Phthalates and Alternative Plasticizers Differentially affect Phenotypic Parameters in Gonadal Somatic and Germ Cell Lines

2021 ◽  
Author(s):  
Abishankari Rajkumar ◽  
Trang Luu ◽  
Marc A Beal ◽  
Tara S Barton-Maclaren ◽  
Barbara F Hales ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Cell Line ◽  
Cell Lines ◽  
Lipid Droplet ◽  
Reproductive Toxicity ◽  
Mitochondrial Activity ◽  
Steroidogenic Cell ◽  
Ethylhexyl Phthalate ◽  
And Oxidative Stress ◽  
Sertoli Cell Line

Abstract The developmental and reproductive toxicity associated with exposure to phthalates has motivated a search for alternatives. However, there is limited knowledge regarding the adverse effects of some of these chemicals. We used high-content imaging to compare the effects of mono (2-ethylhexyl) phthalate (MEHP) with six alternative plasticizers: di-2-ethylhexyl terephthalate (DEHTP); diisononyl-phthalate (DINP); di-isononylcyclohexane-1,2-dicarboxylate (DINCH); 2-ethylhexyl adipate (DEHA); 2,2,4-trimethyl 1,3-pentanediol diisobutyrate (TXIB) and di-iso-decyl-adipate (DIDA). A male germ spermatogonial cell line (C18-4), a Sertoli cell line (TM4) and two steroidogenic cell lines (MA-10 Leydig and KGN granulosa) were exposed for 48h to each chemical (0.001-100 μM). Cell images were analyzed to assess cytotoxicity and effects on phenotypic endpoints. Only MEHP (100 μM) was cytotoxic and only in C18-4 cells. However, several plasticizers had distinct phenotypic effects in all four cell lines. DINP increased Calcein intensity in C18-4 cells, whereas DIDA induced oxidative stress. In TM4 cells, MEHP, and DINCH affected lipid droplet numbers, while DEHTP and DINCH increased oxidative stress. In MA-10 cells, MEHP increased lipid droplet areas and oxidative stress; DINP decreased the number of lysosomes, while DINP, DEHA and DIDA altered mitochondrial activity. In KGN cells, MEHP, DINP and DINCH increased the number of lipid droplets, whereas DINP decreased the number of lysosomes, increased oxidative stress and affected mitochondria. The Toxicological Priority Index (ToxPi) provided a visual illustration of the cell line specificity of the effects on phenotypic parameters. The lowest administered equivalent doses were observed for MEHP. We propose that this approach may assist in screening alternative plasticizers.

Di(2-ethylhexyl)phthalate induces reproductive toxicity via JAZF1/TR4 pathway and oxidative stress in pubertal male rats

2019 ◽  
Vol 35 (3) ◽  
pp. 228-238 ◽  
Author(s):  
Yu-Qin Shi ◽  
Guo-Qing Fu ◽  
Jing Zhao ◽  
Shen-Zhou Cheng ◽  
You Li ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Morphological Changes ◽  
Reproductive Toxicity ◽  
Male Reproduction ◽  
Male Rats ◽  
Terminal Deoxynucleotidyl Transferase ◽  
Cyclin A1 ◽  
Ethylhexyl Phthalate ◽  
Dehp Exposure ◽  
And Oxidative Stress

Di(2-ethylhexyl)phthalate (DEHP) is a typical endocrine-disrupting chemical and reproductive toxicant. Although previous studies have attempted to describe the mechanism by which DEHP exposure results in reproductive dysfunction, few studies focused on puberty, a critical period of reproductive development, and the increased susceptibility to injury in adolescents. To elucidate the mechanism underpinning the testicular effects of DEHP in puberty, we sought to investigate the JAZF1/TR4 pathway in the testes of pubertal rats. Specifically, we focused on the role of the JAZF1/TR4 pathway in male reproduction, including the genes JAZF1, TR4, Sperm 1, and Cyclin A1. In the present study, rats were exposed to increasing concentrations of DEHP (0, 250, 500, and 1000 mg/kg/day) by oral gavages for 30 days. Then we assayed testicular zinc and oxidative stress levels. Our results indicated that DEHP exposure could lead to oxidative stress and decrease the contents of testicular zinc. Additionally, significant morphological changes and cell apoptosis were observed in testes exposed to DEHP, as identified by hematoxylin and eosin staining and the terminal deoxynucleotidyl transferase-mediated nick and labeling assay. By measuring the expression levels of the above relevant genes by qPCR, we found the DEHP-induced increased expression of JAZF1 and decreased expression of TR4, Sperm 1, and Cyclin A1. Therefore, we have demonstrated that in vivo exposure to DEHP might induce reproductive toxicity in pubertal male rats through the JAZF1/TR4 pathway and oxidative stress.

scholarly journals A Comparative Study of the Cytotoxic Effects and Oxidative Stress of Gossypol on Bovine Kidney and HeLa Cell Lines

2021 ◽  
Vol 15 (3) ◽  
pp. 157-164
Author(s):  
Mahsa Daneshmand ◽  
◽  
Jamileh Salar Amoli ◽  
Tahereh Ali Esfahani ◽  
◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Hela Cell ◽  
Cell Line ◽  
Cell Lines ◽  
Toxic Effects ◽  
Cytotoxic Effects ◽  
Bovine Kidney ◽  
Hela Cell Lines ◽  
Significant Difference ◽  
And Oxidative Stress

Background: Cotton seed is one of the main sources of protein in animal feeds, containing gossypol, which has been shown to have toxic effects. Results reported by various studies also indicate the anti-cancer effects of gossypol on various cell types. However, its toxic effects on human and animal cells have not been fully established. This study was planned to investigate, for the first time, the cytotoxic effects and oxidative stress induced by gossypol on normal Bovine Kidney (BK) and HeLa cell lines, representing typical healthy and cancer cells, respectively. Methods: The BK and HeLa cell lines were treated for 24, 48 or 72 hours with 5, 10 or 20 ppm of gossypol (+/-). The cellular bio-availability and cytotoxicity were measured by MTT assay. The catalase and Malondialdehyde (MDA) levels were also measured to represent the oxidative stress parameters. Results: The percentages of cytotoxicity in BK and HeLa cell lines were calculated at a gossypol concentration of 5, 10 and 20 ppm over 24, 48 or 72 hours of incubation, respectively. The Lethal Concentration 50 (lC50) values were also determined for the two cell lines. No changes in the catalase and lipid peroxidase activities were observed in either cell line. Conclusion: The percentage of the gossypol cytotoxicity was concentration-dependent. By comparing the IC50 in both cell lines using one-way Analysis of Variance (ANOVA) analysis, a significant difference was observed, suggesting that Hela cells were less sensitive to gossypol than the BK cells. Lack of changes in the oxidative stress, as tested by catalase and MDA assays, demonstrated that gossypol did not induce oxidative stress in either cell line.

scholarly journals Elucidation of the Effects of Bisphenol A and Structural Analogs on Germ and Steroidogenic Cells using Single Cell High-Content Imaging

2021 ◽  
Author(s):  
Abishankari Rajkumar ◽  
Trang Luu ◽  
Marc A Beal ◽  
Tara S Barton-Maclaren ◽  
Bernard Robaire ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Bisphenol A ◽  
Cell Viability ◽  
Cell Line ◽  
Cell Lines ◽  
Lipid Droplets ◽  
Additional Data ◽  
High Content Imaging ◽  
Steroidogenic Cells ◽  
And Oxidative Stress

Abstract Concerns about the potential adverse effects of bisphenol A (BPA) have led to an increase in the use of replacements, yet the toxicity data for several of these chemicals are limited. Using high-content imaging we compared the effects of BPA, BPAF, BPF, BPS, BPM, and BPTMC in germ (C18-4 spermatogonial) and steroidogenic (MA-10 Leydig and KGN granulosa) cell lines. Effects on cell viability and phenotypic markers were analyzed to determine benchmark concentrations (BMCs) and estimate administered equivalent doses (AEDs). In all three cell lines, BPA was one of the least cytotoxic bisphenol compounds tested, while BPM and BPTMC were the most cytotoxic. Interestingly, BPF and BPS were cytotoxic only in MA-10 cells. Effects on phenotypic parameters, including mitochondria, lysosomes, lipid droplets, and oxidative stress, were both bisphenol- and cell-line specific. BPA exposure affected mitochondria (BMC: 1.2 μM; AED: 0.09 mg/kg/day) in C18-4 cells. Lysosome numbers were increased in MA-10 cells exposed to BPA or BPAF but decreased in KGN cells exposed to BPAF or BPM. Lipid droplets were decreased in C18-4 cells exposed to BPF and in MA-10 cells exposed to BPTMC but increased in BPF, BPM and BPTMC exposed KGN cells. BPA and BPM exposure induced oxidative stress in MA-10 and KGN cells, respectively. In summary, structurally similar bisphenols displayed clear cell-line specific differences in BMC and AED values for effects on cell viability and phenotypic endpoints. This approach, together with additional data on human exposure, may aid in the selection and prioritization of responsible replacements for BPA. 

scholarly journals Genomic Adaption and Mutational Patterns in a HaCaT Subline Resistant to Alkylating Agents and Ionizing Radiation

2021 ◽  
Vol 22 (3) ◽  
pp. 1146
Author(s):  
Reinhard Ullmann ◽  
Benjamin Valentin Becker ◽  
Simone Rothmiller ◽  
Annette Schmidt ◽  
Horst Thiermann ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Cell Line ◽  
Copy Number ◽  
Resistant Cell ◽  
Resistant Cell Line ◽  
Chromosomal Translocations ◽  
Dna Copy Number ◽  
Mutational Signatures ◽  
Copy Number Changes ◽  
And Oxidative Stress

Sulfur mustard (SM) is a chemical warfare agent that can damage DNA via alkylation and oxidative stress. Because of its genotoxicity, SM is cancerogenic and the progenitor of many chemotherapeutics. Previously, we developed an SM-resistant cell line via chronic exposure of the popular keratinocyte cell line HaCaT to increasing doses of SM over a period of 40 months. In this study, we compared the genomic landscape of the SM-resistant cell line HaCaT/SM to its sensitive parental line HaCaT in order to gain insights into genetic changes associated with continuous alkylation and oxidative stress. We established chromosome numbers by cytogenetics, analyzed DNA copy number changes by means of array Comparative Genomic Hybridization (array CGH), employed the genome-wide chromosome conformation capture technique Hi-C to detect chromosomal translocations, and derived mutational signatures by whole-genome sequencing. We observed that chronic SM exposure eliminated the initially prevailing hypotetraploid cell population in favor of a hyperdiploid one, which contrasts with previous observations that link polyploidization to increased tolerance and adaptability toward genotoxic stress. Furthermore, we observed an accumulation of chromosomal translocations, frequently flanked by DNA copy number changes, which indicates a high rate of DNA double-strand breaks and their misrepair. HaCaT/SM-specific single-nucleotide variants showed enrichment of C > A and T > A transversions and a lower rate of deaminated cytosines in the CpG dinucleotide context. Given the frequent use of HaCaT in toxicology, this study provides a valuable data source with respect to the original genotype of HaCaT and the mutational signatures associated with chronic alkylation and oxidative stress.

scholarly journals Hyperbaric Oxygen Treatment: Effects on Mitochondrial Function and Oxidative Stress

Biomolecules ◽  
2021 ◽  
Vol 11 (12) ◽  
pp. 1827
Author(s):  
Nofar Schottlender ◽  
Irit Gottfried ◽  
Uri Ashery
Keyword(s):  
Oxidative Stress ◽  
Hyperbaric Oxygen ◽  
Mitochondrial Function ◽  
Defense Mechanisms ◽  
Term Treatment ◽  
Mitochondrial Activity ◽  
Hyperbaric Oxygen Treatment ◽  
Oxygen Treatment ◽  
Long Term Treatment ◽  
And Oxidative Stress

Hyperbaric oxygen treatment (HBOT)—the administration of 100% oxygen at atmospheric pressure (ATA) greater than 1 ATA—increases the proportion of dissolved oxygen in the blood five- to twenty-fold. This increase in accessible oxygen places the mitochondrion—the organelle that consumes most of the oxygen that we breathe—at the epicenter of HBOT’s effects. As the mitochondrion is also a major site for the production of reactive oxygen species (ROS), it is possible that HBOT will increase also oxidative stress. Depending on the conditions of the HBO treatment (duration, pressure, umber of treatments), short-term treatments have been shown to have deleterious effects on both mitochondrial activity and production of ROS. Long-term treatment, on the other hand, improves mitochondrial activity and leads to a decrease in ROS levels, partially due to the effects of HBOT, which increases antioxidant defense mechanisms. Many diseases and conditions are characterized by mitochondrial dysfunction and imbalance between ROS and antioxidant scavengers, suggesting potential therapeutic intervention for HBOT. In the present review, we will present current views on the effects of HBOT on mitochondrial function and oxidative stress, the interplay between them and the implications for several diseases.

Cartap-induced cytotoxicity in mouse C2C12 myoblast cell line and the roles of calcium ion and oxidative stress on the toxic effects

Toxicology ◽  
2006 ◽  
Vol 219 (1-3) ◽  
pp. 73-84 ◽  
Author(s):  
Jiunn-Wang Liao ◽  
Jaw-Jou Kang ◽  
Chian-Ren Jeng ◽  
Shao-Kuang Chang ◽  
Ming-Jang Kuo ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Cell Line ◽  
Calcium Ion ◽  
Toxic Effects ◽  
C2c12 Myoblast ◽  
Myoblast Cell Line ◽  
Myoblast Cell ◽  
And Oxidative Stress
Sign in / Sign up

Export Citation Format

Share Document