoceptors are detectable in vas deferens from at least the 7th through the 30th day after castration and that testosterone prevents this plasticity.

Histological analysis of effects of 24% EDTA gel for nonsurgical treatment of periodontal tissues

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Diabetes triggers the loss of tooth structure associated to radiographical and histological dental changes and its evolution to progressive pulp and periapical lesions in rats

The aim of this study was to evaluate the putative influence of diabetes without metabolic control in the loss of tooth structure as well as histological changes in dentin and pulp tissue in rats. Diabetes was induced in Wistar rats (n=25) by intravenous administration of alloxan (42mg/kg). Diabetic and non-diabetic control rats were evaluated at 1, 3, 6, 9 and 12 months of follow-up. In order to evaluate the presence and progression of dental caries and periapical lesions, hemimandibles were removed and submitted to radiographical, histological, and morphometrical procedures. Dental caries were detected after radiographical and histological evaluations in diabetic group from the third month of diabetes onset, increasing gradually in frequency and severity in periods. Diabetic rats dental pulps also presented significant reduction in volume density of collagen fibers and fibroblasts at third month, parallel with a trend towards the increase in inflammatory cells volume density. Diabetic rats presented a generalized pulp tissue necrosis after 6 months of diabetes induction. Moreover, periapical lesions were not detected in control group, while these lesions were observed in all rats after 3, 6, 9, and 12 months of diabetes induction. Uncontrolled diabetes seems to trigger the loss of tooth structure, associated to histological dental changes and mediates its evolution to progressive severe pulp and periapical lesions in rats. Therefore, diabetes may be considered a very important risk factor regarding alterations in dental pulp, development of dental caries, and periapical lesions.

Does spinal block through tattooed skin cause histological changes in nervous tissue and meninges?: an experimental model in rabbits

Although there is no documented evidence that tattoo pigments can cause neurological complications, the implications of performing neuraxial anesthesia through tattooed skin are unknown. In this study, we aimed to assess whether spinal puncture performed through tattooed skin of rabbits determines changes over the spinal cord and meninges. In addition, we sought to evaluate the presence of ink fragments entrapped in spinal needles. Thirty-six young male adult rabbits, each weighing between 3400 and 3900 g and having a spine length between 38.5 and 39 cm, were divided by lot into 3 groups as follows: GI, spinal puncture through tattooed skin; GII, spinal puncture through tattooed skin and saline injection; and GIII, spinal puncture through skin free of tattoo and saline injection. After intravenous anesthesia with ketamine and xylazine, the subarachnoid space was punctured at S1-S2 under ultrasound guidance with a 22-gauge 2½ Quincke needle. Animals in GII and GIII received 5 μL/cm of spinal length (0.2 mL) of saline intrathecally. In GI, the needle tip was placed into the yellow ligament, and no solution was injected into the intrathecal space; after tattooed skin puncture, 1 mL of saline was injected through the needle over a histological slide to prepare a smear that was dyed by the Giemsa method to enable tissue identification if present. All animals remained in captivity for 21 days under medical observation and were killed by decapitation. The lumbosacral spinal cord portion was removed for histological analysis using hematoxylin-eosin stain. None of the animals had impaired motor function or decreased nociception during the period of clinical observation. None of the animals from the control group (GIII) showed signs of injuries to meninges. In GII, however, 4 animals presented with signs of meningeal injury. The main histological changes observed were focal areas of perivascular lymphoplasmacyte infiltration in the pia mater and arachnoid. There was no signal of injury in neural tissue in any animal of both groups. Tissue coring containing ink pigments was noted in all GI smears from the spinal needles used to puncture the tattooed skin. On the basis of the present results, intrathecal injection of saline through a needle inserted through tattooed skin is capable of producing histological changes over the meninges of rabbits. Ink fragments were entrapped inside the spinal needles, despite the presence of a stylet.

Pancreatitis subtypes survey in 852 childhood-onset systemic lupus erythematosus patients: a multicenter cohort

Pancreatitis is a rare and a life-threatening SLE manifestation in childhood-onset systemic lupus erythematosus (c-SLE). The objective of this study was to systematically classify pancreatitis in c-SLE according to the International Study Group of Pediatric Pancreatitis (INSPPIRE) and determine the overall prevalence, clinical features, laboratory and first episode outcomes. A multicenter cohort study in 10 Pediatric Rheumatology centers, including 852 cSLE patients. Pancreatitis was diagnosed in 22/852 (2.6%) cSLE patients. It was classified as acute pancreatitis in 20 (91%), acute recurrent pancreatitis in 2 (9%) and none of them had chronic pancreatitis. None of them had gallstones, traumatic pancreatitis or reported alcohol/tobacco use. The comparison of patients with pancreatitis (first episode) and without this complication revealed a shorter disease duration [1(0-10) vs. 4(0-23) years, p < 0.0001] and higher median of SLEDAI-2K [21(0-41) vs. 2(0-45), p < 0.0001]. The frequencies of fever (p < 0.0001), weight loss (p < 0.0001), serositis (p < 0.0001), nephritis (p < 0.0001), arterial hypertension (p < 0.0001), acute renal failure (p < 0.0001), macrophage activation syndrome (p < 0.0001) and death (p = 0.001) were also higher in patients with pancreatitis. The frequencies of intravenous methylprednisolone use (p < 0.0001) and the median of prednisone dose [55(15-60) vs. 11(1-90)mg/day, p < 0.0001] were significantly higher in patients with pancreatitis. Of note, the two patients with acute recurrent pancreatitis had two episodes, with pain-free interval of 1 and 4 years. This was the first study characterizing pancreatitis using the INSPPIRE standardized definitions in patients with cSLE showing that the predominant form is acute pancreatitis seen in association with glucocorticoid treatment and active severe disease.

Homogenous demineralized dentin matrix and platelet-rich plasma for bone tissue engineering in cranioplasty of diabetic rabbits: biochemical, radiographic, and histological analysis

This study evaluated the effects of homogenous demineralized dentin matrix (HDDM) slices and platelet-rich plasma (PRP) in surgical defects created in the parietal bones of alloxan-induced diabetic rabbits, treated with a guided bone regeneration technique. Biochemical, radiographic, and histological analyses were performed. Sixty adult New Zealand rabbits were divided into five groups of 12: normoglycaemic (control, C), diabetic (D), diabetic with a PTFE membrane (DM), diabetic with a PTFE membrane and HDDM slices (DM-HDDM), and diabetic with PTFE membrane and PRP (DM-PRP). The quantity and quality of bone mass was greatest in the DM-HDDM group (respective radiographic and histological analyses: at 15 days, 71.70±16.50 and 50.80±1.52; 30 days, 62.73±16.51 and 54.20±1.23; 60 days, 63.03±11.04 and 59.91±3.32; 90 days, 103.60±24.86 and 78.99±1.34), followed by the DM-PRP group (respective radiographic and histological analyses: at 15 days 23.00±2.74 and 20.66±7.45; 30 days 31.92±6.06 and 25.31±5.59; 60 days 25.29±16.30 and 46.73±2.07; 90 days 38.10±14.04 and 53.38±9.20). PRP greatly enhanced vascularization during the bone repair process. Abnormal calcium metabolism was statistically significant in the DM-PRP group (P<0.001) for all four time intervals studied, especially when compared to the DM-HDDM group. Alkaline phosphatase activity was significantly higher in the DM-HDDM group (P<0.001) in comparison to the C, D, and DM-PRP groups, confirming the findings of intense osteoblastic activity and increased bone mineralization. Thus, HDDM promoted superior bone architectural microstructure in bone defects in diabetic rabbits due to its effective osteoinductive and osteoconductive activity, whereas PRP stimulated angiogenesis and red bone marrow formation.

The Fresh Frozen Bone Allograft (FFBA) to reconstruc the atrophic maxilla: evaluation of the clinical and histological viability at six and nine months in the maxillary sinus

This work aimed to analyze clinically and histologically the allogen bone graft behavior at 6 and 9 months. A leukoderm, female, 55 years old patient sought dental care for oral rehabilitation with dental implants and implant supported fixed prosthesis in the maxilla. Bilateral sinus lifting procedure were performed in a same patient, the analysis were made after 6 and 9 months, respectively. At 6 months, there was lack of a better bone remodeling in the region, associated to the presence of fibrous connective tissue within the collected tissue, showing us that this is not the best stage of tissue maturation to place dental implants. The 9 months period showed an improvement, with a more organized bone tissue surrounding particles of homogenous bone, what possibly had increased implant stability at the time of surgery. There is no doubt about the increase of clinical applications of FFBA, but there are no studies available regarding the standardization of time to load implants inserted in allografts. So papers with long-term monitoring are necessary to eliminate questions that still remain to be answered.