Evaluation of Conventional and Hyaluronic Acid-Coated Thymoquinone Liposomes in an In Vitro Model of Dry Eye

Abstract

Dry eye disease (DED) is a common ocular disorder characterized by an inadequate lubrication of the eye by tears leading to inflammation and the alteration of the ocular surface. Current treatments are often limited due to their side effects and ineffectiveness. Thymoquinone (TQ) is a natural compound present in the essential oil of Nigella sativa L., with anti-inflammatory and antioxidant activities. In this study, conventional and hyaluronic acid-coated liposomes were developed to improve TQ activity at ocular level. In the present study, the cytoprotective effects of TQ or TQ liposomes were assessed against oxidative and inflammatory processes in human corneal epithelial cells (HCE-2). Hyperosmolarity conditions (450 mOsm) were used as a model of DED. Interleukin-1β (IL-1β), Interleukin-6 (IL-6) and tumor necrosis factor (TNFα) were quantified by quantitative real-time polymerase chain reaction (RT-qPCR); COX-2 and Phospho-NF-κB p65 (p-p65) by Western blotting (WB). Moreover, the mitochondrial reactive oxygen species (mtROS) levels were measured by MitoSOX assay. The hyperosmotic treatment induced a significant increase of the proinflammatory genes and proteins expression that were significantly decreased in the liposomes-treated cells. The coincubation with hyaluronic acid-coated liposomes significantly reverted the increase of mtROS production, evidently stimulated by the hyperosmotic stress. Our data suggest that TQ-loaded liposomes have potential as a therapeutic agent in dry eye disease, improving the TQ efficacy.

Antinociceptive Action of Thymoquinone-Loaded Liposomes in an In Vivo Model of Tendinopathy

“…Both coated liposomes maintained good values of sizes, PdI, ZP and EE% ( Table 1 ). The increase in the particle sizes and the change of ZP confirmed the HA deposition [ 39 ].…”

“…There are no examples in the literature of TQ HA-coated liposomes, except for two studies by the authors for the ocular delivery of TQ. Liposomal formulations, and, in particular, HA-coated liposomes, reduced the toxicity of TQ at high doses in HCE-2 and HConEC cells and improved the absorption at the nucleus level, with a more pronounced effect for HA-coated liposomes [ 26 , 39 ]. Furthermore, liposomes were effective in dry eye disease.…”

Anti-Inflammatory Properties of Cannabidiol and Beta-Caryophyllene Alone or Combined in an In Vitro Inflammation Model

“…The viability of HaCaT cells exposed to CBD (0.001–1 µM) or BCP (1–100 µM) for 24 h, and of sister cultures exposed to 5 μg/mL of LPS in the presence or absence of CBD and BCP, was evaluated by MTT assay. The MTT (1 mg/mL) was added to the cells [ 50 ] after taking out 100 µL of medium from each well for the LDH assay. The MTT-containing solution was removed, and dimethyl sulfoxide (DMSO) was added to the wells to dissolve the formazan crystal formations.…”

“…The cytotoxicity in HaCaT cells was measured by the release of LDH into the extracellular fluid, 24 h after drug exposure, as previously described [ 50 ]. The LDH level corresponding to complete cell death was determined for each experiment by analyzing sister cultures exposed to Triton-X 0.01%.…”

Virtual Screening-Accelerated Discovery of a Phosphodiesterase 9 Inhibitor with Neuroprotective Effects in the Kainate Toxicity In Vitro Model

“…1 μg of RNA was retrotranscribed using iScript (Bio-Rad, Milan, Italy). Real-time polymerase chain reaction (RT-PCR) was performed as reported. , The following primers were used: PDE9 rat: forward 5′-TCAGAGCGAACTCCCTACAAGGTGAG-3′ and reverse 5′-CTCCACCACTTTGAGTCCTTCCAATTCC-3′; 18S rat: forward 5′-GGCGGCTTTGGTGACTCTAGATAACC-3′ and reverse 5′-CCTGCTGCCTTCCTTGGATGTGG-3′.…”