Acid and alkaline phosphatase activity in broiler chicks fed with different levels of phytase and non-phytate phosphorus

Tartrate-resistant acid (ACP) and alkaline phosphatase (ALP) activities were evaluated in the serum and bone of broiler chicks fed with various amounts of non-phytate phosphorus (NPP) or phytase. Data were analysed using a 4×3 factorial design containing four NPP levels per period. Analyses were performed in chicks aged 1-21 days (0.21; 0.29; 0.37; 0.45 ppm) and 36-42 days (0.13; 0.21; 0.29; 0.37 ppm) and under three different phytase level treatments (0, 500 and 1000 FTU/kg) for each period. In 42-day-old animals, the serum ACP and ALP activities did not differ in response to NPP and phytase levels and bone ACP activity decreased with increased phosphorus levels. We observed effects on ALP activity by approximately 70% in lower phosphorus (0.13 and 0.21) levels without phytase. The phytase addition decreased (P<0.05) ALP values in lower phosphorus levels. The bone ALP and ACP levels of 21-day-old animals were not affected by phosphorus or phytase. Pi depletion induces a significant increase in alkaline phosphatase synthesis, suggesting that the function of this enzyme is downregulated by phosphorus. © 2013 Copyright Taylor and Francis Group, LLC.

Impact of diabetes mellitus and metabolic control on bone healing around osseointegrated implants: Removal torque and histomorphometric analysis in rats

Objectives: To evaluate bone healing around dental implants with established osseointegration in experimental diabetes mellitus (DM) and insulin therapy by histomorphometric and removal torque analysis in a rat model. Materials and methods: A total of 80 male Wistar rats received a titanium implant in the tibiae proximal methaphysis. After a healing period of 60 days, the rats were divided into four groups of 20 animals each: a 2-month control group, sacrificed at time (group A), a diabetic group (group D), an insulin group (group I), and a 4-month control group (group C), subdivided half for removal torque and half for histomorphometric analysis. In the D and I groups the DM was induced by a single injection of 40 mg/kg body weight streptozotocin (STZ). Two days after DM induction, group I received subcutaneous doses of insulin twice a day, during 2 months. Groups C and D received only saline. Two months after induction of DM, the animals of groups D, C and I were sacrificed. The plasmatic levels of glucose (GPL) were monitored throughout the experiment. Evaluation of the percentages of bone-to-implant contact and bone area within the limits of the implant threads was done by histomorphometric and mechanical torque analysis. Data were analyzed by anova at significant level of 5%. Results: The GPL were within normal range for groups A, C and I and higher for group D. The means and standard deviations (SD) for histomorphometric bone area showed significant difference between group D (69.34 ± 5.00%) and groups C (78.20 ± 4.88%) and I (79.63 ± 4.97%). Related to bone-to-implant contact there were no significant difference between the groups D (60.81 + 6.83%), C (63.37 + 5.88%) and I (66.97 + 4.13%). The means and SD for removal torque showed that group D (12.91 ± 2.51 Ncm) was statistically lower than group I (17.10 ± 3.06 Ncm) and C (16.95 ± 5.39 Ncm). Conclusions: Diabetes mellitus impaired the bone healing around dental implants with established osseointegration because the results presented a lower percentage of bone area in group D in relation to groups C and I resulting in a lowest torque values for implant removal. Moreover, insulin therapy prevents the occurrence of bone abnormalities found in diabetic animals and osseointegration was not compromised. © 2012 John Wiley & Sons A/S.

Bone morphogenetic protein 15 and fibroblast growth factor 10 enhance cumulus expansion, glucose uptake, and expression of genes in the ovulatory cascade during in vitro maturation of bovine cumulus-oocyte complexes

Oocyte-secreted factors (OSFs) regulate differentiation of cumulus cells and are of pivotal relevance for fertility. Bone morphogenetic protein 15 (BMP15) and fibroblast growth factor 10 (FGF10) are OSFs and enhance oocyte competence by unknown mechanisms. We tested the hypothesis that BMP15 and FGF10, alone or combined in the maturation medium, enhance cumulus expansion and expression of genes in the preovulatory cascade and regulate glucose metabolism favouring hyaluronic acid production in bovine cumulus-oocyte complexes (COCs). BMP15 or FGF10 increased the percentage of fully expanded COCs, but the combination did not further stimulate it. BMP15 increased cumulus cell levels of mRNA encoding a disintegrin and metalloprotease 10 (ADAM10), ADAM17, amphiregulin (AREG), and epiregulin (EREG) at 12 h of culture and of prostaglandin (PG)-endoperoxide synthase 2 (PTGS2), pentraxin 3 (PTX3) and tumor necrosis factor alpha-induced protein 6 (TNFAIP6 (TSG6)) at 22 h of culture. FGF10 did not alter the expression of epidermal growth factor-like factors but enhanced the mRNA expression of PTGS2 at 4 h, PTX3 at 12 h, and TNFAIP6 at 22 h. FGF10 and BMP15 stimulated glucose consumption by cumulus cells but did not affect lactate production or levels of mRNA encoding glycolytic enzymes phosphofructokinase and lactate dehydrogenase A. Each growth factor increased mRNA encoding glucosamine:fructose-6-PO4 transaminases, key enzymes in the hexosamine pathway leading to hyaluronic acid production, and BMP15 also stimulated hyaluronan synthase 2 (HAS2) mRNA expression. This study provides evidence that BMP15 and FGF10 stimulate expansion of in vitro-matured bovine COCs by driving glucose metabolism toward hyaluronic acid production and controlling the expression of genes in the ovulatory cascade, the first acting upon ADAM10, ADAM17, AREG, and EREG and the second on downstream genes, particularly PTGS2. © 2013 Society for Reproduction and Fertility.

Organically produced coffee exerts protective effects against the micronuclei induction by mutagens in mouse gut and bone marrow

While researchers have extensively evaluated the beneficial effects of coffee consumption in reducing the frequency of certain diseases, studies examining the differences between organic and conventional coffee intake are still needed. Therefore, this paper aims to investigate the functional effects of organic and conventional coffee by examining both its chemical composition and its mutagenic/antimutagenic properties. Infusions of 10% or 20% (w/v) of organic and conventional coffee were administered by gavage (10 mL/kg b.w., once or twice a day) to male Swiss mice against doxorubicin (DXR) and 1,2-dimethylhydrazine dihydrochloride (DMH)-induced mutagenicity. The levels of chlorogenic acids, caffeine and trigonelline from the coffee infusions and oxidative stress analysis from the liver were measured by HPLC. Gut and bone marrow micronucleus assays were used as mutagenic/antimutagenic endpoints, as well as the crypt measurements and gut apoptosis index. The in vivo tests revealed that only organic coffee exerted protective effects, despite oxidative stress analysis and crypt measurements not showing differences among treatments. Intriguingly, the low dose (10% w/v mL/kg) displayed a robust protective effect that showed a significant reduction in bone marrow micronuclei (26.8%), gut micronuclei (11.5%) and apoptosis (27.8%), whereas the higher coffee dose (2 × 20% w/v) only showed a protective effect against bone marrow micronucleus (43.7%). These results highlight that organic coffee could be considered to have beneficial functional effects, although it is still a challenge to define conclusions from analytical data and all the possible interactions from this complex food matrix. © 2013 Elsevier Ltd. All rights reserved.

Immunological aspects of fresh-frozen allogeneic bone grafting for lateral ridge augmentation

Objectives: To present some immunological aspects of fresh-frozen allogeneic bone grafting for lateral bone augmentation, based on the quantitative evaluation of IL-10, IL-1β, IFN- γ and TNF- α in patients sera. Material and methods: Thirty-three partially or totally edentulous patients received fresh-frozen allogeneic bone (AL - 20 patients) or autologous bone onlay block grafts (AT - 13 patients) prior to oral implant placement. Blood samples were collected from each patient at various time-points during a 6 month-period (baseline, 14, 30, 90 and 180 days postoperatively). Quantitative evaluation of IL-10, IL-1β, IFN- γ and TNF- α was performed by enzyme linked immunosorbent assay (ELISA). Results: For all evaluated markers and at all evaluated periods, inter-group comparisons showed no statistically significant differences between the groups, while the observed values were within normal levels. For AL-treated patients, intra-group evaluation showed statistically significant increase of TNF-α from baseline to 90 (P < 0.001) and 180 (P < 0.01) days, and from 14 to 90 (P < 0.01) and 180 (P < 0.05) days. IFN- γ showed intercalated results, with a decrease from baseline to 14 days (P < 0.05), and increase from 14 to 90 days (P < 0.001) and 180 (P < 0.05) days. No differences between the periods of evaluation were found for the AT group. Conclusions: AL grafting for lateral bone augmentation, similar to AT grafting, does not seem to challenge the immune system significantly. © 2012 John Wiley & Sons A/S.

Orthodontic force increases interleukin-1β and tumor necrosis factor-α expression and alveolar bone loss in periodontitis

Background: The present study aims to evaluate the effects of orthodontic movement (OM) on the periodontal tissues of rats with ligature-induced periodontal disease. Methods: Eighty-eight rats were divided into four groups: 1) negative control (sham operated); 2) periodontal disease; 3) OM; and 4) periodontal disease followed by OM (OMP). Rats were sacrificed 3 hours or 1, 3, or 7 days after OM commencement. Bone volume fraction (BVF) and bone mineral density (BMD) were assessed in hemimaxillae by microcomputed tomography analysis. Expression of the proinflammatory cytokines interleukin (IL)-1β and tumor necrosis factor (TNF)-α were evaluated in gingival samples by quantitative polymerase chain reaction and enzyme-linked immunosorbent assay, and in the furcation region by immunohistochemistry analysis (IHC). Results: The OMP group had lower BVF and BMD levels compared to the other groups at day 7 (P <0.05). Maximum messenger ribonucleic acid expression of both cytokines was observed in the OMP group at day 1 (P <0.05). In the same period, all proteins were expressed in high levels for all test groups compared to the control group. The number of cells positive for IL-1β and TNF-α by IHC was highest in the OMP group at day 1, with progressive reduction thereafter. Conclusion: The results suggest that OM acts synergistically with periodontal disease in periodontal breakdown through upregulation of proinflammatory cytokines.

Effect of physical training on metabolic and bone profile in weaning rats

Introduction: The practice of moderate-intensity exercise can reduce the risk of infections and improve metabolic aspects of the person. Objective: To investigate the effects of aerobic physical training on endocrine and metabolic aspects, bone and immune systems. Methods: Twenty Wistar rats were divided in two groups: sedentary (SG) and trained group (TG). Training program consisted in swimming, 6 weeks, supporting a workload corresponding to 5% of body weight. At the end of the experiment, were performed counting total and differential leukocyte count and hematocrit. After training period, were analyzed glucose, total protein, triglycerides, cholesterol, liver and muscle samples for the determination of the levels of glycogen, and determination of the tibia length and bone area. All dependent variables were analyzed by one-way analysis of variance (ANOVA) and a significance level of P < 0.05 was used for all comparisons. Results: Hematocrit (%) analyzed showed a significant difference, with higher values ffor TG (54.63 ± 1.41) than for the SG (49.5 ± 1.65). The total leukocyte count was not significantly different, as there was no difference in the differential count. Total cholesterol showed significant decrease in TG (TG = 68.27 ± 13.71 mg/dL; SG = 94.44 ± 28.09), the total protein levels also showed significant reduction (TG = 7.3 ± 0.40 g/dL; SG = 7.74 ± 0.36 g/dL) glucose levels and triglyceride showed no significant differences. The bone length showed significant difference (TG = 40±0.14 mm; SG = 42.10 ± 0.12mm). Tibial area showed a lower value for TG (1.53 ± 0.12cm2) than for SG (1.67 ± 0.18cm2); however, the difference was not statistically significant. Conclusion: It can be concluded that aerobic exercise training is able to produce some unique physiological changes in young rats. There is also the need to prescribe exercises that meet the particular maturational stage of development.

Horizontal ridge augmentation of the atrophic anterior maxilla using rhBMP-2/ACS or autogenous bone grafts: A proof-of-concept randomized clinical trial

Aim To compare the effect of recombinant human bone morphogenetic protein-2 (rhBMP-2) in an absorbable collagen sponge carrier (ACS) with autogenous bone graft for augmentation of the edentulous atrophic anterior maxilla. Methods Twenty-four subjects were enrolled in a randomized, controlled, parallel-group, open-label clinical trial. Subjects either received rhBMP-2/ACS (1.5 mg/ml) or particulated autogenous bone harvested from the mandibular retromolar region. A titanium-mesh was used to provide space and wound stability. A guide was used to standardize clinical recordings using an analogue caliper. Alveolar ridge width was also assessed using cone-beam computed tomography. Results rhBMP-2/ACS yielded significantly greater radiographic horizontal bone gain compared with autogenous bone graft at immediate subcrestal levels (1.5 ± 0.7 versus 0.5 ± 0.9 mm; p = 0.01); non-significant differences were observed at mid- (2.9 ± 0.8 versus 2.9 ± 0.9 mm; p = 0.98) and apical (1.7 ± 0.9 versus 1.8 ± 1.1 mm; p = 0.85) crestal levels. No significant differences in clinical horizontal bone gain were observed at 6 months between rhBMP-2/ACS and autogenous bone graft (3.2 ± 0.9 mm versus 3.7 ± 1.4 mm; p = 0.31). Sixty-two implants were placed after 6 month of healing with no significant differences between groups for number of implants, implant size, primary stability and survival. Conclusions rhBMP-2/ACS appears a realistic alternative for augmentation of the edentulous atrophic anterior maxilla. © 2013 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Effect of phytase supplementation on performance, bone densitometry and carcass yield in broilers chicks

An experiment was conducted to evaluate the performance, bone densitometry and carcass yield of broilers chicks, using different levels of phytase enzyme. Nine hundred and sixty male one-day-old broiler chicks were used. The birds were distributed in a completely randomized experimental design, involving five treatments and six replications of 32 chicks each. The treatments consisted of a control diet for each phase, and four other diets were formulated adding growing levels of the phytase enzyme (250, 500, 750 and 1,000 FTU of phytase kg-1 feed). When adding the phytase enzyme, the nutritional matrix was valued to guarantee the same nutritional levels as the control diet. In general, the addition of phytase enzyme determined a linear decrease on the performance of the birds. However, the performance obtained with the level of 250 FTU phytase kg-1 feed were no different from the control treatment. The best bone density results were observed in the control treatment with no phytase, and the highest leg and thigh yield were obtained at the level of 514 FTU phytase kg-1.

Release of Bone Markers in Immediately Loaded and Nonloaded Dental Implants: A Randomized Clinical Trial

The aim of this study was to compare the release of bone markers during osseointegration of immediately loaded and nonloaded implants. Forty patients who were indicated for rehabilitation with dental implants randomly received either implant and prosthesis placement within 72 hours (group IM) or implant insertion and no prosthesis placement (group NL). Peri-implant crevicular fluid was collected immediately after implant insertion and 7, 15, 30, 60, 90, and 120 days after surgery and levels of osteoprotegerin, transforming growth factors, osteocalcin, osteopontin, and parathyroid hormone were evaluated using Luminex assay. Bleeding index and peri-implantar sulcus depth were also evaluated. The data were compared using statistical tests ( = 5%). No statistical difference was found regarding demographic and clinical parameters (p > .05). Transforming growth factors, osteoprotegerin, osteopontin, and parathyroid hormone presented an earlier release peak in group IM than in NL group (p < .05). Osteocalcin achieved higher levels in group IM versus group NL between 7 and 30 days of evaluation (p < .05). It may be concluded that earlier loading positively modulates bone mediators release around immediately loaded implants when compared with nonloaded dental implants (ClinicalTrials.gov NCT01909999).

Resveratrol Decreases Periodontal Breakdown and Modulates Local Levels of Cytokines During Periodontitis in Rats

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

Oral squamous carcinoma cells secrete RANKL directly supporting osteolytic bone loss

Objective: Local invasion of bone is a frequent complication of oral squamous cell carcinoma (OSCC). Development of these osteolytic lesions is mediated by osteoclasts. Receptor activation of NF-kappa B ligand (RANKL) signaling, counteracted by osteoprotegerin (OPG), regulates osteoclastogenesis. Previous studies in rodent models have demonstrated that inhibition of RANKL decreases tumor growth and lesions within bone. However, the contributory role of OSCC cells to this disease process has yet to be defined.Methods: RANKL expression was assessed in a panel of OSCC cell lines by qPCR, flow cytometry, and ELISA. Induction of osteoclastogenesis was assessed by co-culture with macrophages or with OSCC-derived conditioned medium. In an animal model of bone invasion, nude mice were injected intratibially with UMSCC-11B cells expressing a RANKL luciferase promoter to detect tumor-derived RANKL activity. Osteolytic lesions were analyzed by X-ray, micro-CT, and histological methods. RANKL expression was assessed in human OSCC tissues by immunohistochemistry.Results: We demonstrated that OSCCs express varied levels of all RANKL isoforms, both membrane-bound and soluble RANKL. Both co-culture and treatment with OSCC-conditioned media induced osteoclastogenesis. In mice, we demonstrated human RANKL promoter activity during bone invasion. Over the course of the experiment, animals suffered osteolytic lesions as RANKL-driven luciferase expression increased with time. After 8 weeks, human-derived RANKL was detected in areas of bone resorption by immunohistochemistry. Similar epithelial RANKL expression was detected in human OSCC tissues.Conclusion: These data demonstrate the ability of OSCCs to produce RANKL, directly altering the tumor microenvironment to increase osteoclastogenesis and mediate local bone invasion. (C) 2012 Elsevier Ltd. All rights reserved.

Testosterone Regulates Bone Response to Infl ammation

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

Apical periodontitis and periodontal disease increase serum IL-17 levels in normoglycemic and diabetic rats

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

Influence of Delivery Method on Neuroprotection by Bone Marrow Mononuclear Cell Therapy following Ventral Root Reimplantation with Fibrin Sealant

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