Combined hyperosmolarity and inflammatory conditions in stressed human corneal epithelial cells and macrophages to evaluate osmoprotective agents as potential DED treatments
| Autor: | Rocío Herrero-Vanrell, Jose M. Benitez-del-Castillo, Irene T. Molina-Martínez, José Javier López-Cano, M.A. González-Cela-Casamayor, Vanessa Andrés-Guerrero |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
Taurine
Osmotic shock Cell Survival Hypertonic Solutions Apoptosis Pharmacology Flow cytometry Cellular and Molecular Neuroscience chemistry.chemical_compound Mice Tecnología farmaceútica Betaine Hypromellose Derivatives Stress Physiological Carnitine Hyaluronic acid medicine Animals Humans MTT assay Hyaluronic Acid Cells Cultured Osmole Inflammation Mice Inbred BALB C medicine.diagnostic_test Osmotic concentration Tumor Necrosis Factor-alpha Macrophages Osmolar Concentration Epithelium Corneal Flow Cytometry Sensory Systems Ophthalmology chemistry Dry Eye Syndromes |
| Zdroj: | E-Prints Complutense. Archivo Institucional de la UCM instname |
| Popis: | Purpose: To develop an easy-to-perform combined model in human corneal epithelial cells (HCECs) and Balb/c mice macrophages J774.A1 (MP) for preliminary screening of potential ophthalmic therapeutic substances. Methods: HCECs were exposed to different osmolarities (350–500 mOsm/L) and MTT assay was employed for cell survival and flow cytometry to assess apoptosis-necrosis and relative cell size (RCS) distribution. Effectiveness of Betaine, L-Carnitine, Taurine at different concentrations (ranging from 20 mM to 200 mM) was studied. Also, mucoadhesive polymers such as Hyaluronic acid (HA) and Hydroxypropylmethylcellulose (HPMC) (0.4 and 0.8%) were evaluated. Cells were pre-incubated with the compounds (8h) and then exposed to hyperosmotic stress (470 mOsm/L) for 16h. Moreover, anti-inflammatory activity was performed in LPS-stimulated MP. Results: Exposure to hyperosmotic solutions between 450 and 500 mOsm/L promoted the highest cell death after 16h exposures (p < 0.0001) with a drop in viability to 34.96% ± 11.77 for 470 mOsm/L. Pre-incubation with Betaine at 150 mM and 200 mM provided the highest cell survival against hyperosmolarity (66.01% ± 3.65 and 65.90% ± 0.78 respectively) while HA 0.4% was the most effective polymer in preventing cell death (42.2% ± 3.60). Flow cytometry showed that Betaine and Taurine at concentrations between 150-200 mM and 20–80 mM respectively presented the highest anti-apoptotic activity. Also, HA and HPMC polymers reduced apoptoticinduced cell death. All osmoprotectants modified RCS, and polymers increased their value over 100%. LCarnitine 50 mM, Taurine 40 mM and HA 0.4% presented the highest TNF-α inhibition activity (60%) albeit all of them showed anti-inflammatory inhibition percentages higher than 20%. Conclusions: HCECs hyperosmolar model combined with inflammatory conditions in macrophages allows the screening of osmoprotectants by simulating chronic hyperosmolarity (16h) and inflammation (24h). |
| Databáze: | OpenAIRE |
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