Potential of Bioactive Glass Particles of Different Size Ranges to Affect Bone Formation in Interproximal Periodontal Defects in Dogs

Background: The aim of this study was to compare the potential of bioactive glass particles of different size ranges to affect bone formation in periodontal defects, using the guided tissue regeneration model in dogs. Methods: In six dogs, 2-wall intrabony periodontal defects were surgically created and chronified on the mesial surfaces of mandibular third premolars and first molars bilaterally. After 1 month, each defect was randomly assigned to treatment with bioabsorbable membrane in association with bioactive glass with particle sizes between 300 and 355 mu m (group 1) or between 90 and 710 mu m (group 2), membrane alone (group 3), or negative control (group 4). The dogs were sacrificed 12 weeks after surgeries, and histomorphometric measurements were made of the areas of newly formed bone, new mineralized bone, and bioactive glass particle remnants. Results: With regard to the area of bioactive glass particle remnants, there was a statistically significant difference between groups 1 and 2, favoring group 1. There were greater areas of mineralized bone in groups 1 and 2 compared to groups 3 and 4 (P<0.05). Conclusion: The bioactive glass particles of small size range underwent faster resorption and substitution by new bone than the larger particles, and the use of bioactive glass particles favored the formation of mineralized bone. J Periodontol 2009;80:808-815.

Leishmanicidal and cholinesterase inhibiting activities of phenolic compounds of Dimorphandra gardneriana and Platymiscium floribundum, native plants from Caatinga biome

Nos últimos anos, o Ministério da Saúde do Brasil e a Organização Mundial da Saúde tem apoiado a investigação de novas tecnologias que possam contribuir para a vigilância, novos tratamentos e controle da leishmaniose visceral no país. Assim, o objetivo deste trabalho foi isolar compostos de plantas do bioma Caatinga, e investigar a toxicidade destes compostos contra as formas promastigotas e amastigotas de Leishmania infantum chagasi, principal parasita responsável pela leishmaniose visceral na América do Sul, e avaliar a sua capacidade para inibir a enzima acetil-colinesterase (AChE). Após a exposição aos compostos em estudo, foram realizados testes utilizando a forma promastigota que expressa luciferase e ELISA in situ para medir a viabilidade das formas promastigotas e amastigota, respectivamente. O ensaio colorimétrico MTT foi realizado para determinar a toxicidade destas substâncias utilizando células monocíticas murina RAW 264.7. Todos os compostos foram testados in vitro para as sua propriedade anti-colinesterásica. Um cumarina, escoparona, foi isolada a partir de hastes de Platymiscium floribundum, e os flavonóides, rutina e quercetina, foram isolados a partir de grãos de Dimorphandra gardneriana. Estes compostos foram purificados, utilizando cromatografia em coluna gel eluída com solventes orgânicos em misturas de polaridade crescente, e identificados por análise espectral. Nos ensaios leishmanicidas, os compostos fenólicos mostraram eficácia contra as formas extracelulares promastigotas, com EC50 para escoporona de 21.4µg/mL e para quercetina e rutina 26 e 30.3µg/mL, respectivamente. Os flavonóides apresentaram resultados comparáveis à droga controle, a anfotericina B, contra as formas amastigotas com EC50 para quercetina e rutina de 10.6 e 43.3µg/mL, respectivamente. Os compostos inibiram a enzima AChE com halos de inibição variando de 0,8 a 0,6cm, indicando um possível mecanismo de ação para a atividade leishmanicida.

PHOSPHORUS DOSES DETERMINE THE PREVALENCE OF NATIVE ARBUSCULAR MYCORRHIZAL FUNGI IN Araucaria angustifolia

A greenhouse experiment was installed with bait cultures to extract the AMF species present in a rhizosphere soil sample of a native Araucaria angustifolia forest in Campos do Jordao, Brazil. The experimental design was completely randomized, with four increasing phosphorus doses (0, 20, 50, and 150 mg kg(-1), as triple superphosphate), with five replicates, the bait plant was araucaria, and all pots were inoculated with 100 g of rhizospheric soil collected in an araucaria forest. After twelve months the spores were extracted, counted and identified, and the percent root colonization was also determined. When taking all four P doses into account, eleven AMF species could be identified: Acaulospora bireticulata, Acaulospora morrowiae, Acaulospora sp., Entrophospora colombiana, Gigaspora margarita, Glomus diaphanum, Glomus etunicatum, Glomus macrocarpum, Scutellospora calospora, Scutellospora gilmorei, and Scutellospora pellucida. There was no effect of the P dose on the total amount of spores neither on the percent root colonization. However, the correspondence analysis showed that the different AMF species were selectively associated mostly to either one or another P dose.

Bovine Bone Matrix Action Associated With Morphogenetic Protein in Bone Defects in Rats Submitted to Alcoholism

Extended excessive alcohol use causes changes in bone tissue, thus affecting osteogenesis. The objective of this study was to evaluate if demineralized bone matrix (Gen-ox (R)) associated with bone morphogenetic protein (Gen-pro (R)) changes bone neoformation in rats submitted to experimental alcoholism. Forty male rats (Rattus norvegicus) were separated into 2 groups of 20 animals each: Group E1, which received ethyl alcohol at 25% and had the surgical cavity filled in only with blood clot; and Group E2. which received ethyl alcohol at 25% and had the surgical cavity filled in with demineralized bovine cortical bone associated with bone morphogenetic protein. The animals were submitted to a three-week period of gradual adaptation to alcohol, and then continued receiving alcohol at 25% for 90 days, when the surgical cavity was made. After the surgery, the animals continued consuming alcohol until reaching the sacrifice periods of 10, 20, 40, and 60 days, when the tibias were removed for histological processing. Results showed that surgical cavity repair and bone marrow reorganization occurred faster in Group E1 than in Group E2. At the end of the experiment, it was observed that animals in Group E2 had thick bony trabeculae surrounding the implanted material particles and a small area of connective tissue in the surface region. In conclusion, the implanted material did not accelerate bone neoformation, rather it served as a structure for osteogenesis.

High-Performance Liquid Chromatography Under Partially Denaturing Conditions (dHPLC) is a Fast and Cost-Effective Method for Screening Molecular Defects: Four Novel Mutations Found in X-Linked Chronic Granulomatous Disease

Implementing precise techniques in routine diagnosis of chronic granulomatous disease (CGD), which expedite the screening of molecular defects, may be critical for a quick assumption of patient prognosis. This study compared the efficacy of single-strand conformation polymorphism analysis (SSCP) and high-performance liquid chromatography under partially denaturing conditions (dHPLC) for screening mutations in CGD patients. We selected 10 male CGD patients with a clinical history of severe recurrent infections and abnormal respiratory burst function. gDNA, mRNA and cDNA samples were prepared by standard methods. CYBB exons were amplified by PCR and screened by SSCP or dHPLC. Abnormal DNA fragments were sequenced to reveal the nature of the mutations. The SSCP and dHPLC methods showed DNA abnormalities, respectively, in 55% and 100% of the cases. Sequencing of the abnormal DNA samples confirmed mutations in all cases. Four novel mutations in CYBB were identified which were picked up only by the dHPLC screening (c.904 insC, c.141+5 g>t, c.553 T>C, and c.665 A>T). This work highlights the relevance of dHPLC, a sensitive, fast, reliable and cost-effective method for screening mutations in CGD, which in combination with functional assays assessing the phagocyte respiratory burst will contribute to expedite the definitive diagnosis of X-linked CGD, direct treatment, genetic counselling and to have a clear assumption of the prognosis. This strategy is especially suitable for developing countries.

Biomechanical Performance of Flexible Intramedullary Nails With End Caps Tested in Distal Segmental Defects of Pediatric Femur Models

Background: Unstable distal femoral fractures in children are challenging lesions with restricted surgical options for adequate stabilization. Elastic nails have become popular for treating femoral shaft fractures, yet they are still challenging for using in distal fractures. The aim of this study was to test whether end caps (CAP) inserted into the nail extremity improved the mechanical stabilization of a segmental defect at the distal femoral metaphyseal-diaphyseal junction created in an artificial pediatric bone model. Methods: Two 3.5-mm titanium elastic nails (TEN) were introduced intramedullary into pediatric femur models, and a 7.0-mm-thick segmental defect was created at the distal diaphyseal-metaphyseal junction. Nondestructive 4-point bending, axial-bending, and torsion tests were conducted. After this, the end caps were inserted into the external tips of the nails and then screwed into the bone cortex. The mechanical tests were repeated. Stiffness, displacement, and torque were analyzed using the Wilcoxon nonparametric test for paired samples. Results: In the combined axial-bending tests, the TEN + CAP combination was 8.75% stiffer than nails alone (P < 0.01); in torsion tests, the TEN + CAP was 14% stiffer than nails alone (P < 0.01). In contrast, the 4-point bending test did not show differences between the methods (P = 0.91, stiffness; P = 0.51, displacement). Thus, the end caps contributed to an increase in the construct stability for torsion and axial-bending forces but not for 4-point bending forces. Conclusions: These findings indicate that end caps fitted to elastic nails may contribute to the stabilization of fractures that our model mimics (small distal fragment, bone comminution, and distal bone fragment loss).

Extraoral Implants in the Rehabilitation of Craniofacial Defects: Implant and Prosthesis Survival Rates and Peri-Implant Soft Tissue Evaluation

Purpose: Few reports have evaluated cumulative survival rates of extraoral rehabilitation and peri-implant soft tissue reaction at long-term follow-up. The objective of this study was to evaluate implant and prosthesis survival rates and the soft tissue reactions around the extraoral implants used to support craniofacial prostheses. Materials and Methods: A retrospective study was performed of patients who received implants for craniofacial rehabilitation from 2003 to 2010. Two outcome variables were considered: implant and prosthetic success. The following predictor variables were recorded: gender, age, implant placement location, number and size of implants, irradiation status in the treated field, date of prosthesis delivery, soft tissue response, and date of last follow-up. A statistical model was used to estimate survival rates and associated confidence intervals. We randomly selected 1 implant per patient for analysis. Data were analyzed using the Kaplan-Meier method and log-rank test to compare survival curves. Results: A total of 150 titanium implants were placed in 56 patients. The 2-year overall implant survival rates were 94.1% for auricular implants, 90.9% for nasal implants, 100% for orbital implants, and 100% for complex midfacial implants (P = .585). The implant survival rates were 100% for implants placed in irradiated patients and 94.4% for those placed in nonirradiated patients (P = .324). The 2-year overall prosthesis survival rates were 100% for auricular implants, 90.0% for nasal implants, 92.3% for orbital implants, and 100% for complex midfacial implants (P = .363). The evaluation of the peri-implant soft tissue response showed that 15 patients (26.7%) had a grade 0 soft tissue reaction, 30 (53.5%) had grade 1, 6 (10.7%) had grade 2, and 5 (8.9%) had grade 3. Conclusions: From this study, it was concluded that craniofacial rehabilitation with extraoral implants is a safe, reliable, and predictable method to restore the patient's normal appearance. (C) 2012 American Association of Oral and Maxillofacial Surgeons J Oral Maxillofac Surg 70:1551-1557, 2012

Bilateral osteoporotic bone marrow defects of the mandible: a case report

Osteoporotic bone marrow defect of the jaws has been reported as a poorly demarcated radiolucency that affect mainly posterior mandible of middle-aged woman. The incidence of this condition is not exactly established and its pathogenesis remains unknown. An additional unusual case of osteoporotic bone marrow defects occurring bilaterally in the mandibular edentulous regions of a 32-year-old white woman is presented reinforcing its diagnostic criteria and histopathological findings.

Bone healing pattern in surgically created circumferential defects around submerged implants: an experimental study in dog

Objective: To describe the healing of marginal defects below or above 1 mm of dimension around submerged implants in a dog model. Material and methods: In 12 Labrador dogs, all mandibular premolars and first molars were extracted bilaterally. After 3 months of healing, full-thickness flaps were elevated in the edentulous region of the right side of the mandible. Two recipient sites were prepared and the marginal 5mm were widened to such an extent to obtain, after implant installation, a marginal gap of 0.5mm at the mesial site (small defect) and of 1.25mm at the distal site (large defect). Titanium healing caps were affixed to the implants and the flaps were sutured allowing a fully submerged healing. The experimental procedures were subsequently performed in the left side of the mandible. The timing of the experiments and sacrifices were planned in such a way to obtain biopsies representing the healing after 5, 10, 20 and 30 days. Ground sections were prepared and histomorphometrically analyzed. Results: The filling of the defect with newly formed bone was incomplete after 1 month of healing in all specimens. Bone formation occurred from the base and the lateral walls of the defects. A larger volume of new bone was formed in the large compared with the small defects. Most of the new bone at the large defect was formed between the 10- and the 20-day period of healing. After 1 month of healing, the outline of the newly formed bone was, however, located at a similar distance from the implant surface (about 0.4mm) at both defect types. Only minor newly formed bone in contact with the implant, starting from the base of the defects, was seen at the large defects (about 0.8mm) while a larger amount was detected at the small defects (about 2.2 mm). Conclusion: Marginal defects around titanium implants appeared to regenerate in 20-30 days by means of a distance osteogenesis. The bone fill of the defects was, however, incomplete after 1 month.

Early peri-implant endosseous healing of two implant surfaces placed in surgically created circumferential defects. A histomorphometric and fluorescence study in dogs

Objective Several implant surfaces are being developed, some in the nanoscale level. In this study, two different surfaces had their early healing properties compared in context of circumferential defects of various widths. Material and methods Six dogs had the mandibular premolars extracted. After 8weeks, four implants were placed equicrestally in each side. One acted as control, while the others were inserted into sites with circumferential defects of 1.0, 1.5 and 2.0mm wide and 5mm deep. A nano-modified surface was used on one side and a micro-rough on the other. Bone markers were administered on the third day after implant placement and then after 1, 2, 4weeks to investigate the bone formation dynamic through fluorescence analysis. Ground sections were prepared from 8-week healing biopsies and histomorphometry was performed. Results The fluorescence evaluation of the early healing showed numerically better results for the nano-modified group; however this trend was not followed by the histomorphometric evaluation. A non-significant numerical superiority of the micro-rough group was observed in terms of vertical bone apposition, defect bone fill, bone-to-implant contact and bone density. In the intra-group analysis, the wider defects showed the worse results while the control sites showed the best results for the different parameters, but without statistical relevance. Conclusion Both surfaces may lead to complete fill of circumferential defects, but the gap width has to be considered as a challenge. The nano-scale modification was beneficial in the early stages of bone healing, but the micro-rough surface showed numerical better outcomes at the 8-week final period.

Responses of transplanted native tree species to invasive alien grass removals in an abandoned cattle pasture in the Lacandon region, Mexico

We tested the early performance of 16 native early-, mid-, and late-successional tree species in response to four intensities of grass removal in an abandoned cattle pasture dominated by the introduced, invasive African grass, Cynodon plectostachyus, within the Lacandon rainforest region, southeast Mexico. The increase in grass removals significantly improved the performance of many species, especially of early-and mid-successional species, while performance of late-successional species was relatively poor and did not differ significantly among treatments. Good site preparation and at least one additional grass removal four months after seedling transplant were found to be essential; additional grass removals led to improved significantly performance of saplings in most cases. In order to evaluate the potential of transplanting tree seedlings successfully in abandoned tropical pastures, we developed a "planting risk index", combining field performance measurements and plantation cost estimations. Our results showed a great potential for establishing restoration plantings with many early-and mid-successional species. Although planting risk of late-successional species was considered high, certain species showed some possibilities of acclimation after 18 months and should be considered in future plantation arrangements in view of their long-term contributions to biodiversity maintenance and also to human welfare through delivery of ecosystem services. Conducting a planting risk analysis can help avoid failure of restoration strategies involving simultaneous planting of early-, mid-, and late-successional tree species. This in turn will improve cost-effectiveness of initial interventions in large-scale, long-term restoration programs.

Retrospective Clinical Study of 988 Dual Acid-Etched Implants Placed in Grafted and Native Bone for Single-Tooth Replacement

Purpose: To evaluate the influence of sex, implant characteristics, and bone grafting on the survival rate of dual acid-etched (DAE) implants. Materials and Methods: Patients treated with internal-hex DAE implants for single-tooth replacement in a military dental clinic between January 2005 and December 2010 were included in this study. Clinical data related to implant characteristics, implant location, presence of grafted bone, and implant failures were collected. The primary outcome was implant loss. The survival rate was analyzed using the Kaplan-Meier method. Cox regression modeling was used to determine which factors would predict implant failure. Results: DAE implants were evaluated in a total of 988 patients (80.3% men). Twenty-four (2.4%) implants failed, most were cylindric (54.2%) with regular platforms (70.8%) and were 10 mm long (58.3%). The failure rate was 2.4% for the anterior maxilla, 3.3% for the posterior maxilla, 1.6% for the anterior mandible, and 2.0% for posterior mandible. The cumulative survival rate was 97.6%. The failure rate was 8.8% in implants placed after sinus augmentation, 7.3% in bone block-grafted areas, and 1.6% in native bone. Based on multivariable analysis (Cox regression), sinus augmentation and bone block grafting had a statistically significant effect on implant failure; the hazard ratios were 5.5 and 4.6, respectively. Conclusion: The results revealed that DAE implants had high survival rates, and no influence of sex, location, shape, diameter, or length on failure rates could be observed. However, a significant association was observed between failure and presence of bone graft in the implant area. Int J Oral Maxillofac Implants 2012;27:1243-1248

Enzyme replacement prevents enamel defects in hypophosphatasia mice

Hypophosphatasia (HPP) is the inborn error of metabolism characterized by deficiency of alkaline phosphatase activity, leading to rickets or osteomalacia and to dental defects. HPP occurs from loss-of-function mutations within the gene that encodes the tissue-nonspecific isozyme of alkaline phosphatase (TNAP). TNAP knockout (Alpl-/-, aka Akp2-/-) mice closely phenocopy infantile HPP, including the rickets, vitamin B6-responsive seizures, improper dentin mineralization, and lack of acellular cementum. Here, we report that lack of TNAP in Alpl-/- mice also causes severe enamel defects, which are preventable by enzyme replacement with mineral-targeted TNAP (ENB-0040). Immunohistochemistry was used to map the spatiotemporal expression of TNAP in the tissues of the developing enamel organ of healthy mouse molars and incisors. We found strong, stage-specific expression of TNAP in ameloblasts. In the Alpl-/- mice, histological, mu CT, and scanning electron microscopy analysis showed reduced mineralization and disrupted organization of the rods and inter-rod structures in enamel of both the molars and incisors. All of these abnormalities were prevented in mice receiving from birth daily subcutaneous injections of mineral-targeting, human TNAP at 8.2?mg/kg/day for up to 44 days. These data reveal an important role for TNAP in enamel mineralization and demonstrate the efficacy of mineral-targeted TNAP to prevent enamel defects in HPP. (C) 2012 American Society for Bone and Mineral Research.

Testing the Performance of Fourteen Native Tropical Tree Species in Two Abandoned Pastures of the Lacandon Rainforest Region of Chiapas, Mexico

The rainforest of Mexico has been degraded and severely fragmented, and urgently require restoration. However, the practice of restoration has been limited by the lack of species-specific data on survival and growth responses to local environmental variation. This study explores the differential performance of 14 wet tropical early-, mid- or late-successional tree species that were grown in two abandoned pastures with contrasting land-use histories. After 18 months, seedling survival and growth of at least 7 of the 14 tree species studied were significantly higher in the site with a much longer history of land use (site 2). Saplings of the three early-successional species showed exceptional growth rates. However, differences in performance were noted in relation to the differential soil properties between the experimental sites. Mid-successional species generally showed slow growth rates but high seedling survival, whereas late-successional species exhibited poor seedling survival at both the study sites. Stepwise linear regressions revealed that the species integrated response index combining survivorship and growth measurements, was influenced mostly by differences in soil pH between the two abandoned pastures. Our results suggest that local environmental variation among abandoned pastures of contrasting land-use histories influences sapling survival and growth. Furthermore, the similarity of responses among species with the same successional status allowed us to make some preliminary site and species-specific silvicultural recommendations. Future field experiments should extend the number of species and the range of environmental conditions to identify site generalists or more narrowly adapted species, that we would call sensitive.

Aggressiveness Overcomes Body-Size Effects in Fights Staged between Invasive and Native Fish Species with Overlapping Niches

Approximately 50 years ago, Nile tilapia were accidentally introduced to Brazil, and the decline of pearl cichlid populations, which has been intensified by habitat degradation, in some locations has been associated with the presence of Nile tilapia. There is, however, little strong empirical evidence for the negative interaction of non-native fish populations with native fish populations; such evidence would indicate a potential behavioural mechanism that could cause the population of the native fish to decline. In this study, we show that in fights staged between pairs of Nile tilapia and pearl cichlids of differing body size, the Nile tilapia were more aggressive than the pearl cichlid. Because this effect prevailed over body-size effects, the pearl cichlids were at a disadvantage. The niche overlap between the Nile tilapia and the pearl cichlid in nature, and the competitive advantage shown by the Nile tilapia in this study potentially represent one of several possible results of the negative interactions imposed by an invasive species. These negative effects may reduce population viability of the native species and cause competitive exclusion.