Evaluation of the anti-resorptive ability of an experimental acetazolamide paste for the treatment of late replanted teeth: A study in rats

When late replantation is performed, the root surface and root canal should be treated. Notwithstanding failures still occur, because of the high rates of root resorption, evidencing the need to search for substances that may inhibit root resorption. The acetazolamide is a known anti-resorptive agent, and its use as root canal dressing may increase the success rates in the treatment of root resorption. Therefore, this study evaluated the effect of an acetazolamide paste used as root canal dressing in late replanted teeth. The study was conducted on 24 maxillary right incisors of rats, which were avulsed and divided in two groups. In group I, the teeth were kept dry for 30min, had their root surfaces rubbed with a blade, and were treated with 2% sodium fluoride at pH 5.5 for 20min; the root canals were instrumented and filled with acetazolamide paste; and then the teeth were replanted. In group II, the treatment was similar to group I, except for the root canal dressing, with utilization of calcium hydroxide in group II. At 15 and 60days after replantation, the animals were killed and the specimens were processed in a histotechnical laboratory for microscopic and morphometric analysis. The results demonstrated the ability of both intracanal substances to limit root resorption, yet they were unable to completely inhibit the root resorption. Replacement resorption lacunae were present in greater proportion in group II, at 60days. It was concluded that the acetazolamide paste was effective to limit the root resorption, being more effective in limiting the replacement resorption compared with calcium hydroxide. © 2012 John Wiley & Sons A/S.

Increased apoptosis in osteoclasts and decreased RANKL immunoexpression in periodontium of cimetidine-treated rats

It has been demonstrated that histamine interferes with the recruitment, formation and activity of osteoclasts via H1- and H2-receptors. Cimetidine is a H2-receptor antagonist used for treatment of gastric ulcers that seems to prevent bone resorption. In this study, a possible cimetidine interference was investigated in the number of alveolar bone osteoclasts. The incidence of osteoclast apoptosis and immunoexpression of RANKL (receptor activator of nuclear factor κB ligand) was also evaluated. Adult male rats were treated with 100mg kg-1 of cimetidine for 50days (CimG); the sham group (SG) received saline. Maxillary fragments containing the first molars and alveolar bone were fixed, decalcified and embedded in paraffin. The sections were stained by H&E or submitted to tartrate-resistant acid phosphatase (TRAP) method. TUNEL (terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling) method and immunohistochemical reactions for detecting caspase-3 and RANKL were performed. The number of TRAP-positive osteoclasts, the frequency of apoptotic osteoclasts and the numerical density of RANKL-positive cells were obtained. Osteoclast death by apoptosis was confirmed by transmission electron microscopy (TEM). In CimG, TRAP-positive osteoclasts with TUNEL-positive nuclei and caspase-3-immunolabeled osteoclasts were found. A significant reduction in the number of TRAP-positive osteoclasts and a high frequency of apoptotic osteoclasts were observed in CimG. Under TEM, detached osteoclasts from the bone surface showed typical features of apoptosis. Moreover, a significant reduction in the numerical density of RANKL-positive cells was observed in CimG. The significant reduction in the number of osteoclasts may be due to cimetidine-induced osteoclast apoptosis. However, RANKL immunoexpression reduction also suggests a possible interference of cimetidine treatment in the osteoclastogenesis. © 2012 The Authors Journal of Anatomy © 2012 Anatomical Society.

Anterior pericranial flap for frontal sinus duct obliteration: Is it a valuable resource?

Inappropriate treatments of frontal sinus fractures may lead to serious complications, such as mucopyocele, meningitis, and brain abscess. Assessment of nasofrontal duct injury is crucial, and nasofrontal duct injury requires sinus obliteration, which is often accomplished by autogenous grafts such as fat, muscle, or bone. These avascular grafts have an increased risk of resorption and infection and donor site morbidity. For these reasons, pericranial flap, which is vascular, should be used for frontal sinus obliteration. The pericranial flap presented with less morbidity procedure and has decreased infection rates, which justifies its use in frontal sinus obliteration. This study aimed to report a case of a comminuted frontal sinus fracture with a brief literature review, regarding the use of pericranial flap. The authors report a case of a 23-year-old male subject with a severely comminuted fracture of the anterior and posterior walls of the frontal sinus. The patient was successfully treated by cranialization with frontal sinus duct obliteration, using anterior pericranial flap. The patient was followed up for 16 months with no postoperative complication, such as infection. Pericranial flap is a good resource for frontal sinus duct obliteration because it is a durable and well-vascularized flap, which determines low rates of postoperative complications. Copyright © 2013 by Mutaz B. Habal, MD.

Influence of presence or absence of teeth adjacent to implants installed immediately into extraction sockets on peri-implant hard tissue levels: An experimental study in the dog

Aim: To evaluate the influence of the presence or absence of adjacent teeth on the level of the mesial and distal alveolar bony crest following healing at sites where implants were installed immediately into extraction sockets. Material and methods: Six Labrador dogs were used. In the right side of the mandible, full-thickness flaps were elevated, and the second, third, and fourth premolars and first molars were extracted. In the left side of the mandible, endodontic treatments of the mesial roots of the third and fourth premolars as well as of the first molars were performed. Full-thickness flaps were elevated, the teeth were hemi-sected, and the distal roots were removed. The second premolars were extracted as well. Subsequently, implants were bilaterally installed with the implant shoulder flush with the buccal bony crest. Implants were placed in the center of the alveoli, but at the fourth premolars, they were placed toward the lingual bony plate of the alveoli. After 3 months of healing, the animals were euthanized and histological sections of the sites prepared. Results: Larger bony crest resorption was observed at the test compared with the control sites, both at the bucco-lingual and mesio-distal aspects. The differences between test and controls for the coronal level of osseointegration were smaller than those for resorption. When data from all mesial and distal sites facing an adjacent tooth were collapsed and compared with those opposing an edentulous zone, lower bony crest resorption and deeper residual marginal defects were found at the sites with neighboring teeth. Conclusion: The extraction of teeth adjacent to a socket into which implants were installed immediately after tooth extraction caused more alveolar bone resorption both for the bucco-lingual and at the mesio-distal aspects compared with sites adjacent to a maintained tooth. © 2012 John Wiley & Sons A/S.

Influence of buccal bony crest width on marginal dimensions of peri-implant hard and soft tissues after implant installation. An experimental study in dogs

Aim: To evaluate the influence of the width of the buccal bony wall on hard and soft tissue dimensions following implant installation. Material and methods: Mandibular premolars and first molars of six Labrador dogs were extracted bilaterally. After 3 months of healing, two recipient sites, one on each side of the mandible, were prepared in such a way as to obtain a buccal bony ridge width of about 2 mm in the right (control) and 1 mm in the left sides (test), respectively. Implants were installed with the coronal margin flush with the buccal alveolar bony crest. Abutments were placed and the flaps were sutured to allow a non-submerged healing. After 3 months, the animals were euthanized and ground sections obtained. Results: All implants were completely osseointegrated. In respect to the coronal rough margin of the implant, the most coronal bone-to-implant contact was apically located 1.04 ± 0.91 and 0.94 ± 0.87 mm at the test and control sites, respectively, whereas the top of the bony crest was located 0.30 ± 0.40 mm at the test and 0.57 ± 0.49 mm at the control sites. No statistically significant differences were found. A larger horizontal bone resorption, however, evaluated 1 mm apically to the rough margin, was found at the control (1.1 ± 0.7 mm) compared to the test (0.3 ± 0.3 mm) sites, the difference being statistically significant. A thin peri-implant mucosa (2.4-2.6 mm) was found at implant installation while, after 3 months of healing, a biological width of 3.90-4.40 mm was observed with no statistically significant differences between control and test sites. Conclusions: A width of the buccal bony wall of 1or 2 mm at implant sites yielded similar results after 3 months of healing in relation of hard tissue and soft tissues dimensions after implant installation. © 2012 John Wiley & Sons A/S.

Ridge preservation at implants installed immediately after molar extraction. An experimental study in the dog

Aim: To evaluate the influence of deproteinized bovine bone mineral (DBBM), in conjunction with a collagen membrane, on bone resorption at implants installed in a lingual position immediately into extraction sockets with horizontal residual buccal defects >2.0 mm. Material & methods: The pulp tissue of the mesial roots of 1M1 was removed in six Labrador dogs, and the root canals were filled with gutta-percha and cement. Flaps were elevated. The molars were hemi-sectioned and the distal roots removed. Implants were installed in a lingual position and with the shoulder flush with the buccal bony crest. After installation, defects of about 2.5 and 2.7 mm in width resulted at the buccal aspects of the test and control sites, respectively. Only in the left site (test), deproteinized bovine bone mineral (DBBM) particles were placed into the defect concomitantly with the placement of a collagen membrane. On the control sites, no biomaterials were applied. A non-submerged healing was allowed. Results: After 3 months of healing, one control implant was not integrated and was excluded from the analysis, together with the contralateral test implant. All remaining implants were integrated into mature bone. The buccal alveolar bony crest was resorbed more at the test compared with the control sites, 2.2 ± 0.9 mm and 1.5 ± 1.3 mm, respectively. The vertical resorption of the lingual plate was 1.6 ± 1.5 mm and 1.5 ± 1.1 mm at the test and control sites, respectively. Only small residual DBBM particles were found at the test sites (1.4%). Conclusion: The use of DBBM particles to fill buccal defects of ≥2.5 mm at implants installed immediately into alveolar extraction sockets did not preserve the buccal bony wall. © 2012 John Wiley & Sons A/S.

A influência do uso e ocupação do solo na qualidade e genotoxicidade da água no reservatório de itupararanga, São Paulo, Brasil

The Brazilian reservoirs have been under great anthropogenic pressure due to continued population growth, which in turn places great pressures on the environment. Given this issue, this study aimed to analyze the influence of land use on the quality and genotoxicity of water in the Itupararanga reservoir, São Paulo, Brazil. Water and sediment samples were analyzed for their quality, genotoxicity and benthic diversity, as well as their interrelationship with the landscape. The reservoir was found to be eutrophic in most sampling stations, and genetic al terations were found in radicular cells of exposed Allium cepa in some sampling stations, besides a low diversity of benthic organisms. Among the variables that most contributed to the genetic alterations and water quality are the presence of riparian vegetation, the bare soil and urban occupation. With the results obtained in this study it was concluded that the quality of water and sediment at the Itupararanga reservoir is compromised, requiring emergency managements to ensure the sustainability of water resources.

Magnesium deficiency results in an increased formation of osteoclasts

Magnesium (Mg2+) deficiency is a frequently occurring disorder that leads to loss of bone mass, abnormal bone growth and skeletal weakness. It is not clear whether Mg2+ deficiency affects the formation and/or activity of osteoclasts. We evaluated the effect of Mg2+ restriction on these parameters. Bone marrow cells from long bone and jaw of mice were seeded on plastic and on bone in medium containing different concentrations of Mg2+ (0.8 mM which is 100% of the normal value, 0.4, 0.08 and 0 mM). The effect of Mg2+ deficiency was evaluated on osteoclast precursors for their viability after 3 days and proliferation rate after 3 and 6 days, as was mRNA expression of osteoclastogenesis-related genes and Mg2+-related genes. After 6 days of incubation, the number of tartrate resistant acid phosphatase-positive (TRACP+) multinucleated cells was determined, and the TRACP activity of the medium was measured. Osteoclastic activity was assessed at 8 days by resorption pit analysis. Mg2+ deficiency resulted in increased numbers of osteoclast-like cells, a phenomenon found for both types of marrow. Mg2+ deficiency had no effect on cell viability and proliferation. Increased osteoclastogenesis due to Mg2+ deficiency was reflected in higher expression of osteoclast-related genes. However, resorption per osteoclast and TRACP activity were lower in the absence of Mg2+. In conclusion, Mg2+ deficiency augmented osteoclastogenesis but appeared to inhibit the activity of these cells. Together, our in vitro data suggest that altered osteoclast numbers and activity may contribute to the skeletal phenotype as seen in Mg2+ deficient patients. © 2012 Elsevier Inc. All rights reserved.

IL-4 and IL-13 inhibit IL-1β and TNF-α induced kinin B 1 and B2 receptors through a STAT6-dependent mechanism

Background and Purpose Bone resorption induced by interleukin-1β (IL-1β) and tumour necrosis factor (TNF-α) is synergistically potentiated by kinins, partially due to enhanced kinin receptor expression. Inflammation-induced bone resorption can be impaired by IL-4 and IL-13. The aim was to investigate if expression of B1 and B2 kinin receptors can be affected by IL-4 and IL-13. Experimental Approach We examined effects in a human osteoblastic cell line (MG-63), primary human gingival fibroblasts and mouse bones by IL-4 and IL-13 on mRNA and protein expression of the B1 and B2 kinin receptors. We also examined the role of STAT6 by RNA interference and using Stat6-/- mice. Key Results IL-4 and IL-13 decreased the mRNA expression of B1 and B2 kinin receptors induced by either IL-1β or TNF-α in MG-63 cells, intact mouse calvarial bones or primary human gingival fibroblasts. The burst of intracellular calcium induced by either bradykinin (B2 agonist) or des-Arg10-Lys-bradykinin (B1 agonist) in gingival fibroblasts pretreated with IL-1β was impaired by IL-4. Similarly, the increased binding of B1 and B2 ligands induced by IL-1β was decreased by IL-4. In calvarial bones from Stat6-deficient mice, and in fibroblasts in which STAT6 was knocked down by siRNA, the effect of IL-4 was decreased. Conclusions and Implications These data show, for the first time, that IL-4 and IL-13 decrease kinin receptors in a STAT6-dependent mechanism, which can be one important mechanism by which these cytokines exert their anti-inflammatory effects and impair bone resorption. © 2013 The Authors. British Journal of Pharmacology © 2013 The British Pharmacological Society.

Variação entre a lâmina de irrigação aplicada e armazenada no solo sob condições de irrigação com baixa uniformidade de distribuição de água

In sprinkler irrigation is important to have a good uniformity of application so that the water in the root zone does not show areas with little available water while others have percolating beyond the plant roots, even though the soil allows a lateral redistribution of water. A usual way to obtain the uniformity of irrigation is by measures of dispersion. The aim of this work was to evaluate the uniformity of water stored in a sand soil before and after non uniformity depth irrigation applied from one sprinkler, as well the variation of storage in a 0-0,45 m layer soil using a neutron probe. The statistical design was completely randomized, and for the variable Christiansen uniformity coefficient modified (CUCHH) were 10 treatments (the irrigation depth, 8 intervals of readings storage, more the existing one before irrigation added to the irrigation depth). For the variable soil water, the treatments were the same, excepting the irrigation depth. Despite the low surface uniformity (16.3%), there was not significantly difference between the storage uniformity before, after irrigation and the potential, however there was from these to the surface uniformity. From de irrigation depth, 15.3 mm, only 6.1 mm remained in the layer from 0 to 0.45 m. There was not significantly difference between the water stored in the soil before and after irrigation within a period of up to 134 hours, being the increase in storage due to irrigation was just 11.7%.

Healing of fresh frozen bone allograft with or without platelet-rich plasma: A histologic and histometric study in rats

Purpose: This study histomorphometrically analyzed the effect of autogenous platelet-rich plasma (PRP) on healing of fresh frozen bone allograft (FFBA) in bony defects in rat calvaria. Materials and Methods: A 5mm-diameter defect was created in the calvarium of 30 rats. Animals were divided into three groups: C (defect was filled by blood clot only), FFBA (defect was filled with 0.01mL of FFBA), and FFBA/PRP (defect was filled with 0.01mL of FFBA combined with 100μL of PRP). All animals were euthanized at 30 days postoperatively. Histomorphometry and histology analyses were performed. Data were statistically analyzed (analysis of variance, Tukey, p<.05). Results: FFBA had a statistically smaller new bone area than groups FFBA/PRP and C. No statistically significant differences were observed between groups FFBA and FFBA/PRP with regard to remaining bone graft particle area. Conclusion: It can be concluded that (1) PRP improved the incorporation of FFBA, increasing the amount of new bone formed; (2) PRP has not influenced the resorption of nonviable particles of the FFBA; and (3) presence of remaining FFBA particles might have accounted for the smaller amount of new bone observed in group FFBA when compared with control group. © 2011 Wiley Periodicals, Inc.

Cysteine proteinase inhibitors regulate human and mouse osteoclastogenesis by interfering with RANK signaling

The cysteine proteinase inhibitor cystatin C inhibited RANKL-stimulated osteoclast formation in mouse bone marrow macrophage cultures, an effect associated with decreased mRNA expression of Acp5, Calcr, Ctsk, Mmp9, Itgb3, and Atp6i, without effect on proliferation or apoptosis. The effects were concentration dependent with half-maximal inhibition at 0.3 μM. Cystatin C also inhibited osteoclast formation when RANKL-stimulated osteoclasts were cultured on bone, leading to decreased formation of resorption pits. RANKL-stimulated cells retained characteristics of phagocytotic macrophages when cotreated with cystatin C. Three other cysteine proteinase inhibitors, cystatin D, Z-RLVG-CHN2 (IC50 0.1 μM), and E-64 (IC 50 3 μM), also inhibited osteoclast formation in RANKL-stimulated macrophages. In addition, cystatin C, Z-RLVG-CHN2, and E-64 inhibited osteoclastic differentiation of RANKL-stimulated CD14+ human monocytes. The effect by cystatin C on differentiation of bone marrow macrophages was exerted at an early stage after RANKL stimulation and was associated with early (4 h) inhibition of c-Fos expression and decreased protein and nuclear translocation of c-Fos. Subsequently, p52, p65, IκBα, and Nfatc1 mRNA were decreased. Cystatin C was internalized in osteoclast progenitors, a process requiring RANKL stimulation. These data show that cystatin C inhibits osteoclast differentiation and formation by interfering intracellularly with signaling pathways downstream RANK. © FASEB.

Correlation between hand/wrist and panoramic radiographs in severe secondary hyperparathyroidism

Objectives: Hand/wrist and dental radiographs are important for osteoporosis analysis in secondary hyperparathyroidism (SHPT). This study evaluated whether a correlation exists between the effects of the disease on the hands and jaws, and investigated the association between osteoporosis progression in the hands and parathyroid hormone (PTH) levels in chronic kidney disease (CKD) patients. Materials and methods: Four panoramic radiographic parameters (mental index, mandibular cortical index, trabecular bone pattern, and calcification/resorption) and four corresponding hand/wrist radiographic parameters (metacarpal cortical thickness, phalangeal cortical index, trabecular bone pattern, and calcification/resorption) were applied to investigate possible correlation between the effects of SHPT on the jaws and hands/wrists, by Spearman's correlation coefficient. PTH levels and the hand/wrist radiographic parameters were also tested by spearman's correlation coefficient (p < 0.05). The presence of brown tumors, vascular calcifications, and acroosteolysis on the hands was also evaluated. Results: Mandibular cortical index was strongly correlated with the phalangeal cortical index (p = 0.000). Phalangeal cortical index and trabecular bone pattern of hand/wrist correlated with PTH levels (0.002 and 0.000, respectively). Brown tumors occurred in four CKD patients, while both vascular calcifications and acroosteolysis were observed in 19 patients. Conclusion: There is a significant correlation between the morphological changes caused by secondary hyperparathyroidism in hand and jaw bones. The morphological status can be assessed using the mandibular cortical index, besides the phalangeal cortical index. The latter correlates well with parathyroid hormone levels of advanced chronic kidney disease. Clinical relevance: Panoramic images reveal morphological changes in the jaw bone, indicating likewise changes in the hand/wrist in severe secondary hyperparathyroidism. The severity of the bone changes may be a reflection of the parathyroid hormone levels in advanced chronic kidney disease. © 2012 Springer-Verlag.

Source-driven remobilizations of nutrients within stem wood in Eucalyptus grandis plantations

Nutrient remobilizations in tree ligneous components have been little studied in tropical forests. A complete randomized block design was installed in Brazilian eucalypt plantations to quantify the remobilizations of phosphorus (P), potassium (K), calcium (Ca), magnesium (Mg), and sodium (Na) within stem wood. Three treatments were studied: control with neither K nor Na addition (C), 3 kmol ha-1 K applied (+K), and 3 kmol ha-1 Na applied (+Na). Biomass and nutrient contents were measured in the stem wood of eight trees destructively sampled at 1, 2, 3 and 4 years after planting in each treatment and annual rings were localized on discs of wood sampled every 3 m in half of the trees. Chemical analyses and wood density measurements were performed individually for each ring per level and per tree sampled. Nutrient remobilizations in annual rings were calculated through mass balance between two successive ages. Our results show that nutrient remobilizations within stem wood were mainly source-driven. Potassium and Na additions largely increased their concentration in the outer rings as well as the amounts remobilized in the first 2 years after the wood formation. The amount of Na remobilized in annual rings was 15 % higher in +Na than in +K the fourth year after planting despite a 34 % higher production of stem wood in +K leading to a much higher nutrient sink. A partial substitution of K by Na in the remobilizations within stem wood might contribute to enhancing Eucalyptus grandis growth in K-depleted soils. © 2013 Springer-Verlag Berlin Heidelberg.

Influence of presence or absence of keratinized mucosa on the alveolar bony crest level as it relates to different buccal marginal bone thicknesses. an experimental study in dogs

Objective: To investigate the influence of the presence or absence of keratinized mucosa on the alveolar bony crest level as it relates to different buccal marginal bone thicknesses. Material and methods: In six beagle dogs, the mandibular premolars and first molars were extracted bilaterally. In the right side of the mandible (test), flaps were elevated, and the buccal as well as part of the lingual masticatory mucosa was removed. The flap was released coronally to allow a primary wound closure. In the left side, the wounds were left unsutured with the keratinized mucosa remaining (control). After 3 months of healing, a complete absence of keratinized mucosa was found at the test sites. Two recipient sites were prepared at each side of the mandible, one in the premolar and one in the molar region. A buccal bony ridge width of approximately 1 and 2 mm was obtained at the premolar and molar region, respectively. Implants were installed with the shoulder flush with the buccal alveolar bony crest, and abutments were connected to allow a nonsubmerged healing. At least 2 mm of keratinized mucosa was surrounding the control sites, while at the test sites, the implants were bordered by alveolar mucosa. After 3 months, the animals were euthanized and ground sections obtained. Results: A higher vertical bony crest resorption was observed at the test compared with the control sites both at the premolar and molar regions, the differences being statistically significant. The top of the peri-implant mucosa was located more coronally at the control compared with the test sites. The horizontal resorption measured 1 mm below the implant shoulder was similar at the test and control sites. Only limited differences were found between premolar and molar sites, with the exclusion of the horizontal resorption that was higher at the test compared with the control sites. Conclusions: A higher alveolar buccal bony crest resorption and a more apical soft tissue marginal position should be expected, when implants are surrounded with thin alveolar mucosa at the time of installation, independently of the thickness of the buccal bony crest. © 2013 John Wiley & Sons A/S.