In Vivo Comparison of Hard Tissue Regeneration with Human Mesenchymal Stem Cells Processed with Either the FICOLL Method or the BMAC Method

Objective: To compare new bone formation in maxillary sinus augmentation procedures using biomaterial associated with mesenchymal stem cells (MSCs) separated by two different isolation methods. Background: In regenerative medicine open cell concentration systems are only allowed for clinical application under good manufacturing practice conditions. Methods: Mononuclear cells, including MSCs, were concentrated with either the synthetic poylsaccharid (FICOLL) method (classic open system-control group, n = 6 sinus) or the bone marrow aspirate concentrate (BMAC) method (closed system-test group, n = 12 sinus) and transplanted in combination with biomaterial. A sample of the cells was characterized by their ability to differentiate. After 4.1 months (SD +/- 1.0) bone biopsies were obtained and analyzed. Results: The new bone formation in the BMAC group was 19.9% (90% confidence interval [CI], 10.9-29), and in the FICOLL group was 15.5% (90% CI, 8.6-22.4). The 4.4% difference was not significant (90% CI, -4.6-13.5; p = 0.39). MSCs could be differentiated into osteogenic, chondrogenic, and adipogenic lineages. Conclusion: MSCs harvested from bone marrow aspirate in combination with bovine bone matrix particles can form lamellar bone and provide a reliable base for dental implants. The closed BMAC system is suited to substitute the open FICOLL system in bone regeneration procedures.

Histological Outcomes on the Development of New Space-making Devices for Maxillary Sinus Floor Augmentation

Background: Previous studies have pointed out that the mere elevation of the maxillary sinus membrane promotes bone formation without the use of augmentation materials. Purpose: This experimental study aimed at evaluating if the two-stage procedure for sinus floor augmentation could benefit from the use of a space-making device in order to increase the bone volume to enable later implant installation with good primary stability. Materials and Methods: Six male tufted capuchin primates (Cebus apella) were subjected to extraction of the three premolars and the first molar on both sides of the maxilla to create an edentulous area. The sinuses were opened using the lateral bone-wall window technique, and the membrane was elevated. One resorbable space-making device was inserted in each maxillary sinus, and the bone window was returned in place. The animals were euthanatized after 6 months, and biopsy blocks containing the whole maxillary sinus and surrounding soft tissues were prepared for ground sections. Results: The histological examination of the specimens showed bone formation in contact with both the schneiderian membrane and the device in most cases even when the device was displaced. The process of bone formation indicates that this technique is potentially useful for two-stage sinus floor augmentation. The lack of stabilization of the device within the sinus demands further improvement of space-makers for predictable bone augmentation. Conclusions: It is concluded that (1) the device used in this study did not trigger any important inflammatory reaction; (2) when the sinus membrane was elevated, bone formation was a constant finding; and (3) an ideal space-making device should be stable and elevate the membrane to ensure a maintained connection between the membrane and the secluded space.

The Thyroid Hormone Receptor (TR) beta-Selective Agonist GC-1 Inhibits Proliferation But Induces Differentiation and TR beta mRNA Expression in Mouse and Rat Osteoblast-Like Cells

Previous studies showed anabolic effects of GC-1, a triiodothyronine (T3) analogue that is selective for both binding and activation functions of thyroid hormone receptor (TR) beta 1 over TR alpha 1, on bone tissue in vivo. The aim of this study was to investigate the responsiveness of rat (ROS17/2.8) and mouse (MC3T3-E1) osteoblast-like cells to GC-1. As expected, T3 inhibited cellular proliferation and stimulated mRNA expression of osteocalcin or alkaline phosphatase in both cell lineages. Whereas equimolar doses of T3 and GC-1 equally affected these parameters in ROS17/2.8 cells, the effects of GC-1 were more modest compared to those of T3 in MC3T3-E1 cells. Interestingly, we showed that there is higher expression of TR alpha 1 than TR beta 1 mRNA in rat (similar to 20-90%) and mouse (similar to 90-98%) cell lineages and that this difference is even higher in mouse cells, which highlights the importance of TR alpha 1 to bone physiology and may partially explain the modest effects of GC-1 in comparison with T3 in MC3T3-E1 cells. Nevertheless, we showed that TR beta 1 mRNA expression increases (similar to 2.8- to 4.3-fold) as osteoblastic cells undergo maturation, suggesting a key role of TR beta 1 in mediating T3 effects in the bone forming cells, especially in mature osteoblasts. It is noteworthy that T3 and GC-1 induced TR beta 1 mRNA expression to a similar extent in both cell lineages (similar to 2- to 4-fold), indicating that both ligands may modulate the responsiveness of osteoblasts to T3. Taken together, these data show that TR beta selective T3 analogues have the potential to directly induce the differentiation and activity of osteoblasts.

Histological evaluation of chitosan-based biomaterials used for the correction of critical size defects in rat`s calvaria

Chitosan, a biopolymer obtained from chitin, and its derivates, such as chitosan hydrochloride, has been reported as wound healing accelerators and as possible bone substitutes for tissue engineering, and therefore these Substances could be relevant in dentistry and periodontology. The purpose of this investigation was to make a histological evaluation of chitosan and chitosan hydrochloride biomaterials (gels) used in the correction of critical size bone defects made in rat`s calvaria. Bone defects of 8 mm in diameter were surgically created in the calviria of 50 Holtzman (Rattus norvegicus) rats and filled with blood clot (control), low molecular weight chitosan, high molecular weight chitosan, low molecular weight chitosan hydrochloride, and high molecular weight chitosan hydrochloride, numbering 10 animals, divided into two experimental periods (15 and 60 days), for each biomaterial. The histological evaluation was made based on the morphology of the new-formed tissues in defect`s region, and the results indicated that there was no statistical difference between the groups when the new bone formation in the entire defect`s area were compared (p > 0.05) and, except in the control groups, assorted degrees of inflammation Could be Seen. In Conclusion, chitosan and chitosan hydrochloride biomaterials used in this study were not able to promote new bone formation in critical size defects made in rat`s calvaria. (C) 2009 Wiley Periodicals, Inc. J Biomed Mater Res 93A: 107-114, 2016

Avaliação da presença de osteopatia decorremte do diabetes tipo 1 em criança e adolecentes do Rio Grande do Norte

Diabetes Mellitus (DM) and osteoposes are chronic diseases with great socioeconomic consequences, mainly due to the late complications and consequent disabilities. The potential effects of DM on bone metabolism remain a very conroversial issue, and disagreement exists with regard to the clinical implications of diabetic osteopenia and the mechanism of its ocurrence. The issue is further complicated by the contribuicion of the especific factors, such as duration of disease an dthe degree of metabolic control. The objective of this study is to identify the osteopathy in children and adolescents with DM 1 assisted in the hospital of pediatrics, UFRN, through biochemical markers of bone and mineral metabolism and the extent of bone mineral density. The study was composed by 74 diabetics type 1 patients (DM1) of both gender and aged 6 to 20 yars. Normoglicêmic group was composed by 97 healthy subjects of both genders, which showed the same age range of DM1, in addition to same socioeconomic class. These individuals qere students from the networks of public education in the city of Natal-RN, randomly invited to paticipate in our study. Both groups DM1 and NG were divided intofour subgroups, according to the classification of tanner , T1, T2, T3, T4 for achieving a benchmark. Diabetic individuals showed up with a poor glycemic control. the group DN1 T4 showed an incresead value for total protein, albumin, urea and microalbumiuria are predictors of grumelura injury in DM1 patients . The total alkaline phosphatase activitywas kept on high levels for both groups because they are in a stature development age. For osteocalcin there were decreased levels for groups Dm1 T1, T2, and T3 when compared to their NG (s), suggesting that this decrease could be associated with reduction in the number and/or differentiation os osteoblasts thereby contributing to reducing bone formation. There were no changes in the activity of TRAP. The serum concentrations of total and ionized calcium, phosphorus and magnesium were included within the RV. It was observed that the BMD (Z- SCORE ) has always been within the RV for both groups, despite to DM1 T4. Taking all together, our results support the hypothesis that children and adolescents with type 1 DM present the risk in the long run to suffer a reduction in the bone mass, associated to poor glicemic control and disease duration. It could limit the bone growth and increase the probality of development of osteopenia, as well as other complications surch as retinopathy and renal failure

Análise macroscópica e histológica do emprego da poliuretana derivada do óleo de mamona (Ricinus communis) aplicada na tíbia de cães em fase de crescimento

OBJETIVO: Estudar o efeito da poliuretana de mamona aplicada ao osso de cães em crescimento. MÉTODOS: Foram utilizados 12 cães subdivididos aleatoriamente em 3 grupos, os quais receberam o implante de mamona na face medial proximal da tíbia, com análise macroscópica e histopatológica aos 30 (GIII), 60 (GII) e 90 (GI) dias. RESULTADOS: A poliuretana foi recoberta por uma cápsula conjuntiva fibrosa, não ocorrendo proliferação óssea ao seu redor. CONCLUSÃO: A poliuretana é biocompatível, comportando-se como espaçador biológico em cães. Nesta espécie não ocorre a osteointegração.

Kinetic Analysis of Substrate Utilization by Native and TNAP-, NPP1-, or PHOSPHO1-Deficient Matrix Vesicles

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

Ação local do alendronato sódico na reparação óssea de ratos espontaneamente hipertensos (SHR)

FUNDAMENTO: A hipertensão arterial é uma desordem caracterizada por alterações relevantes no tecido ósseo. O alendronato sódico tem indicação no tratamento de doenças ósseas, por causa de sua afinidade pela hidroxiapatita, inibindo as reabsorções ósseas. OBJETIVO: Analisar a ação local do alendronato sódico na reparação óssea de ratos espontaneamente hipertensos (SHR). MÉTODOS: Um defeito ósseo foi criado no fêmur esquerdo de 80 ratos. de acordo com o material utilizado no local, criaram-se quatro grupos: controle (C), amido (Am), alendronato 1 mol (A1) e alendronato 2 mol (A2). Após 7 e 21 dias, os animais foram sacrificados. Foram realizadas análises histológicas e histomorfométricas e os dados foram submetidos a análise de variância (ANOVA) e teste de Tukey (5%). RESULTADOS: Aos 7 dias, observou-se, na área do defeito, tecido conjuntivo com hemorragia e inflamação em todos os grupos. Alguns apresentavam matriz osteóide. Os grupos A1 e A2 apresentaram, ainda, uma rede de fibrina. Aos 21 dias, as trabéculas ósseas fechavam praticamente a extensão do defeito nos grupos C e Am. No grupo A1 de animais machos, observaram-se trabéculas que se irradiavam do canal medular até a área do defeito. Nos grupos A1 e A2, constatou-se apenas a presença de tecido conjuntivo com mínima deposição de osteóide. Um achado histológico marcante foi a formação de tecido ósseo extracortical subperiosteal nos animais dos grupos A1 e A2. CONCLUSÃO: Concluiu-se que a administração do alendronato sódico não contribuiu para o reparo ósseo nos ratos SHR, mas possivelmente tenha sido responsável pelas formações ósseas extracorticais observadas.

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.

Ectopic ossification presenting as osteoid metaplasia in a salivary mucocele in a Shih Tzu dog

Background: Salivary mucocele is an accumulation of saliva in a single or multiloculated cavity lined by connective tissue that is contiguous to a salivary gland-duct complex and is the most common condition affecting the salivary glands in dogs. Occasionally, different types of metaplastic lesions, such as squamous and osseous metaplasia - which are rare lesions in animals - can be observed in association with salivary mucocele.Case presentation: A right facial enlargement was suddenly observed in a 4-year-old non-spayed female Shih-Tzu dog. The lesion presented itself as a soft and fluctuant mass located in the right side of the face near to the neck. Histologically, the mass consisted of a cavitary formation without an epithelial lining. Additionally, microscopic examination revealed the presence of osteoid-producing cells which gave rise to areas of bone formation, probably induced by irritation due to the presence sialoliths. Such cells and bone formations were also present in the cavity wall, consequently leading us to classify the condition as a salivary mucocele with osseous metaplasia.Conclusions: In the present case, the pathogenesis was probably associated with the presence of sialoliths, which can behave as etiological agents for the metaplastic lesion. The occurrence of osteoid metaplasia is a rare peculiar condition in the canine salivar y gland, and due to the rarity and lack of information about this specific disease, no clinical data can yet be associated with the development of salivary mucocele with osseous metaplasia in dogs.

Marcadores do metabolismo ósseo em gatos

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

Alveolar osseous defect in rat for cell therapy. Preliminary report

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

Análise da influência do ultrassom de baixa intensidade na região de reparo ósseo em ratos sob ausência de carga

Há evidências de que o ultrassom (US) de baixa intensidade pode acelerar a regeneração óssea. Este trabalho objetivou verificar a ação do US no defeito ósseo, criado experimentalmente em tíbias de ratos sob ausência de carga. Vinte Rattus novergicus albinus, Wistar adultos, divididos em: G1 (n=10), grupo experimental de 15 dias sem suspensão, e G2 (n=10), grupo experimental de 15 dias suspenso pela cauda, foram submetidos à osteotomia em ambas as tíbias e à aplicação do US, frequência de 1,5 MHz, ciclo de trabalho 1:4, 30 mW/cm², nas tíbias direitas por 12 sessões de 20 minutos. Após o sacrifício, as tíbias foram submetidas à análise da Densidade Mineral Óssea (DMO). Os resultados demonstraram DMO de 0,139±0,018 g/cm² para tíbia tratada; 0,131±0,009 g/cm² para tíbia controle no G1; e no G2 registrou-se 0,120±0,009 g/cm² para tíbia tratada e 0,106±0,017 g/cm² para tíbia controle. Houve diferença significante entre os grupos nos quais o G2 apresentou menor DMO, o que demonstra que a suspensão prejudica a manutenção das propriedades ósseas, e entre as tíbias tratadas e controles do G2, demonstrando que o US acelerou o processo de reparo, concluindo que a impossibilidade do estímulo mecânico causada pela não deambulação em um processo de reparo ósseo pode ser minimizada pela ação do US. No G1, a aplicação do US não teve influência significante no aumento da DMO, talvez pelo fato dos animais já terem estímulo mecânico suficiente à formação óssea.

Early healing after elevation of the maxillary sinus floor applying a lateral access: a histological study in monkeys

AimTo describe the early healing within the void obtained after the elevation of the sinus mucosa and simultaneous implant installation without the use of any grafting material in monkeys.Material and methodsImplants were installed simultaneously with the elevation of the maxillary sinus using the lateral approach in eight monkeys without the use of grafting material. The healing after 4, 10, 20 and 30 days was evaluated in the area distal to the implants. Paraffin sections were prepared and analyzed using qualitative histological methods.ResultsThe healing process was initiated by the formation of a coagulum and followed by a provisional matrix and woven bone. Subsequently, a parallel-fiber bone replaced woven bone. The dimension of the elevated area shrank during the healing process. Sprouts of woven bone, present to a moderate extent after 4 days, were more numerous after 10 and 20 days. Newly formed bone originated from the sinus walls and septa, while there was no evidence of participation of the Schneiderian membrane in this process. After 30 days, the window access appeared to be closed by a layer of newly formed trabecular bone.ConclusionsThe coagulum that filled the void distal to the implant after simultaneous elevation of the sinus floor gave rise to newly formed bone. However, the void occupied by the coagulum shrank substantially. The Schneiderian membrane did not provide a basis for new bone formation in the early phase of healing.To cite this article:Scala A, Botticelli D, Rangel IG Jr, de Oliveira JA, Okamoto R, Lang NP. Early healing after elevation of the maxillary sinus floor applying a lateral access: a histological study in monkeys.Clin. Oral Impl. Res. 21, 2010; 1320-1326.doi: 10.1111/j.1600-0501.2009.01964.x.

Osteocalcin immunolabeling during the alveolar healing process in ovariectomized rats treated with estrogen or raloxifene

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