Collagen membranes at immediate implants: a histomorphometric study in dogs

Aim To evaluate the influence of resorbable membranes on hard tissue alterations and osseointegration at implants placed into extraction sockets in a dog model. Material and methods In the mandibular premolar region, implants were installed immediately into the extraction sockets of six Labrador dogs. Collagen-resorbable membranes were placed at the test sites, while the control sites were left uncovered. Implants were intended to heal in a submerged mode. After 4 months of healing, the animals were sacrificed, and ground sections were obtained for histomorphometric evaluation. Results After 4 months of healing, a control implant was not integrated (n=5). Both at the test and at the control sites, bone resorption occurred. While the most coronal bone-to-implant contact was similar between the test and the control sites, the alveolar bone crest outline was maintained to a higher degree at the buccal aspect of the test sites (loss: 1.7 mm) compared with the control sites (loss: 2.2 mm). Conclusions The use of collagen-resorbable membranes at implants immediately placed into extraction sockets contributed to a partial (23%) preservation of the buccal outline of the alveolar process. To cite this article:Caneva M, Botticelli D, Salata LA, Souza SLS, Carvalho Cardoso L, Lang NP. Collagen membranes at immediate implants: a histomorphometric study in dogs.Clin. Oral Impl. Res. 21, 2010; 891-897.doi: 10.1111/j.1600-0501.2010.01946.x.

Influence of implant positioning in extraction sockets on osseointegration: histomorphometric analyses in dogs

Aim To evaluate the influence of implant positioning into extraction sockets on osseointegration. Material and methods Implants were installed immediately into extraction sockets in the mandibles of six Labrador dogs. In the control sites, the implants were positioned in the center of the alveolus, while in the test sites, the implants were positioned 0.8 mm deeper and more lingually. After 4 months of healing, the resorptive patterns of the alveolar crest were evaluated histomorphometrically. Results All implants were integrated in mineralized bone, mainly composed of mature lamellar bone. The alveolar crest underwent resorption at the control as well as at the test sites. After 4 months of healing, at the buccal aspects of the control and test sites, the location of the implant rough/smooth limit to the alveolar crest was 2 +/- 0.9 mm and 0.6 +/- 0.9 mm, respectively (P < 0.05). At the lingual aspect, the bony crest was located 0.4 mm apically and 0.2 mm coronally to the implant rough/smooth limit at the control and test sites, respectively (NS). Conclusions From a clinical point of view, implants installed into extraction sockets should be positioned approximately 1 mm deeper than the level of the buccal alveolar crest and in a lingual position in relation to the center of the alveolus in order to reduce or eliminate the exposure above the alveolar crest of the endosseous (rough) portion of the implant. To cite this article:Caneva M, Salata LA, de Souza SS, Baffone G, Lang NP, Botticelli D. Influence of implant positioning in extraction sockets on osseointegration: histomorphometric analyses in dogs.Clin. Oral Impl. Res. 21, 2010; 43-49.

Acellular dermal matrix as a barrier in guided bone regeneration: a clinical, radiographic and histomorphometric study in dogs

Objectives The purpose of this study was to evaluate the effectiveness of the acellular dermal matrix (ADM) as a membrane for guided bone regeneration (GBR), in comparison with a bioabsorbable membrane. Material and methods In seven dogs, the mandibular pre-molars were extracted. After 8 weeks, one bone defect was surgically created bilaterally and the GBR was performed. Each side was randomly assigned to the control group (CG: bioabsorbable membrane made of glycolide and lactide copolymer) or the test group (TG: ADM as a membrane). Immediately following GBR, standardized digital X-ray radiographs were taken, and were repeated at 8 and 16 weeks post-operatively. Before the GBR and euthanasia, clinical measurements of the width and thickness of the keratinized tissue (WKT and TKT, respectively) were performed. One animal was excluded from the study due to complications in the TG during wound healing; therefore, six dogs remained in the sample. The dogs were sacrificed 16 weeks following GBR, and a histomorphometric analysis was performed. Area measurements of new tissue and new bone, and linear measurements of bone height were performed. Results Post-operative healing of the CG was uneventful. In the TG membrane was exposed in two animals, and one of them was excluded from the sample. There were no statistically significant differences between the groups for any histomorphometric measurement. Clinically, both groups showed an increase in the TKT and a reduction in the WKT. Radiographically, an image suggestive of new bone formation could be observed in both groups at 8 and 16 weeks following GBR. Conclusion ADM acted as a barrier in GBR, with clinical, radiographic and histomorphometric results similar to those obtained with the bioabsorbable membrane. To cite this article:Borges GJ, Novaes AB Jr, de Moraes Grisi MF, Palioto DB, Taba M Jr, de Souza SLS. Acellular dermal matrix as a barrier in guided bone regeneration: a clinical, radiographic and histomorphometric study in dogs.Clin. Oral Impl. Res. 20, 2009; 1105-1115.

In vitro osteogenesis on fluorcanasite glass-ceramic with three different chemical compositions

This study aimed at investigating in vitro osteogenesis on three fluorcanasite glass-ceramic compositions with different solubilities (K3, K5, and K8). Osteoblastic cells were obtained from human alveolar bone fragments and cultured under standard osteogenic condition until subconfluence. First passage cells were cultured on K3, K5, and K8 and on Bioglass (R) 45S5 (45S5-control). Cell adhesion was evaluated at 24 h. For proliferation and viability, cells were cultured for 1, 4, and 10 days. Total protein content and alkaline phosphatase (ALP) activity were measured at 7, 14, and 21 days. Cultures were stained with Alizarin red at 21 days, for detection of mineralized matrix. Data were compared by ANOVA followed by Duncan`s test. Cell adhesion, cell proliferation, viability, total protein content, and ALP activity were not affected by fluorcanasite glass-ceramic composition and solubility. Bone-like formation was similar on all fluorcanasite-glass ceramics and was reduced compared to 45S5. The changes in the chemical composition and consequently solubility of the fluorcanasite glass-ceramics tested here did not significantly alter the in vitro osteogenesis. Further modifications of the chemical composition of the fluorcanasite glass-ceramic would be required to improve bone response, making this biomaterial a good candidate to be employed as a bone substitute.

Flap vs. ""flapless"" surgical approach at immediate implants: a histomorphometric study in dogs

Aim To compare the remodeling of the alveolar process at implants installed immediately into extraction sockets by applying a flap or a ""flapless"" surgical approach in a dog model. Material and methods Implants were installed immediately into the distal alveoli of the second mandibular premolars of six Labrador dogs. In one side of the mandible, a full-thickness mucoperiosteal flap was elevated (control site), while contra-laterally, the mucosa was gently dislocated, but not elevated (test site) to disclose the alveolar crest. After 4 months of healing, the animals were sacrificed, ground sections were obtained and a histomorphometric analysis was performed. Results After 4 months of healing, all implants were integrated (n=6). Both at the test and at the control sites, bone resorption occurred with similar outcomes. The buccal bony crest resorption was 1.7 and 1.5 mm at the control and the test sites, respectively. Conclusions ""Flapless"" implant placement into extraction sockets did not result in the prevention of alveolar bone resorption and did not affect the dimensional changes of the alveolar process following tooth extraction when compared with the usual placement of implants raising mucoperiosteal flaps. To cite this article:Caneva M, Botticelli D, Salata LA, Souza SLS, Bressan E, Lang NP. Flap vs. ""flapless"" surgical approach at immediate implants: a histomorphometric study in dogs.Clin. Oral Impl. Res. 21, 2010; 1314-1319.doi: 10.1111/j.1600-0501.2009.01959.x.

Influence of lateral pressure to the implant bed on osseointegration: an experimental study in dogs

Aim To study osseointegration and bone-level changes at implants installed using either a standard or a reduced diameter bur for implant bed preparation. Material and methods In six Labrador dogs, the first and second premolars were extracted bilaterally. Subsequently, mesial roots of the first molars were endodontically treated and distal roots, including the corresponding part of the crown, were extracted. After 3 months of healing, flaps were elevated and recipient sites were prepared in all experimental sites. The control site was prepared using a standard procedure, while the test site was prepared using a drill with a 0.2 mm reduced diameter than the standard one used in the contra-lateral side. After 4 months of healing, the animals were euthanized and biopsies were obtained for histological processing and evaluation. Results With the exception of one implant that was lost, all implants were integrated in mineralized bone. The alveolar crest underwent resorption at control as well as at test sites (buccal aspect similar to 1 mm). The most coronal contact of bone-to-implant was located between 1.2 and 1.6 mm at the test and between 1.3 and 1.7 mm at the control sites. Bone-to-implant contact percentage was between 49% and 67%. No statistically significant differences were found for any of the outcome variables. Conclusions After 4 months of healing, lateral pressure to the implant bed as reflected by higher insertion torques (36 vs. 15 N cm in the premolar and 19 vs. 7 N cm in the molar regions) did not affect the bone-to-implant contact. To cite this article:Pantani F, Botticelli D, Garcia IR Jr., Salata LA, Borges GJ, Lang NP. Influence of lateral pressure to the implant bed on osseointegration: an experimental study in dogs.Clin. Oral Impl. Res. 21, 2010; 1264-1270.doi: 10.1111/j.1600-0501.2009.01941.x.

Influence of energy and pulse repetition rate of Er : YAG laser on enamel ablation ability and morphological analysis of the laser-irradiated surface

Objective: To assess the influence of energy and pulse repetition rate of Er:YAG laser on the enamel ablation ability and substrate morphology. Methods: Fifteen crowns of molars were sectioned in four fragments, providing 60 samples, which were ground to flatten the enamel surface. The initial mass was obtained by weighing the fragments. The specimens were hydrated for I h, fixed, and a 3-mm-diameter area was delimited. Twelve groups were randomly formed according to the combination of laser energies (200, 250, 300, or 350 mJ) and pulse repetition rates (2, 3, or 4 Hz). The final mass was obtained and mass loss was calculated by the difference between the initial and final mass. The specimens were prepared for SEM. Data were submitted to ANOVA and Scheffe test. Results: The 4 Hz frequency resulted in higher mass loss and was statistically different from 2 and 3 Hz (p < 0.05). The increase of frequency produced more melted areas, cracks, and unselective and deeper ablation. The 350 mJ energy promoted greater mass loss, similar to 300 mJ. Conclusions: The pulse repetition rate influenced more intensively the mass loss and morphological alteration. Among the tested parameters, 350 mJ/3 Hz improved the ability of enamel ablation with less surface morphological alterations. (C) 2007 Wiley Periodicals, Inc. J Biomed Mater Res.

Effect of Two Restorative Materials on Root Dentine Erosion

This study sought to evaluate the microhardness of root dentine adjacent to glass-ionomer and composite resin restorations after erosive challenge. A crossover study was performed in two phases of 4 consecutive days each. One hundred twelve bovine root dentine slabs were obtained, and standardized box-shaped cavities were prepared at center of each specimen. The prepared cavities were randomly restored with glass-ionomer cement or composite resin. The slabs were randomly assigned among 14 volunteers, which wore intraoral palatal device containing four restored root dentin slabs. Starting on the second day, half of the palatal acrylic devices were immersed extraorally in a lemonade-like carbonated soft drink for 90 s, four times daily for 3 days. Alter 3-day wash-out, dentine slabs restored with the alternative material were placed into palatal appliance and the volunteers started the second phase of this study. After erosive challenges. microhardness measurements were performed. Regardless of the restorative material employed, eroded specimens demonstrated lower microhardness value (p < 0.0001). At eroded condition examined in this study, dentine restored with glass-ionomer cement showed higher microhardness values (p < 0.0001). It may be concluded that the glass-ionomer cement decreases the progression of root dentine erosion at restoration margin. (C) 2010 Wiley Periodicals, Inc J Biomed Mater Res Part B Appl Biomater 93B 304-305, 2010

Immunohistochemical, tomographic and histological study on onlay bone graft remodeling. Part II: calvarial bone

Objectives Little information is available on the molecular events that occur during graft incorporation over time. The calvarial bone (Cb) grafts have been reported to produce greater responses compared with other donor regions in maxillofacial reconstructions, but the scientific evidences for this are still lacking. The objectives of this study are (1) to study the morphological pattern of Cb onlay bone grafts and compare them with the biological events through immunohistochemical responses and (2) to establish the effects of perforations in maintaining the volume and bone density of the receptor bed. Material and methods Sixty New Zealand White rabbits were submitted to Cb onlay bone grafts on the mandible. In 30 rabbits, the receptor bed was perforated (perforated group), while for the remaining animals the bed was kept intact (non-perforated group). Six animals from each group were sacrificed at 5, 7, 10, 20 and 60 days after surgery. Histological sections from the grafted area were prepared for immunohistochemical and histological analyses. Immuno-labeling was found for proteins Osteoprotegerin (OPG), receptor activator of nuclear factor-kappa beta ligand (RANKL), alkaline phosphatase (ALP), osteopontin (OPN), vascular endothelial growth factor (VEGF), tartrate-resistant acid phosphatase (TRAP), Type I collagen (COL I) and osteocalcin (OC). The tomography examination [computerized tomography (CT) scan] was conducted just after surgery and at the sacrifice. Results The histological findings revealed that the perforations contributed to higher bone deposition during the initial stages at the graft-receptor bed interface, accelerating the graft incorporation process. The results of the CT scan showed lower resorption for the perforated group (P < 0.05), and both groups showed high bone density rates at 60 days. This set of evidences is corroborated by the immunohistochemical outcomes indicating that proteins associated with revascularization and osteogenesis (VEGF, OPN, TRAP and ALP) were found in higher levels in the perforated group. Conclusions These findings indicate that the bone volume of calvarial grafts is better maintained when the receptor bed is perforated, probably resulting from more effective graft revascularization and greater bone deposition. The process of bone resorption peaked between 20 and 60 days post-operatively in both groups although significantly less in the perforated group. To cite this article:Pedrosa Jr WF, Okamoto R, Faria PEP, Arnez MFM, Xavier SP, Salata LA. Immunohistochemical, tomographic and histological study on onlay bone grafts remodeling. Part II: calvarial bone.Clin. Oral Impl. Res. 20, 2009; 1254-1264.doi: 10.1111/j.1600-0501.2009.01747.x.

Immunohistochemical, tomographic and histological study on onlay iliac grafts remodeling

The information concerning the molecular events taking place in onlay bone grafts are still incipient. The objective of the present study is to correlate the effects of perforation of resident bone bed on (1) the timing of onlay autogenous graft revascularization; (2) the maintenance of volume/density of the graft (assessed through tomography); and (3) the occurrence of bone remodeling proteins (using immunohistochemistry technique) delivered in the graft. Thirty-six New Zealand White rabbits were subjected to iliac crest onlay bone grafting on both sides of the mandible. The bone bed was drill-perforated on one side aiming at accelerating revascularization, whereas on the other side it was kept intact. After grafts fixation and flaps suture all animals were submitted to tomography on both mandible sites. Six animals were sacrificed, respectively, at 3, 5, 7, 10, 20 and 60 days after surgery. A second tomography was taken just before sacrifice. Histological slides were prepared from each grafted site for both immunohistochemistry analysis [osteopontin, osteocalcin, type I collagen and vascular endothelial growth factor (VEGF) anti-bodies] and histometric analysis. The values on bone volume measured on tomography showed no statistic significance (P >= 0.05) between perforated and intact sites. Grafts placed on perforated beds showed higher bone density values compared with non-perforated ones at 3 days (P <= 0.05). This correlation was inverted at 60 days postoperatively. The findings from VEGF labeling revealed a tendency for earlier revascularization in the perforated group. The early revascularization of bone grafts accelerated the bone remodeling process (osteocalcin, type I collagen and osteopontin) that led to an increased bone deposition at 10 days. The extended osteoblast differentiation process at intermediate stages in the perforated group cooperated for a denser bone at 60 days.

An alternative method to study the cooling of glasses from melt to solid phase

The aim of this preliminary work was to present a novel method, suitable to investigate the glass cooling, from melt to solid state, based on a fast, non-usual and easy microwave method. The following glass system xBaO . (100-x)B(2)O(3) (x = 0% and 40%) was selected as an example for this study. The melt was poured inside a piece of waveguide and then, its cooling was monitored by the microwave signal as a function of time. The variations in the signal can provide valuable informations about some structural changes that take place during the cooling stages, such as relaxation processes. This method can be useful to investigate the cooling and heating of other materials, opening new possibilities for investigation of dielectric behavior of materials under high temperatures. (C) 2008 Elsevier Inc. All rights reserved.

Thiol and Sulfenic Acid Oxidation of AhpE, the One-Cysteine Peroxiredoxin from Mycobacterium tuberculosis: Kinetics, Acidity Constants, and Conformational Dynamics

Drug resistance and virulence of Mycobacterium tuberculosis are partially related to the pathogen`s antioxidant systems. Peroxide detoxification in this bacterium is achieved by the heme-containing catalase peroxidase and different two-cysteine peroxiredoxins. M. tuberculosis genome also codifies for a putative one-cysteine peroxiredoxin, alkyl hydroperoxide reductase E (MtAhpE). Its expression was previously demonstrated at a transcriptional level, and the crystallographic structure of the recombinant protein was resolved under reduced and oxidized states. Herein, we report that the conformation of MtAhpE changed depending on its single cysteine redox state, as reflected by different tryptophan fluorescence properties and changes in quaternary structure. Dynamics of fluorescence changes, complemented by competition kinetic assays, were used to perform protein functional studies. MtAhE reduced peroxynitrite 2 orders of magnitude faster than hydrogen peroxide (1.9 x 10(7) M(-1) s(-1) vs 8.2 x 10(4) M(-1) s(-1) at pH 7.4 and 25 degrees C, respectively). The latter also caused cysteine overoxidation to sulfinic acid, but at much slower rate constant (40 M(-1) s(-1)). The pK(a) of the thiol in the reduced enzyme was 5.2, more than one unit lower than that of the sulfenic acid in the oxidized enzyme. The pH profile of hydrogen peroxide-mediated thiol and sulfenic acid oxidations indicated thiolate and sulfenate as the reacting species. The formation of sulfenic acid as well as the catalytic peroxidase activity of MtAhpE was demonstrated using the artificial reducing substrate thionitrobenzoate. Taken together, our results indicate that MtAhpE is a relevant component in the antioxidant repertoire of M. tuberculosis probably involved in peroxide and specially peroxynitrite detoxification.

The effect of DMSA-functionalized magnetic nanoparticles on transendothelial migration of monocytes in the murine lung via a beta(2) integrin-dependent pathway

Magnetic nanoparticles surface-functionalized with meso-2,3-dimercaptosuccinic acid (MNPs-DMSA) constitute an innovative and promising approach for tissue- and cell-targeted delivery of therapeutic drugs in the lung. Transendothelial migration of leukocytes in the lung is a side effect of endovenous administration of MNPs-DMSA. Using cytologic and phenotypic analysis of murine bronchoalveolar lavage cells, we identified monocytes/macrophages as the main subpopulation of leukocytes involved in this process. Moreover, ultrastructural analysis revealed the presence of nanoparticles inside of numerous macrophages from bronchoalveolar lavage. MNPs-DMSA at concentrations as high as 1 X 10(15) nanoparticles/mL had no toxic effects on macrophages, as evidenced by 3-(4, 5-dimethylthiazolyi-2)-2,5-diphenyltetrazolium bromide (MTT) assay. Notably, MNPs-DMSA up-regulated the mRNA expression of E, L- and P-selectin and macrophage-1 antigen in the murine lung. Upregulation of these cell adhesion molecules was associated with an increased concentration of tumor necrosis factor-alpha in lung. Finally, the critical relevance of the beta(2) integrin-dependent pathway in leukocyte transmigration elicited by MNPs-DMSA was demonstrated by use of knockout mice. Our results characterize mechanisms of the pro-inflammatory effects of MNPs-DMSA in the lung, and identify beta(2) integrin-targeted interventions as promising strategies to reduce pulmonary side effects of MNPs-DMSA during biomedical applications. (C) 2009 Elsevier Ltd. All rights reserved.

Tissue response around silicon nitride implants in rabbits

The chemical and dimensional stability associated with suitable fracture toughness and propitious tribological characteristics make silicon nitride-based ceramics potential candidates for biomedical applications, mainly as orthopedic implants. Considering this combination of properties, silicon nitride components were investigated in relation to their biocompatibility. For this study, two cylindrical implants were installed in each tibia of five rabbits and were kept in the animals for 8 weeks. During the healing time, tissue tracers were administrated in the animals so as to evaluate the bone growth around the implants. Eight weeks after the surgery, the animals were euthanized and histological analyses were performed. No adverse reactions were observed close to the implant. The osteogenesis process occurred during the entire period defined by the tracers. However, this process occurred more intensely 4 weeks after the surgery. In addition, the histological analyses showed that bone growth occurred preferentially in the cortical areas. Different kinds of tissue were identified on the implant surface, characterized by lamellar bone tissue containing osteocytes and osteons, by a noncalcified matrix containing osteoblasts, or by the presence of collagen III, which may change to collagen I or remain as a fibrous tissue. The results demonstrated that silicon nitride obtained according to the procedure proposed in this research is a biocompatible material. (c) 2007 Wiley Periodicals, Inc.

Exercise-associated thermographic changes in young and elderly subjects

This study aimed at evaluating the thermographic changes associated with localized exercise in young and elderly subjects. An exercise protocol using 1 kg load was applied during 3 min to the knee flexors of 14 elderly (67 +/- 5 years) and 15 young (23 +/- 2 years) healthy subjects. The posterior thigh`s skin temperature of the exercised limb and contralateral limb were measured by infrared thermography on pre-exercise, immediately post-exercise, and during the 10-min period post-exercise. Difference (p < 0.01) between elderly and young subjects was observed on pre-exercise temperature. Although differences were not observed between pre-exercise and immediately post-exercise temperature in the exercised limb, thermographic profile displayed heat concentration in exercised areas for both groups. Temperature reduction was only observed for the young group on the 10-min post-exercise (p < 0.05) in the exercised limb (30.7 +/- 1.7 to 30.3 +/- 1.5 degrees C). In contrast, there was a temperature reduction post-exercise (p < 0.01) in the contralateral limb for both groups. These results present new evidences that elderly and young subjects display similar capacity of heat production; however, the elderly subjects presented a lower resting temperature and slower heat dissipation. This work contributes to improve the understanding about temperature changes in elderly subjects and may present implications to the sports and rehabilitation programs.