Cytotoxicity and Dentin Permeability of Carbamide Peroxide and Hydrogen Peroxide Vital Bleaching Materials, in vitro

There has been recent concern about the inadvertent exposure of dentin with patent tubules as well as gingiva to bleaching systems containing 10-15% carbamide peroxide or 2-10% hydrogen peroxide for more than a few minutes. The aims of the present study were: (1) to determine the cytotoxicity of dilutions of hydrogen peroxide in cell culture; (2) to measure hydrogen peroxide diffusion from bleaching agents through dentin in vitro; and (3) to determine the risk of hydrogen peroxide-induced cytotoxicity from exposure of dentin to these vital bleaching agents. The 50% inhibitory dose (ID50) of hydrogen peroxide to succinyl dehydrogenase activity in cultured cells was found to be 0.58 mmol/L after 1 h. All bleaching materials demonstrated diffusion of hydrogen peroxide through dentin in an "in vitro pulp chamber" device. The one- and six-hour diffusates of all bleaching agents through 0.5-mm dentin exceeded the ID50 in monolayer cultures. Inhibition of succinyl dehydrogenase activity corresponded to the amount of hydrogen peroxide that can rapidly diffuse through dentin in vitro and reach concentrations which are toxic to cultured cells in less than 1 h.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/66707/2/10.1177_00220345930720051501.pd

The effect of repeated stretching on the force decay and compluance of vulcanized cis-polyisoprene orthodontic elastics

Compliance measurements, used in the past to measure the viscoelastic properties of dental impression materials, were used to assess these properties in vulcanized cis-polyisoprene orthodontic elastics, and the results were compared with traditional force decay measurements. Both methods were also used to evaluate the effect of repeated stretching on these elastics. Compliance measurements successfully characterized the viscoelastic behavior of the elastics, and the results agreed with force decay measurements. Repeated stretching significantly reduced the force and the compliance of the elastics. There was no statistical difference in the force or compliance measurements after the elastics were stretched more than 200 times. Stretching for 1000 cycles of 400% extension reduced elastic force by approximately 12%.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/31027/1/0000704.pd

The effect of cell monolayer density on the cytotoxicity metal ions which are released from dental alloys

The effect of cell density (number of cells per unit area of a monolayer culture) on the in vitro cytotoxicity of metal ions which are known to be released from dental materials was investigated. The effects of cell density (1) may explain previous discrepancies in in vitro tests, (2) may be important in wound healing where cell density changes over time, and (3) may help clarify the mechanisms of cytotoxicity of metal ions. Balb/c 3T3 fibroblasts were plated at cell densities ranging from 10,000-80,000 cells/cm2 and were exposed to 8 concentrations of 10 different metal ions. After 24 h, the succinic dehydrogenase activity and DNA synthesis were measured to quantify the cytotoxic effect. Higher cell densities markedly reduced the sensitivity of these fibroblasts to all metal ions except Al+3 and Zn+2, but the magnitude of the reduction was metal dependent. In addition, the DNA synthesis was inhibited more than the succinic dehydrogenase activity for all metal ions except Zn+2. The unique effect of cell density on each metal ion supported the hypothesis that the effect was not simply caused by a dilution of the number of metal ions per cell. Given these results, the effect of cell density should be carefully selected in in vitro cytotoxicity tests.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/30808/1/0000466.pd

Precision of and new methods for testing in vitro alloy cytotoxicity

Previous studies have utilized in vitro alloy cytotoxicity tests to evaluate dental casting alloys. The purposes of this study were to: (1) evaluate the precision of the optical density and visual tests previously used, (2) evaluate a new test measuring absorbance of solubilized formazan dyes, and (3) test the correlation between these tests for cytotoxicity. Balb/c 3T3 cells were plated in 24-well culture trays at 25,000 cells/cm2 around ten types of dental casting alloy (six samples/alloy) and incubated for 72 h. Cells were histochemically stained with MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide) succinate for 2 h, then fixed, washed, and dried. Toxicity was measured by optical densitometer (OD) scanning, visual assessment, and 560-nm absorbance of DMSO-solubilized dyes. Measurements of rings of inhibition were not used, because they did not provide precise data, and correlated poorly with the other methods. The results were analyzed by ANOVA, Tukey intervals, and coefficients of variation (CV's). MTT required shorter incubation times for adequate staining, allowed for solubilization of the monolayers, and was less expensive than NBT (2,2'-di-p-nitro-phenyl-5,5'-diphenyl-3,3'-dimethoxy-[3,3'-dimethoxy-4,4'-biphenylene] ditetrazolium chloride). Results showed that all three methods ranked alloy toxicities similarly (p = 0.05). The solubilization method was most discriminating due to lower CV's. Correlation between densitometer and solubilization methods was excellent (R2 = 0.96). Between-experiment CV's were generally less than 20%, and often less than 10%. Between-sample CV;s were generally less than 20%. CV's were consistently lower for the solubilization method. Thus, all methods were repeatable and correlated well, but the solubilization method was more precise and discriminating.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/30285/1/0000687.pd

Correlation of cytotoxicity with elemental release from mercury- and gallium-based dental alloys in vitro

Objectives. An in vitro screening test was used to compare the cytotoxicity and elemental release from mercury- and gallium-based dental restorative materials. Methods. The test employed three sequential extractions of the samples into cell-culture medium which were then used to evaluate the cytotoxicity of the samples and the release of elements from the samples. Cytotoxicity was measured by placing the extract in contact with Balb/c mouse fibroblasts for 24 h and measuring the succinic dehydrogenase activity of the cells. The release of elements was measured by means of atomic absorption spectrophotometry. Results. Samples of Tytin (Kerr) showed no cytotoxicity compared to Teflon controls. Dispersalloy (Johnson and Johnson) was severely cytotoxic initially when Zn release was greatest, but was less toxic between 48 and 72 h as Zn release decreased. Gallium Alloy GF (Tokuriki Honten) was moderately cytotoxic after 8 h, and increased in cytotoxicity thereafter, which correlated with a substantial and persistent release of Ga from this material. Significance. The results of the current study concurred with in vivo assessments of these materials, and the use of sequential extractions was useful in determining trends in the cytotoxicity and elemental release from these materials.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/31360/1/0000272.pd

Cytotoxicity of Titanate-Calcium Complexes to MC3T3 Osteoblast-Like Cells

Monosodium titanates (MST) are a relatively novel form of particulate titanium dioxide that have been proposed for biological use as metal sorbents or delivery agents, most recently calcium (II). In these roles, the toxicity of the titanate or its metal complex is crucial to its biological utility. The aim of this study was to determine the cytotoxicity of MST and MST-calcium complexes with MC3T3 osteoblast-like cells; MST-Ca(II) complexes could be useful to promote bone formation in various hard tissue applications. MC3T3 cells were exposed to native MST or MST-Ca(II) complexes for 24-72 h. A CellTiter-Blue5 assay was employed to assess the metabolic activity of the cells. The results showed that MST and MST-Ca(II) suppressed MC3T3 metabolic activity significantly in a dose-, time-, and cell-density-dependent fashion. MST-Ca(II) suppressed MC3T3 metabolism in a statistically identical manner as native MST at all concentrations. We concluded that MST and MST-Ca(II) are significantly cytotoxic to MC3T3 cells through a mechanism yet unknown; this is a potential problem to the biological utility of these complexes

Bone Marrow Derived Mesenchymal Stem Cells Inhibit Inflammation and Preserve Vascular Endothelial Integrity in the Lungs after Hemorrhagic Shock

Hemorrhagic shock (HS) and trauma is currently the leading cause of death in young adults worldwide. Morbidity and mortality after HS and trauma is often the result of multi-organ failure such as acute lung injury (ALI) and acute respiratory distress syndrome (ARDS), conditions with few therapeutic options. Bone marrow derived mesenchymal stem cells (MSCs) are a multipotent stem cell population that has shown therapeutic promise in numerous pre-clinical and clinical models of disease. In this paper, in vitro studies with pulmonary endothelial cells (PECs) reveal that conditioned media (CM) from MSCs and MSC-PEC co-cultures inhibits PEC permeability by preserving adherens junctions (VE-cadherin and β-catenin). Leukocyte adhesion and adhesion molecule expression (VCAM-1 and ICAM-1) are inhibited in PECs treated with CM from MSC-PEC co-cultures. Further support for the modulatory effects of MSCs on pulmonary endothelial function and inflammation is demonstrated in our in vivo studies on HS in the rat. In a rat “fixed volume” model of mild HS, we show that MSCs administered IV potently inhibit systemic levels of inflammatory cytokines and chemokines in the serum of treated animals. In vivo MSCs also inhibit pulmonary endothelial permeability and lung edema with concurrent preservation of the vascular endothelial barrier proteins: VE-cadherin, Claudin-1, and Occludin-1. Leukocyte infiltrates (CD68 and MPO positive cells) are also decreased in lungs with MSC treatment. Taken together, these data suggest that MSCs, acting directly and through soluble factors, are potent stabilizers of the vascular endothelium and inflammation. These data are the first to demonstrate the therapeutic potential of MSCs in HS and have implications for the potential use of MSCs as a cellular therapy in HS-induced lung injury

Dental Material: Properties and Manipulation

x. 373 hal.; 23 c

Effect of alloy surface composition on release of elements from dental casting alloys

The release of elements from dental casting alloys is a continuing concern because of the potentially harmful biological effects the elements may have on local tissues. The surfaces of the alloys appear to be most important in controlling the release of these elements. In the current study, the surfaces of high-, reduced-, and no-gold dental alloys were analysed by X-ray photoelectron spectroscopy before and after they were exposed to a biological medium for up to 96 h. The goal was to relate the release of elements from these alloys to their surface composition, and to determine the depth of the effect of the medium. The depth of the effect of the exposure was determined by argon milling of the alloy surface after exposure to the medium. Elements that were released into the medium were measured by means of atomic absorption spectroscopy. The release of elements from alloys was greater when the atomic ratio of noble to non-noble elements at the surface was less than 1. The depth of the effect of the medium varied with the alloy, but was always less than 100 Å. The surface composition was significantly different from layers only 5 Å below. It was concluded that the surface concentration of noble elements is important in controlling the release of non-noble elements from these alloys, and the surface composition appeared to be only one or two atomic layers thick. of the three types of alloys, the high-gold alloy appeared to develop the most stable surface composition which released the lowest levels of elements.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/72575/1/j.1365-2842.1996.tb00896.x.pd

Effect of cell line on in vitro metal ion cytotoxicity

Objectives. The choice of cell line for in vitro biological tests which assess the cytotoxicity of dental materials remains controversial, yet this issue is important because these tests are widely used to rate the biocompatibility of new and existing materials, and many different cell lines are commonly used. The Purpose of the current study was to quantify the responses of four cell lines (Balb/c 3T3, L929, ROS 17/2.8 and WI-38) to 14 metal ions which are released from dental materials, and relate these responses to the metabolic activity and population doubling times of these cells.Methods. Succinic dehydrogenase (SDH) activity was used to monitor metabolic activity and cytotoxic response.Results. The cell lines responded differently to most metal ions. In general, the Balb/c 3T3 line was the most sensitive, and the WI-38 line was the least sensitive. However, there were many exceptions depending on the metal ion. The passage number of the cells also affected the cytotoxic response. It was concluded that the cytotoxicity of materials which release metal ions will be significantly different depending on which cell line is selected and its passage number.Significance. Based on the findings that cell lines ranked the toxicities of the metal ions similarly, it seems reasonable to use these types of in vitro tests to rank the cytotoxicities of materials. However, if these types of tests are used to predict in vivo cytotoxicity, care should be taken to choose conditions and cells which are relevant.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/31603/1/0000532.pd