Antitumor and cytotoxic activity of Kielmeyera coriacea mart. Zucc. and Pyrostegia venusta (ker-gawl.) Miers extracts

This study aimed to investigate the antitumor and cytotoxicity activities of Kielmeyera coriacea and Pyrostegia venusta extracts. Therefore, the hydroalcoholic extracts of P. venusta flowers and K. coriacea leaves were prepared. The extracts were evaporated and the dry extracts were diluted at concentrations of 1.0, 0.1, 0.01 and 0.001 mg/ml for carrying out the bioassays. Artemia salina eggs were incubated in saline solution at 28°C for 24 h. The larvae were treated with different extracts concentrations and the mortality was evaluated after 24 and 48 h. Five discs of potato were placed in Petri dishes and 50 μl of inoculum of Agrobacterium tumefaciens were added to it at 28°C for 24 h incubation. So, 50 μl of the extracts in different concentrations were added. Positive and negative controls were made. The P. venusta and K. coriacea extracts did not show statistically significant acute toxicity. K. coriacea extract showed (mean% of tumor ± standard deviation) 15.30 ± 3.24, 6.34 ± 3.82, 7.57 ± 2.92 and 5.77 ± 2.85 and P. venusta showed 25.82 ± 5.15, 38.40 ± 8.28, 15.75 ± 4.44 and 13.38 ± 7.92, with their concentrations for the antitumor bioassay, and the positive control showed 25.80 ± 6.14. According to the obtained results it was established that the K. coriacea and P. venusta extracts showed antitumor activity but did not show significant cytotoxic activity in A. salina test.

Use of HEPG2 cells to assay the safety of cosmetic active substances

The importance of this study is based on the need to obtain simple and efficient in vitro models to predict the in vivo toxicity of cosmetics, aiming not to use animals as experimental model. Here, we proposed the use of HepG2 cells, which are widely applied to simulate the hepatic function of the human organism in vitro. This cell line was chose since recent studies have shown that the liver is potentially the most frequently targeted organ by cosmetic ingredients, and beyond that, considering the widely application of in vitro assays to test the cutaneous permeation of cosmetic products, including the assays applying modified Franz cells, this technique becomes indispensable. Three different cosmetic active substances were used, and the toxicity to HepG2 cells was assessed by the MTT method. The treatment with hyaluronic acid showed no toxicity to HepG2 cells. Treating the cells with P. guajava L. extract were verified that increasing the amount of the extract in the media, the cellular viability decreased, and finally, the treatment of alpha-lipoic acid showed a cytoprotective effect in relation to the treatment with propylene glycol. The study demonstrated the suitability in using HepG2 cells to assess the safety of cosmetic active substances, helping in the prediction of if the substance could be hepatotoxic if could reach the bloodstream

Antitumor and Anti-Inflammatory Effects of Palladium(II) Complexes on Ehrlich Tumour

Palladium(II) complexes are an important class of cyclopalladated compounds that play a pivotal role in various pharmaceutical applications. Here, we investigated the antitumour, anti-infl ammatory, and mutagenic effects of two complexes: [Pd(dmba)(Cl)tu] (1) and [Pd(dmba)(N3)tu] (2) (dmba = N,N-dimethylbenzylamine and tu = thiourea), on Ehrlich ascites tumour (EAT) cells and peritoneal exudate cells (PECs) from mice bearing solid Ehrlich tumour. The cytotoxic effects of the complexes on EAT cells and PECs were assessed using the 3-(4,5-dimethylthiazol-3-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay. The effects of the complexes on the immune system were assessed based on the production of nitric oxide (NO) (Griess assay) and tumour necrosis factor-Į (TNF-Į), interleukin-12 (IL-12), and interleukin-10 (IL-10) (ELISA). Finally the mutagenic activity was assessed by the Ames test using the Salmonella typhimurium strain TA 98. Cisplatin was used as a standard. The IC50 ranges for the growth inhibition of EAT cells and PECs were found to be (72.8 ± 3.23) µM and (137.65 ± 0.22) µM for 1 and (39.7 ± 0.30) µM and (146.51 ± 2.67) µM for 2, respectively. The production of NO, IL-12, and TNF-Į, but not IL-10, was induced by both complexes and cisplatin. The complexes showed no mutagenicity in vitro, unlike cisplatin, which was mutagenic in the strain. These results indicate that the complexes are not mutagenic and have potential immunological and antitumour activities. These properties make them promising alternatives to cisplatin.

Cytotoxicity of dimethyl sulfoxide (DMSO) in direct contact with odontoblast-like cells

To evaluate the cytotoxicity of dimethyl sulfoxide (DMSO) on the repair-related activity of cultured odontoblast-like MDPC-23 cells. Methods Solutions with different concentrations of DMSO (0.05, 0.1, 0.3, 0.5 and 1.0 mM), diluted in culture medium (DMEM), were placed in contact with MDPC-23 cells (5 × 104 cells/cm2) for 24 h. Eight replicates (n = 8) were prepared for each solutions for the following methods of analysis: violet crystal dye for cell adhesion (CA), quantification of total protein (TP), alizarin red for mineralization nodules formation (MN) and cell death by necrosis (flow cytometry); while twelve replicates (n = 12) were prepared for viable cell number (Trypan Blue) and cell viability (MTT assay). Data were analyzed by ANOVA and Tukey or Kruskal–Wallis and Mann–Whitney's tests (p < 0.05). Results Cell viability, adhesion and percentage of cell death by necrosis were not affected by DMSO at any concentration, with no statistical significant difference among the groups. A significant reduction in total protein production was observed for 0.5 and 1.0 mM of DMSO compared to the control while increased mineralized nodules formation was seen only for 1.0 mM DMSO. Significance: DMSO caused no or minor cytotoxic effects on the pulp tissue repair-related activity of odontoblast-like cells.

Protective effect of sodium ascorbate on odontoblast-like cells MDPC-23 exposed to a bleaching agent

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Transdentinal cytotoxicity of carbodiimide (EDC) and glutaraldehyde on odontoblast-like cells

Objective: To evaluate the transdentinal cytotoxicity of three different concentrations of carbodiimide (EDC) or 5% glutaraldehyde (GA) on MDPC-23 cells. Methods: Seventy 0.4-mm-thick dentin disks obtained from human molars were adapted to artificial pulp chambers. MDPC-23 cells were seeded on the pulpal surface of the disks. After 48 hours, the occlusal dentin was acid-etched and treated for 60 seconds with one of the following solutions (n=10): no treatment (negative control); 0.1 M, 0.3 M, or 0.5 M EDC; 5% GA; Sorensen buffer; or 29% hydrogen peroxide (positive control). Cell viability and morphology were assessed by methyltetrazolium assay and scanning electron microscopy (SEM), respectively. The eluates were collected after the treatments and applied on MDPC-23 seeded in a 24-well plate to analyze cell death, total protein (TP), and collagen production. The last two tests were performed 24 hours and seven days after the challenge. Data were analyzed by Kruskal-Wallis and Mann-Whitney tests (p<0.05). Results: EDC at all test concentrations did not reduce cell viability, while 5% GA did increase cell metabolism. Cell death by necrosis was not elicited by EDC or 5% GA. At the 24-hour period, 0.3 M and 0.5 M EDC reduced TP production by 18% and 36.8%, respectively. At seven days, increased TP production was observed in all groups. Collagen production at the 24-hour period was reduced when 0.5 M EDC was used. After seven days, no difference was observed among the groups. SEM showed no alteration in cell morphology or number, except in the hydrogen peroxide group. Conclusions: Treatment of acid-etched dentin with EDC or GA did not cause transdentinal cytotoxic effects on odontoblast-like cells.

Efeito citotóxico de agentes clareadores a base de peróxido de hidrogênio a 20% e 38% sobre células odontoblastóides

The aim of this in vitro study was to evaluate the trans-enamel and transdentinal cytotoxic effects of two in-office tooth bleaching techniques that employ bleaching gels containing 20% and 38% of H2 O2 on cultured odontoblast-like cell line (MDPC-23). Sixty enamel/dentin discs were obtained from bovine central incisors and placed individually in artificial pulp chambers. Six groups were formed according to the following enamel treatments: G1- 20% H2 O2 (1 application); G2- 20% H2 O2 (2 applications); G3- 38% H2 O2 (1 application); G4- 38% H2 O2 (2 applications); G5- 38% H2 O2 (3 applications); and G6- control (no treatment). In G1 and G2, the bleaching gel was left in contact with the enamel surface for 45 min in each application. However, in G3, G4, and G5 the bleaching gel was applied for only 10 min per application. After the last application, the extracts were collected and applied on previously cultured cells (30.000 cells/cm2 ) for 24 h. Cell metabolism was evaluated by the MTT assay and cell morphology was analysed by scanning electron microscopy. Cell metabolism decreased by 96.29%; 96.11%; 96.42%; 95.62%; and 97.18% in G1, G2, G3, G4, and G5, respectively. All treated groups differed significantly from non-treated control group (G6) (p < 0.05). However, the difference in cell metabolism among treated groups was not significant statistically. In addition, significant morphological cell alterations were observed in all treated groups. Under the tested experimental conditions, the extracts collected after both tooth bleaching techniques evaluated in this study caused severe toxic effects on cultured odontoblast-like cell MDPC-23.

Efeito antimicrobiano e citotóxico do óleo essencial de Cymbopogon citratus sobre células odontoblastóides

The aim of this study was to evaluate the antimicrobial and cytotoxic effect of essential oil (EO) of lemon grass (Cymbopogon citratus). From the agar diffusion method, different concentrations of EO (0.135%, 0.2% and 1%), and control solutions (chlorhexidine (Chx), distilled water (Ad) and cereal alcohol (Ac)) were applied on cultures of Candida albicans (C.a), Streptococcus mutans (S.m), Streptococcus sobrinus (S.sob) and Lactobacillus acidophilus (L.a). For C.a, S.m and S.sob, the largest inhibition zones in descending order were: Chx, Ac and EO 1%, while the latter two were statistically similar (Mann-Whitney, p> 0.05). For L.a, the largest inhibition halo was observed for the Chx, followed by EO at 1%, 0.2%, 0.135% and Ac. For evaluation of cytotoxicity, the following groups were set: G1: 0,1% EO; G2: pure EO; G3 (positive control): H2 O2 ; G4: cereal alcohol; and G5 (negative control): culture medium – DMEM. The solutions were applied on the cultured MDPC-23 cells, which were plated (30,000 cells/cm2 ) in wells of 24 well-dishes. Cell metabolism was evaluated by MTT assay. Considering G5 (negative control) as 100% of cell metabolism, it was observed for G1, G2, G3 and G4 a percentage reduction in cell metabolism of 29.6%, 82%, 81.2% and 33.4%, respectively. It was concluded that the low concentration of 0,1% OE (C. citratus) was able to inhibit the growth of the strains tested as well as caused mild cytotoxicity to the cultured MDPC-23 cells.

Color alteration, hydrogen peroxide diffusion, and cytotoxicity caused by in-office bleaching protocols

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Effect of different periods of preconditioning with saliva on adhesion of C. albicans to a denture base resin by crystal violet staining and XTT assay.

Aim: The role of saliva on Candida adhesion to biomaterials has not been clearly defined. The present study investigates whether different periods of preconditioning with saliva would influence the adhesion of Candida albicans to a denture base resin. Methods: Ninety samples of acrylic resin with smooth surfaces were made and then divided into five groups: one control without saliva, and four experimental groups conditioned in saliva for periods of 30 min, 1, 3, or 12 h. Candida adhesion was evaluated by crystal violet staining and 2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-5-([phenylamino] carbonyl)-2H-tetrazolium-hydroxide assay. Results: The one-way analysis of variance revealed that there were no significant differences among the mean number of adherent cells or among the mean absorbance for all groups. No significant correlation was found between the two methods used for assessing Candida albicans adhesion. Conclusion: The different periods of preconditioning with saliva had no significant influence on the adhesion of Candida albicans to the denture base acrylic resin.

Metabolism of L929 cells after contact with acrylic resins. Part 1: Acrylic denture base resins

The purpose of this study was to evaluate the effectiveness of complementary heat treatment and water storage in reducing cytotoxicity of acrylic resins denture bases used in Brazil by the MTT assay. Material and Methods: First, nine specimens were fabricated from metal matrix in the form of discs with 14 mm in diameter and 1.2 mm of thick. Immediately after making, 24 or 48 hours after storage in distilled water, the samples of heat-polymerized resins were divided into 3 groups (n = 3) according to the type of thermal treatment: Group 1: samples were individually exposed to microwave energy (500 W for 3 minutes); Group 2: samples were immersed in water at 550 C for 60 minutes; Group 3: samples did not receive heat treatment. To prepare the extracts, 3 samples of each group were placed into vials containing 3 mL of culture medium and stored at 37°C for 24 hours. L929 cells were used and the MTT assay was performed to analyze the cellular metabolism. Two-factor analysis of variance was used to detect significant among groups at 5% significance. Results: After statistical analysis, the materials were classified according to the cytotoxic effect: non-cytotoxic, slightly cytotoxic; moderately cytotoxic; and strongly cytotoxic. The results showed that the resins ranged from moderately cytotoxic to non-cytotoxic, but no statistically significant difference among experimental groups. Furthermore, the water storage and thermal treatments reduced the cytotoxicity of the resins. Conclusions: It was concluded that the resins studied are potentially toxic and that treatments can decrease their cytotoxicity.

Dose-Response and Time-Course of a-Tocoferol Mediating the Cytoprotection Of Dental Pulp Cells Against Hydrogen Peroxide

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Estudo das alterações bioquímicas em sangue e diferentes órgãos de Phrynops geoffroanus (Schweigger, 1812) (Testudines: Chelidae) coletados em ambiente contaminado ou expostos ao benzo[a]pireno

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Chronic reparative changes in medium-sized vessels in a case of primary cutaneous anaplastic large-cell lymphoma with angioinvasive features and cytotoxic phenotype: new histopathological findings in line with indolent clinical behavior

Angioinvasion/angiodestruction has been reported in a small subset of primary cutaneous anaplastic large-cell lymphomas (PCALCL). Recently, PCALCL with angioinvasive features and cytotoxic phenotype has been characterized as a variant associated with good clinical outcomes despite worrisome histopathologic features. We report a case of PCALCL with angioinvasive features and cytotoxic phenotype associated with reparative changes on the wall of medium-sized vessels involved by the neoplasm, including intimal fibroblastic proliferation and luminal obliteration. This vascular pattern, although previously unreported in PCALCL, is in accordance with the indolent behavior observed in this entity and provides a further link with lymphomatoid papulosis type E.

Evaluation of pH effects on genomic integrity in adipose-derived mesenchymal stem cells using the comet assay

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)