Fraser McDonald

Physical and chemical properties of a new root-end filling material.

This study determined the chemical composition, pH, and radiopacity of mineral trioxide aggregate (MTA), and also compared the setting time, compressive strength, and solubility of this material with those of amalgam, Super-EBA, and Intermediate Restorative Material (IRM). X-ray energy dispersive spectrometer in conjunction with the scanning electron microscope were used to determine the composition of MTA, and the pH value of MTA was assessed with a pH meter using a temperature-compensated electrode. The radiopacity of MTA was determined according to the method described by the International Organization for Standardization. The setting time and compressive strength of these materials were determined according to methods recommended by the British Standards Institution. The degree of solubility of the materials was assessed according to modified American Dental Association specifications. The results showed that the main molecules present in MTA are calcium and phosphorous ions. In addition, MTA has a pH of 10.2 initially, which rises to 12.5 three hours after mixing. MTA is more radiopaque than Super-EBA and IRM. Amalgam had the shortest setting time (4 min) and MTA the longest (2 h 45 min). At 24 h MTA had the lowest compressive strength (40 MPa) among the materials, but it increased after 21 days to 67 MPa. Finally, except for IRM, none of the materials tested showed any solubility under the conditions of this study.

Cellular response to mineral trioxide aggregate

This investigation studied the cytomorphology of osteoblasts in the presence of Mineral Trioxide Aggregate (MTA) and examined cytokine production. MTA and Intermediate Restorative Material (IRM) were prepared and placed in separate Petri dishes. Osteoblasts (cell-line MG-63), grown to confluence in Hams F12/Dulbeccos modified Eagles medium, were seeded into the dishes, which were incubated for 1 to 7 days. The specimens were viewed by scanning electron microscopy. For cytokine evaluation, cells were grown either alone or in other dishes containing the test materials for 1 to 144 h. Media were removed for ELISA analysis of interleukin (IL)-1 alpha, IL-1 beta, IL-6, and macrophage colony-stimulating factor. Scanning electron microscopy revealed healthy cells in contact with MTA at 1 and 3 days; in contrast, cells in the presence of IRM appeared rounded. The ELISA assays revealed raised levels of all ILs at all periods when cells were grown in the presence of MTA; in contrast, cells grown alone or with IRM produced undetectable amounts. The macrophage colony-stimulating factor was produced by cells irrespective of the group. It seems that MTA offers a biologically active substrate for bone cells and stimulates IL production.

Mineral trioxide aggregate stimulates a biological response in human osteoblasts

We report a novel material that appears to stimulate cytokine production in human osteoblasts and allow good adherence of the cells to the material. We have examined cultured osteoblasts (MG-63) in the presence of mineral trioxide aggregate (MTA) as set in moist conditions; secondly, we examined the behavior of these MG-63 cells with respect to cytokine and osteocalcin production and alkaline phosphatase activity. Standard ELISA assays were used for assessment of interleukin (IL)-1 alpha, IL-1 beta, IL-6, macrophage colony stimulating factor (M-CSF), and osteocalcin. Furthermore the levels of alkaline phosphatase were measured to establish the level of differentiation of the cells. Cells without MTA served as controls. Cells also were grown in the presence of polymethylmethacrylate (PMA), the commonly used orthopedic cement. In all dishes cells were seen adhering to the base and MTA at 6 h and had increased to confluence at 144 h. IL-1 alpha (175.1 +/- 32.6 pg/mL), IL-1 beta (154.0 +/- 26.7 pg/mL), and IL-6 (214.7 +/- 21.8 pg/mL) were raised when the cells were grown in the presence of MTA at 144 h, with raised values at all time intervals. M-CSF appeared to be unaffected although the overall value was high (7,045.0 +/- 89.5 pg/mL). In contrast, cells grown in the absence of MTA produced negligible amounts of these cytokines (< pg/mL) as did those cells grown in the presence of PMA. Osteocalcin production increased when cells were grown on MTA from 3.8 +/- 0.87 ng/mL to 19.7 +/- 2.8 ng/mL. No osteocalcin could be detected with PMA. Cells in contact with MTA also appeared to have levels of alkaline phosphatase similar to those reported elsewhere (4.3 +/- 0.21 mumol/mg protein/min). No cells could be found attached to PMA and so no alkaline phosphatase activity could be measured.

Osteoblast biocompatibility of mineral trioxide aggregate

This study investigated the biocompatibility of variants of mineral trioxide aggregate (MTA), by culturing human MG63 osteosarcoma cells in the presence of materials, observing cytomorphology and cell growth, and then assaying cytokine expression from the cells. Reference materials were employed. Cell growth was quantified by preparing samples (n = 6) at 2, 4 and 7 days, for viewing by scanning electron microscopy and then scoring the amount of material that was covered by healthy cells. Subsequently, samples of culture media were tested using ELISA assays for expression of Interleukin (IL)-1alpha, IL-6, IL-8, IL-11 and macrophage colony stimulating factor (M-CSF). These assays were compared with controls where no material was present, and where media and fetal calf serum had not been exposed to cells. Results showed good cell growth on MTA. Expression of IL-6 from cells was only evident in the presence of MTA and Interpore 200. Interleukin-8 was expressed in high concentrations only in the presence of MTA. There was no evidence of expression of IL-1alpha or IL-11 with any material. Production of M-CSF was high for all materials. It appears that the variants of MTA are biocompatible and suitable for use in clinical trials.

An evaluation of accelerated Portland cement as a restorative material

Biocompatibility of two variants of accelerated Portland cement (APC) were investigated in vitro by observing the cytomorphology of SaOS-2 osteosarcoma cells in the presence of test materials and the effect of these materials on the expression of markers of bone remodelling. Glass ionomer cement (GIC), mineral trioxide aggregate (MTA) and unmodified Portland cement (RC) were used for comparison. A direct contact assay was undertaken in four samples of each test material, collected at 12, 24, 48 and 72 h. Cell morphology was observed using scanning electron microscopy (SEM) and scored. Culture media were collected for cytokine quantification using enzyme-linked immunosorbent assay (ELISA). On SEM evaluation, healthy SaOS-2 cells were found adhering onto the surfaces of APC variant, RC and MTA. In contrast, rounded and dying cells were observed on GIC. Using ELISA, levels of interleukin (IL)-1beta, IL-6, IL-18 and OC were significantly higher in APC variants compared with controls and GIC (p<0.01), but these levels of cytokines were not statistically significant compared with MTA. The results of this study provide evidence that both APC variants are non-toxic and may have potential to promote bone healing. Further development of APC is indicated to produce a viable dental restorative material and possibly a material for orthopaedic

Regulation of InsP3 receptor activity by neuronal Ca2+‐binding proteins

Inositol 1,4,5‐trisphosphate receptors (InsP3Rs) were recently demonstrated to be activated independently of InsP3 by a family of calmodulin (CaM)‐like neuronal Ca2+‐binding proteins (CaBPs). We investigated the interaction of both naturally occurring long and short CaBP1 isoforms with InsP3Rs, and their functional effects on InsP3R‐evoked Ca2+ signals. Using several experimental paradigms, including transient expression in COS cells, acute injection of recombinant protein into Xenopus oocytes and 45Ca2+ flux from permeabilised COS cells, we demonstrated that CaBPs decrease the sensitivity of InsP3‐induced Ca2+ release (IICR). In addition, we found a Ca2+‐independent interaction between CaBP1 and the NH2‐terminal 159 amino acids of the type 1 InsP3R. This interaction resulted in decreased InsP3 binding to the receptor reminiscent of that observed for CaM. Unlike CaM, however, CaBPs do not inhibit ryanodine receptors, have a higher affinity for InsP3Rs and more potently inhibited IICR. We also show that phosphorylation of CaBP1 at a casein kinase 2 consensus site regulates its inhibition of IICR. Our data suggest that CaBPs are endogenous regulators of InsP3Rs tuning the sensitivity of cells to InsP3.

Endothelin-1-Stimulated InsP3-Induced Ca2+ Release Is a Nexus for Hypertrophic Signaling in Cardiac Myocytes

Ca(2+) elevations are fundamental to cardiac physiology-stimulating contraction and regulating the gene transcription that underlies hypertrophy. How Ca(2+) specifically controls gene transcription on the background of the rhythmic Ca(2+) increases required for contraction is not fully understood. Here we identify a hypertrophy-signaling module in cardiac myocytes that explains how Ca(2+) discretely regulates myocyte hypertrophy and contraction. We show that endothelin-1 (ET-1) stimulates InsP(3)-induced Ca(2+) release (IICR) from perinuclear InsP(3)Rs, causing an elevation in nuclear Ca(2+). Significantly, we show that IICR, but not global Ca(2+) elevations associated with myocyte contraction, couple to the calcineurin (CnA)/NFAT pathway to induce hypertrophy. Moreover, we found that activation of the CnA/NFAT pathway and hypertrophy by isoproterenol and BayK8644, which enhance global Ca(2+) fluxes, was also dependent on IICR and nuclear Ca(2+) elevations. The activation of IICR by these activity-enhancing mediators was explained by their ability to stimulate secretion of autocrine/paracrine ET-1.

An evaluation of slot size in orthodontic brackets--are standards as expected?

The slots of five upper left central incisor brackets from 11 commercially available bracket systems (3M Unitek, Monrovia, Calif: Twin Torque Roth, Clarity MBT, and Victory Series MBT; Dentarum, Pforzheim, Germany: Discovery Roth (0.56 mm) and Elegance Plastic Roth; Forestadent, Pforzheim, Germany: Mini Mono MBT; TP LaPorte, Indiana: Nu-Edge Roth and Mxi Advant-Edge Roth; Ormco Corp., Orange, Calif: Damon II SL Roth; Ortho Organizers, San Marcos, Calif: Elite Mini Opti-MIM Roth and Elite Mini Opti-MIM MBT) were measured in the 0.022-inch (0.5588 mm) dimension. Measurements were taken after operator calibration, and a digital readout was produced. Results indicate that all bracket slots are oversized. Three bracket systems slots (Twin Torque, Clarity, and Mini Mono) were within 5% (+/-1.08, 1.655, 1.75) of their stated dimensions with essentially parallel slot walls. The Elegance Plastic slot was parallel sided but oversized by 12% (+/-1.15). The geometry of bracket slots was also variable. The Victory Series slot was slightly divergent with the top oversized by 6% (+/-1.035). The Nu-Edge slot was divergent and slot top oversized by 14% (+/-1.32). The Mxi Advant-Edge, Damon II SL, Elite Mini Opti-MIM Roth, and MBT were all convergent, and the base of the Damon slot was oversized by 17% (+/-1.79). The Discovery bracket was convergent, and the slot base was oversized by 24% (+/-1.255), which was the largest recorded variance. This bracket also had a 7% difference between the widths of the slot top and the base. Inaccurate machining of bracket slot dimensions and the use of undersized archwires may directly and adversely affect three-dimensional tooth positioning.

Analysis of the vertical facial form in patients with severe hypodontia

We examined the lateral cephalograms of Russian patients in the following categories: control with acceptable occlusions (group 1); severe hypodontia with absence of six or more teeth (group 2); and severe hypodontia associated with hypohidrotic ectodermal dysplasia (HED) (group 3). Analysis was in a cross-sectional manner, comparing dimensions at the start of the mixed dentition phase (age 6-10) and in the permanent dentition (age 12-18). The groups were matched for age and sex. Thirty-one hard- and soft-tissue landmarks were traced, and 35 linear, 19 angular, and 7 ratioed measurements were taken and compared, using analysis of variance to compare the means of each group. A reduced anterior face height was found in groups 2 and 3 as a consequence of a reduced anterior lower face height. In group 2 in the mixed dentition, the posterior face height was also reduced. The inclination of the mandible (<Se S Go Gn) was significantly reduced to 28.22 degrees +/- 0.71 degrees in group 2 and to 24.07 degrees +/- 0.97 degrees in group 3. The facial profile appeared flat or concave (<se pn pg was increased up to 8.42 degrees +/- 1.56 degrees in children and 16.81 degrees +/- 2.18 degrees in adolescents). The subnasion point was behind the aesthetic line (EL), and in group 2 patients the naso-labial angle was obtuse when compared to nonaffected patients. In group 3 patients, the naso-labial angle became acute and lips were protuberant and everted as a consequence of the reduced vertical height. Groups 2 and 3 have the typical facial characteristics unique to hypodontia, with reduced vertical dimensions as a consequence of limited alveolar bone growth. However, group 3 patients have a unique abnormal craniofacial development.

Static magnetic field effects on the sagittal suture in Rattus Norvegicus

Twenty-day-old Wistar albino rats were exposed to static magnetic fields by placing a neodymium-iron-boron magnetic over their sagittal suture. Cellular activity was monitored by the uptake of tritiated thymidine in control, north, south, and unoperated animals at 1, 3, 5, and 10 days (n = 10 per group). A total of 160 animals were used for this part of the study, with the animals examined 1, 3, 5, and 10 days after surgery. Bone remodeling was examined by tetracycline fluorescence with 10 animals allocated to 5- and 10-day periods for north and south poles (n = 10 per group) and control experiments. This consisted of the placement of unmagnetized alloy, similar in size and shape to the magnets, and also included unoperated animals (n = 5 per group). A total of 60 animals were used for the tetracycline study and were examined at 5 and 10 days after surgery. While the tetracycline examination revealed very little change, the thymidine reflected a reduction in thymidine uptake subsequent to placement of the magnet, reaching a maximal effect at 3 days and returning to a normal value thereafter. This questions the potential of static magnetic fields affecting cell mitotic activity as previously reported.