Additional Insights Into the Role of Osteocalcin in Osteoblast Differentiation and in the Early Steps of Developing Alveolar Process of Rat Molars.

This study investigated whether osteocalcin (OCN) is present in osteoblast precursors and its relationship with initial phases of alveolar process formation. Samples of maxillae of 16-, 18-, and 20-day-old rat embryos (E16, E18, and E20, respectively), and 05-, 10-, and 15-day-old postnatal rats (P05, P10, and P15, respectively) were fixed and embedded in paraffin or araldite. Immunohistochemistry for osterix (Osx), alkaline phosphatase (ALP), and OCN detection was performed and the number of immunolabelled cells was computed. Non-decalcified sections were subjected to the von Kossa method combined with immunohistochemistry for Osx or OCN detection. For OCN immunolocalization, samples were fixed in 0.5% glutaraldehyde/2% formaldehyde and embedded in LR White resin. The highest number of ALP- and OCN-immunolabelled cells was observed in dental follicle of E16 specimens, mainly in basal portions of dental alveolus. In corresponding regions, osteoblasts in differentiation adjacent to von Kossa-positive bone matrix exhibited Osx and OCN immunoreactivity. Ultrastructural analysis revealed OCN immunoreactive particles inside osteoblast in differentiation, and in bone matrix associated with collagen fibrils and within matrix vesicles, at early stages of alveolar process formation. Our results indicate that OCN plays a role in osteoblast differentiation and may regulate calcium/phosphate precipitation during early mineralization of the alveolar process.

Use of corticocancellous allogeneic bone blocks impregnated with bone marrow aspirate: a clinical, tomographic, and histomorphometric study.

The aim of this study was to evaluate the efficacy of allogeneic block grafts impregnated with autologous bone marrow in horizontal ridge augmentation therapy. Ten patients with severe ridge volume deficiency in the anterior maxilla were treated with horizontal ridge augmentation. The patients were randomized into two groups: Five patients, using two allogeneic block grafts, were in the control group, and five patients, using two allogeneic block grafts impregnated with autologous bone marrow, were in the test group. Hematologists collected 4 mL of bone marrow from the iliac crest of the patients in the test group immediately prior to the surgeries. The blocks were fixed using titanium screws to obtain rigid fixation and to standardize the reference points for measurement purposes. CT scans were obtained both preoperatively and six months postoperatively to allow evaluation of horizontal bone gain. After a healing period of six months, the sites were reopened and the screws were removed. Before implant placement, bone cores were harvested and prepared for histologic and histomorphometric evaluation. Tomographic and histomorphometric measurements were recorded. The test group demonstrated better tomographic results (P < 0.05) in augmenting alveolar thickness, with a mean value of 4.60 ± 1.43 mm (118.23 ± 56.93%), while the control group had bone gain of 2.15 ± 0.47 mm (49.91 ± 20.24%). Despite the different results in alveolar thickness gained between groups, all sites received dental implants. The histomorphometric analysis also showed better results (P < 0.05) in the amount of vital mineralized bone in the test group as compared to the control group. The findings of this study suggest that an autologous bone marrow aspirate can increase the regenerative potential of corticocancellous allogeneic bone grafts.

Effects of estrogen deficiency followed by streptozotocin-induced diabetes on periodontal tissues of female rats.

Although both estrogen deficiency and diabetes contribute to periodontal tissue deterioration, the combined effects of these conditions on periodontium is unknown. Thus, we analyzed the combined effects of ovariectomy followed by streptozotocin (STZ)-induced diabetes on periodontal tissues of rats. Twenty adult rats were ovariectomized (OVX) or SHAM-operated (SHAM). After 3 weeks, the rats received an intraperitoneal injection of STZ (60 mg/kg/body weight) to induce diabetes or vehicle (blank) solution. The groups were assigned as follows (n = 5): SHAM-vehicle (SHAM), OVX-vehicle (OVX), SHAM + STZ (SHAM-Di), and OVX + STZ (OVX-Di). Seven weeks post-diabetes induction, the rats were euthanized. Blood samples were collected for glucose measurements and maxillae were processed for paraffin embedding. Sections stained with hematoxylin/eosin, Masson's trichrome, and picrosirius-red were used for alveolar bone loss and collagen fiber analysis in the lamina propria. Immunohistochemistry was performed for runt-related transcription factor 2 (Runx2), matrix metalloproteinase 9 (MMP-9), and tryptase detection. Alveolar bone loss and fewer collagen fibers were observed in the OVX-Di group, collagen fibers with irregular organization, and MMP-9 immunoreactivity were more evident in diabetic groups, and MMP-9-positive osteoclasts on alveolar bone surface were noticed in all groups. The OVX-Di group showed lower Runx2 immunoreactivity (osteoblast formation marker), and more tryptase-positive cells (mast cell marker) in the alveolar bone marrow. Our results indicate that estrogen depletion, followed by STZ-induced diabetes, promotes periodontal tissue deterioration that is more evident than both interventions applied alone. Furthermore, our results points to a possible participation of bone-derived mast cells in this process.

Hysteroscopic sterilization with occlusion of sheep uterine tube using n-butyl-2-cyanoacrylate adhesive.

PURPOSE: To evaluate the fertility and analyze the macroscopic, microscopic and morphometric aspects of sheep uterine tube sterilization with a hysteroscopically insert of n-butyl-2-cyanoacrylate adhesive. METHODS: 12 adult sheep, with one previous pregnancy, were distributed as follows: group L (n=3) subjected to laparotomy and Pomeroy uterine tube ligation, group S (n=3) subjected to hysteroscopic application of saline solution in tube isthmus and group AD(n=6), that was subjected to hysteroscopic application of 0.5 ml of n-2-butil-cyanoacrylate in tube isthmus. They were mated with fertile males for ninety days. The non pregnant sheep, at the 90th day, were subjected to laparotomy with uterus and tubes uterine resection. The fragments of uterine tubes were fixated in 10% formalin and processes for histology evaluated, and slices dyes for H.E. Data were evaluated by Wilcoxon and Mann-Whitney and Fisher's exact test. RESULTS: All sheep from groups L and AD did not get pregnant (0%) in contrast with sheep from group S (100%); the adhesive remained integral in the uterine tube lumen. The percentual of adherences (66.6%) and fibrosis responses (100%) was significantly higher in the group L than group AD (0%) (p<0.01). The diameter of the caudal tube in group AD (2652.15 +/- 45.76 mm) was significantly wider than that of the group L (1868.27 +/- 56.11* microm) (p < 0.05). CONCLUSION: The hysteroscopic insertion of cyanoacrylate in the uterine tube lumen of sheep was effective to obstruct the uterine tube and to promote the sterilization.

The role of bovine preserved peritoneum in rats ventral hernia: a histological evaluation.

PURPOSE: To assess the role of preserved bovine parietal peritoneum as a material for hernia repair in a rat ventral hernia model. METHODS: An abdominal wall defect (15mm x 25mm) was created in Wistar male rats (n=40). Control animals (n=20) had the polypropylene (PP) mesh sutured into the defect, whereas bovine preserved peritoneum (BPP) was used in experimental group (n=20). After 7 and 28 days, the abdominal wall was taken off and histological studies of the amount of collagen by Sirius Red stain and morphometric evaluation consisted in quantitative analysis of the collagen by using specific software (Imagelab). The Mann-Whitney, Kruskal-Wallis and ANOVA tests were applied for statistical analysis (pd"0.05). RESULTS: Histological examination revealed no difference between the BPP and PP groups (p = 0.55 NS). CONCLUSION: BPP is suitable for the closure of ventral hernias in rat model as shown by its morphological properties.

Rat pinealectomy: a modified direct visual approach.

PURPOSE: To report a new, direct visual approach for rat pinealectomy. METHODS: Eighty adult female rats (Rattus norvegicus albinus EPM-1 strain) were weighted and anesthetized intraperitoneally with 15 mg/kg xylazine and 30 mg/kg ketamine. The animal was fastened to a dissection table, an incision was made in the skin and the subcutaneous tissue, bringing the lambda into view. The skullcap was opened with a dental drill, bringing the cerebral hemispheres and the superior sagittal sinus into view. The pineal gland, located under the venous sinus, was removed in a single piece using tweezers. Next, the bone fragment was returned to its place and the surgical layers were sutured. RESULTS: This new technique is easy to be done, avoids bleedings and removes only the pineal gland without damage to the remaining encephalon. In addition it makes possible the achievement of a sham surgery, allowing the pineal gland to remain intact. CONCLUSION: The proposed technique intends to facilitate studies aiming to better understanding the complexity and importance of the pineal gland on reproductive and other body systems.

Rat pinealectomy: a modified direct visual approach / Pinealectomia em ratos: técnica modificada com visualização direta

PURPOSE: To report a new, direct visual approach for rat pinealectomy. METHODS: Eighty adult female rats (Rattus norvegicus albinus EPM-1 strain) were weighted and anesthetized intraperitoneally with 15 mg/kg xylazine and 30 mg/kg ketamine. The animal was fastened to a dissection table, an incision was made in the skin and the subcutaneous tissue, bringing the lambda into view. The skullcap was opened with a dental drill, bringing the cerebral hemispheres and the superior sagittal sinus into view. The pineal gland, located under the venous sinus, was removed in a single piece using tweezers. Next, the bone fragment was returned to its place and the surgical layers were sutured. RESULTS: This new technique is easy to be done, avoids bleedings and removes only the pineal gland without damage to the remaining encephalon. In addition it makes possible the achievement of a sham surgery, allowing the pineal gland to remain intact. CONCLUSION: The proposed technique intends to facilitate studies aiming to better understanding the complexity and importance of the pineal gland on reproductive and other body systems.

OBJETIVO: Apresentar nova técnica para pinealectomia em ratos. MÉTODOS: 80 ratos adultos fêmeas (Rattus norvegicus albinus) foram pesados e em seguida anestesiados por via intraperitoneal com xilazina e cetamina. Em seguida os animais foram fixados em uma prancha de cortiça e feita uma incisão na pele e no tecido subcutâneo, na região superior da cabeça, evidenciando a junção dos ossos parietais e occipital. Na região do lambda, realizou-se uma perfuração circular, na calota craniana, com o auxilio de uma broca (4 mm) acoplada a um micromotor. Nesse orifício, após a dissecação da dura-mater visibiliza-se a confluência dos seios venosos longitudinal e transverso. Com o auxilio de uma pinça curva esses seios são deslocados, ligados e identificada a glândula pineal, que pode ser removida em peça única. Em seguida, o fragmento ósseo é devolvido ao seu lugar e as camadas cirúrgicas suturadas. RESULTADOS: Esta nova técnica é fácil de ser feita, evita hemorragias e só remove a glândula pineal, sem prejuízos para o restante do encéfalo. Além disso, possibilita a realização de uma cirurgia sham, visto que a glândula pineal permanece sem alterações. CONCLUSÃO: Espera-se que a técnica proposta facilite estudos visando a uma melhor compreensão da complexidade e importância da glândula pineal sobre a reprodução e outros sistemas do organismo.

Hysteroscopic sterilization with occlusion of sheep uterine tube using n-butyl-2-cyanoacrylate adhesive

PURPOSE: To evaluate the fertility and analyze the macroscopic, microscopic and morphometric aspects of sheep uterine tube sterilization with a hysteroscopically insert of n-butyl-2-cyanoacrylate adhesive. METHODS: 12 adult sheep, with one previous pregnancy, were distributed as follows: group L (n=3) subjected to laparotomy and Pomeroy uterine tube ligation, group S (n=3) subjected to hysteroscopic application of saline solution in tube isthmus and group AD(n=6), that was subjected to hysteroscopic application of 0.5 ml of n-2-butil-cyanoacrylate in tube isthmus. They were mated with fertile males for ninety days. The non pregnant sheep, at the 90th day, were subjected to laparotomy with uterus and tubes uterine resection. The fragments of uterine tubes were fixated in 10 percent formalin and processes for histology evaluated, and slices dyes for H.E. Data were evaluated by Wilcoxon and Mann-Whitney and Fisher's exact test. RESULTS: All sheep from groups L and AD did not get pregnant (0 percent) in contrast with sheep from group S (100 percent); the adhesive remained integral in the uterine tube lumen. The percentual of adherences (66.6 percent) and fibrosis responses (100 percent) was significantly higher in the group L than group AD (0 percent) (p<0.01). The diameter of the caudal tube in group AD (2652.15 ± 45.76 mm) was significantly wider than that of the group L (1868.27 ± 56.11* μm) (p < 0.05). CONCLUSION: The hysteroscopic insertion of cyanoacrylate in the uterine tube lumen of sheep was effective to obstruct the uterine tube and to promote the sterilization.

OBJETIVO: Avaliar a fertilidade e aspectos macroscópicos, microscópicos e morfométricos da esterilização histeroscópica de tubas uterinas de ovelhas com o adesivo de n-butil-2-cianoacrilato. MÉTODOS: 12 ovelhas adultas, com uma prenhez anterior, foram distribuídas como segue: o grupo L (n=3) submetidas à laparotomia e laqueadura tipo Pomeroy, grupo S (n=3) submetidas à aplicação histeroscópica de solução salina no istmo tubário e grupo AD (n=6), com aplicação histeroscópica de 0,5 ml de cianoacrilato. As ovelhas foram acasaladas com machos de comprovada fertilidade por noventa dias. As ovelhas não prenhes aos 90 dias, foram submetidas à laparotomia com ressecção do útero e tubas uterinas, que foram fixadas em formalina 10 por centos e os cortes histológicos corados em hematoxilina/eosina. Os resultados foram avaliados pelo teste de Wilcoxon e teste exato de Fisher. RESULTADOS: Todas as ovelhas dos grupos L e AD não ficaram prenhes (0 por cento) ao contrário das ovelhas do grupo S (100 por cento); o adesivo permaneceu íntegro no lúmen tubário. O percentual de aderências (66.6 por cento) e de fibrose (100 por cento) foi significativamente maior no grupo L do que no grupo AD (0 por cento) (p<0,01). O diâmetro da porção caudal no grupo AD (2652,15 ± 45,76 μm) foi significativamente maior do que grupo L (1868,27 ± 56.11 mm) (p<0,05). CONCLUSÃO: A inserção histeroscópica do cianoacrilato no lúmen tubário de ovelhas foi eficaz para obstruir a tuba uterina e promover a esterilização.

The role of bovine preserved peritoneum in rats ventral hernia: a histological evaluation / Aspectos histológicos do uso do peritônio bovino conservado na correção de hérnia ventral em ratos

PURPOSE: To assess the role of preserved bovine parietal peritoneum as a material for hernia repair in a rat ventral hernia model. METHODS: An abdominal wall defect (15mm x 25mm) was created in Wistar male rats (n=40). Control animals (n=20) had the polypropylene (PP) mesh sutured into the defect, whereas bovine preserved peritoneum (BPP) was used in experimental group (n=20). After 7 and 28 days, the abdominal wall was taken off and histological studies of the amount of collagen by Sirius Red stain and morphometric evaluation consisted in quantitative analysis of the collagen by using specific software (Imagelab®). The Mann-Whitney, Kruskal-Wallis and ANOVA tests were applied for statistical analysis (pd"0.05). RESULTS: Histological examination revealed no difference between the BPP and PP groups (p = 0.55 NS). CONCLUSION: BPP is suitable for the closure of ventral hernias in rat model as shown by its morphological properties.

OBJETIVO: Estudar os aspectos histológicos do uso de uma prótese de peritônio bovino na correção de hérnia ventral em um modelo animal de doença. MÉTODOS: Utilizando 40 ratos machos Wistar, comparou-se o implante do peritônio bovino com a tela de polipropileno na correção de um defeito provocado na parede abdominal do animal. Após 7 (sub-grupo A) e 28 (sub-grupo B) dias de observação, as peças foram retiradas e procedeu-se o estudo histológico através da quantificação de colágeno pelo método de captação e processamento digital de imagens, sob a coloração do Sirius Red. Os testes de Mann-Whitney, de Kruskal-Wallis e ANOVA foram utilizados e estabeleceu-se em 0,05 o nível para rejeição da hipótese de nulidade (pd"0.05). RESULTADOS: A quantificação do colágeno na interface do implante mostrou equivalência entre os grupos Peritônio e Polipropileno (p=0,55 NS). CONCLUSÃO: O peritônio parietal bovino apresentou infiltração de tecido fibrocolágeno semelhante a da tela de polipropileno na correção de hérnia ventral em ratos.

Experimental model of double wounds on the rats back, in order to study the skin cicatrization process on rat treated with cellulose coat

To the present study were used males rats (Rattus Norvegicus, var. Albinus, Rodentia Mammalia) EPM-1 Wistar, which came from Biotério Central da UNIFESP-EPM. Each animal was submitted to two wound of circular shape, made with a ôpunchö, with superficial cuts of 2 cm diameter; located on medium line of dorsal region. In the present study, one wound was re-covered with the cellulose coat and the other serves as a control. This experimental form, makes it possible to us realize different studies such as macroscopic aspects of the wounds as well morphometric and morphological parameters analysis.

Morphological aspects of the lamina propria of gingiva of persistent oestrous rats, or submitted to oophorectomy and treated with estrogen or dexamethasone

Los autores realizaron un estudio ultraestructural de la gingiva de ratas en estro permanente, ooforectomizadas y tratadas con estrógenos, con el objetivo de evaluar si esta estructura es un órgano blanco de las hormonas sexuales. Los resultados mostraron que la lámina propia de la gingiva de los animales en estro permnente y, en los ooforectomizados tratados con estrógenos, era rica en células y en fibras colágenas. En estos dos grupos se observaron grandes concentraciones de eosinófilos y fibroblastos activos, como también, de macrófagos y linfocitos, observándose además en el estroma, áreas electrotranslúcidas. En el grupo ooforectomizado se observó gran concentración de células y, el estroma denso contenía inmumerables fibras colágenas y substancias intercelular amorfa. En este grupo, los fibroblastos mostraron señales de atrofia y algunos macrófagos y leucocitos, señales degenerativas. El estroma gingival del grupo tratado con dexametasona, se mostró atrofiado, con gran número de células y fibras colágenas como de fibroblastos y fibrocitos. Fue observado un reducido número de macrófagos y de leucocitos. Se concluyó que la gingiva debe ser considerada como un órgano blanco de los estrógenos y que la ooforectomía no altera esa característica

Estudo morfológico das entero-anastomoses com suturas em pontos separados complementados com adesivo sintético ou biológico em coelho / Morphologic study of enteroanastomosis with sutures in separated stitches supplementeds with synthetical adhesive or biological adhesive

OBJETIVO: Verificar a eficácia dos adesivos de fibrina e cianoacrilato como coadjunvantes nas anastomoses de intestino delgado de coelhos. MÉTODOS: Oitenta coelhos, linhagem Nova Zelândia, machos, adultos, foram submetidos a enterectomia de 3cm e enterorrafia e distribuídos em quatro grupos: Grupo A (dois pontos eqüidistantes); Grupo B (quatro pontos eqüidistantes); Grupo C (seis pontos eqüidistantes); Grupo D (oito pontos eqüidistantes). No 14º dia de observação, os animais foram submetidos a uma laparotomia mediana para estudo macroscópico da cavidade abdominal para identificação e avaliação de aderências, deiscências e/ou fístulas. O segmento intestinal anastomosado foi retirado e feito um molde de alginato para posterior avaliação do índice de estenose. As peças retiradas foram processadas para estudo histológico e avaliação de fibras colágenas, macrófagos, neovascularização e granuloma tipo corpo estranho. RESULTADOS: Não houve diferença significante na avaliação de deiscência e fístulas com os dois adesivos entre os quatros sub-grupos. A aderência ocorreu em maior número nos sub-grupos do adesivo sintético. O índice de estenose foi maior nos animais do adesivo sintético somente nos sub-grupos com dois e oito pontos. A análise histológica mostrou que em todos os parâmetros avaliados (macrófago, neovascularização e granuloma) não houve diferença significante entre os dois tipos de adesivos. Em relação a porcentagem de colágeno verificou-se que houve diferença significante somente no grupo B em que o adesivo sintético foi menor que o biológico. CONCLUSAO: O adesivo biológico de fibrina mostra mais eficácia que o adesivo sintético como complemento de suturas nas anastomoses intestinais.

Observações ultra-estruturais da lâmina própria da gengiva da rata albina ooferectomizada / Ultrastructural observations on the gingival lamina propria in oophorectomized albino rats

Através da microscopia eletrônica os autores estudaram alguns elementos constitutivos da mucosa gengival de rata (Rattus Norvergicus Albinus) para verificar a ação gengival como órgão-alvo dos hormônios ovarianos. Para tanto, selecionaram 3 ratas em fase de estro para grupo controle e 6 ratas submetidas à ooforectomia bilateral. Com base nos resultados obtidos, os autores observaram modificações significativas de algumas estruturas, fato indicativo de ser a lâmina própria sensível à ação dos hormônios ovarianos. Chegaram à conclusão de que a gengiva pode ser incluída no grupo dos órgãos-alvo da ação dos hormônios sexuais