Contents of major components, U, and Th in bones and bone debris

Uranium and thorium contents, as well as their distribution patterns have been studied in biogenic phosphates from the Atlantic and Pacific Oceans. Differently lithified fish remains (bones, scales, teeth) and marine mammal bones (ribs, vertebras, earbones) collected from both reduced shelf sediments and oxidized pelagic ones have been analyzed. U content in the material varies from 0.7 to 700 ppm, and Th content - from <0.5 to 14 ppm. U/Th ratio varies from 0.16 to 400. Contents of both elements increase with lithification of biogenic phosphates. U concentration is more intense on shelves, whereas thorium concentration increases in pelagic areas. Partial positive correlation of U and Th with Fe and negative correlation of U with organic carbon are noted. The latter corresponds to higher lithification of biogenic phosphates. Calcium phosphate transformed from hydroxyapatite to fluorcarbonate-apatite is the main carrier of U, while transformed organic matter is a minor agent. Thorium is mainly bound with Fe.

The role of bone debris in early healing adjacent to hydrophilic and hydrophobic implant surfaces in man

To evaluate morphologically and morphometrically the sequential healing and osseointegration events at moderately rough implant surfaces with and without chemical modification. Particularly the role of bone debris in initiating bone formation was emphasized.

Avaliação dos enxertos ósseos autógenos triturados manualmente ou coletados durante osteotomia: análise histológica e histométrica em coelhos

Pós-graduação em Odontologia - FOA

Bone-healing pattern at the surface of titanium implants: An experimental study in the dog

Objective: To study the early sequential stages of osseointegration at implants installed in alveolar bony. Materials 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. Implants were installed, and the flaps were sutured to allow a fully submerged healing. The timing of the installations in the left side of the mandible and of sacrifices were performed with a schedule that various observation periods to sacrifice from 5, 10, 20, and 30 days were available so that n = 6 was obtained per each healing period. Ground sections were prepared and analyzed. Results: Newly formed bone in contact with the implant surface was found after 10 days of healing and the percentage increased up to 50% after 1 month of healing. A higher percentage was found in the trabecular compared with the cortical bony compartment. Old bone decreased by about 50% during healing, being still present after 1 month (16%). The proportions of bone debris and bone particles were at 27% after 5 days and decreased during healing to 6% after 1 month. Conclusion: Osseointegration (new bone-to-implant contact) developed at various rates for cortical and trabecular compartments, respectively. In the trabecular region, mesenchymal cells were identified, subsequently developing into new bone in contact with the implant surface. In the cortical compartment, however, resorptive processes were observed throughout all periods of healing. The proportion of newly formed bone percentage was lower compared with that of the trabecular area. Old bone was still present after 1 month of healing in both compartments. Bone debris and small bone particles appeared to be involved in initial bone formation. © 2013 John Wiley & Sons A/S.

Preparation of intact bovine tail intervertebral discs for organ culture

The intervertebral disc (IVD) is the joint of the spine connecting vertebra to vertebra. It functions to transmit loading of the spine and give flexibility to the spine. It composes of three compartments: the innermost nucleus pulposus (NP) encompassing by the annulus fibrosus (AF), and two cartilaginous endplates connecting the NP and AF to the vertebral body on both sides. Discogenic pain possibly caused by degenerative intervertebral disc disease (DDD) and disc herniations has been identified as a major problem in our modern society. To study possible mechanisms of IVD degeneration, in vitro organ culture systems with live disc cells are highly appealing. The in vitro culture of intact bovine coccygeal IVDs has advanced to a relevant model system, which allows the study of mechano-biological aspects in a well-controlled physiological and mechanical environment. Bovine tail IVDs can be obtained relatively easy in higher numbers and are very similar to the human lumbar IVDs with respect to cell density, cell population and dimensions. However, previous bovine caudal IVD harvesting techniques retaining cartilaginous endplates and bony endplates failed after 1-2 days of culture since the nutrition pathways were obviously blocked by clotted blood. IVDs are the biggest avascular organs, thus, the nutrients to the cells in the NP are solely dependent on diffusion via the capillary buds from the adjacent vertebral body. Presence of bone debris and clotted blood on the endplate surfaces can hinder nutrient diffusion into the center of the disc and compromise cell viability. Our group established a relatively quick protocol to "crack"-out the IVDs from the tail with a low risk for contamination. We are able to permeabilize the freshly-cut bony endplate surfaces by using a surgical jet lavage system, which removes the blood clots and cutting debris and very efficiently reopens the nutrition diffusion pathway to the center of the IVD. The presence of growth plates on both sides of the vertebral bone has to be avoided and to be removed prior to culture. In this video, we outline the crucial steps during preparation and demonstrate the key to a successful organ culture maintaining high cell viability for 14 days under free swelling culture. The culture time could be extended when appropriate mechanical environment can be maintained by using mechanical loading bioreactor. The technique demonstrated here can be extended to other animal species such as porcine, ovine and leporine caudal and lumbar IVD isolation.

Rare earth and other element contents in surface layer bottom sediments and ferromanganese nodules along the Transatlantic profile

Behavior of rare earth elements (REE) and Th is studied along the Transatlantic transect at 22°N. It is shown that both REE and Th contents relative to Al (the most lithogenic element) increase toward the pelagic region. The increasing trend becomes more complicated due to variations in content of biogenic calcium carbonate that acts as a diluting component in sediments. REE composition varies symmetrically relative to the Mid-Atlantic Ridge (MAR) emphasizing weak hydrothermal influence on sediments of the ridge axis, although the well-known criteria for hydrothermal contribution, such as Al/(Al+Mn+Fe) and (Fe+Mn)/Ti, do not reach critical values. Variations in REE content and composition allowed to distinguish the following five sediment zones in the transect: (I) terrigenous sediments of the Nares abyssal plain; (II) pelagic sediments of the North American Basin; (III) carbonate ooze of the MAR axis; (IV) pelagic sediments of the Canary Basin; and (V) terrigenous clay and calcareous mud of the African continental slope and slope base. Ferromanganese nodules of the hydrogenous type with extremely high Ce (up to 1801 ppm) and Th (up to 138 ppm) contents occur in pelagic sediments. It is ascertained that P, REE, and Th contents depend on Fe content in Atlantic sediments. Therefore, one can suggest that only minor amount of phosphorus is bound with bone debris. Low concentration of bone debris phosphorus is a result of relatively high sedimentation rates in the Atlantic Ocean, as compared with those in pelagic regions of the Pacific Ocean.

(Table 1) Results of X-ray radiographic analyses of authigenic phosphorites from the Southwest Africa shelf

In the biologically productive area on the Southwest Africa shelf a current process of diagenetic phosphorite formation occurs. Remains of phytoplankton are the main source of phosphorus in host bottom sediments. Phosphorus concentrates in nodules due to its redistribution in bottom sediments and precipitation from interstitial waters. Accumulation of the phosphate nodules, as well as of scattered fish bone debris in sediments occurs at sudden change in hydrological conditions and removal of fine sediment fractions due to increased bottom currents or during transgressions.

Evaluation of autogenous bone grafts, particulate or collected during osteotomy with implant burs: Histologic and histomorphometric analysis in rabbits

Purpose: the aim of this study was to evaluate bone regeneration in bone cavities filled with particulate autogenous bone either harvest in blocks and subjected to milling procedures or collected during osteotomy with implant burs. Materials and Methods: In 12 rabbits, 3 noncritical unicortical cavities 7 mm in diameter were prepared with a trephine drill on the right tibia. The cavities were filled respectively with particulate autogenous bone achieved with a manual bone crusher ( particulate group), with particulate autogenous bone obtained using bone collector during osteotomy ( collected group), and with blood clot ( control group). Animals were sacrificed at 7, 15, and 30 days after surgery ( 4 animals for each time period). The sections were examined by histologic and histomorphometric analysis. Results: At 7 days, the samples were filled by coagulum, and bone particles were observed only in the collected (24%) and particulate groups (44.75%). At 15 days, there was connective differentiation in all groups, with presence of grafted bone particles and onset of newly formed bone in the collected (38.88%) and particulate groups (46.0%). At 30 days, there was bone fill ( immature trabecular bone) of the cavities in the control (50%), collected (64.63%) and particulate groups (66%). Conclusion: No significant difference was demonstrated between noncritical unicortical bone defects in rabbit tibiae filled with particulate bone harvested as a block and subjected to milling and those filled with bone collected during osteotomy with implant drills when the defects were observed up to 30 days following their creation.

Results of analyses of unmelted meteoritic debris collected from Eltanin ejecta in three sediment core from POLARSTERN cruise ANT-XII/4

A total of 1.7 g of unmelted meteorite particles have been recovered from FS Polarstern piston cores collected on expedition ANT XII/4 that contain ejecta from the Eltanin impact event. Most of the mass (1.2 g) is a large, single specimen that is a polymict breccia, similar in mineralogy and chemistry to howardites or the silicate fraction of mesosiderites. Most of the remaining mass is in several large individual pieces (20-75mg each) that are polymict breccias, fragments dominated by pyroxene, and an igneous rock fragment. The latter has highly fractionated REE, similar to those reported in mafic clasts from mesosiderites. Other types of specimens identified include fragments dominated by maskelynite or olivine. These pieces of the projectile probably survived impact by being blown off the back surface of the Eltanin asteroid during its impact into the Bellingshausen Sea.

Bone response to unloaded titanium implants in the fibula, iliac crest, and scapula: an animal study in the Yorkshire pig

The reconstruction of extended maxillary and mandibular defects with prefabricated free flaps is a two stage procedure, that allows immediate function with implant supported dentures. The appropriate delay between prefabrication and reconstruction depends on the interfacial strength of the bone–implant surface. The purpose of this animal study was to evaluate the removal torque of unloaded titanium implants in the fibula, the scapula and the iliac crest. Ninety implants with a sandblasted and acid-etched (SLA) surface were tested after healing periods of 3, 6, and 12 weeks, respectively. Removal torque values (RTV) were collected using a computerized counterclockwise torque driver. The bicortical anchored 8 mm implants in the fibula revealed values of 63.73 Ncm, 91.50 Ncm, and 101.83 Ncm at 3, 6, and 12 weeks, respectively. The monocortical anchorage in the iliac crest showed values of 71.40 Ncm, 63.14 Ncm, and 61.59 Ncm with 12 mm implants at the corresponding times. The monocortical anchorage in the scapula demonstrated mean RTV of 62.28 Ncm, 97.63 Ncm, and 99.7 Ncm with 12 mm implants at 3, 6, and 12 weeks, respectively. The study showed an increase of removal torque with increasing healing time. The interfacial strength for bicortical anchored 8 mm implants in the fibula was comparable to monocortical anchored 12 mm implants in the iliac crest and the scapula at the corresponding times. The resistance to shear seemed to be determined by the type of anchorage (monocortical vs. bicortical) and the length of the implant with greater amount of bone–implant interface.

Cross-talk of subchondral bone osteoblasts and articular cartilage chondrocytes : a new insight in understanding osteoarthritis pathogenesis

Osteoarthritis (OA) is the most common musculoskeletal disorder and represents a major health burden to society. In the course of the pathological development of OA, articular cartilage chondrocytes (ACCs) undergo atypical phenotype changes characterized by the expression of hypertrophic differentiation markers. Also, the adjacent subchondral bone shows signs of abnormal mineral density and enhanced production of bone turnover markers, indicative of osteoblast dysfunction. Collectively these findings indicate that the pathological changes typical of OA, involve alterations of the phenotypic properties of cells in both the subchondral bone and articular cartilage. However, the mechanism(s) by which these changes occur during OA development are not completely understood. The purpose of this project was to address the question of how subchondral bone osteoblasts (SBOs) and ACCs interact with each other with respect to regulation of respective cells’ phenotypic properties and in particular the involvement of mitogen activated protein kinase (MAPK) signalling pathways under normal and OA joint condition. We also endeavoured to test the influence of cross-talk between SBOs and ACCs isolated from normal and OA joint on matrix metalloproteinase (MMP) expression. For this purpose tissues from the knees of OA patients and normal controls were collected to isolate SBOs and ACCs. The cellular cross-talk of SBOs and ACCs were studied by means of both direct and indirect co-culture systems, which made it possible to identify the role of both membrane bound and soluble factors. Histology, immunohistochemistry, qRT-PCR, zymography, ELISA and western blotting were some of the techniques applied to distinguish the changes in the co-cultured vs. non co-cultured cells. The MAPK signalling pathways were probed by using targeted MAPK inhibitors, and their activity monitored by western blot analysis using phospho MAPK specific antibodies. Our co-culture studies demonstrated that OA ACCs enhanced the SBOs differentiation compared to normal ACCs. We demonstrated that OA ACCs induced these phenotypic changes in the SBOs via activating an ERK1/2 signalling pathway. The findings from this study thus provided clear evidence that OA ACCs play an integral role in altering the SBO phenotype. In the second study, we tested the influence of normal SBOs and OA SBOs on ACCs phenotype changes. The results showed that OA SBOs increased the hypertrophic gene expression in co-cultured ACCs compared to normal SBOs, a phenotype which is considered as pathological to the health and integrity of articular cartilage. It was demonstrated that these phenotype changes occurred via de-activation of p38 and activation of ERK1/2 signaling pathways. These findings suggest that the pathological interaction of OA SBOs with ACCs is mediated by cross-talking between ERK1/2 and p38 pathways, resulting in ACCs undergoing hypertrophic differentiation. Subsequent experiments to determine the effect on MMP regulation, of SBOs and ACCs cross-talk, revealed that co-culturing OA SBOs with ACCs significantly enhanced the proteolytic activity and expression of MMP-2 and MMP-9. In turn, co-culture of OA ACCs with SBOs led to abundant MMP-2 expression in SBOs. Furthermore, we showed that the addition of ERK1/2 and JNK inhibitors reversed the elevated MMP-2 and MMP-9 production which otherwise resulted from the interactions of OA SBOs-ACCs. Thus, this study has demonstrated that the altered interactions between OA SBOs-ACCs are capable of triggering the pathological pathways leading to degenerative changes seen in the osteoarthritic joint. In conclusion, the body of work presented in this dissertation has given clear in vitro evidence that the altered bi-directional communication of SBOs and ACCs may play a role in OA development and that this process was mediated by MAPK signalling pathways. Targeting these altered interactions by the use of MAPK inhibitors may provide the scientific rationale for the development of novel therapeutic strategies in the treatment and management of OA.

Serum bone formation marker correlation with improved osseointegration in osteoporotic rats treated with simvastatin

Objective: Simvastatin has been shown to enhance osseointegration of pure titanium implants in osteoporotic rats. This study aimed to evaluate the relationship between the serum level of bone formation markers and the osseointegration of pure titanium implants in osteoporotic rats treated with simvastatin. Materials and methods: Fifty-four female Sprague Dawley rats, aged 3 months old, were randomly divided into three groups: Sham-operated group (SHAM; n=18), ovariectomized group (OVX; n=18), and ovariectomized with Simvastatin treatment group (OVX+SIM; n=18). Fifty-six days after ovariectomy, screw-shaped titanium implants were inserted into the tibiae. Simvastatin was administered orally at 5mg/kg each day after the placement of the implant in the OVX+SIM group. The animals were sacrificed at either 28 or 84 days after implantation and the undecalcified tissue sections were processed for histological analysis. Total alkaline phosphatase (ALP), bone specific alkaline phosphatase (BALP) and bone Gla protein (BGP) were measured in all animal sera collected at the time of euthanasia and correlated with the histological assessment of osseointegration. Results: The level of ALP in the OVX group was higher than the SHAM group at day 28, with no differences between the three groups at day 84. The level of BALP in the OVX+SIM group was significantly higher than both OVX and SHAM groups at days 28 and 84. Compared with day 28, the BALP level of all three groups showed a significant decrease at day 84. There were no significant differences in BGP levels between the three groups at day 28, but at day 84 the OVX+SIM group showed significantly higher levels than both the OVX and SHAM groups. There was a significant increase in BGP levels between days 28 and 84 in the OVX+SIM group. The serum bone marker levels correlated with the histological assessment showing reduced osseointegration in the OVX compared to the SHAM group which is subsequently reversed in the OVX+SIM group.