Influence of simvastatin on bone regeneration of tibial defects and blood cholesterol level in rats

The purpose of this study was to evaluate the effects of simvastatin, by oral or subcutaneous administration, on tibial defects regeneration and blood cholesterol level in rats. A surgical defect was made on the right tibia of 40 male animals assigned to 4 groups (n=10), based on two routes of administration and on the use or not of simvastatin: subcutaneous injection of simvastatin (7 mg/kg) (group AT) or only the vehicle of drug suspension (group AC), above the defect area, for 5 days; and 20 mg/kg of simvastatin macerated on water (group BT) or only water (group BC), orally, daily, during the whole observation period. The animals were sacrificed after 15 or 30 days, when blood samples were analyzed to check plasma cholesterol levels. Tibiae were removed and, after decalcification and routine laboratorial processing, histological and histomorphometrical analyses were carried out. ANOVA was used for statistical analysis at 5% signficance level. The histological and histomorphometrical analyses showed significant differences only between the experimental periods (p<0.05). Animals sacrificed after 30 days showed better bone repair (p<0.05). There was no statistically significant difference (p>0.05) for blood cholesterol levels between the groups. In conclusion, simvastatin administration either orally or subcutaneously did not improve bone repair of experimental tibial defects and did not alter blood cholesterol levels in rats.

Bone mineralization in Brazilian adolescents: The years of maximum bone mass incorporation

Puberty is the fundamental period for bone mass (BM) acquisition. In this period mineralization is found to increase with levels of high bone formation. The critical years of intense bone anabolism deserve special attention, as adequate gain could minimize fracture risk in later years. The objective of this work was to study bone mineral content (BMC) and bone mineral density (BMD) in male adolescents with age bracket and maturation level. Sixty-one healthy male 10 to 19 year-olds were evaluated for calcium intake, weight, stature, BMI, puberty stage and BMC and BMD in the lumbar spine and femur. BM was measured by bone densitometry (DXA). Calcium intake was calculated by recording 3 days diet. Puberty stage was defined as per Tanner. Descriptive statistics was used with means and standard deviations, linear correlation, and analysis of variance for comparison between age groups, and the Tukey test (p<0.05). Linear correlation was positive and indicated body weight as the main correlation variable with BMD in both studied locations (p<0.01). BMC and BMD increased with age, differences were significant from 14 to 15 years, and when adolescents reached Tanner stage G4. These results showed a pronounced increase in bone mineralization, with the years after 14 to 15 being critical for BM acquisition in Brazilian adolescents.

Role of annexin 1 gene expression in mouse craniofacial bone development

BACKGROUND: Annexin 1 is a 37-kDa protein that has complex intra- and extracellular effects. To discover whether the absence of this protein alters bone development, we monitored this event in the annexin-A1 null mice in comparison with littermate wild-type controls. METHODS: Radiographic and densitometry methods were used for the assessment of bone in annexin-A1 null mice at a gross level. We used whole-skeleton staining, histological analysis, and Western blotting techniques to monitor changes at the tissue and cellular levels. RESULTS: There were no gross differences in the appendicular skeleton between the genotypes, but an anomalous development of the skull was observed in the annexin-A1 null mice. This was characterized in the newborn annexin-A1 null animals by a delayed intramembranous ossification of the skull, incomplete fusion of the interfrontal suture and palatine bone, and the presence of an abnormal suture structure. The annexin-A1 gene was shown to be active in osteocytes during this phase and COX-2 was abundantly expressed in cartilage and bone taken from annexin-A1 null mice. CONCLUSIONS: Expression of the annexin-A1 gene is important for the normal development of the skull in mice, possibly through the regulation of osteoblast differentiation and a secondary effect on the expression of components of the cPLA2-COX-2 system. © 2007 Wiley-Liss, Inc.

The influence of ovariectomy, simvastatin and sodium alendronate on alveolar bone in rats

Bisphosphonates are currently used in the treatment of many diseases involving increased bone resorption such as osteoporosis. Statins have been widely used for the treatment of hypercholesterolemia and recent studies have shown that these drugs are also capable of stimulating bone formation. The purpose of this study was to evaluate thel influence of an estrogen deficient state and the effects of simvastatin and sodium alendronate therapies on alveolar bone in female rats. Fifty-four rats were either ovariectomized (OVX) or sham operated. A month later, the animals began to receive a daily dose of simvastatin (SIN - 25 mg/kg), sodium alendronate (ALN - 2 mg/kg) or water (control) orally. Thirty-five days after the beginning of the treatment, the rats were sacrificed and their left hemimandibles were removed and radiographed using digital X-ray equipment. The alveolar radiographic density under the first molar was determined with gray-level scaling and the values were submitted to analysis of variance (α = 5%). Ovariectomized rats gained more weight (mean ± standard deviation: 20.06 ± 6.68%) than did the sham operated animals (12.13 ± 5.63%). Alveolar radiographic density values, expressed as gray levels, were lowest in the OVX-water group (183.49 ± 6.47), and differed significantly from those observed for the groups receiving alendronate (sham-ALN: 193.85 ± 3.81; OVX-ALN: 196.06 ± 5.11) and from those of the sham-water group (193.66 ± 4.36). Other comparisons between groups did not show significant differences. It was concluded that the ovariectomy reduced alveolar bone density and that alendronate was efficient for the treatment of this condition.

Reconstruction of Mandibular Segmental Defects Using the Guided-Bone Regeneration Technique With Polylactide Membranes and/or Autogenous Bone Graft: A Preliminary Study on the Influence of Membrane Permeability

Purpose: Bone maintenance after mandibular reconstruction with autogenous iliac crest may be disappointing due to extensive resorption in the long term. The potential of the guided-bone regeneration (GBR) technique to enhance the healing process in segmental defects lacks comprehensive scientific documentation. This study aimed to investigate the influence of polylactide membrane permeability on the fate of iliac bone graft (BG) used to treat mandibular segmental defects. Materials and Methods: Unilateral 10-mm-wide segmental defects were created through the mandibles of 34 mongrel dogs. All defects were mechanically stabilized, and the animals were divided into 6 treatment groups: control, BG alone, microporous membrane (poly L/DL-lactide 80/20%) (Mi); Mi plus BG; microporous laser-perforated (15 cm2 ratio) membrane (Mip), and Mip plus BG. Calcein fluorochrome was injected intravenously at 3 months, and animal euthanasia was carried out at 6 months postoperatively. Results: Histomorphometry showed that BG protected by Mip was consistently related to larger amounts of bone compared with other groups (P ≤ .0001). No difference was found between defects treated with Mip alone and BG alone. Mi alone rendered the least bone area and reduced the amount of grafted bone to control levels. Data from bone labeling indicated that the bone formation process was incipient in the BG group at 3 months postoperatively regardless of whether or not it was covered by membrane. In contrast, GBR with Mip tended to enhance bone formation activity at 3 months. Conclusions: The use of Mip alone could be a useful alternative to BG. The combination of Mip membrane and BG efficiently delivered increased bone amounts in segmental defects compared with other treatment modalities. © 2008 American Association of Oral and Maxillofacial Surgeons.

Effects of Long-Term FK 506 Therapy on the Alveolar Bone and Cementum of Rats

Cyclosporine (CsA) and tacrolimus (FK 506) exert complex, incompletely understood actions on bone. The objective of the study was to evaluate the effects of long-term tacrolimus therapy on the periodontium. Rats were treated for 60, 120, 180, and 240 days with daily subcutaneous injections of 1 mg/kg body weight of FK 506. After the experimental period, we obtained serum levels of calcium and alkaline phosphatase (ALP). After histological processing, the alveolar bone and cementum, as well as volume densities of bone (Vb) and osteoclasts (Vo), were assessed at the regions of the lower first molar. There was a tendency toward a statistically significant decrease in ALP levels with FK 506; however, serum calcium levels increased during the long periods. At 60, 180, and 240 days of treatment with FK 506, we did not observe Vb and Vo alterations. At 120 days of treatment, there was an evident decrease in Vb, but it did not show alveolar bone loss. We did not observe any alterations of cementum among rats treated with FK 506. It may be concluded that FK 506 administration did not induce side effects on the periodontium. © 2009 Elsevier Inc. All rights reserved.

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-κβ 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. © 2009 John Wiley & Sons A/S.

Bone structure, strength and microhardness resulting after hindlimb unloading in rats

Introduction: Bone strength is influenced by a number of different determinants, such as mass, size, geometry and also by the intrinsic material properties of the tissue. Aims: The structure and mechanical properties of the femur were analyzed in aged (14 mo-old) animals submitted to hindlimb unloading (HU) for 21 days. Methods: Twenty Wistar rats were randomly divided into Control and HU groups and the femur was submitted to dual X ray absorptiometry (DXA), DIGORA radiographic density, mechanical compression testing and Knoop microhardness analyse in cortical and cancellous bone. Results: Femurs from HU group presented significantly lower failure load, decreased bone mineral density (BMD)/bone mineral content (BMC) in whole bone; proximal/distal epiphysis and middiaphyseal cortical bone measured by DXA were similar in the two groups; radiographic density from areas in proximal epiphysis was significantly lower in HU group, and microhardness measured at periosteal and endosteal levels were also signifcantly diminished in HU compared with Control group. Conclusion: Disuse induced damage in the trabecular femoral bone architecture with decisive effect on the head and trochanteric fossa, which became weaker. Bone diaphyseal cortical hardness also suffered effect of unloading, probably related to osteocyte/osteoblast activity.

In vitro analysis with human bone marrow stem cells on Ti-15Mo alloy for dental and orthopedic implants application

Aim: Nowadays, research on orthopedic and dental implants is focused on titanium alloys for their mechanical properties and corrosion resistance in the human body environment. Another important aspect to be investigated is their surface topography, which is very important to osseointegration. With laser beam irradiation for roughening the implants surface an easier control of the microtopography is achieved, and surface contamination is avoided. The aim of this study was to assess human bone marrow stem cells response to a newly developed titanium alloy, Ti-15Mo, with surface topography modified by laser beam irradiation. Materials and methods: A total of 10 Ti machined disks (control), 10 Ti-15Mo machined disks and 10 Ti-15Mo disks treated by laser beam-irradiation were prepared. To study how Ti-15Mo surface topografy can induce osteoblast differentiation in mesenchymal stem cells, the expression levels of bone related genes and mesenchymal stem cells marker were analyzed, using real time Reverse Transcription-Polymerase Chain Reaction. Results: In Test 1 (comparison between Ti-15Mo machined disks and Ti-machined disks) quantitative real-time RT-PCR showed a significant induction of ALPL, FOSL1 and SPP1, which increase 20% or more. In Test 2 (comparison between Ti-15Mo laser treated disks and Ti-machined disks) all investigated genes were up-regulated. By comparing Test 1 and Test 2 it was detected that COL1A1, COL3A1, FOSL1 and ENG sensibly increased their expression whereas RUNX2, ALPL and SPP1 expression remained substantially unchanged. Conclusion: The present study demonstrated that laser treated Ti-15Mo alloys are promising materials for implants application.

Factors that contribute to low bone density in postmenopausal women in different Amazonian communities

Background: The aim of this study was to verify socioeconomic differences, nutrition, body balance and quality of life (QoL) in postmenopausal women with low bone mineral density (BMD) in two Amazonian communities. Methods: A total of 42 female volunteers participated in the study. The volunteers were separated into two groups: Villa (n= 20; 53±5.5 years) and City (n= 22; 56±7.9 years). The following evaluation instruments were used: dual energy X-ray absorptiometry (DXA); a socioeconomic questionnaire; a QoL questionnaire; a dietary habits questionnaire; and a balance test. Parametric and nonparametric tests were used. Results: The data showed significant differences in socioeconomic level (Δ%=+15.9%, p=0.000),lumbar spine L2-L4 (Δ%=+0.10%,p=0.007), balance(Δ%=+4.3%,p=0.03)and some important aspects of nutrition, such as the consumption of milk (Δ%=+34%, p=0.01) and alcohol (+14.8%, p=0.0001). These significant differences also contributed to the total QoL score (Δ%=+76.2%, p=0.000) and the majority of the QoL-related functions. Conclusion: This study verified that socioeconomic level, nutritional status, physical activity levels and QoL can influence the BMD of postmenopausal women. The study suggests new strategies for official health organizations to use in order to prevent and treat osteoporosis. In addition, this study can provide an orientation to physical activity, nutrition and medical professionals. © The Author(s), 2011.

Physiologic values of broiler femurs at different growth phases using Bone densitometry and Bone Breaking Strength

The objective of this experiment was to determine the normal values of Bone Radiographic Density (BRD) by using the optical densitometry in radiographic images and the Bone Breaking Strength (BBS) of broiler femurs at different ages (8, 22 and 42 d of age). A total of 60 Cobb male broilers were distributed in three age groups of 20 birds. The BRD and the BBS (maxim force and rigidity) values increased (p<0.01) over the course of ages, presenting greater values at 42 d of age when comparing to 8 and 22 d of age, evidencing a biomechanical adaptation of femur to growth. This experiment offers results that can be used in other experiments of broilers fed with different nutritional levels and they can also be related to pathological values, allowing the diagnosis of diseases that affect the integrity of the poultry leg. © Asian Network for Scientific Information, 2011.

Muscular strength, bone density and body composition of a woman with systemic lupus erythematosus submitted to a resistance training program: A case report

Introduction. Physical activity can provide long-term benefits for systemic lupus erythematosus (SLE). Objective. This study sought to demonstrate the effects of progressive resistance training on the muscular strength, bone mineral density (BMD) and body composition of pre-menopausal women with SLE undergoing glucocorticoid (GC) treatment. Materials and Methods. This is the case report of a 43-year-old African-South American premenopausal woman with non-extensive SLE and low bone density. A six-month program with three bimonthly cycles of 70%, 80%, and 90% intensity according to the 10 maximum-repetition test was used. Dual-energy X-ray absorptiometry (DXA) was used to measure the BMD, T-scores and body composition, and indirect fluorescence was used to measure the levels of antinuclear antibodies. Student's t-test was used. Results. Statistical improvement was noted in all strength exercises, including the 45° leg press (Δ%=+50%, p<0.001) and knee extension (Δ%=+15%, p=0.003) to maintain the BMD of the L2-L4 lumbar (Δ%=+0.031%; p=0.46) as well as the trochanter (Δ%=+0.037%; p=0.31) and BMI (Δ%=-0.8, p=0.54). Conclusion. In this case study, the presented methodology had a positive effect on strength and contributed to the maintenance of BMD and body composition in a woman with SLE undergoing GC treatment. © 2012 Revista Andaluza de Medicina del Deporte.

Arterial hypertension perpetuates alveolar bone loss

Few studies have focused on the impact of hypertension on the progression of periodontitis (PD). The purpose of this study was to evaluate whether hypertension affects PD by enhancing bone loss even after the stimulus for PD induction is removed. Ligature-induced PD was created on the first mandibular molars of spontaneously hypertensive rats (SHR) and normotensive rats (Wistar Kyoto-WKY). The animals were assigned to non-ligated controls (C) and PD groups: WKY-C, WKY-PD, SHR-C, and SHR-PD. After 10 days, five animals of each group were killed and the ligatures of the other animals were removed. On the 21st day (11 days without PD induced), the remaining animals were killed. The jaws were defleshed and the amount of bone loss was measured. After 10 days, the PD groups showed more bone loss than its controls (P < .05); SHR-PD = 0.72 ± 0.05 mm, SHR-C = 0.39 ± 0.04 mm, WKY-PD = 0.75 ± 0.04 mm, and WKY-C = 0.56 ± 0.04 mm. The cumulative bone loss on day 21 (0.94 ± 0.13 mm) was significantly worse than on day 10 only in SHR-PD group (P < .05). The final bone loss differences between PD and C groups accounted for 102% (SHR) and 26% (WKY) increase in comparison with the initial control levels. Hypertension is associated with progressive alveolar bone loss even when the stimulus for PD induction is removed and it may be speculated that host condition perpetuates alveolar bone loss. © 2013 Informa Healthcare USA, Inc.

Autogenous bone combined with anorganic bovine bone for maxillary sinus augmentation: Analysis of the osteogenic potential of cells derived from the donor and the grafted sites

Objectives: This study aimed to comparatively evaluate the in vitro osteogenic potential of cells obtained from the mandibular ramus (MR, autogenous bone donor site) and from the maxillary sinus (MS) bone grafted with a mixture of anorganic bovine bone (ABB) and MR prior to titanium implant placement (MS, grafted implant site). Material and methods: Cells were obtained from three patients subjected to MS floor augmentation with a 1: 1 mixture of ABB (GenOx Inorg®) and MR. At the time of the sinus lift procedure and after 8 months, prior to implant placement, bone fragments were taken from MR and MS, respectively, and subjected to trypsin-collagenase digestion for primary cell culturing. Subcultured cells were grown under osteogenic condition for up to 21 days and assayed for proliferation/viability, osteoblast marker mRNA levels, alkaline phosphatase (ALP) activity and calcium content/Alizarin red staining. ALP activity was also determined in primary explant cultures exposed to GenOx Inorg® (1: 1 with MR) for 7 days. Data were compared using either the Mann-Whitney U-test or the Kruskal-Wallis test. Results: MS cultures exhibited a significantly lower osteogenic potential compared with MR cultures, with a progressive increase in cell proliferation together with a decrease in osteoblast markers, reduced ALP activity and calcium content. Exposure of MR-derived primary cultures to GenOx Inorg® inhibited ALP activity. Conclusion: These results suggest that the use of GenOx Inorg® in combination with MR fragments for MS floor augmentation inhibits the osteoblast cell differentiation at the implant site in the long term. © 2013 John Wiley & Sons A/S.

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.