Browsing by Subject "Cytotoxicity"

Now showing 1 - 19 of 19
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    Antifungal effects and mechanism of action of viscotoxin A3
    (Federation of European Biochemical Societies [Society Publisher], 2006-01-01) Guidici Besseghini, Ana Marcela; Poveda Larrosa, José Antonio; Molina Gallego, María Luisa; Canal, Laura de la; González Ros, José Manuel; Pfüller, Karola; Pfüller, Uwe; Villalaín Boullón, José; Bioquímica y Biología Molecular B e Inmunología
    Viscotoxins are cationic proteins, isolated from different mistletoe species, that belong to the group of thionins, a group of basic cysteine-rich peptides of ≈ 5 kDa. They have been shown to be cytotoxic to different types of cell, including animal, bacterial and fungal. The aim of this study was to obtain information on the cell targets and the mechanism of action of viscotoxin isoform A3 (VtA3). We describe a detailed study of viscotoxin interaction with fungal-derived model membranes, its location inside spores of Fusarium solani, as well as their induced spore death. We show that VtA3 induces the appearance of ion-channel-like activity, the generation of H2O2, and an increase in cytoplasmic free Ca2+. Moreover, we show that Ca2+ is involved in VtA3-induced spore death and increased H2O2 concentration. The data presented here strongly support the notion that the antifungal activity of VtA3 is due to membrane binding and channel formation, leading to destabilization and disruption of the plasma membrane, thereby supporting a direct role for viscotoxins in the plant defence mechanism.
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    Are denture adhesives safe for oral cells?
    (Wiley, 2020-07-12) López García, Sergio; Pecci Lloret, María Pilar; García Bernal, David; Guerrero Gironés, Julia; Pecci Lloret, Miguel Ramón; Rodríguez Lozano, Francisco Javier; Dermatología, Estomatología, Radiología y Medicina Física
    Purpose: To compare the cytotoxicity of six commercially available denture adhesives on human gingival cells: Poligrip Flavour Free Fixative Cream, Fixodent Pro Duo Protection, Novafix cream, FittyDent, Polident Total Action, and Fixodent ProPlus Duo Protection. Material and Methods: Eluates of denture adhesives were brought into contact with human gingival cells and compared to untreated cells (w/o any dental adhesive elute). Cell toxicity was assessed by measuring cell viability (3-(4,5-imethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) (MTT) assays), cell morphology (immunofluorescenceassays), induction of apoptosis/necrosis and production of reactive oxygen species (ROS) (flow cytometry assays). In addition, the pH of each sample was determined. Data were analyzed using one-way analysis of variance (ANOVA) followed by Dunnett’s multiple comparisons test. Results: All denture adhesives tested led to a reduction in pH, especially Fixodent Pro Duo Protection and Fixodent Pro Plus Duo Protection. The cell viability assays showed that Fixodent Pro Duo Protection (1:1 72 hours, p = 3.04 × 10−6; 1:2 72 hours, p = 2.07 × 10−6; 1:4 72 hours, p = 2.04 × 10−6) and Fixodent Pro Plus Duo Protection (1:1 72 hours, p = 2.01 × 10−6; 1:2 72 hours, p = 3.03 × 10−6; 1:4 72 hours, p = 2.02 × 10−6) significantly decreased cell viability at all dilutions. Compared to the control group and the rest of the adhesives, Poligrip Flavour Free Fixative Cream (PFF 1:1 72 hours, p = 2.24 × 10−6; 1:2 72 hours, p = 2.44 × 10−6; 1:4 72 hours, p = 2.04 × 10−6) showed a significantly higher cell viability score at all dilutions. Fixodent Pro Duo Protection and Fixodent Pro Plus Duo Protection, both adhesives containing zinc salts in their composition, were responsible for necrosis, and the number of cells was much reduced, with aberrant morphology and pyknotic nucleus. Finally, Fixodent (1:2, p = 2.04 × 10−6, 1:4, p = 0.00036; 1:2, p = 8.82 × 10−6, 1:4, p = 2.30 × 10−6) products significantly promoted ROS production in gingival cells. Conclusions: The results suggest that denture adhesives containing zinc in their composition could be responsible of the decrease of cell viability, ROS production, aberrant cell morphology, and induction of apoptosis and cell death. However, other possible additional cytotoxic factors must be considered. Thus, more studies are necessary to confirm this hypothesis.
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    Biocompatibility Evaluation of Four Dentin Adhesives Used as Indirect Pulp Capping Materials
    (2017) Cortés, Olga; Alcaina, Antonia; Bernabe, Antonia; Dermatología, Estomatología, Radiología y Medicina Física
    In many cases, the indirect pulp treatment (IPT) is an acceptable treatment for deciduous teeth with reversible pulp inflammation. Various medicaments have been used for IPT, ranging from calcium hy- droxide and glass ionomers to dentin adhesives. Objective: This in vitro trial aimed to measure cyto- toxicity in a cell culture, comparing the following four adhesives: Xeno® V (XE), Excite® F DSC (EX), Adhese® OneF (AD) and Prime & Bond NT (PB). Materials and methods: The adhesives were pre- pared according to the manufacturer’s instructions. After 24 hours of exposure, the cell viability was evaluated using a photometrical test (MTT test). Data were subjected to analysis of variance (ANO- VA). Results: Adhesives, the main component of which was 2-hydroxyethyl methacrylate (HEMA), were found to be less cytotoxic, while those that included the monomer urethane dimethacrylate (UDMA were the most cytotoxic ) in their composition. The effects on cell viability assay varied be- tween the adhesives assayed with statistically significant differences. Conclusions: The results may support the argument that Adhese® OneF is the least cytotoxic of the adhesives assayed, and may be considered as an adhesive agent for indirect pulp treatment. However, Prime and Bond NT showed a reduced biocompatibility under the same conditions.
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    Biocompatibility of new pulp-capping materials NeoMTA Plus, MTA Repair HP and Biodentine on human dental pulp stem cells
    (Elsevier Science, 2017-11-01) Tomás Catalá, Christopher Joseph; Collado-González, Mar; García Bernal, David; Oñate Sánchez, Ricardo Elías; Forner, Leopoldo; Llena, Carmen; Lozano, Adrián; Moraleda Jiménez, José María; Rodríguez Lozano, Francisco Javier; Biología Celular e Histología
    Introduction: The aim of the present study was to evaluate the in vitro cytotoxicity of MTA Repair HP, Neo-MTA Plus, and Biodentine, new bioactive materials used for dental pulp capping, on human dental pulp stem cells (hDPSCs). Methods: Biological testing was carried out in vitro on hDPSCs. Cell viability and cell migration assays were performed using eluates of each capping material. To evaluate cell morphology and cell attachment to the different materials, hDPSCs were directly seeded onto the material surfaces and analyzed by scanning electron microscopy. The chemical composition of the pulp-capping materials was determined by energy-dispersive X-ray and eluates were analyzed by inductively coupled plasma-mass spectrometry. Statistical differences were assessed by analysis of variance and Tukey test (P < .05). Results: Cell viability was moderate after 24 and 48 hours in the presence of MTA Repair HP and NeoMTA Plus, whereas at 48 and 72 hours, Biodentine showed higher rates of cell viability than MTA Repair HP and NeoMTA Plus (P < .001). A cell migration assay revealed adequate cell migration rates for MTA Repair HP and NeoMTA Plus, both similar to the control group rates, meanwhile the highest cell migration rate was observed in the presence of Biodentine (P < .001). Scanning electron microscope studies showed a high degree of cell proliferation and adhesion on Biodentine disks but moderate rates on MTA Repair HP and NeoMTA Plus disks. Energydispersive X-ray pointed to similar weight percentages of C, O, and Ca in all 3 materials, whereas other elements such as Al, Si, and S were also found. Conclusions: The new pulp-capping materials MTA Repair HP, NeoMTA Plus, and Biodentine showed a suitable degree of cytocompatibility with hDPSCs, and good cell migration rates, although Biodentine showed higher rates of proliferation time-dependent.
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    Biological effects of acid-eroded MTA Repair HP and ProRoot MTA on human periodontal ligament stem cells
    (2019) Rodríguez Lozano, Francisco Javier; Oñate Sánchez, Ricardo Elías; Collado-González, Mar; López-García, Sergio; García Bernal, David; Tomás Catalá, Christopher Joseph; Moraleda Jiménez, José María; Lozano, Adrián; Forner, Leopoldo; Dermatología, Estomatología, Radiología y Medicina Física
    Objective The aim of this study was to analyze the biological effects of MTA Repair HP and ProRoot MTA on human periodontal ligament stem cells (hPDLSCs) after exposure to acidic and neutral environments. Materials and methods Discs of each material (n = 30) were exposed to phosphate buffered saline (pH = 7.4) or butyric acid (pH = 5.2) for 7 days, and biological testing was carried out in vitro on hPDLSCs. Cell viability and apoptosis assays were performed using eluates of each root-end filling material. To evaluate cell attachment to the different materials, hPDLSCs were directly seeded onto the material surfaces and analyzed by scanning electron microscopy. The chemical composition of the rootend filling materials was determined by energy-dispersive x-ray and eluates were analyzed by inductively coupled plasma-mass spectrometry. Statistical differences were assessed by ANOVA and Tukey test (p < 0.05). Results Under an acidic environment, both materials displayed similar ion release abilities, with the increased release of Si and Ca ions. Substantial changes in microstructure were observed for both materials after exposure to acidic pH. In addition, material exposure to an acidic environment showed a similar degree of cell adherence, and, surprisingly, MTA Repair HP exhibited higher cell viability rates at pH 5.2 than ProRoot MTA. Conclusions Exposure to an acidic environment promoted Si and Ca ion release from ProRoot MTA and MTA Repair HP. Moreover, we observed optimal biological properties of ProRoot MTA and MTA Repair HP in terms of cell viability, cell death, and cell attachment in both environments. Clinical relevance These results may suggest that MTA Repair HP and ProRoot exhibited optimal biological properties in terms of cell viability, cell death and cell attachment in acidic environment, being considered as materials for root-end filling and perforations.
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    CD33 (Siglec-3) Inhibitory Function: Role in the NKG2D/DAP10 Activating Pathway.
    (Hindawi, 2019) Hernández-Caselles, Trinidad; Corral-San Miguel, Rubén; Ruiz-Alcaraz, Antonio José; García-Peñarrubia, Pilar; Bioquímica y Biología Molecular B e Inmunología
    CD33 (siglec-3), a well-known target in leukemia therapy, is an inhibitory sialoadhesin expressed in human leukocytes of the myeloid lineage and some lymphoid subsets, including NK cells. It may constitute a control mechanism of the innate immune system; nevertheless, its role as an inhibitory receptor remains elusive. Using human NK cells as a cellular model, we analyzed CD33 inhibitory function upon different activating receptors. In high-cytotoxicity NKL cells, CD33 displayed a prominent inhibition on cytotoxicity triggered by the activating receptors NKG2D and, in a lower extent, 2B4, whereas it did not inhibit NKp46-induced cytotoxicity. NKp46 was partially inhibited by CD33 only when low-cytotoxicity NKL cells were tested. CD33 triggering did not inhibit IFN-γ secretion, contrasting with ILT-2 and CD94/NKG2A inhibitory receptors that inhibited cytotoxicity and IFN-γ secretion induced by all activating receptors tested. CD33-mediated inhibition of NKG2D-induced triggering involved Vav1 dephosphorylation. Our results support the role of CD33 as an inhibitory receptor preferentially regulating the NKG2D/DAP10 cytotoxic signaling pathway, which could be involved in self-tolerance and tumor and infected cell recognition.
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    Comparison of cytopathological changes induced by mercury chloride exposure in renal cell lines (VERO and BGM)
    (Elsevier, 2004-03-31) Romero García, Diego; Gómez Zapata, Maximiliano; Luna, Aurelio; Ciencias Sociosanitarias
    The response to mercury chloride was assessed in two cell lines of renal origin, determining the range of toxic concentrations by Neutral Red assay after 24-h of exposure. Morphological changes in the Buffalo Green Monkey (BGM) and VERO cell lines after exposure to subcytotoxic doses (0.045 and 0.038 mM, respectively) equivalent to EC10 (effective concentrations 10%) of mercury chloride were evaluated at the structural and ultrastructural level by optic, transmission and scanning microscopy. Using transmission electron microscopy, the most notable findings in treated cells were the presence of intracytoplasmic inclusion bodies and apoptotic bodies. Scanning microscopy pointed to a cell with a disrupted perinuclear region and a decreased number of surface microvilli. Similar alterations in both in vivo and in vitro experiments have been described by other authors. We conclude that BGM and VERO renal cell lines can be considered as useful tools for toxicological studies involving mercury chloride.
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    Cytotoxic effect of Ochratoxin A on the renal corpuscles of rat kidney: could Ochratoxin A cause kidney failure?
    (Murcia: F. Hernández, 2011) Abdu, Suzan; Ali, Awatif; Ansari, Shatha
    To demonstrate that Ochratoxin A can cause kidney failure as the kidney is the primary target for OTA cytotoxicity. Ochratoxin A (OTA) is a mycotoxin found in our food. The cytotoxic effect of a low cumulative dose of OTA on the renal corpuscles of the kidney tissue has been investigated in this report. This study was based on two groups in which weaning albino rats were used: (1) control; (2) OTA-treated rats (289 µg/kg/day). After 28 days of treatment, a significant decrease in body weight, kidney weight and relative weight were detected in OTA treated rats. Serum creatinine and urea level were slightly elevated. These results revealed significant histological as well as ultrastructral lesions in the OTA treated group. The lesions included global congestion in the renal tissue and loss of demarcation between the cortex and medulla. The normal architecture of the renal corpuscles was destroyed and most of the corpuscles lost their ordinary look. The most apparent histopathological changes were urinary space disappearance and hypercellularity. In addition, congested, undifferentiated, atrophied, hypertrophied, fragmented, sclerotic, degenerated, and obliterated renal corpuscles were distinct. The ultrastructural lesions observed in the renal corpuscles in OTA on treated rats included; proliferation and swelling of the endothelial cells with occasional loss of fenestrae; narrowing of the capillary lumen; damaged podocytes with deteriorated secondary foot processes, hypertrophied and proliferated mesangial cells with expanded mesangial matrix. The endothelium was clearly defected and vacuolated, and lost its fenestrations in many glomerular capillaries. In addition, the glomerular basement membrane (GBM) became visibly thickened and tortuous. Necrotic glomerular cells were frequently observed. Pre-apoptotic cells were also seen. It was concluded that the exposure to relatively low OTA concentrations induced significant lesions to the renal corpuscles. Moreover, it activated oxidative damage and necrosis which can cause extensive damage to the kidney and ultimately kidney failure.
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    Cytotoxic effects of 3-methylindole on alveolar epithelial cells and macrophages: with special reference to microtubular and filamentous assemblies in alveolar type I cells of bovine lung
    (Murcia : F. Hernández, 1988) Atwal, Onkar S.; Minhas, Kanwal J.; Perry, Michael S.
    The alveolar type I cell is a major permeability barrier between the pulmonary interstitium and alveolar spaces and its thin cytoplasmic processes are greatly susceptible to injury. These cells are often observed to undergo progressive vesiculation, vacuolization and desquamation during 3-methylindole (3MI)-induced acute pulmonary edema after oral administration in goats and cattle. The present study describes proliferation of SER and the presence of polymerized tubulin in the form of microtubules arranged in large bundles shown at ultrastructural level as well as with immunofluorescence staining for tubulin in alveolar type I cells 72 hours after 3MI treatment. Such changes were not seen in pulmonary endothelial cells, alveolar type I1 cells, alveolar macrophages and neutrophils. The possible role of microtubules in alveolar type I cell as a mechanistic support to resist disruption against the forces of interstitial and alveolar edema is compared with alveolar type I1 cells, alveolar macrophages and neutrophils. The latter cells undergo dynamic movements in response to inflammatory stimuli and therefore did not show microtubules in their cytoplasm
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    Human dental Pulp stem cells exhibit different biological behaviours in response to commercial bleaching products
    (MDPI, 2018-06-27) Llena, Carmen; Collado-González, Mar; Tomás Catalá, Christopher Joseph; García Bernal, David; Oñate Sánchez, Ricardo Elías; Rodríguez Lozano, Francisco Javier; Forner, Leopoldo; Biología Celular e Histología
    The purpose of this study was to evaluate the diffusion capacity and the biological effects of different bleaching products on human dental pulp stem cells (hDPSCs). The bleaching gel was applied for 90, 30 or 15 min to enamel/dentine discs that adapted in an artificial chamber. The diffusion of hydrogen peroxide (HP) was analysed by fluorometry and the diffusion products were applied to hDPSCs. Cell viability, cell migration and cell morphology assays were performed using the eluates of diffusion products. Finally, cell apoptosis and the expression of mesenchymal stem cell markers were analysed by flow cytometry. Statistical analysis was performed using analysis of variance and Kruskal–Wallis or Mann–Whitney tests ( < 0.05). Significant reductions of approximately 95% in cell viability were observed for the 3 x 15 min groups (p < 0.001), while 1 x 30 min of PerfectBleach and 1 x 90 min of PolaNight resulted in reductions of 50% and 60% in cell viability, respectively (p < 0.001). Similar results were obtained in the migration assay. Moreover, the 3 x 15 min group was associated with cell morphology alterations and reductions of >70% in cell live. Finally, hDPSCs maintained their mesenchymal phenotype in all conditions. Similar concentrations of carbamide peroxide (CP) and HP in different commercial products exhibited different biological effects on hDPSCs.
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    Improving anticancer therapy with naringenin-loaded silk fibroin nanoparticles
    (MDPI, 2020-04-10) Fuster, Marta G.; Carissimi, Guzmán; García Montalbán, Mercedes; Víllora Cano, Gloria; Ingeniería Química
    Naringenin (NAR), a flavonoid present in a variety of fruits, vegetables and herbs, exhibits a wide range of pharmacological effects, including anticancer activity. Nevertheless, its application in cancer therapy is limited due to its low bioavailability at the tumour site because of its poor solubility in water and slow dissolution rate. To improve the therapeutic efficacy of NAR, emergent research is looking into using nanocarriers. Silk fibroin (SF), from the Bombyx mori silkworm, is a biocompatible and biodegradable polymer with excellent mechanical properties and an amphiphilic chemistry that make it a promising candidate as a controlled release drug system. The aim of this work is to synthesize naringenin-loaded silk fibroin nanoparticles (NAR-SFNs) by dissolving the SF in the ionic liquid 1-ethyl-3-methylimidazolium acetate, using high-power ultrasounds and rapid desolvation in methanol followed by the adsorption of NAR. The NAR-SFNs were characterized by dynamic light scattering, Fourier transform infrared spectroscopy and thermogravimetric analysis. The drug loading content and encapsulation efficiency were calculated. The drug release profile best fitted a first order equation. The cytotoxicity effects of free NAR, bare silk fibroin nanoparticles (SFNs) and NAR-SFNs were assessed on HeLa and EA.hy926 cells via 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The results demonstrated the higher in vitro anticancer potential of synthesized NAR-SFNs than that of free NAR in HeLa cancer cells.
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    Influence of dual-cure and self-cure abutment cements for crown implants on human gingival fibroblasts biological properties
    (Elsevier, 2021-09-06) Guerrero Gironés, Julia; López García, Sergio; Pecci Lloret, Miguel Ramón; Pecci Lloret, María Pilar; García Bernal, David; Dermatología, Estomatología, Radiología y Medicina Física
    Aims: To analyze the biological effects of the cements Relyx Unicem 2, Panavia V5, Multilink Hybrid Abutment and SoloCem on human gingival fibroblast cells (HGFs). Materials and methods: HGFs were exposed to different eluates (n = 40) of the studied resin-based cements. Their cytotoxic effects and influence on cell migration were assessed using MTT and wound-healing assays, respectively. Level of HGF attachment, cell morphology and F-actin cytoskeleton content after exposition to the different eluates were analyzed by scanning electron microscopy (SEM) and confocal microscopy analysis, respectively. The levels of intracellular reactive oxygen species (ROS) produced by the eluates of the different cements were also determined by flow cytometry. Data were analyzed by one-way analysis of variance (ANOVA) followed by Tukey´s test. Results: Eluates of SoloCem significantly reduces the viability of HGFs (69% reduction compared to control at 48 h). Cell migration of HGFs in presence of undiluted SoloCem eluates was significantly lower than in the control (88% open wound area at 24 h). Contrarily, migration speed with Multilynk eluates was similar to that of the control group at all periods of time and all dilutions studied. SEM analysis showed very few cells in SoloCem group, and a moderate cell growth in Multilink, Panavia and Relyx groups were detected. Finally, ROS levels detected in HGFs treated with the more concentrated SoloCem and Relyx dilutions were significantly enhanced compared with that in the control cells or the other groups (44% and 11% ROS positive cells, respectively). Conclusions: The results obtained in the present work suggest that Multilink hybrid abutment has better biological properties and lower cytotoxicity for cementing implant crowns on abutments.
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    Morphological characterisation of BGM (Buffalo Green Monkey) cell line exposed to low doses of cadmium chloride
    (Elsevier, 2003-01-23) Romero García, Diego; Gómez Zapata, M.; Luna, A.; García Fernández, A.J.; Ciencias Sociosanitarias
    Morphological changes in the Buffalo Green Monkey (BGM) cell line after exposure to a subcytotoxic dose (0.062 mm, equivalent to EC10—effective concentration 10%) of cadmium chloride have been evaluated. Cells were exposed for 24 h and the effects observed at the ultrastructural level by transmission and scanning microscopy. Using transmission electron microscopy, the most notable findings in treated cells were the presence of intranuclear inclusion bodies and thin intracytoplasmic granules associated to myelin figures and the presence of apoptotic bodies. Other morphological alterations included cell vacuolisation and a reduced cytoplasm volume, condensation of the mitochondria and a decreased number of cytoplasmic organelles, except lysosomes and autophagic vacuoles, which increased in number. Scanning electron microscopy pointed to a cell with a disrupted perinuclear region and a decrease in the number of surface microvilli. We conclude that the BGM cell line may be considered an useful tool for toxicological studies involving cadmium.
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    Morphological characterization of renal cell lines (BGM and VERO) exposed to low doses of lead nitrate
    (Murcia : F. Hernández, 2004) Romero, D.; Gómez Zapata., Maximiliano; Luna, A.; García-Fernández, A.J.
    The response to lead nitrate has been assessed in two cell lines of renal origin. The range of toxic concentrations was determined by Neutral Red assay after 24-h of exposure. Morphological changes in the Buffalo Green Monkey (BGM) and VERO cell lines after exposure to subcytotoxic doses (1.38 mM and 1.04 mM, respectively) equivalent to EC10 (effective concentrations 10%) of lead nitrate were evaluated at the ultrastructural level by transmission microscopy. The most notable finding in treated cells was the presence of inclusion bodies in the form of irregular granules of varying size in both cytoplasm and lysosomes. Cell membrane integrity was not affected. The number of phagolysosomes and myeline figures associated to the inclusion bodies was higher than in the control cultures. We conclude that the phagolysosomic mechanism fails to digest this metal ion and the BGM and VERO renal cell lines can be considered as useful tools for toxicological studies involving lead nitrate.
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    Morphological characterization of renal cell lines (BGM and VERO) exposed to low doses of lead nitrate.
    (Sociedad Española de Histología e Ingeniería Tisular, 2004) Romero García, Diego; Gómez Zapata, Maximiliano; Luna, A.; García Fernández, Antonio Juan; Ciencias Sociosanitarias
    The response to lead nitrate has been assessed in two cell lines of renal origin. The range of toxic concentrations was determined by Neutral Red assay after 24-h of exposure. Morphological changes in the Buffalo Green Monkey (BGM) and VERO cell lines after exposure to subcytotoxic doses (1.38 mM and 1.04 mM, respectively) equivalent to EC10 (effective concentrations 10%) of lead nitrate were evaluated at the ultrastructural level by transmission microscopy. The most notable finding in treated cells was the presence of inclusion bodies in the form of irregular granules of varying size in both cytoplasm and lysosomes. Cell membrane integrity was not affected. The number of phagolysosomes and myeline figures associated to the inclusion bodies was higher than in the control cultures. We conclude that the phagolysosomic mechanism fails to digest this metal ion and the BGM and VERO renal cell lines can be considered as useful tools for toxicological studies involving lead nitrate.
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    Pulp cell cultures obtained with two different methods for in vitro cytotoxicity tests.
    (2006) Cortés, Olga; García, Carlos; Pérez, Leonor; Boj, Juan R.; Alcaina, Antonia; Dermatología, Estomatología, Radiología y Medicina Física
    Aim: To describe two different protocols for obtaining pri- mary pulp cell cultures, one derived from explants and the other following dissociation into single cell suspension by enzyme digestion. Methods: Human pulp tissue was obtained from three healthy premolars. The harvested pulp tissue was prepared for culture using physical methods (one of the premolars) and enzyme, type XI collagenase, (the two remaining premolars). Results: In the case of explant based culture, only limited growth was observed in some cases. However, by enzyme digestion, after two weeks cell growth was evident, and differences in cell type were observed according to the tooth involved. Conclusion: It has been possible to obtain abundant biological material using an enzyme digestion-based protocol for testing purposes, with low experimental variability, as all cells originated from the same individual.
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    Synthesis and Characterization of New Ruthenium (II) Complexes of Stoichiometry [Ru(p-Cymene)Cl2L] and Their Cytotoxicity against HeLa-Type Cancer Cells
    (MDPI, 2022-10-26) Fuster, M. G.; Moulefera, I.; Montalbán, M. G.; Pérez, J.; Víllora Cano, Gloria; García, G.; Ingeniería Química; Facultad de Química
    When the [Ru(p-cymene)( -Cl)Cl]2 complex is made to react, in dichloromethane, with the following ligands: 2-aminobenzonitrile (2abn), 4-aminobenzonitrile (4abn), 2-aminopyridine (2ampy) and 4-aminopyridine (4ampy), after addition of hexane, the following compounds are obtained: [Ru(p-cymene)Cl2(2abn)] (I), [Ru(p-cymene)Cl2(4abn)] (II), [Ru(p-cymene)Cl2(2ampy] (III) and [Ru(pcymene) Cl2( -(4ampy)] (IV). All the compounds are characterized by elemental analysis of carbon, hydrogen and nitrogen, proton nuclear magnetic resonance, COSY 1H-1H, high-resolution mass spectrometry (ESI), thermogravimetry and single-crystal X-ray diffraction (the crystal structure of III is reported and compared with the closely related literature of II). The cytotoxicity effects of complexes were described for cervical cancer HeLa cells via 3-(4.5-dimethylthiazol-2-yl)-2.5-diphenyltetrazolium bromide (MTT) assay. The results demonstrate a low in vitro anticancer potential of the complexes.
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    Thermo-setting glass ionomer cements promote variable biological responses of human dental pulp stem cells
    (2018) Collado-González, Mar; Pecci Lloret, Miguel Ramón; Tomás Catalá, Christopher Joseph; García Bernal, David; Oñate Sánchez, Ricardo Elías; Llena, Carmen; Forner, Leopoldo; Rosa, Vinicius; Rodríguez Lozano, Francisco Javier; Dermatología, Estomatología, Radiología y Medicina Física
    Objective: To evaluate the in vitro cytotoxicity of Equia Forte (GC, Tokyo, Japan) and Ionostar Molar (Voco, Cuxhaven, Germany) on human dental pulp stem cells (hDPSCs). Methods: hDPSCs isolated from third molars were exposed to several dilutions of Equia Forte and Ionostar Molar eluates (1/1, 1/2 and 1/4). These eluates were obtained by storing material samples in respective cell culture medium for 24 h (n = 40). hDPSCs in basal growth culture medium were the control. Cell viability and cell migration assays were performed using the MTT and wound-healing assays, respectively. Also, induction of apoptosis and changes in cell phenotype were evaluated by flow cytometry. Changes in cell morphology were analysed by immunocytofluorescence staining. To evaluate cell attachment to the different materials, hDPSCs were directly seeded onto the material surfaces and analyzed by scanning electron microscopy (SEM). The chemical composition of the materials was determined by energy dispersive X-ray (EDX) and eluates were analyzed by inductively coupled plasma-mass spectrometry (ICP-MS). Statistical analysis was performed with analysis of variance (ANOVA) and Student's t-test (α < 0.05). Results: Undiluted Equia Forte extracts led to a similar cell proliferation rates than the control group from 72 h onwards. There were no significance differences between Equia Forte and Ionostar Molar in terms of cell apoptosis and phenotype. However, in presence of Equia extracts the migration capacity of hDPSCs was higher than in presence of Ionostar Molar (p < 0.05). Also, SEM studies showed a higher degree of cell attachment when Equia Forte extracts were used. Finally, EDX analysis pointed to different weight percentages of C, O and Ca ions in glass ionomer cements, while other elements such as La, Al, Si, W, Mo and F were also detected. Significance: In summary, Equia Forte promoted better biological responses in hDPSCs than Ionostar Molar.
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    Topical fluoride varnishes promote several biological responses on human gingival cells
    (Elsevier, 2021-04-18) López García, Sergio; Pecci Lloret, María Pilar; Pecci Lloret, Miguel Ramón; Guerrero Gironés, Julia; Rodríguez Lozano, Francisco Javier; García Bernal, David; Dermatología, Estomatología, Radiología y Medicina Física
    Objective: To compare the in vitro cytotoxicity of four commercial topical fluoride varnishes widely used in daily dental practice for the prevention of caries on human fibroblasts: Cervitec F, Fixofluor, Fluor Protector S and Duraphat. Material and methods: Human gingival fibroblasts (hGF) were exposed to different concentrations of fluoride varnishes extracts. Biological assays, including MTT and IC50 value determination, annexin-V/7- AAD staining, cell migration and F-actin staining with phalloidin were carried out. Statistical analyses were performed using one-way ANOVA and Tukey’s post hoc test. Results: At 4% concentration, all of the fluoride varnishes extracts affected fibroblasts metabolic activity, exhibiting ahighdegree of cytotoxicity at allmeasured time points.At 0.1%and 1%, Duraphat and Fixofluor or Fluor Protector S and Cervitec F exerted the lowest or highest cytotoxic effects, respectively. Similar effects were evidenced when induction of apoptosis/necrosis and cell migration assays were analyzed. Immunocytochemical assays revealed a similarnumber offibroblasts, without changes inthemorphology and F-actin content at 0.1% concentration of alltested materials, while at 1% concentration, Fluor Protector S and Cervitec F showed few cells with aberrant morphology or non-adhered cells, respectively. Conclusions: Different commercial topical fluoride varnishes with the same therapeutic indication may exhibit different biological effects and cytotoxicity on fibroblasts.