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

Abstract

Rutin (RUT) is considered one the most attractive flavonoids from a therapeutic perspective due to its multispectral pharmacological activities including antiradical, anti-inflammatory, antiproliferative, and antimetastatic among others. Still, this compound presents a low bioavailability what narrows its clinical applications. To overcome this inconvenience, the current paper was focused on the synthesis, characterization, and toxicological assessment of two RUT bioconjugates obtained by enzymatic esterification with oleic acid (OA) and linoleic acid (LA)—rutin oleate (RUT-O) and rutin linoleate (RUT-L), as flavonoid precursors with improved physicochemical and biological properties. Following the enzymatic synthesis in the presence of Novozyme® 435, the two bioconjugates were obtained, their formation being confirmed by RAMAN and FT-IR spectroscopy. The in vitro and in ovo toxicological assessment of RUT bioconjugates (1–100 µM) was performed using 2D consecrated cell lines (cardiomyoblasts - H9c2(2-1), hepatocytes—HepaRG, and keratinocytes—HaCaT), 3D reconstructed human epidermis tissue (EpiDerm™), and chick chorioallantoic membranes, respectively. The results obtained were test compound, concentration—and cell-type dependent, as follows: RUT-O reduced the viability of H9c2(2-1), HepaRG, and HaCaT cells at 100 µM (to 77.53%, 83.17%, and 78.32%, respectively), and induced cell rounding and floating, as well as apoptotic-like features in the nuclei of all cell lines, whereas RUT-L exerted no signs of cytotoxicity in all cell lines in terms of cell viability, morphology, and nuclear integrity. Both RUT esters impaired the migration of HepaRG cells (at 25 µM) and lack irritative potential (at 100 µM) in vitro (tissue viability >50%) and in ovo (irritation scores of 0.70 for RUT-O, and 0.49 for RUT-L, respectively). Computational predictions revealed an increased lipophilicity, and reduced solubility, drug-likeness and drug score of RUT-O and RUT-L compared to their parent compounds—RUT, OA, and LA. In conclusion, we report a favorable toxicological profile for RUT-L, while RUT-O is dosage-limited since at high concentrations were noticed cytotoxic effects.

In Vitro Evaluation of Alpha-Tocopherol as Potential Agent in Combating the Hepatotoxicity Induced by Ethinylestradiol

“…The current in vitro study aimed to investigate the potential hepatoprotective effect of AT against EEinduced damage in HepaRG cells, considering the following aspects: (i) DIH represents one of the main causes leading to drug withdrawal from therapeutic use [14]; (ii) many drugs have been already classified as hepatotoxic, among which OCs were associated with more than 100 cases of drug-induced liver damage, being included in the category A of hepatotoxicity according to the LiverTox database [3]; (iii) EE is the main estrogenic component of modern OCs which was previously shown to exert hepatotoxicity in vivo [22]; (iv) an effective solution for ameliorating the hepatotoxicity induced by EE might be the use of antioxidants since oestrogens in excess were previously showed to enhance oxidative stress in hepatic cells through various mechanisms such as the depletion of cellular antioxidants (e.g., glutathione) [34]; and (v) AT is a potent radical scavenger with hepatoprotective activity [25]. The HepaRG cells were selected as an experimental model for this study because they represent a highly available alternative to primary human hepatocytes, retain the major functions of the liver, and are widely used in toxicological evaluations [4,10]. The impact of AT, EE, and AT associated with EE on HepaRG cells' viability after a 24 h treatment was evaluated by applying the MTT assay (Figure 1).…”

Eugenol Induces Apoptosis in Tongue Squamous Carcinoma Cells by Mediating the Expression of Bcl-2 Family

“…Cell viability was assessed using the MTT (3-(4,5 dimethylthiazol2-yl)-2,5- diphenyltetrazolium bromide) colorimetric assay, in a similar manner as described before, according to the following protocol [ 18 , 19 ]. Briefly, after reaching 80% confluence, the cells were washed with PBS, detached from the culture flasks using trypsin-EDTA solution, sedimented by centrifugation and, by using trypan blue, they were counted and seeded in 96-well plates at a concentration of 1 × 10 4 /200 µL medium/well.…”

From polyphenol to o‐quinone: Occurrence, significance, and intervention strategies in foods and health implications

“…There are some reports on the safety of specific modified polyphenols. The enzymatic esterification products of rutin (rutin oleate and rutin linoleate) were presumed to be nonirritant, non‐tumorigenic, non‐mutagenic, and non‐reproductively toxic (Dehelean et al., 2022). Among them, rutin linoleate has enhanced in vitro safety even at high concentrations, whereas high‐dose administration of rutin oleate probably triggers toxic events.…”