- Email: [email protected]
Bioinformatic analysis of mast cell degranulation by silver nanoparticles
S190
Abstracts / Toxicology Letters 259S (2016) S73–S247
PP19.9 Cytotoxicity of bismuth subsalicylate nanoparticles in human gingival fibroblast A.L. Vega-Jiménez 1 , A. Almaguer-Flores 1 , E. Camps 2 , M. Uribe-Ramírez 3 , A. Vizcaya-Ruiz 3 1 Laboratorio de Biología Periodontal, Facultad de Odontología UNAM, Ciudad de México, Mexico 2 Departamento de Física, Instituto Nacional de Investigaciones Nucleares, Ciudad de México, Mexico 3 Departamento de Toxicología, Cinvestav-IPN, Ciudad de México, Mexico
Introduction: The use of nanoparticles (NPs) is among the most promising strategies to overcome infections within the medical and dental area. Bismuth subsalicylate (BSS) has been used as an antimicrobial agent against H. pylori infections. However, there is not enough information regarding the antibacterial effect of BBS-NPs in other relevant bacteria strains and their toxic effects in mammalian cells. Objective: The aim of this study was to evaluate the in vitro cytotoxicity of BSS-NPs against cultures of four strains of bacteria of medical importance and in cell cultures of human gingival fibroblast. Materials and methods: BSS-NPs were synthesized by laser ablation and the physico-chemical characterization was performed by UV–Vis spectroscopy, Dynamic light scattering (LDS), Laser Doppler Anemometer (LDA) and Transmission Electron Microscopy (TEM). To evaluate the antibacterial effect, BSS-NPs were tested against Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus and Staphylococcus epidermidis using XTT assay in 24 h. Cell cultures of human gingival fibroblast (HGF-1) were used to measure cell viability by MTS and LDH assays for 24 h. Results: The UV–Vis spectra indicated the presence of salicylic acid functional groups that are present in the BSS chemical structure. LDS measurement reported a diameter in water of between 1.2 and 220 nm and the zeta potential of −9.09 which was obtained by LDA. The use of TEM allowed to observe nanoparticles, with a crystal structure and primary particle size of 10 nm. The results of antibacterial effect indicated a significant growth inhibition in all strains tested (∼89%), being more effective against P. aeruginosa with a 90% at a concentration of 60 g/mL of BSS-NPs. Cell viability assays showed that BSS-NPs did not compromise HGF-1 viability, 96% of mitochondrial activity and 5% of LDH release was observed at 60 g/mL. Conclusions: BSS-NPs showed an efficient antibacterial effect in bacteria strains of dental importance and a low toxicity in cell mammalian cultures; thus BBS-NPs could be a good antimicrobial agent for medical-dental applications. http://dx.doi.org/10.1016/j.toxlet.2016.07.453 PP19.10 Bioinformatic analysis of mast cell degranulation by silver nanoparticles M. Johnson, L. Saba, J.M. Brown Department of Pharmaceutical Sciences, University of Colorado, Aurora, CO, USA Introduction: Silver nanoparticles (AgNP) are the most widely manufactured engineered nanomaterial (ENM) due to their antimicrobial properties. However, with their increased use, there is concern that human and environmental exposures may lead to
adverse outcomes. Our laboratory previously determined that mast cells, which are responsible for allergies and earliest sensors of environmental insult, are activated following AgNP exposure. Genetics are a major contributing factor in many toxicological outcomes, however to date, few studies have examined the contribution of genetics to ENM toxicity. Objective: We hypothesized that genetic factors contribute to regulation of mast cell degranulation following AgNP exposure. Materials and methods: We grew bone marrow-derived mast cells from 23 genetically diverse mouse strains, exposed them to AgNP (25 g/mL, 1 h) and assessed degranulation. Quantitative trait loci (QTL) mapping was performed to identify single nucleotide polymorphisms (SNPs) associated with variation in degranulation following AgNP exposure. Results: Mast cells from different strains of mice displayed a range of degranulation following AgNP exposure, suggesting that multiple genes are likely regulating this response. QTL mapping identified 3 statistically significant (p > 1 × 10−6 ) loci associated with mast cell degranulation; rs32615733 located on chromosome 1 encodes the gene Bzw1, a nuclease that functions in translation initiation; rs27379157 located on chromosome 2 encodes the gene Creb3l1, a transcription factor involved in cell-cell communication; rs300077945 located on chromosome 18 encoding a non-annotated gene. Conclusions: These results provide evidence that a complex set of novel genes regulate mast cell responses to ENM exposure. We are currently integrating transcriptomic analysis (RNAseq) with the genes identified in the QTL plot to comprehensively identify novel candidate susceptibility genes and associated biological pathways of toxicity. Overall, the proposed research will contribute to the field of toxicology by identifying genetic targets that play a role in adverse immune responses to further understand underlying mechanisms of ENM toxicity. Financial support: NIH R01 ES019311. http://dx.doi.org/10.1016/j.toxlet.2016.07.454 PP19.11 Toxicity of silver nanoparticles in biological systems: Does the complexity of biological systems matter? ˜ 1 , B. Borrego 2 , K.O. Juárez-Moreno 3 , M. R. Vázquez-Munoz García-García 3 , J. Mota-Morales 3 , N. Bogdanchikova 1 , A. Huerta-Saquero 1 1 Centro de Nanociencias y Nanotecnología, Universidad Nacional Autónoma de México, Ensenada, BC, Mexico 2 Centro de Investigación en Sanidad Animal, INIA, Valdeolmos, Madrid, Spain 3 CONACyT Research Fellow at Centro de Nanociencias y Nanotecnología, Universidad Nacional Autónoma de México, Ensenada, BC, Mexico
Silver nanoparticles (AgNPs) are one of the most studied nanomaterials, but little is known about their adverse effects. Taking into account their differential toxicity between microorganisms and higher organisms, it is assumed that its effect depends on the complexity of biological systems. Unfortunately, current studies are not comparable due to the limited taxonomic diversity. This study evaluated the effect of AgNPs on biological systems of different levels of complexity. The studied organisms include bacteria, protists, fungi, animal and human cancer cell lines, and viruses. PVP-AgNPs used were subjected to optical and FT-IR analysis. Morphology characterization was performed by TEM and SEM. AgNPs mean hydrodynamic diameter and stability (zeta potential)
Abstracts / Toxicology Letters 259S (2016) S73–S247
PP19.9 Cytotoxicity of bismuth subsalicylate nanoparticles in human gingival fibroblast A.L. Vega-Jiménez 1 , A. Almaguer-Flores 1 , E. Camps 2 , M. Uribe-Ramírez 3 , A. Vizcaya-Ruiz 3 1 Laboratorio de Biología Periodontal, Facultad de Odontología UNAM, Ciudad de México, Mexico 2 Departamento de Física, Instituto Nacional de Investigaciones Nucleares, Ciudad de México, Mexico 3 Departamento de Toxicología, Cinvestav-IPN, Ciudad de México, Mexico
Introduction: The use of nanoparticles (NPs) is among the most promising strategies to overcome infections within the medical and dental area. Bismuth subsalicylate (BSS) has been used as an antimicrobial agent against H. pylori infections. However, there is not enough information regarding the antibacterial effect of BBS-NPs in other relevant bacteria strains and their toxic effects in mammalian cells. Objective: The aim of this study was to evaluate the in vitro cytotoxicity of BSS-NPs against cultures of four strains of bacteria of medical importance and in cell cultures of human gingival fibroblast. Materials and methods: BSS-NPs were synthesized by laser ablation and the physico-chemical characterization was performed by UV–Vis spectroscopy, Dynamic light scattering (LDS), Laser Doppler Anemometer (LDA) and Transmission Electron Microscopy (TEM). To evaluate the antibacterial effect, BSS-NPs were tested against Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus and Staphylococcus epidermidis using XTT assay in 24 h. Cell cultures of human gingival fibroblast (HGF-1) were used to measure cell viability by MTS and LDH assays for 24 h. Results: The UV–Vis spectra indicated the presence of salicylic acid functional groups that are present in the BSS chemical structure. LDS measurement reported a diameter in water of between 1.2 and 220 nm and the zeta potential of −9.09 which was obtained by LDA. The use of TEM allowed to observe nanoparticles, with a crystal structure and primary particle size of 10 nm. The results of antibacterial effect indicated a significant growth inhibition in all strains tested (∼89%), being more effective against P. aeruginosa with a 90% at a concentration of 60 g/mL of BSS-NPs. Cell viability assays showed that BSS-NPs did not compromise HGF-1 viability, 96% of mitochondrial activity and 5% of LDH release was observed at 60 g/mL. Conclusions: BSS-NPs showed an efficient antibacterial effect in bacteria strains of dental importance and a low toxicity in cell mammalian cultures; thus BBS-NPs could be a good antimicrobial agent for medical-dental applications. http://dx.doi.org/10.1016/j.toxlet.2016.07.453 PP19.10 Bioinformatic analysis of mast cell degranulation by silver nanoparticles M. Johnson, L. Saba, J.M. Brown Department of Pharmaceutical Sciences, University of Colorado, Aurora, CO, USA Introduction: Silver nanoparticles (AgNP) are the most widely manufactured engineered nanomaterial (ENM) due to their antimicrobial properties. However, with their increased use, there is concern that human and environmental exposures may lead to
adverse outcomes. Our laboratory previously determined that mast cells, which are responsible for allergies and earliest sensors of environmental insult, are activated following AgNP exposure. Genetics are a major contributing factor in many toxicological outcomes, however to date, few studies have examined the contribution of genetics to ENM toxicity. Objective: We hypothesized that genetic factors contribute to regulation of mast cell degranulation following AgNP exposure. Materials and methods: We grew bone marrow-derived mast cells from 23 genetically diverse mouse strains, exposed them to AgNP (25 g/mL, 1 h) and assessed degranulation. Quantitative trait loci (QTL) mapping was performed to identify single nucleotide polymorphisms (SNPs) associated with variation in degranulation following AgNP exposure. Results: Mast cells from different strains of mice displayed a range of degranulation following AgNP exposure, suggesting that multiple genes are likely regulating this response. QTL mapping identified 3 statistically significant (p > 1 × 10−6 ) loci associated with mast cell degranulation; rs32615733 located on chromosome 1 encodes the gene Bzw1, a nuclease that functions in translation initiation; rs27379157 located on chromosome 2 encodes the gene Creb3l1, a transcription factor involved in cell-cell communication; rs300077945 located on chromosome 18 encoding a non-annotated gene. Conclusions: These results provide evidence that a complex set of novel genes regulate mast cell responses to ENM exposure. We are currently integrating transcriptomic analysis (RNAseq) with the genes identified in the QTL plot to comprehensively identify novel candidate susceptibility genes and associated biological pathways of toxicity. Overall, the proposed research will contribute to the field of toxicology by identifying genetic targets that play a role in adverse immune responses to further understand underlying mechanisms of ENM toxicity. Financial support: NIH R01 ES019311. http://dx.doi.org/10.1016/j.toxlet.2016.07.454 PP19.11 Toxicity of silver nanoparticles in biological systems: Does the complexity of biological systems matter? ˜ 1 , B. Borrego 2 , K.O. Juárez-Moreno 3 , M. R. Vázquez-Munoz García-García 3 , J. Mota-Morales 3 , N. Bogdanchikova 1 , A. Huerta-Saquero 1 1 Centro de Nanociencias y Nanotecnología, Universidad Nacional Autónoma de México, Ensenada, BC, Mexico 2 Centro de Investigación en Sanidad Animal, INIA, Valdeolmos, Madrid, Spain 3 CONACyT Research Fellow at Centro de Nanociencias y Nanotecnología, Universidad Nacional Autónoma de México, Ensenada, BC, Mexico
Silver nanoparticles (AgNPs) are one of the most studied nanomaterials, but little is known about their adverse effects. Taking into account their differential toxicity between microorganisms and higher organisms, it is assumed that its effect depends on the complexity of biological systems. Unfortunately, current studies are not comparable due to the limited taxonomic diversity. This study evaluated the effect of AgNPs on biological systems of different levels of complexity. The studied organisms include bacteria, protists, fungi, animal and human cancer cell lines, and viruses. PVP-AgNPs used were subjected to optical and FT-IR analysis. Morphology characterization was performed by TEM and SEM. AgNPs mean hydrodynamic diameter and stability (zeta potential)