Delayed tooth replantation following root canal filling with calcium hydroxide and MTA: Histomorphometric study in rats

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

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.

Avaliação da biocompatibilidade de dois adesivos dentinários pelo estudo das reações teciduais em conjuntivo de ratos

The biocompatibility of two dentin adhesives (Scotchbond MultiPurpose- 3M and Optibond Multi-Use - Kerr) was evaluated in rats subcutaneous connective tissue implants. Polyethylene tubes filled with the adhesives were surgically implanted into the dorsal subcutaneous tissues of rats. The tubes were left implanted for periods of 14, 30, 60 and 84 days, following wich the animais were killed and the implants, together with the surrounding tissue, were excised. The specimens were then fixed in 10 per cent formalin, sectioned serially and stained with hematoxylin and eosin and then, examined by light microscopy. After 14 and 30 days the tubes were surrounded by a mild to moderate inflammatory infiltration. After 60 and 84 days microscopic examination revealed the formation of a connective tissue capsule around each tube. The microscopic examination of the biopsy specimens allowed to conclude that: a) both dentin adhesives are considered biologically acceptable when placed in contact with rats subcutaneous connective tissue; b) in the observation period of 30 days there were moderate tissue reaction to the Optibond Multi-Use -light cure- (Kerr); c) after 60 and 84 days the use of both dentine adhesives did not interfere in the healing of the surrounding connective tissue

Avaliação do selante de fibrina como arcabouço para células tronco em fígados cirróticos

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Experimental glaucoma model (ischemia and reperfusion): histology, morphometry, protein and gene espression of apoptosis pathway

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

Histologia e histomorfometria do retículo de bovino no período pré-natal

The development of the stomach of ruminants requires further studies because it is a complex process. Therefore, was analyzed the histology and histomorphometry of reticulum at each gestation period. Samples of Nelore fetuses (Bos taurus indicus) was divided in 5 groups: 1 - fetuses at 9 to 15 weeks (8 to 21cm) of gestation, 2 - fetuses at 16 to 22 weeks (23 to 37cm) 3 - fetuses at 23 to 29 weeks (40 to 58cm), 4 - fetuses at 30 to 36 weeks (61 to 77cm) and 5 - fetuses at 37-43 weeks (79 to 88cm). The histological sections were stained with hematoxylin and eosin and Mallory's trichrome and examined by light microscopy. In group 1, the fetuses with 11 cm, showed mucosal projections that eventually become primary crests, lamina propria and submucosa are fused and external and internal muscular and similar. At 16.5 inches, the primary crests had lamina propria and at 18.5 cm, appeared connections between the ridges. In group 2, at 31 cm started epithelial involvement around the crests. In group 3, the fetuses of 42 cm revealed the first secondary papillae and muscular mucosa in the upper portion of the primary crests. Thereafter, it was increased epithelial due to its positioning around the crests, thickening of the lamina propriasubmucosa, muscular layer, especially the internal muscular and serosa. It was concluded that the most visible changes occur in fetuses belonging to the first 3 groups, measurements for the epithelium, muscle layer and total wall were increasing and the other analyzed layers showed variations during fetal development.

Desenvolvimento do rúmen de bovinos no período pré-natal

The development of the domestic mammals’ stomach is a complex process, especially in ruminants. The objective of this research was to perform the analysis of structures of this organ during prenatal period. Samples of Nelore fetuses rumen (Bos Taurus indicus) were divided into five groups: 1 - fetuses with 9 to 15 weeks (8 to 21cm) of gestation, 2 - fetuses with 16 to 22 weeks (23 to 37cm), 3 - fetuses at 23 to 29 weeks (40 to 58cm) 4 - fetuses with 30 to 36 weeks (61 to 77cm) and 5 - fetuses with 37 to 43 weeks (79 to 88cm). Sections were stained with Hematoxylin and eosin and Mallory's trichrome and examined at light microscope. In group 1, in fetuses with 11 cm, could be seen all the layers, mucosa with high epithelium and light cells, lamina propria fused with submucosa, the muscular layer was with two thin sublayers similar in thickness and serosa was thin too. With 13.5 cm, was observed the first irregularities in the mucosa with epithelium projections and lamina propria that will lead rumen papillae. With 16.5 cm there was differentiation of the lamina propria to the submucosa. In group 2, in fetuses with 28 cm were observed complete papillae. From this stage, the remaining groups, there was an increase in thickness of the epithelium, lamina propria, submucosal and muscular layer whit the inner sublayer more thicker than the external and serosa. It was concluded that the main variations occur in fetuses of groups 1 and 2, and after this phase, there is increased in thickness of all layers until the end of the pregnancy period.

Morphological, immunohistochemical and molecular study of renal lesions in canine visceral leishmaniasis (CVL)

Canine visceral leishmaniasis (CVL) is an anthropozoonosis characterized by a clinically chronic progressive disease. Non lymphoid organs are also affected, especially the kidneys. Dogs with leishmaniasis usually die with renal failure despite treatment. Haematoxylin-eosin (HE) staining in kidney tissue sections has low sensitivity for parasite identification. Immunohistochemistry (IHC) and polymerase chain reaction (PCR) are efficient methods for Leishmania sp. antigen and DNA detection in cases of low parasite burden. The present study aims to identify renal lesions of CVL and correlate them with microscopic findings determined by histochemistry, IHC and PCR. Both IHC and PCR provided similar positivity for amastigote identification, 3/20 animals (15%), thus increasing detection of the parasite in renal tissues when compared with histopathologic examination. The lesion most commonly observed with visceral leishmaniasis-positive canine kidney tissue was membranoproliferative glomerulonephritis, followed by interstitial nephritis without correlation to the number of amastigotes.

Caracterização histomorfológica de lesões granulomatosas sugestivas de tuberculose ganglionar e pulmonar em bovinos abatidos na região noroeste do estado de SãoPaulo

Tuberculosis infection among humans transmitted by products of animal origin can be caused by Mycobacterium bovis, a concern in developing countries, because the number of human cases is relatively high compared with countries that have implemented programs to eradicate bovine tuberculosis for many years. Because it is a chronic disease with subclinical evolution, it is transmitted to other animals in the herd livestock resulting in high loss losses resulting high (10 to 25%), as well as the zoonotic nature of the disease on public health. This work aimed to study the histomorphology of granulomas in samples of lymph nodes and lungs of animals with suggestive lesions of tuberculosis, as well as those similar to the sanitary inspection. The animals clinically healthy ante-mortem, were slaughtered in a slaughterhouse in the northwestern state of Sao Paulo between April 2008 and April 2009, considering that carcasses had lymph nodes with lesions typical of tuberculosis, exhibiting nodular and hemorrhagic appearance. The caseous or calcified lesions, purulent or not, of various sizes and shapes, were also evaluated. Of the 307,661 animals slaughtered in the mentioned period, 494 gross suggestive lesions of tuberculosis and from 29 cities in the state of Sao Paulo, 16 from Mato Grosso do Sul, Goias 15, one from Mato Grosso and Paraná. Among these, 200 samples (40,5%) were processed for histopathological evaluation by hematoxylin-eosin (HE) and 90 (45%) were subjected to Ziehl-Neelsen (ZN) and examined by light microscopy. In HE staining, histopathological changes of tuberculosis were found in 197 (98,5%), classified according to the stage of development adopted by Wango et al.(31), where 29 samples were included in stage I (14,5%), 53 in stage II (26,5%), 87 in stage III (43,5%), 28 in stage IV (14%). In the ZN staining Mycobacterium sp was found in 89 (99%) of the specimens. The rate of tuberculosis in the herd studied was 0,16%. We conclude that the identification of the disease in slaughterhouses and meat companies from significant gross lesions, enables the implementation of effective measures, specifically the origin of the disease in cattle herds, providing control measures to prevent its spread.

Cytopathology in the diagnosis of oral trichomoniasis in a rock pigeon (columba livia)

A Rock Pigeon (Columba livia) submitted for necropsy had an oral white-yellow mass with a friable surface and putrid odor. Impression smears were performed and tissue samples were collected for histopathology. Cytopathological analysis revealed numerous pyriform protozoa, compatible with Trichomonas gallinae. Protozoans were not evident within the lesions by histopathology after staining the samples with Hematoxylin and Eosin or Gomori methenamine silver (GMS) stain. We conclude that impression smears from avian oral trichomoniasis suspects, even during post-mortem evaluation, can be a useful technique for the diagnosis of this disease.

Reconstruction of the anterior wall of the frontal sinus using bovine bone matrix: histological study with monkeys

The purpose of this study was to evaluate the repair process in the reconstruction of the anterior wall of the frontal sinus of monkeys with bovine bone matrix. Four adult Cebus apella monkeys underwent an ostectomy of the anterior wall of the frontal sinus. The frontal sinus mucosa and the nasofrontal duct were not manipulated. Reconstruction occurred with implants of bovine bone matrix laminae measuring 2.0 x 2.5 cm and 0.4 mm thick, stabilized under pressure in the lateral wall of the frontal sinus. The monkeys were sacrificed over a period of 150 days and routine laboratory procedures were followed for hematoxylin-eosin staining and histologic evaluation of the specimens. Neoformed bone tissue was observed in contact with the frontal sinus mucosa and the bovine bone matrix. The frontal sinus mucosa remained whole without fibrous tissue or cystic formations. There was no occurrence of cellularization as well as revascularization of the bovine bone matrix, though it has permitted bone conduction on this surface. It was possible to conclude that the demineralized bovine bone matrix was biotolerable, being incorporated into the bone without the presence of inflammatory cells with characteristics of inertness and antigenicity and behaved as an osteoconductive material.

Efeito do ácido tranexâmico associado à cola de fibrina sobre o reparo ósseo: estudo histológico em ratos

The bone repair process is controlled by complex molecular mechanisms that involve systemic and local factors. Fibrin glue is derived from human plasma and mimics the final pathway of coagulation network. Tranexamic acid inhibits fibrinolysis and prevents or decreases the formation of degradation products of fibrin and fibrinogen. The purpose of this study was to evaluate histologically in rats the effect of tranexamic acid associated with the fibrin glue on bone healing. The experiment used 60 (n = 5) male rats in: GI: Control, GII: fibrin glue, GIII tranexamic acid and GIV /fibrin glue/tranexamic acid. Bone defect (2.5mm diameter) was created in right tibia. The animals were euthanized at 7,14 and 30 days postoperatively, and the pieces were processed with hematoxylin and eosin. The results showed at 7 days post-operative surgical cavity filled with dense connective tissue rich in fibroblasts, permeated by delicate neoformed bone trabeculae in percentage of 70-80% for GI, GII and GIII and GIV to 94.8%. At 14 days post-operative newly formed bone was found between 75-85% for GI, GII and GIII and percentage above 95% for GIV. At 30 days postoperative GI and GIV showed 95-100% of mature bone tissue; GII and GIII in percentage close to 80-90%. Based on the results and methodology is concluded that fibrin / tranexamic acid glue association has positive action on bone repair.

Development of bovine abomasum in prenatal period

For this purpose, samples of Nelore fetus abomasum (Bos taurus indicus) were classified into five groups: 1 – fetuses with 9 to 15 weeks (8 to 21 cm) of gestation, 2 – fetuses with 16 to 22 weeks (23 to 37 cm), 3 – fetuses at 23 to 29 weeks (40 to 58 cm) 4 – fetuses with 30 to 36 weeks (61 to 77 cm) and 5 – fetuses with 37 to 43 weeks (79 to 88 cm). Histologic sections were stained with Hematoxylin and eosin, picrosirius and Mallory’s trichrome methods and examined under light microscopy. In fetus with 11 cm of length showed deep mucosa, wide folds and villi lined by single cylindrical epithelium, lamina propria, submucosa, muscular sublayers and serosa. Fetus with 13.5 cm, villi were evident and muscular layer of the mucosa was formed. At 16.5 cm, could be seen an increase in mucosa glands size and number. In fetus of 26 cm, showed gastric mucous glands at great number and ramifications. At 29 cm large folds, were observed. At 37 cm, showed glandular epithelium, muscular layer of the mucosa and muscular layer well development. Fetus with 42 cm, showed deep glands with corresponding mucous cells surrounding by thin lamina propria. It was concluded that the histomorphometric values of muscular layer and total abomasum wall were increased for all groups except for groups 4 and 5 which there was a decline of values, without presenting a significant difference and there was no continuous pattern of growth for other components of abomasum wall.

Histomorfometria do útero de gatas submetidas à ovariosalpingohisterectomia

Histomorphometric data were obtained from cats uterine horns: either nulliparous (n = 6), primiparous (n = 6), multiparous (n = 6) and treated with contraceptive (n = 6). The material was collected after surgery, fixed in paraformaldehyde and embedded in paraplast® resin to be sliced in a microtome. The obtained sections were stained with hematoxylin - eosin and measured under a light microscope: uterine wall total thickness (μm), endometrium total height, endometrial glands diameter and glandular epithelia height, total myometrium, internal and external myometrium and vascular layer thickness. It was concluded that: 1 - contraceptive use and number of pregnancies altered the uterine structure, 2 - one pregnancy does not appear to affect the uterine lining structures as occurs in multiparous cats, 3 – there was no variation on the evaluated structures between nulliparous and primiparous cats except for inner myometrium, 4 - the total myometrium hight and the endometrium hight showed similar variations except for the contraceptive treated cats group, 5 - the outer myometrium showed marked changes in the treated cats, 6 – the less marked morphological variations were for the endometrial glands diameter and glandular epithelium hight, 7 - the presence of dilated endometrial glands was found only in treated cats.

Cytopathology in the diagnosis of oral trichomoniasis in a Rock Pigeon (Columba livia)

A Rock Pigeon (Columba livia) submitted for necropsy had an oral white-yellow mass with a friable surface and putrid odor. Impression smears were performed and tissue samples were collected for histopathology. Cytopathological analysis revealed numerous pyriform protozoa, compatible with Trichomonas gallinae. Protozoans were not evident within the lesions by histopathology after staining the samples with Hematoxylin and Eosin or Gomori methenamine silver (GMS) stain. We conclude that impression smears from avian oral trichomoniasis suspects, even during post-mortem evaluation, can be a useful technique for the diagnosis of this disease.