Abstract
Background: DNA-RNA compounds have shown promising protection against cell oxidative stress. This study aimed to assess the cytotoxicity, protective, or preventive effect of different experimental formulations on oral epithelia’s oxidative stress in vitro. Methods: Reconstituted human oral epithelia (RHOE) were grown air-lifted in a continuous-flow bioreactor. Mouthwashes and gels containing DNA-RNA compounds and other bioactive molecules were tested on a model of oxidative stress generated by hydrogen peroxide treatment. Epithelia viability was evaluated using a biochemical MTT-based assay and confocal microscopy; structural and ultrastructural morphology was evaluated by light microscopy and TEM. Results: DNA-RNA showed non-cytotoxic activity and effectively protected against oxidative stress, but did not help in its prevention. Gel formulations did not express adequate activity compared to the mouthwashes. Excipients played a fundamental role in enhancing or even decreasing the bioactive molecules’ effect. Conclusion: A mouthwash formulation with hydrolyzed DNA-RNA effectively protected against oxidative stress without additional enhancement by other bioactive molecules. Active compounds, such as hyaluronic acid, β-Glucan, allantoin, bisabolol, ruscogenin, and essential oils, showed a protective effect against oxidative stress, which was not synergistic with the one of DNA-RNA. Incorporation of surfactant agents showed a reduced, yet significant, cytotoxic effect.
Highlights
Reactive oxygen species (ROS) include oxidants of natural origin involved as signaling molecules in many cellular pathways that, under normal circumstances, are essential to life
A bioreactor (Figure 1) was used to grow 3D reconstituted human oral epithelia (RHOE) on coupons lifted at the air/liquid interface
The results described in the present study demonstrated, for the first time in vitro, that a mouthwash formulation containing hydrolyzed deoxyribonucleic acid (DNA) and ribonucleic acid (RNA) effectively protected against oxidative stress
Summary
Reactive oxygen species (ROS) include oxidants of natural origin involved as signaling molecules in many cellular pathways that, under normal circumstances, are essential to life. An imbalance between ROS production and the antioxidant defenses that protect cells is referred to as oxidative stress. This imbalance initiates the disruption of cellular redox signaling and control, and leads to molecular damage [1]. An overproduction of ROS or, especially, their exogenous presence can cause harmful oxidative stress that can disrupt normal physiology [2]. ROS have been shown to damage biomolecules essential to cell functioning, such as DNA, proteins, and membrane lipids. ROS have been shown to contribute to cancer development by promoting potentially mutagenic DNA changes and damaging lipids in cell membranes [3,4,5,6,7]
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