In Vitro Toxicological Profile of Labetalol-Folic Acid/Folate Co-Administration in H9c2(2-1) and HepaRG Cells

Abstract

Background and Objectives: The consumption of dietary supplements has increased over the last decades among pregnant women, becoming an efficient resource of micronutrients able to satisfy their nutritional needs during pregnancy. Furthermore, gestational drug administration might be necessary to treat several pregnancy complications such as hypertension. Folic acid (FA) and folate (FT) supplementation is highly recommended by clinicians during pregnancy, especially for preventing neural tube birth defects, while labetalol (LB) is a β-blocker commonly administered as a safe option for the treatment of pregnancy-related hypertension. Currently, the possible toxicity resulting from the co-administration of FA/FT and LB has not been fully evaluated. In light of these considerations, the current study was aimed at investigating the possible in vitro cardio- and hepato-toxicity of LB-FA and LB-FT associations. Materials and Methods: Five different concentrations of LB, FA, FT, and their combination were used in myoblasts and hepatocytes in order to assess cell viability, cell morphology, and wound regeneration. Results: The results indicate no significant alterations in terms of cell viability and morphology in myoblasts (H9c2(2-1)) and hepatocytes (HepaRG) following a 72-h treatment, apart from a decrease in the percentage of viable H9c2(2-1) cells (~67%) treated with LB 150 nM–FT 50 nM. Additionally, LB (50 and 150 nM)–FA (0.2 nM) exerted an efficient wound regenerating potential in H9c2(2-1) myoblasts (wound healing rates were >80%, compared to the control at 66%), while LB-FT (at all tested concentrations) induced no significant impairment to their migration. Conclusions: Overall, our findings indicate that LB-FA and LB-FT combinations lack cytotoxicity in vitro. Moreover, beneficial effects were noticed on H9c2(2-1) cell viability and migration from LB-FA/FT administration, which should be further explored.

Rutin bioconjugates as potential nutraceutical prodrugs: An in vitro and in ovo toxicological screening

“…These results could be explained by the different features of each cell-type tested and of each test compound. H9c2(2-1) cells are morphologically close to immature embryonic cardiomyocytes, possess a good proliferation in vitro, and are applied as reliable tools in cardiotoxicity investigations (Rednic et al, 2022;Witek et al, 2016;Zordoky and EL-Kadi, 2007). These cells present a spindle shape, sugar residues on the cell surface, parallel arrangement, and the capacity to differentiate into multinuclear myotubes.…”

“…Furthermore, H9c2 cells have been widely employed as a model system for in vitro investigations regarding the protective mechanisms associated with flavonoids (Daubney et al, 2015). Similarly, the human immortalized hepatocytes-HepaRG are widely employed as models for various in vitro assays, including toxicity evaluations (Klein et al, 2014;Rednic et al, 2022). Advantageously, the HepaRG cell line maintains the major hepatic-like functions (as drug transporters and enzymes involved in xenobiotics metabolism), have the ability to differentiate into hepatocyte-like and biliary-like cells (Tascher et al, 2019;Young and Young, 2019) and represent a powerful alternative to primary human hepatocytes with higher availability and reproducibility (Rednic et al, 2022;Hu et al, 2019;Blidisel et al, 2021).…”

“…Similarly, the human immortalized hepatocytes-HepaRG are widely employed as models for various in vitro assays, including toxicity evaluations (Klein et al, 2014;Rednic et al, 2022). Advantageously, the HepaRG cell line maintains the major hepatic-like functions (as drug transporters and enzymes involved in xenobiotics metabolism), have the ability to differentiate into hepatocyte-like and biliary-like cells (Tascher et al, 2019;Young and Young, 2019) and represent a powerful alternative to primary human hepatocytes with higher availability and reproducibility (Rednic et al, 2022;Hu et al, 2019;Blidisel et al, 2021). HaCaT cells are spontaneous immortalized human keratinocytes with a long life in culture that express basal properties and could be differentiated under the effects of different inducers as Ca 2+ and high cell density.…”