Osseointegration of zirconia and titanium implants in a rabbit tibiae model evaluated by microtomography, histomorphometry and fluorochrome labeling analyses.

OBJECTIVES: This study compares the osseointegration of machined-zirconia implants containing yttria (M-Y-TZP) with machined (M-Ti) and resorbable blast media (RBM-Ti) titanium implants. MATERIAL AND METHODS: M-Y-TZP, M-Ti and RBM-Ti implants were randomly placed in rabbit tibiae. Fluorochrome bone labels (tetracycline, alizarin and calcein) were administered at different time periods. After 8 weeks, osseointegration was evaluated in terms of bone-to-implant contact (BIC), new bone area (nBA), remaining cortical bone area (rBA) and temporal quantification of fluorochromes, using micro-CT and histomorphometric analyses. RESULTS: RBM-Ti implants showed higher resorption of the remaining cortical bone and bone formation (rBA = 36.9% and nBA = 38.8%) than M-Y-TZP implants (rBA = 48% and nBA = 26.5%). The BIC values showed no differences among the groups in the cortical region (mean = 52.2%) but in the medullary region, they were 0.45-fold higher in the RBM-Ti group (51.2%) than in the M-Y-TZP group (35.2%). In all groups, high incorporation of tetracycline was observed (2nd to 4th weeks), followed by alizarin (4th to 6th weeks) and calcein (6th to 8th weeks). In the cortical region, incorporation of tetracycline was similar between RBM-Ti (49.8%) and M-Y-TZP (35.9%) implants, but higher than M-Ti (28.2%) implants. Subsequently, alizarin and calcein were 1.1-fold higher in RBM-Ti implants than in the other implants. In the medullary region, no significant differences were observed for all fluorochromes. CONCLUSION: All implants favored bone formation and consequently promoted primary stability. Bone formation around the threads was faster in RBM-Ti and M-Y-TZP implants than in M-Ti implants, but limited bone remodeling with M-Y-TZP implants over time can have significant effects on secondary stability, suggesting caution for its use as an alternative substitute for titanium implants.

Tooth morphology, implantation and replacement system of Hoplias malabaricus (Teleostei, Characiformes, Erythrinidae) / Morfologia, implantação e sistema de substituição dentária do Hoplias malabaricus (Teleostei, Characiformes, Erythrinidae)

The oropharyngeal cavity of Hoplias malabaricus, an ichthyophagous freshwater fish, is anatomically adapted to predation. Macroscopic and microscopic analyses were conducted in order to study the morphology and system of implantation and replacement of teeth. The results showed that this teleost has conical and caniniform teeth, with an orthodentin crown covered by an enameloid cap and a vascularised orthodentin in the root. With regard to the implantation system, there is a junction between the tooth and the bone tissue, as a typical physiological dental ankylosis. The teeth are replaced by a resorption process of multinucleated giant cells that actively eliminate the dentin and bone tissue.

A cavidade orofaríngea do Hoplias malabaricus, um peixe de água doce ictiófago, é anatomicamente adaptado à predação. Análises macroscópica e microscópica foram realizadas com o objetivo de estudar a morfologia e o sistema de implantação e substituição dentária. Os resultados mostraram que este teleósteo apresenta dentes cônicos e caniniformes, com coroa de ortodentina coberta por capuz enamelóide e ortodentina vascularizada na raiz. Em relação ao Sistema de implantação, existe uma junção entre o dente e o tecido ósseo, semelhante a uma anquilose dentária fisiológica típica. Os dentes são substituídos por um processo de reabsorção por células gigantes multinucleadas que ativamente eliminam a dentina e o tecido ósseo.

Tooth morphology, implantation and replacement system of Hoplias malabaricus (Teleostei, Characiformes, Erythrinidae).

The oropharyngeal cavity of Hoplias malabaricus, an ichthyophagous freshwater fish, is anatomically adapted to predation. Macroscopic and microscopic analyses were conducted in order to study the morphology and system of implantation and replacement of teeth. The results showed that this teleost has conical and caniniform teeth, with an orthodentin crown covered by an enameloid cap and a vascularised orthodentin in the root. With regard to the implantation system, there is a junction between the tooth and the bone tissue, as a typical physiological dental ankylosis. The teeth are replaced by a resorption process of multinucleated giant cells that actively eliminate the dentin and bone tissue.

Reduced pancreatic β-cell mass is associated with decreased FoxO1 and Erk1/2 protein phosphorylation in low-protein malnourished rats

A low-protein diet leads to functional and structural pancreatic islet alterations, including islet hypotrophy. Insulin-signaling pathways are involved in several adaptive responses by pancreatic islets. We determined the levels of some insulin-signaling proteins related to pancreatic islet function and growth in malnourished rats. Adult male Wistar rats (N = 20 per group) were fed a 17 percent protein (normal-protein diet; NP) or 6 percent protein (low-protein diet; LP), for 8 weeks. At the end of this period, blood glucose and serum insulin and albumin levels were measured. The morphometric parameters of the endocrine pancreas and the content of some proteins in islet lysates were determined. The β-cell mass was significantly reduced (≅65 percent) in normoglycemic but hypoinsulinemic LP rats compared to NP rats. Associated with these alterations, a significant 30 percent reduction in insulin receptor substrate-1 and a 70 percent increase in insulin receptor substrate-2 protein content were observed in LP islets compared to NP islets. The phosphorylated serine-threonine protein kinase (pAkt)/Akt protein ratio was similar in LP and NP islets. The phosphorylated forkhead-O1 (pFoxO1)/FoxO1 protein ratio was decreased by 43 percent in LP islets compared to NP islets (P < 0.05). Finally, the ratio of phosphorylated-extracellular signal-related kinase 1/2 (pErk1/2) to total Erk1/2 protein levels was decreased by 71 percent in LP islets compared to NP islets (P < 0.05). Therefore, the reduced β-cell mass observed in LP rats is associated with the reduction of phosphorylation in mitogenic-related signals, FoxO1 and Erk proteins. The cause/effect basis of this association remains to be determined.

Reduced pancreatic beta-cell mass is associated with decreased FoxO1 and Erk1/2 protein phosphorylation in low-protein malnourished rats.

A low-protein diet leads to functional and structural pancreatic islet alterations, including islet hypotrophy. Insulin-signaling pathways are involved in several adaptive responses by pancreatic islets. We determined the levels of some insulin-signaling proteins related to pancreatic islet function and growth in malnourished rats. Adult male Wistar rats (N = 20 per group) were fed a 17% protein (normal-protein diet; NP) or 6% protein (low-protein diet; LP), for 8 weeks. At the end of this period, blood glucose and serum insulin and albumin levels were measured. The morphometric parameters of the endocrine pancreas and the content of some proteins in islet lysates were determined. The beta-cell mass was significantly reduced ( congruent with 65%) in normoglycemic but hypoinsulinemic LP rats compared to NP rats. Associated with these alterations, a significant 30% reduction in insulin receptor substrate-1 and a 70% increase in insulin receptor substrate-2 protein content were observed in LP islets compared to NP islets. The phosphorylated serine-threonine protein kinase (pAkt)/Akt protein ratio was similar in LP and NP islets. The phosphorylated forkhead-O1 (pFoxO1)/FoxO1 protein ratio was decreased by 43% in LP islets compared to NP islets (P < 0.05). Finally, the ratio of phosphorylated-extracellular signal-related kinase 1/2 (pErk1/2) to total Erk1/2 protein levels was decreased by 71% in LP islets compared to NP islets (P < 0.05). Therefore, the reduced beta-cell mass observed in LP rats is associated with the reduction of phosphorylation in mitogenic-related signals, FoxO1 and Erk proteins. The cause/effect basis of this association remains to be determined.

Fluoride effects on ectopic bone formation in young and old rats.

This study evaluated the effect of fluoride on bone fluoride levels and on ectopic bone formation in young and old rats. Eighty male Wistar rats were assigned to four groups (n = 20/g), which differed according to the fluoride concentration in their drinking water (0, 5, 15 and 50 mg/l). When half of the rats were 90 days old, demineralized bone matrix (DBM) was implanted. The other rats received DBM implants when they were 365 days old. The animals were killed 28 days after. Fluoride in the femur surface, whole femur and plasma was analyzed with an electrode. The implants were analyzed histomorphometrically. Data were tested for statistically significant differences by ANOVA, Tukey's test, t-test and linear regression (p < 0.05). Increases in plasma, femur surface and whole femur fluoride concentrations were observed as water fluoride levels increased. There was also a trend for increase in plasma and femur fluoride concentrations as age increased. Significant positive correlations were found between plasma and femur surface, plasma and femur and femur surface and femur fluoride concentrations. The morphometric analyses indicated an increase in bone formation for younger rats that received 5 mg/l of fluoride in the drinking water. However, this was not statistically significant. The younger rats that received 50 mg/l of fluoride showed impairment in bone formation. Bone formation was not significantly affected among the older rats. The results suggest that lower doses of fluoride in the drinking water, which slightly increase plasma fluoride levels, may have an anabolic effect on bone formation in younger rats.

Tibial segmental bone defect treated with bone plate and cage filled with either xenogeneic composite or autologous cortical bone graft. An experimental study in sheep.

Tibia segmental defect healing in sheep were clinically, radiographically and histologically evaluated. Twelve young sheep aged four to five months were divided into two groups, G1 and G2. A 3.5 cm long segmental defect was created in the right tibial diaphysis with maintenance of the periosteum. The bone defects in both groups were stabilized with a bone plate combined with a titanium cage. In G1 the cage was filled with pieces of autologous cortical bone graft. In G2 it was filled with a composite biomaterial which consisted of inorganic bovine bone, demineralized bovine bone, a pool of bovine bone morphogenetic proteins bound to absorbable ultra-thin powdered hydroxyapatiteand bone-derived denaturized collagen. Except for one G1 animal, all of them showed normal limb function 60 days after surgery. Radiographic examination showed initial formation of periosteal callus in both groups at osteo-tomy sites, over the plate or cage 15 days postoperatively. At 60 and 90 days callus remodeling occurred. Histological and morphometric analysis at 90 days after surgery showed that the quantity of implanted materials in G1 and G2 were similar, and the quantity of new bone formation was less (p = 0.0048) and more immature in G1 than G2, occupying 51 +/- 3.46% and 62 +/- 6.26% of the cage space, respectively. These results suggest that the composite biomaterial tested was a good alternative to autologous cortical bone graft in this experimental ovine tibial defect. However, additional evaluation is warranted prior to its clinical usage.

Mistura de proteínas morfogenéticas ósseas, hidroxiapatita, osso inorgânico e colágeno envolta por membrana de pericárdio no preenchimento de defeito ósseo segmentar em coelhos / Mixture of bone morphogenetic protein, hydroxyapatite, inorganic bone and collagen interposed by pericardium barrier membrane in the filling of the segmental bone defect in rabbits

Avaliou-se o uso de biomaterial de origem bovina na regeneração de defeitos ósseos segmentares empregando-se 12 coelhos, fêmeas, da raça Norfolk, com idade de seis meses e pesos entre 3 e 4,5kg. Realizou-se falha segmentar bilateral de um centímetro de comprimento na diáfise do rádio, com inclusão do periósteo. No membro direito, o defeito foi delimitado por membrana de pericárdio liofilizada, contendo em seu interior mistura de proteínas morfogenéticas ósseas adsorvidas a hidroxiapatita, colágeno liofilizado e osso inorgânico. No membro esquerdo, o defeito não recebeu tratamento. Radiografias foram obtidas ao término do procedimento cirúrgico e aos sete, 30, 60, 90, 120 e 150 dias de pós-operatório. Após eutanásia de seis coelhos aos 60 dias e seis aos 150 dias de pós-cirúrgico, os resultados radiográficos e histológicos mostraram que a regeneração óssea foi inibida nos defeitos segmentares tratados com o biomaterial.

Biomaterials of bovine origin in regenerating segmental bone defects were evaluated. Twelve six-month old Norfolk rabbits, weighting 3 to 4.5kg were used. A 1cm long segmental defect was created in the radial diaphysis, including the periosteum, of both forelimbs. In the right forelimb, the defect was filled using a mixture of bone morphogenic proteins adsorbed to hydroxyapatite, agglutinant of lyophilized collagen in granules and anorganic cortical bone in granules delimited by a pericardial membrane. In the left forelimb, the defect did not receive treatment and served as a control. Radiographies were taken immediately after surgery and at seven, 30, 60, 90, 120 and 150 days post-operatively. Six rabbits were euthanized at 60 days and the other six at 150 days post-surgery for histological evaluation. Radiographic and histological results revealed that bone regeneration was inhibited in the segmental defects receiving biomaterials.

Post-natal maturation of acinar cells of the guinea pig pancreas: an ultrastructural morphometric study.

The morphological maturation of the acinar cells of the guinea pig pancreas during post-natal development was characterized morphometrically by determining the intracytoplasmic accumulation of rough endoplasmic reticulum (RER) and zymogen granules. The following results were obtained for the period analysed, i.e., from 2 to 70 days of post-natal life: (a) the acinar cell volume increased by 210% (P < 0.01); (b) the mostly cisternal RER occupied more than 30% of the cytoplasm at any age studied and their total volume and surface in the cell were increased by 300 and 534% (P < 0.01), respectively; (c) maturation in the morphological pattern of the RER was observed; (d) the mean number of zymogen granules per cell increased from 261 at 2 days to 422 at 70 days (P < 0.01), while their mean diameter increased from 0.52 to 0.94 micron (P < 0.01) during the same period; (e) these increases in granule number and size were responsible for a 500% (P < 0.01) increase in total volume from 2 to 70 days and for a 304% increase (P < 0.01) in total surface from 2 to 35 days; (f) the RER and the zymogen granules together occupied 44, 54, 55 and 57% of the cytoplasm at 2, 14, 35 and 70 days of age, respectively. We conclude that although the pancreatic acinar cells of the guinea pig are morphologically well differentiated at 2 days of age, with the cytoplasm already showing a large amount of RER and zymogen granules, they are still immature. Morphological maturation of the acinar cell occurs during the first months of post-natal life and is characterized by a substantial gain in cell volume and intracytoplasmic accumulation of RER and zymogen granules, which significantly increase of both their absolute volume and total surface, with a higher growth rate being observed during the period from 2 to 14 days of post-natal life.

Allometric analysis of the postnatal development of the Syrian golden hamster pancreas.

The objective of the present study was to analyse allometrically the growth of the Syrian golden hamster pancreas during days 2 to 70 of postnatal development. Body and pancreatic mass were determined, followed by stereological determination of the absolute volume of each morphological compartment of the pancreas. The marked pancreatic growth, by 4360%, was due to an increase in the absolute volume of all morphological compartments, mainly the acini which showed an increase of 10 431%. Bivariate allometric analysis of pancreatic mass and morphological compartmental volume in relation to body mass gain showed: (1) a biphasic pattern for pancreatic mass, acinar volume, excretory duct volume and stromal volume, with the first phase being observed from 2 to 21 days of age and the second from 21 to 70 days of age, with allometry coefficients of 1.537-0.513, 1.770-0.543, 1.651-0.506 and 0.967-0.258, respectively, and (2) a monophasic pattern from 2 to 70 days for intercalated duct volume and islet volume, with allometry coefficients of 0.913 and 1.727, respectively. These results show that during the growth of the pancreas in relation to that of the body some structures - acini and excretory ducts - follow the growth pattern of the organ, while others - intercalated ducts and islets - show a different pattern. This may be related to the genetic growth characteristics of each compartment itself or to some relationship between compartments during some stage of the ontogenetic development of this organ.

Morphometric dimensions of the mouse parotid glands of both sexes.

The goal of this research was to evaluate the morphometric dimensions of the different structures of male and female albino mouse parotid glands. The following morphometric dimensions were evaluated for the acini, intercalated ducts, striated ducts, excretory ducts and stroma: volume density, total compartmental volume, surface density, total external surface, surface-to-volume ratio, cell volume and absolute number of cells. Analysis of the results showed that the parotid gland mass was 43.7% greater (P < 0.01) in the male mice than in the females. This difference was due to the fact that the compartmental volumes of the acini, intercalated ducts and striated ducts were markedly higher in the male mice, 57.6% (P < 0.01), 253.1% (P < 0.01) and 91.1% (P < 0.05), respectively. The higher volume of the acinar morphological compartment was due to the total number of cells and average cell volume being higher in the male mice, 24.8% P < 0.01) and 47.7% (P < 0.01), respectively. Based on the results obtained, it was concluded that there are morphological differences between male and female parotid glands. These differences are detectable through morphometry, mainly in the morphological acinar and intercalated ducts compartments, which are more developed in male mice.

Morphometric dimensions of Syrian golden hamster pancreas of both sexes.

The morphometric dimensions of the various structures of the pancreas of adult Syrian golden hamsters, of both sexes, were evaluated using stereological methods. The average body mass of the animals used was 133.8 +/- 2.45 g and 140.6 +/- 7.98 g for the males and females, respectively, and the pancreatic mass, 389.9 +/- 14.88 and 409.7 +/- 21.42 mg, respectively. The analysis of variance of the obtained data showed that: a) the acini, intercalated ducts and stroma did not present statistically significant differences in any of the dimensions evaluated, with the exception of the nucleus volume of the acinar cells which was 8.5% larger in the female (P < 0.05); b) the excretory ducts exhibited surface density, total external surface, surface-to-volume ratio, and absolute cell number, 18%, 33%, 14%, and 44%, respectively, larger in the females (P < 0.05); and c) the pancreatic islets of the females exhibited volume density, total volume and absolute cell number, 20%, 27% and 27%, respectively, larger than those of the males (P < 0.05).