Adesão de células mesenquimais da medula óssea de camundongos e do ligamento periodontal de ratos a diferentes superfícies de titânio: estudo comparativo in vitro

The present experiment used cell culture to analyze the adhesion capacity of mouse mesenchymal bone marrow cells and rat periodontal ligament to different titanium surfaces. Grade II ASTM F86 titanium discs 15mm in diameter and 1.5mm thick were used and received 2 distinct surface treatments (polished and cathodic cage plasma nitriding). The cells were isolated from the mouse bone marrow and rat periodontal ligament and cultured in α-MEM basic culture medium containing antibiotics and supplemented with 10% FBS and 5% CO2, for 72 hours at 37ºC in a humidified atmosphere. Subculture cells were cultured in a 24-well plate with a density of 1 x 104 cells per well. The titanium discs were distributed in accordance with the groups, including positive controls without titanium discs. After a 24-hour culture, the cells were counted in a Neubauer chamber. The results show that both the mouse mesenchymal bone marrow cells and rat periodontal ligament cells had better adhesion to the control surface. The number of bone marrow cells adhered to the polished Ti surface was not statistically significant when compared to the same type of cell adhered to the Ti surface treated by cathodic cage plasma nitriding. However a significant difference was found between the control and polished Ti groups. In relation to periodontal ligament cell adhesion, a significant difference was only found between the control and plasma-treated Ti surfaces. When comparing equal surfaces with different cells, no statistically significant difference was observed. We can therefore conclude that titanium is a good material for mesenchymal cell adhesion and that different material surface treatments can influence this process

Avaliação microscópica dos fragmentos ósseos obtidos por diferentes métodos de osteotomia e de irrigação em aloenxertos irradiados e congelados de coelho

Oral and facial bone defects can undertake appearance, psychosocial well-being and stomathognatic function of its patients. Over the yerars several strategies for bone defect regeneration have arised to treat these pathologies, among them the use of frozen and irradiated bone allograft. Manipulation of bone grafts it s not determined yet, and several osteotomy alternatives can be observed. The present work evaluated with a microscope the bone fragments obtained from different osteotomy methods and irrigation on rings and blocks allografts irradiated and frozen at 80° negative in a rabbit model. The study is experimental in vitro and it sample was an adult male New Zealand rabbit. The animal was sacrificed to obtain long bones, that were submitted to freezing at 80º negative and irradiated with Cobalt- 60. Then the long bones were sectioned into 24 bone pieces, divided into 4 groups: G1 (n=06) osteotomy was performed with bur No. 6 forming rings with 5 mm thickness with high-speed handpiece with manual irrigation; G2 (n=06) osteotomy was performed with bur No. 6 forming rings with 5 mm thick with surgical motor with a manual irrigation rotation 1500 rpm; GA (n=06), osteotomy with trephine using manual irrigation with saline; and GB (n=06), osteotomy with trephine using saline from peristaltic pumps of surgical motor. Five bone pieces of each group were prepared for analysis on light microscopy (LM) and one on electronic scan electronic microscopy (SEM). On the SEM analysis edges surface, presence of microcracks and Smear Layer were evaluated. Analyzing osteotomy technics on SEM was observed: increased presence of microcracks cutting with high speed; increased presence of areas covered by Smear Layer when cutting with motor implant. The irrigation analysis with SEM was observed: that the presence of microcracks does not depend on the type of irrigation; on manual irrigation, there was greater discrepancy between the cutting lines. The descriptive analysis of the osteotomy and irrigation process on LM showed: histological analysis showing the bony margins with clear tissue changed layer, composed of blackened tissue of charred appearance near to the cortical bone; on the edges of the bony part, bone fragments that were displaced during the bone cut and bone irregularities were observed. After analysis of results we can conclude: that there was greater regularity of the bone cut using high-speed handpiece than using motor implant; the cut with trephine using saline irrigated from peristaltic pumps of surgical motor showed greater homogeneity when compared with manual irrigation; charred tissue was found in all obtained bone samples, whit no significant statistically difference on the proportion of carbonization of the two analysed technics

Avaliação de alterações orais em pacientes submetidos a transplante de medula óssea

Bone marrow transplantation (BMT) is currently the best therapeutic option for patients with hematologic diseases, solid tumors or autoimmune disorders. It is characterized by intravenous infusion of hematopoietic stem cells in order to restore marrow function. However, this procedure requires concomitant immunosuppression treatment, which favors the development of certain complications, often manifested in the oral cavity. This study aimed to evaluate the incidence of oral changes in patients undergoing BMT and to correlate these results with clinical aspects related to the patients and the transplants performed. This is a prevalence study, with cross-sectional design, carried out in a BMT service at the Institute of Onco-Hematology of Natal (ION) and Natal Hospital Center. Data collection was based on questionnaires, clinical examination of the oral cavity and consultation in the medical records. The sample consisted of 51 patients undergoing BMT. After the analysis, was found a general status with good health conditions and presence of oral changes in about half of patients who composed the sample. The manifestations observed were, in decreasing order of frequency: mucositis; gingival alteration and thrombocytopenic purpura; mucosal pigmentation; lichenoid reaction and candidiasis. The oral changes were observed more frequently in cases of allogeneic TMO, in different periods post-transplant, without significant differences related to the source of cells. It was found statistically significant association between the presence of graft-versus-host disease (GVHD) and oral changes (p < 0,001). Therefore, it is concluded that there is a relatively high incidence of changes in oral cavity of patients receiving bone marrow transplantation, a fact which confirms the need to consider this site for examination, diagnosis, treatment and prognosis of possible complications of BMT

Exprassão imuno-histoquímica dos fatores de reabsorção óssea em lesões centrais e periféricas de células gigantes

The Giant Cell Lesions, both the Central Giant Cells Lesions (CGCL) as the Peripheral Giant Cells Lesions (PGCL), correspond to a group of oral lesions that are histologically similar entities; however they show a variable clinical behaviour. The purpose of this study was to compare the immunohistochemical expression of bone resorption factors RANK (Receptor Activator of Nuclear Factor kappa B), RANKL (Receptor Activator of Nuclear Factor kappa B Ligand) and OPG (Osteoprotegerin) between CGCL and PGCL. Additionally, these bone resorption factors were examined in terms of aggressiveness of these lesions. The sample consisted of 61 cases, 30 cases of PGCL and 31 CGCL (16 non-aggressive and 15 aggressive). The analysis was performed by quantification of mononuclear cells (MO) and giant multinucleated cells (CG) immunopositive to anti-RANK, anti-RANKL and anti-OPG antibodies in 10 fields. Moreover, according to the proportion between the amount of cells positive for RANKL and OPG, the cases were categorized into: RANKL>OPG, OPG>RANKL e RANKL=OPG. CGCL showed a higher amount of MO (p=0.002) and total cells (p=0.003) both positives to RANKL compared with the PGCL. Additionally, the CGCL revealed a significant association with the ratio of RANKL>OPG (p=0.001). Analysis of the bone resorption factors revealed no significant differences between aggressive and non-aggressive CGCL (p>0.05). It was observed a positive correlation between the markers themselves, and a negative correlation between lesion size and quantity of OPG positive MO cells (p=0,004) and total cells (p=0,009). Through these results, we suggest that the greatest CGCL resorptive potential compared to the PGCL, may have occurred to the high expression of RANKL. Furthermore differences in the biological behavior of aggressive and non-aggressive CGCL appear to be related to the expression of these bone resorption factors

Expressão imuno-histoquimica de proteínas relacionadas com a reabsorção óssea em carcinoma epidermoide de palato duro e língua

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior

Plasticidade de células tronco mesenquimais de medula óssea de ratos na presença de meio condicionado do nervo facial e do fator de crescimento fibroblástico 2

A number of evidences show the influence of the growth of injured nerve fibers in Peripheral Nervous System (PNS) as well as potential implant stem cells (SCs) to make it more suitable for nerve regeneration medium. In this perspective, this study aimed to evaluate the plasticity of mesenchymal stem cells from bone marrow of mice in the presence of culture medium conditioned with facial nerve explants (D-10) and fibroblast growth factor-2 (FGF-2). In this perspective, the cells were cultivated only with DMEM (group 1), only with D-10(group 2), only with FGF-2(group 3) or with D-10 and FGF-2(group 4). The growth and morphology were assessed over 72 hours. Quantitative phenotypic analysis was taken from the immunocytochemistry for GFAP, OX-42, MAP-2, β-tubulin III, NeuN and NF-200 on the fourth day of cultivation. Cells cultured with conditioned medium alone or combined with FGF-2 showed distinct morphological features similar apparent at certain times with neurons and glial cells and a significant proliferative activity in groups 2 and 4 throughout the days. Cells cultived only with conditioned medium acquired a glial phenotype. Cells cultured with FGF-2 and conditioned medium expressed GFAP, OX-42, MAP-2, β-tubulin III, NeuN and NF-200. On average, area and perimeter fo the group of cells positive for GFAP and the área of the cells immunostained for OX-42 were higher than those of the group 4. This study enabled the plasticity of mesenchymal cells (MCs) in neuronal and glial nineage and opened prospects for the search with cell therapy and transdifferentiation

Avaliação histológica da regeneração óssea do rádio e ulna em cães submetidos ao alongamento com o fixador de Ilizarov

O objetivo do estudo foi analisar histologicamente a qualidade, tipo e orientação dos tecidos formados na área de regeneração óssea produzida durante alongamento simultâneo do rádio e da ulna, com fixador externo de Ilizarov, a partir de osteotomia subperiosteal diafisária distal, com dois incrementos diários de 0,5 mm. A montagem foi composta de dois anéis e quatro hastes telescópicas, sendo a distração óssea iniciada no sexto dia de pós-operatório. Utilizaram-se 15 cães, sem raça definida, adultos, com peso corpóreo entre 17 e 30 kg, divididos por sorteio em subgrupos (A, B, C, D e E) compostos por três animais, que foram submetidos a eutanásia após os seguintes procedimentos: A- oito dias de alongamento, B- 15 dias de alongamento, C- 22 dias de alongamento, D- 28 dias de alongamento e oito dias de fase neutra com o fixador, E- 28 dias de alongamento, 60 dias de fase neutra com fixador e 45 dias sem fixador. Os resultados obtidos mostram que o tipo e localização da osteotomia, bem como o ritmo de distração utilizado, não interferiram sobremaneira na regeneração óssea. A reparação óssea foi eficiente, notadamente intramembranosa, contudo nos subgrupos B e C foi observada a presença de cartilagem.

Efeito de diferentes doses de ácido retinoico sobre a resistência óssea de ratos jovens

OBJETIVO: Avaliar os efeitos da suplementação de diferentes doses de todo-trans ácido retinóico sobre a resistência óssea, por meio de ensaio biomecânico de flexão, em tíbia de ratos jovens. MÉTODOS: Foram estudados 58 ratos jovens, com quatro diferentes doses de vitamina A em suas dietas, sendo divididos em 4 grupos: grupo-controle (n=15), sem acréscimo de todo-trans ácido retinoico; grupo com acréscimo de 0,3mg de todo-trans ácido retinoico por kg de ração (n=13); grupo com 10mg de todo-trans ácido retinoico por kg de ração (n=15); e grupo com 50mg de todo-trans ácido retinoico por kg de ração (n=15). O estudo durou 30 dias. Após o sacrifício dos animais, suas patas esquerdas foram congeladas, dissecadas e as tíbias submetidas ao ensaio de flexão. Foram avaliados a carga máxima e o coeficiente de rigidez. Foi aplicada análise de variância one-way. O nível de significância estatístico adotado foi p<0,05. RESULTADOS: Os valores médios de carga máxima (em Newton) foram: grupo-controle =37,94, DP=4,76; grupo todo-trans ácido retinoico 0,3=36,49, DP= 4,38; grupo todo-trans ácido retinoico 10=40,12, DP=6,03; grupo todo-trans ácido retinoico 50=35,68, DP=5,22 (p=0,107). Os valores médios de coeficiente de rigidez (em Newton/milímetros) foram: grupo-controle =31,84 DP=6,75; grupo todo-trans ácido retinoico 0,3=29,18, DP=4,35; grupo todo-trans ácido retinoico 10=35,48, DP=8,14; grupo todo-trans ácido retinoico 50=30,31, DP=7,14 (p=0,85). CONCLUSÃO: Conclui-se que a exposição a diferentes doses de todo-trans ácido retinoico, em ratos, durante 30 dias, não exerce efeito sobre a resistência óssea, quando avaliada por ensaios biomecânicos.

Qualidade de vida de mulheres com baixa massa óssea na pós-menopausa

OBJETIVO: avaliar a qualidade de vida de pacientes com osteoporose e osteopenia, acompanhadas em ambulatórios especializados em osteoporose e climatério, comparando-as com pacientes com densidade mineral óssea (DMO) normal. MÉTODOS: estudo de série de casos transversal, observacional, que se propôs a analisar, por meio do questionário Medical Outcomes Study 36 Short-Form Health Survey (SF-36), a qualidade de vida de mulheres com osteopenia e osteoporose. Foram avaliadas 124 mulheres na pós-menopausa divididas em três grupos: 55 pacientes com diagnóstico densitométrico de osteoporose, 35 com o de osteopenia e 34 que apresentavam DMO normal. Os três grupos foram comparados com relação aos dados demográficos, características clínicas e de estilo de vida e aos diferentes domínios do SF-36. RESULTADOS: as pacientes dos grupos osteopenia e DMO normal apresentaram menor idade média (56,7±7,1 e 52,9±5,4 anos), maior índice de massa corpórea (IMC) (28,6±3,7 e 30,9±5,1 kg/m²) e menor tempo de menopausa (8,4±5,9 e 5,8±4,5 anos) quando comparadas ao grupo osteoporose (61,8±10,1 anos, IMC de 25,7±5,3 kg/m², 15,5±7,5 anos, respectivamente; p<0,05). de acordo com o SF-36, não houve diferença significativa entre os grupos com relação aos domínios, à exceção do domínio vitalidade, que se mostrou superior no grupo osteoporose. Com relação à impressão pessoal sobre seu estado de saúde, das pacientes que o consideraram bom, um maior percentual pertencia ao grupo osteoporose, e entre aquelas que o consideraram ruim, um percentual menor pertencia ao grupo osteopenia. CONCLUSÃO: a qualidade de vida foi similar em mulheres com osteoporose e osteopenia, em relação às com DMO normal, à exceção do domínio vitalidade, que foi superior, paradoxalmente, nas pacientes com osteoporose.

Influência do tamoxifeno associado ao diabete melito na densidade mineral óssea

Considering that osteopenia and osteoporosis are diabetes mellitus complications, and that tamoxifen (TAM) is an anti-estrogenic drug used in breast cancer treatment, this drug may have a beneficial action preventing accentuaded bone loss associated to diabetes. Female Wistar rats (n=60) weighting 180-250g were divided in four groups: Group C, control animals (n=5); Group T, animals treated with TAM (n=5); Group D, diabetic animals (n=5); and Group DT, diabetic animals treated with TAM (n=5). Oestrus cycle was evaluated before the beggining of experimental period to select the animals with regular cycle. This evaluation continued throughout the study period and for all studied groups. Diabetes was induced by a intra perithoneal injection of streptozotocin (STZ) in a concentration of 45 mg/Kg of body weight. Those animals with serum glicose levels 250 mg/dL were considered diabetics. Animals were sacrificed in the periods of 30, 60 and 90 days after diabetes onset. Left femur histomorphometric measurements and serum biochemical analysis (glycemia, alkaline phosphatase, tartaric-resistant acid phosphatase, calcium, phosphorous, magnesium, total proteins, albumin, globulins, urea and creatinine) were done. Histomorphometric results showed a progressive bone loss in Group D animals when compared to those from Group C all over the experimental period, becoming accentuaded in the 90 days period. In relation to Groups T and DT, values approcimated to those obtained for control group were found during the whole period of study. Those data may indicate a bone mass recovery or a diminished bone loss due to diabetes when animals were treated with TAM. During the whole experimental period animals of groups D and DT maintained glycemic levels above 250 mg/dL whereas animals of groups C and T maintained those levels below 150mg/dL. Alkaline phosphatase activity was increased in all study periods for groups D and DT when compared to group C animals over the 90 days period. Tartarate-resistant acid phosphatase activity was showed unaltered in all periods of study and for all groups. Calcium and magnesium results were also unaltered, maintaining reference levels for all groups in all experimental periods. Phosphorous levels were increased in groups D and DT when compared to groups C and T in the 30 days period. However no difference was found in the periods of 60 and 90 days for this test. No difference was found for total proteins levels for groups C, T, D and DT over the study period. Albumin levels were reduced in DT group in the 60 days period and in D and DT groups in the 90 days period. Urea levels were significantly increased in the 30, 60 and 90 days study periods in groups D and DT when compared to groups C and T. Creatinine results showed a significantly increase in the 90 days period for groups D and DT when compared to groups C and T, and maintaining unaltered in the 30 and 60 days periods. These results suggest that the treatment with TAM may reduce bone loss caused by diabetes mellitus

Associação entre espessura da pele e densidade óssea em mulheres adultas

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

O esporte e suas implicações na saúde óssea de atletas adolescentes

A adolescência é um período fundamental para a aquisição da massa óssea. em adolescentes atletas, o pico de massa óssea pode apresentar maior incremento, em virtude do estresse mecânico imposto aos ossos pelo exercício físico praticado. O objetivo desta revisão foi investigar o papel do treinamento esportivo vigoroso e precoce sobre a saúde óssea de atletas adolescentes. Através da revisão da literatura científica, envolvendo adolescentes atletas de diferentes modalidades e de ambos os sexos, é possível inferir que a densidade mineral óssea é potencializada pelos exercícios, quando grupos de atletas são comparados com grupos de controle. Entretanto, muito se discute na literatura quanto à recomendação da intensidade adequada da prescrição de exercício físico para população adolescente, uma vez que, caso o treinamento se torne muito extenuante, os benefícios gerados pela atividade sobre a saúde dos ossos podem ser minimizados ou anulados. Embora muita controvérsia ainda envolva o tema, independente do tipo de esporte praticado, o aumento de intensidade do treinamento deve ser razoável e coerente com as metas, sendo enfatizado treinamento seguro e eficaz para cada uma das faixas de idade e momentos da maturação biológica, independente dos calendários competitivos.

Impacto da ingestão de cálcio sobre a mineralização óssea em adolescentes

A puberdade destaca-se como período fundamental para a aquisição de massa óssea. Durante essa fase da vida, a mineralização encontra-se aumentada com taxas de formação óssea superior às de reabsorção. Nesse sentido, o objetivo desta revisão foi investigar a inter-relação da ingestão dietética de cálcio com a mineralização óssea, durante a puberdade. Entre os fatores influenciadores nutricionais, merecem destaque o fosfato e o magnésio, que, juntamente com o cálcio, mantêm a integridade estrutural do esqueleto. A revisão de literatura indicou que os excessos de proteína e de refrigerantes, na alimentação dos jovens, acarretam comprometimento na mineralização óssea, embora os dados sejam ainda contraditórios. Conclui-se que, durante o período da puberdade, é indicado manter o aporte de cálcio em níveis adequados, na perspectiva de maximizar o pico da massa óssea.

Iatrogenic cortical aneurysm post-craniotomy

Taking into account the number of craniotomies performed every day around the world, iatrogenic aneurysm post-craniotomy is extremely rare with only anecdotal cases reported in literature. We report an iatrogenic aneurysm affecting a cortical vessel which probably developed during dural closure of a conventional craniotomy. The aneurysm was discovered 6 months after surgery on a routine control angiography. The patient was successfully treated by trapping the parent vessel and excising the aneurysm. Histopathological findings were compatible with a true type of traumatic aneurysm. The possibility of this rare condition occurring highlights the risk of arterial injury during craniotomy.

Análise preditiva da densidade mineral óssea em adolescentes brasileiros eutróficos do sexo masculino

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