Differentiated mitochondrial function in mouse 3T3 fibroblasts and human epithelial or endothelial cells in response to chemical exposure

Abstract

Mouse 3T3 fibroblasts are commonly used for in vitro toxicity testing; however, their sensitivity to stimuli is not well defined. To assess the sensitivity of the 3T3 cell line, the study compared the changes in mitochondrial membrane potential (MMP) occurring after exposure to eight chemicals known to demonstrate pro-apoptotic activity (glycerol, isopropanol, ethanol, paracetamol, propranolol, cobalt chloride, formaldehyde and atropine). Five cell lines were used as follows: mouse 3T3 fibroblasts, human epithelial cells (A549, Caco-2 and HepG2) and human endothelial cells (HMEC-1). Cell sensitivity was assessed based on the total area under and over the dose-response curves (AUOC) in relation to baselines. The 3T3 fibroblasts had the highest AUOC values and were the most sensitive to the action of all the examined chemicals, with the exception of formaldehyde. Significant changes in MMP between the 3T3 cell line and other cells were observed after cell treatment with atropine (A549, Caco-2 or HMEC-1 cells vs 3T3 cells, P < 0.05), propranolol (A549 vs 3T3 cells, P < 0.01; HepG2 vs 3T3 cells, P < 0.05), cobalt chloride (A549 cells vs 3T3 cells, P < 0.01) or ethanol (HMEC-1 vs 3T3, P < 0.05). Formaldehyde appeared the most toxic compound for Caco-2 cells (Caco-2 vs 3T3 cells, P < 0.05). The surface areas (AUOC) calculated for each other chemical and obtained for HepG2, Caco-2, A549 and HMEC-1 did not differ significantly between cell lines. We postulate that mouse 3T3 fibroblasts demonstrate significantly higher relative sensitivity to many agents with toxic potential.

Transcriptomic‐based toxicological investigations of ethanol to human umbilical vein endothelial cells

“…HUVECs were seeded at a density of 4 × 10 4 per well in 24‐well plates and incubated with 5‐, 10‐, 25‐, 50‐, or 100‐mM ethanol (purchased from Sigma‐Aldrich, USA). The highest concentration (100 mM) is comparable with previous in vitro studies (Boncler, Lukasiak, Dastych, Golanski & Watala, 2019; Fisher, Thomas & Abdul‐Rasool, 2020; Mishra, Agas, Schuetz, Kalluru & Haorah, 2020; Xue et al, 2018). Assuming the highest ethanol consumption as 9.8 L per year (World Health Organisation, 2018) and blood volume of adults as 5 L, the ethanol level in blood every day is approximately 92 mM, close to the highest concentration used in this study.…”

“…We used two concentrations. The concentration of 50 mM has been used in previous toxicological studies (Boncler, Lukasiak, Dastych, Golanski & Watala, 2019; Devall et al, 2020; Fisher, Thomas & Abdul‐Rasool, 2020; Mishra, Agas, Schuetz, Kalluru & Haorah, 2020; Xue et al, 2018; Zhu et al, 2017) and could be reached after daily consumption of ethanol according to the calculation described in Section 2. A much lower concentration 5 mM was also used, because researchers suggested that low levels of ethanol could induce hormesis (Calabrese & Baldwin, 2003; Hayes, 2007).…”

“…Interestingly, numerous studies found that environmental toxicants, for example, mercury (Farkhondeh, Afshari, Mehrpour & Samarghandian, 2020), coronavirus (Hayden, 2020), and nanomaterials (Cao et al, 2017), induced toxicological responses to endothelial cells leading to CVD development. Previous studies also revealed that high levels of ethanol induced toxic effects in endothelial cells (Boncler, Lukasiak, Dastych, Golanski & Watala, 2019; Fisher, Thomas & Abdul‐Rasool, 2020; Mishra, Agas, Schuetz, Kalluru & Haorah, 2020; Xue et al, 2018). However, most of the studies were based on traditional toxicological methods, which focused on a specific signaling pathway but may fail to provide the information about the systemic changes in endothelial cells after ethanol exposure.…”

Antimicrobial Activity of Lipopeptide Biosurfactants Against Foodborne Pathogen and Food Spoilage Microorganisms and Their Cytotoxicity

“…that make them advantageous in food and food packaging applications Bumbudsanpharoke et al, 2015), that's why it seemed necessary to add them here. Data of these preservatives/antioxidants such as formaldehyde (E240) (Marcsek et al, 2007;Boncler et al, 2019), Salvia rosmarinus extract (E392) (Aherne et al, 2007;Waller et al, 2016), butylhydroxyanisol (BHA) (E320) (Labrador et al, 2007) and biological extracts such as carvacol (the main component of many essential oil) (Llana- Ruiz-Cabello et al, 2014;Mooyottu et al, 2014), iturin A (another antifungal lipopeptide produced by B. subtilis) (EFSA Panel on Additives and Products or Substances used in Animal Feed [FEEDAP], 2017;Zhao et al, 2018), Thymus vulgaris oil extract (Pérez-González et al, 2019) and nanoparticles such as ZnO NP (Song et al, 2014;Venkateasan et al, 2017) and Ag NP (Song et al, 2014;Kumar et al, 2017) have been collected from literature. In conclusion, as shown in Figure 4, lipopeptides had low toxicity on human cell lines.…”

Construction of Sol–Gel Phase-Reversible Hydrogels with Tunable Properties with Native Nanofibrous Protein as Building Blocks

“…We chose Caco-2 and NIH/ 3T3 to conduct proliferation and cytotoxicity assay because the former has been widely used as a model of the intestinal epithelial barrier to measure interaction, uptake, and cellular transport of drugs and food components, 50 and the latter has been shown in many other biological and toxicological studies (wound healing, angiogenesis, and tissue regeneration). 51 Hydrogels with different protein concentrations at the same molar ratio of AbPM/zinc ions as 1/2.5 were tested shown in Figure 6. No significant cytotoxic effects on Caco-2 cells were observed for 60−100 μM AbPM gels at 6 h with over 90% of cells remaining viable.…”