Irritation and cytotoxic potential of denture adhesives

Objective: The present study aimed to examine the in vitro biocompatibility of denture adhesives. Background: Denture adhesives absorb water to become viscous and sticky, and by this process, other constituents like colouring, flavouring, wetting and preserving agents may be released. Some of these constituents may induce adverse reactions among users of denture adhesives. Materials and methods: Five commercially available denture adhesives; three different creams, a powder, and a cushion were included in the study. The irritation and cytotoxic potential was evaluated using the Hen's Egg Test Chorioallantoic Membrane (HET‐CAM) method and three cell culture methods; filter diffusion, dimethylthiazol diphenyltetrazolium bromide (MTT) assay and agar diffusion. Results: None of the tested denture adhesives showed a noteworthy acute irritation as evaluated by the HET‐CAM method. None of the tested denture adhesives induced cytotoxicity in the filter diffusion test. One of the denture adhesives induced a severe cytotoxic reaction in both the MTT and agar diffusion assays. These tests employ longer exposure times than in both the filter diffusion and the HET‐CAM test. Conclusion: Denture adhesives are commonly used throughout the day, and our results raise the concern that denture adhesives may contribute to mucosal inflammation in denture wearers.     

Oral health related quality of life of edentulous patients after denture relining with a silicone-based soft liner

doi: 10.1111/j.1741‐2358.2011.00503.x Oral health related quality of life of edentulous patients after denture relining with a silicone‐based soft liner Background: Knowledge of benefits caused by a treatment on quality of life is very relevant. Despite the wide use and acceptance of soft denture liners, it is necessary to evaluate the patient’s response about the use of these materials with regard to improvement in oral health related quality of life (OHRQoL). Objectives: The aim of this study was to evaluate the influence of denture relining in the OHRQoL of edentulous patients. Materials and methods: Thirty‐two complete denture wearers had their lower dentures relined with a silicone‐based material (Mucopren soft, Kettenbach, Germany) according to chairside procedures. OHRQoL was assessed before and after 3 months of relining by means of OHIP‐EDENT, and the median scores were compared by Wilcoxon test (p ≤ 0.05). Results: After 3 months of relining, participants reported significant improvement of their OHRQoL (p ≤ 0.01). Conclusion: Denture relining with a soft liner may have a positive impact on the perceived oral health of edentulous patients.     

Generation of toxic phospholipid(s) during oxyhemoglobin-induced peroxidation of phosphatidylcholines

When egg yolk diacylglycerophosphocholine (PC) liposomes were incubated with human oxyhemoglobin, peroxidation of liposomal lipid was induced, as monitored by an increase of thiobarbituric acid (TBA)-reactive substances, an increase of lipid hydroperoxides and the generation of chemiluminescence in the presence of luminol. During the reaction, cytotoxic substance(s), which induced shedding of acetylcholinesterase-enriched vesicles from human erythrocytes, were produced. Formation of TBA-reactive substances and lipid hydroperoxides preceded generation of chemiluminescence, conversion of oxyhemoglobin to methemoglobin and production of the toxic substances. Either superoxide dismutase or catalase could suppress generation of chemiluminescence, but not other events. Methemoglobin or ferrous ion plus ascorbate could induce peroxidation of the liposomes without production of the cytotoxic substance(s). Synthetic PCs containing both saturated and polyunsaturated fatty acyl chains caused the production of cytotoxic products which induced shedding of vesicles from erythrocytes, whereas those containing only polyunsaturated fatty acyl chains did not, suggesting that the molecular species which can produce cytotoxic products may be phospholipids containing both saturated and polyunsaturated fatty acids. The mechanism of oxyhemoglobin-induced peroxidation of lipids will be also discussed.     

华中五味子抗氧化和细胞毒活性研究

我们研究了华中五味子的根、茎、叶、果实乙醇提取物对脂性自由基DPPH及超氧阴离子的清除作用,同时观察了四种不同提取物在体外对肿瘤细胞株的细胞毒作用。我们发现,华中五味子根的乙醇提取物清除自由基活性和对肿瘤细胞杀伤作用比茎、叶和果实的提取物都强,研究结果提示华中五味子的根可以作为抗氧化剂的新来源。     

藏药砂生槐种子提取物抑菌和细胞毒活性的初步研究

砂生槐是我国西藏高原一种特有植物,是一种极为宝贵的药用植物资源.我们首次从砂生槐种子中获得氯仿、95%乙醇、75%乙醇和水提取物,用琼脂扩散实验测定其抑菌活性和用MTT法测定其对肿瘤细胞的细胞毒效应,表明氯仿提取物和95%乙醇提取物具有广谱的抑菌活性,抑菌活性较强的是氯仿提取物.各种提取物显示出浓度依赖性的细胞毒效应,氯仿、95%乙醇、75%乙醇和水提取物对胃癌SGC-7901细胞系的LC50分别是4.5、1.4、1.9和41.7 mg/mL,抑制胃癌SGC-7901细胞增殖作用较强的成分是砂生槐种子95%乙醇提取物.这一发现为开发砂生槐这一宝贵植物资源的药用价值提供了重要的依据.     

Effect of water storage and heat treatment on the cytotoxicity of soft liners

doi: 10.1111/j.1741‐2358.2011.00463.x Effect of water storage and heat treatment on the cytotoxicity of soft liners Objective: To evaluate the effect of water storage time on the cytotoxicity of soft liners. Methods: Sample discs of soft liners Dentusoft, Dentuflex, Trusoft, Ufi‐Gel‐P and denture base acrylic resin Lucitone‐550 were prepared and divided into four groups: GN: No treatment, G24: Stored in water at 37°C for 24 h; G48: Stored in water at 37°C for 48 h, GHW: Immersed in water at 55°C for 10 min. To analyse the cytotoxic effect, three samples of each group were placed in tubes with Dubelcco’s Modified Eagle Mediums and incubated at 37°C for 24 h. During this period, the toxic substances were leached to the culture medium. The cytotoxicity was analysed quantitatively by the incorporation of radioactivity ³H‐thymidine checking the number of viable cells (synthesis of DNA). The data were statistically analysed using two‐way anova and Tukey’s honestly significant difference tests (α = 0.05). Results: Treatments did not reduce the cytotoxicity effect of the soft liners (p > 0.05). It was found that Ufi‐Gel‐P had a non‐cytotoxic effect, Trusoft had a slightly cytotoxic effect, Dentuflex had a moderated cytotoxic effect, Dentusoft alternated between slightly and non‐cytotoxic effect, and Lucitone‐550 had non‐cytotoxic effect when stored in water for 48 h. Conclusion: The effect of water storage and the heat treatment did not reduce the cytotoxicity of the soft liners.     

The toxic effect of dermotrophic substances in tests in vitro

In the model system of tissue culture, the effect of dermatotrophic substances, such as the tenzides and conservatory substances, was tested. The cytotoxic effect of the substances was evaluated according to the vitality of the cells and the lactate dehydrogenase (LDH) activity in the suspension culture of human lymphocytes and further according to the migration inhibition of cells from the animal spleen fragments (SF). The cells obtained from rabbits, guinea pigs, mice and man were used. The highest correlation between the concentration of the substance in the cultivation medium and its toxic effect in the culture has been proved in the tests of LDH activity from the supernatant of the human peripheral lymphocytes culture and further in the migration inhibition test from rabbit SF. Both the tests are in mutual correlation. The determination of LDH activity represents a sensitive marker of the cell damage, the least sensitive is the estimation of the cell vitality. It has been proved that the system of suspension culture of human peripheral lymphocytes and the migration inhibition test from rabbit SF is suitable for the screening determination of the effect of dermatotrophic substances. The system of tissue culture compared to classical tests of skin irritation performed in vivo secures overall objectivity in evaluating the results and the possibility of making a number of parallels without any ethical limits.     

Neurotoxic,cytotoxic, apoptotic and antiproliferative effects of some marine algae extracts on the NA2B cell line

Oxidative stress contributes to cancer pathologies and to apoptosis. Marine algae exhibit cytotoxic, antiproliferative and apoptotic effects; their metabolites have been used to treat many types of cancer. We investigated in culture extracts of Petalonia fascia, Jania longifurca and Halimeda tuna to determine their effects on mouse neuroblastoma cell line, NA2B. NA2B cells were treated with algae extracts, and the survival and proliferation of NA2B cells were assessed using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The effects of algae extracts on oxidative stress in NA2B cells also were investigated using nitric oxide synthase (NOS) immunocytochemistry and apoptosis was assessed using terminal deoxynucleotidyl transferase dUTP nick end labeling. We observed significant neurite inhibition with moderate damage by the neurotoxicity-screening test (NST) at IC₅₀ dilutions of the extracts. MTT demonstrated that J. longifurca extracts were more toxic than P. fascia and H. tuna extracts. We found an increase of endothelial and inducible NOS immunostaining for oxidative stress and TUNEL analysis revealed increased apoptosis after application of extract. Our findings suggest that the algae we tested may have potential use for treatment of cancer.     

Tests with Daphnia magna: a new approach to prescreen toxicity of newly synthesized acetylcholinesterase reactivators

Reactivators of phosphorylated acetylcholinesterase (oximes) are substances used as a human antidotal therapy for organophosphate poisoning. The objective of our study was to examine if juveniles of freshwater microcrustacean Daphnia magna could be employed as test animals in early screen toxicity tests of those substances as a first step for further experiments with daphnids intoxicated by organophosphates. For this purpose, seven different oximes were investigated. It was found that toxicity of all tested oximes increased with time. Mono-quaternary oximes were approximately ten fold (EC50, 14.9 mg.l(-1)) more toxic in 24 hour tests and five fold (EC50 was 79.46 mg.l(-1)) more toxic in 48 hour tests than bis-quaternary oximes. Tests with daphnids were shown to be easy to carry out at low cost and provided valuable results which could be used as a starting point for further research.     

Antibacterial and anti-inflammatory effects of Jeju medicinal plants against acne-inducing bacteria

Propionibacterium acnes and Staphylococcus epidermidis are pus-forming bacteria that trigger inflammation in acne. The present study was conducted to evaluate the antimicrobial activities of Jeju medicinal plants against these etiologic agents of acne vulgaris. Ethanol extracts of Jeju plants were tested for antimicrobial activities by disc diffusion and broth dilution methods. The results from the disc diffusion assays revealed that four medicinal plants, Mollugo pentaphylla, Angelica anomala, Matteuccia orientalis, and Orixa japonica inhibited the growth of both pathogens. Among these, A. anomala had strong inhibitory effects. Its MIC values were 15.6 microg/ml and 125 microg/ml against P. acnes and S. epidermidis, respectively. The cytotoxic effects of the four extracts were determined by colorimetric MTT assays using two animal cell lines: human dermal fibroblasts and HaCaT cells. Although the M. orientalis root extract had moderate cytotoxicity in HaCaT cells at 200 microg/ml, most extracts exhibited low cytotoxicity at 200 microg/ml in both cell lines. In addition, the extracts reduced the P. acnes-induced secretion of interleukin-8 and tumor necrosis factor-alpha (TNF-alpha) in THP-1 cells, an indication of their anti-inflammatory effects. Based on these results, we suggest that M. pentaphylla, A. anomala, M. orientalis, and O. japonica are attractive acne-mitigating candidates for topical application.     

DNA protecting and genotoxic effects of olive oil related components in cells exposed to hydrogen peroxide

In search for compounds, able to protect nuclear DNA in cells exposed to oxidative stress, extracts from olive leaves, olive fruits, olive oil and olive mill waste water were tested by using the “single cell gel electrophoresis” methodology (comet assay). Jurkat cells in culture were exposed to continuously generated hydrogen peroxide (11.8±1.5 μM per min) by direct addition into the growth medium of the appropriate amount of the enzyme “glucose oxidase” in the presence or absence of the tested total extracts. The protective effects of the tested extracts or isolated compounds were evaluated from their ability to decrease hydrogen peroxide-induced formation of single strand breaks in the nuclear DNA, while the toxic effects were estimated from the increase of DNA damage when the extracts or isolated compounds were incubated directly with the cells. Significant protection was observed in extracts from olive oil and olive mill waste water. However, above a concentration of 100 μg/ml olive oil extracts exerted DNA damaging effects by themselves in the absence of any H₂O₂. Extracts from olive leaves and olive fruits although protective, were also able to induce DNA damage by themselves. Main compounds isolated from the above described total extracts, like oleuropein glucoside, tyrosol, hydroxytyrosol and caffeic acid, were tested in the same experimental system and found to exert cytotoxic (oleuropein glucoside), no effect (tyrosol) or protective effects (hydroxytyrosol and caffeic acid). In conclusion, cytoprotective as well as cytotoxic compounds with potential pharmaceutical properties were detected in extracts from olive oil related sources by using the comet assay methodology.     

Effect of reline material and denture base surface treatment on the impact strength of a denture base acrylic resin

doi:10.1111/j.1741‐2358.2009.00292.x
Effect of reline material and denture base surface treatment on the impact strength of a denture base acrylic resin Objective: In this study, the effect of relining and surface treatment on the impact strength (IS) of a heat‐polymerising denture base acrylic resin (Lucitone 550‐L) was evaluated. Materials and methods: Rectangular bars of L were made (60 × 6 × 2 mm) and relined (2 mm) with the relining resins Ufi Gel Hard (UH) and Tokuso Rebase Fast (TR). Specimens relined with L and intact L, TR and UH specimens were also made (60 × 6 × 4 mm), for comparison. Before relining, the L surface was left untreated or wetted with methyl methacrylate monomer and/or the bonding agents (BA) supplied by manufacturers of the reline resins. V‐notches were machined at the midpoint of the length of all specimens. The notches were made either across the width (Nw) or across the thickness of the specimens (Nth). The Charpy impact test was performed using a 0.5‐J pendulum, which had been specially designed and constructed. Data were analysed separately for each notch position using one‐way analysis of variance and Tukey honestly significant difference post‐hoc test (p = 0.05). Results: The IS of L was similar to that of L/L. For the Nw notch, treating the denture base L with TR BA and relining with TR reline material produced the highest IS. Conclusion: The IS of specimens made from heat polymerising acrylic resin Lucitone 550 was increased after relining using the hard chairside reline resin TR with its proprietary BA.     

Effect of<Emphasis Type="Italic">Pleurotus ferulae</Emphasis> extracts on viability of human lung cancer and cervical cancer cell lines

When SiHa cells were incubated for varying periods of time with extracts of PFF and PFM, the cytotoxicity of the ethanol extracts of PFF was higher than those of the other extracts. These results indicated that the extracts from fruiting bodies ofP. ferulae contain antitumor substances. When A549, SiHa and HeLa cells were incubated with different concentrations of PFF and PFM extracts, the ethanol extracts of PFF showed strong cytotoxicity against A549 cells at concentrations over 10 μg/mL and against SiHa and HeLa cells at concentrations over 40 μg/mL. However, the differences in the cytotoxic effects of the hot water and ethanol extracts of PFM and the hot water extracts of PFF on all 3 cancer cells were not significant. Also, the PFF ethanol extracts induced synergistic effects on the TRAIL-induced apoptosis in A549 cells, which were strongly resistant to TRAIL. These results indicated that ethanol extracts of PFF were the most prominent antitumor agents toward lung cancer cells (A549).     

Cytotoxic Effects of 6-Hydroxydopamine,Merocyanine-540 and Related Compounds on Human Neuroblastoma-and Hematopoietic Stem Cells

6-Hydroxydopamine(6-OHDA) and Merocyanine-540(MC-540) have been used clinically for purging of neuroblastoma cells prior to autologous bone marrow transplantation. Both substances were found to be more toxic against neuroblastoma cells than against hematopoietic stem cells. The more pronounced cytotoxic effects of 6-OHDA against neuroblastoma cells were not caused by its selective uptake; the rapid autooxidation at physiological pH leads to the formation of H,O, already in the incubation medium. Cytotoxic effects were not detected in short-time test systems (4 hour chromium-51 release assay) but only after longer incubation periods. In contrast, MC-540 proved to be toxic almost equally in short- and long-time test systems. 4-Hydroxynonenal(4-HNE) that may be formed in the plasma membrane subsequently to photoactivation of MC-540 was only slightly more toxic to neuroblastoma cells than to hematopoietic cells. Although the use of 6-OHDA and MC-540 in bone marrow purging has some limitations, the sensitivity of neuroblastoma cells against reactive oxygen compounds may be exploited more generally for therapy of this tumor.     

Cell cultures in the biocompatibility study of synthetic materials

In vitro cytotoxicity (Neutral Red uptake, Kenacid Blue and MTT) and cytocompatibility (cell adhesion and proliferation) tests were applied to the biocompatibility study of a series of poly(ester-ether-ester) block copolymers of potential interest as biomaterials. Our results indicate that the copolymer extracts after 72 hours incubation with a 3T3 mouse fibroblast cell line do not induce significant toxic effects. Furthermore, human umbilical vein endothelial cells seeded on thin copolymer films show a normal pattern of growth. We conclude that thein vitro tests used are a valid instrument to evaluate the potential toxic action of synthetic materials on different cell compartments and that the tested materials seem to be promising for future applications in the field of biomedical devices.     

Cytotoxic Activity and Related Mechanisms of Prenylflavonoids Isolated from Mallotus conspurcatus Croizat

Five prenylflavonoids, 6‐prenylnaringenin ( 1 ), 8‐prenylnaringenin ( 2 ), 7‐O‐methyl‐8‐prenylnaringenin ( 3 ), 7‐O‐methyl‐6‐prenylnaringenin ( 4 ), and 4′‐O‐methyl‐6‐prenylnaringenin ( 5 ), were isolated from the traditional herb Mallotus conspurcatus Croizat (Euphorbiaceae). Compounds 1 – 5 revealed cytotoxic activity against cervical cancer (HeLa) cells with IC₅₀ values ranging from 10.08 to 60.16 μm by MTT method, and interestingly, these prenylflavonoids were less toxic to normal HL‐7702 cells. Furthermore, compounds 1 and 5 could inhibit the c‐myc expression and telomerase activity and cause mitochondrial dysfunction. These findings might contribute to a better understanding of the biological activities of prenylflavonoids and lay the foundation for further studies on the cytotoxic activity of natural products isolated from M. conspurcatus.     

Flexural and bond strengths of relined denture polymers assessed by four‐point bending tests and Weibull analysis

doi: 10.1111/j.1741‐2358.2011.00515.x Flexural and bond strengths of relined denture polymers assessed by four‐point bending tests and Weibull analysis Objectives: The aim of this study was to (1) investigate the flexural strengths of three denture resins i.e. heat, photopolymerised and microwaved and how it was affected by relining with auto‐ and visible light–polymerised hard reliners, (2) investigate the bond strengths between denture resins and hard reliners and (3) interpret the results of both tests by utilising Weibull analysis. Materials and methods: Specimens (65 × 10 × 2.5 mm) from denture resins, relined and bonded combinations were tested using a four‐point bending test in a universal testing machine and a crosshead speed of 5 mm/min. Ten specimens for each bulk resin and denture resin–reliner combination for a total of 150 were tested. Results: Statistical analysis indicated significant differences between bulk materials (p < 0.001) and between reliners (p < 0.001) for flexural and bond strength tests. Conclusion: It was concluded that (1) the four‐point flexural strength was different between the denture base materials, (2) flexure strength between bulk and relined or between relined with autopolymerised and photopolymerised bases was different, (3) flexural strength among relined denture bases was different and (4) bond strengths among relined denture bases were different.     

Tests with Daphnia magna: A new approach to prescreen toxicity of newly synthesized acetylcholinesterase reactivators

Reactivators of phosphorylated acetylcholinesterase (oximes) are substances used as a human antidotal therapy for organophosphate poisoning. The objective of our study was to examine if juveniles of freshwater microcrustacean Daphnia magna could be employed as test animals in early screen toxicity tests of those substances as a first step for further experiments with daphnids intoxicated by organophosphates. For this purpose, seven different oximes were investigated. It was found that toxicity of all tested oximes increased with time. Mono-quaternary oximes were approximately ten fold (EC₅₀, 14.9 mg.l^{? 1}) more toxic in 24 hour tests and five fold (EC₅₀ was 79.46 mg.l^{? 1}) more toxic in 48 hour tests than bis-quaternary oximes. Tests with daphnids were shown to be easy to carry out at low cost and provided valuable results which could be used as a starting point for further research.     

Ichtyotoxic activity of extracts from Mexican marine macroalgae

Seventy-three species of macroalgae from the Mexican Pacific, Atlantic and Caribbean coast were screened for ichtyotoxic activity. Ethanolic, acetonic and aqueous extracts were prepared and tested against the fish Carassius auratus. The extracts were classified on the basis of their effects as: toxic if the fish died in two hours or less; moderately toxic, if the organism behaved abnormally but death did notoccur, and non-toxic if the fish did not display any change. 79% species were ichtyotoxic to some degree. Extracts of 39 species were toxic, with at least one extract with lethal effects, 19 were moderately toxic and 15 species were non-toxic. Only the extracts ofDictyota bartayresiana, Dictyota cervicornis,Lobophora variegata, Bryothamnion triquetrum and Laurencia obtusa were toxic in all three solvents. The acetone and ethanol extracts were more active, and therefore are more suitable for extraction of toxic substances. This revised version was published online in August 2006 with corrections to the Cover Date.     

Microplate screening for apoptosis with antibody to single-stranded DNA distinguishes anticancer drugs from toxic chemicals

The effect of anticancer drugs and toxic compounds on cultures of human leukemic cells was evaluated by an enzyme-linked immunosorbent assay (Apoptosis ELISA) that uses a monoclonal antibody against single-stranded DNA to quantitate the apoptotic cells. The concentrations of 13 anticancer drugs, which increased Apoptosis ELISA absorbance, were close to the cytotoxic concentrations determined by the long-term cell survival assay. Short-term tetrazolium-based microculture tetrazolium (MTT) assay was significantly less sensitive than the Apoptosis ELISA and the cell survival assay for all anticancer drugs. For 6 drugs, cytotoxic concentrations measured by the MTT assay were at least 1 log higher than the concentrations inducing apoptosis. Importantly, in contrast to the anticancer drugs, 14 toxic chemicals did not increase the Apoptosis ELISA absorbance at cytotoxic concentrations. The difference in apoptosis induction by the anticancer drugs and the toxic chemicals was especially large in cultures treated with drug concentrations 2-fold higher than the IC(50) dose. Although all of the anticancer drugs tested induced intense ELISA reaction (mean absorbance 2.0), all toxic chemicals tested did not induce apoptosis. The Apoptosis ELISA assay could have useful applications in drug development as it can distinguish between clinically useful anticancer drugs and toxic compounds, has sensitivity similar to that of the long-term cell survival assay, and provides insight into the mechanism of drug cytotoxicity by differentiating between compounds killing cells by apoptosis and necrosis.