Evelyn Komaratih and Yuyun Rindiastuti and Yohanes Widodo Wirohadidjojo and Fedik Abdul Rantam and Aristika Dinaryati and Ni Made Inten Lestari and Cita Rosita Sigit Prakoeswa (2019) The Resveratrol Increase of Hepatocyte Growth Factor (HGF) and Epidermal Growth Factor (EGF) Levels in Wharton's Jelly Mesenchymal Stem Cells (WJ-MSCS) Secretome: Toward Cell Free Therapy in Dry Eye Disease (DED). Biochemical and Cellular Archives, 19 (Supp-2). pp. 4737-4743. ISSN 09725075

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Abstract

ABSTRACT : This study aims to explore and determine the effect of resveratrol at various doses on WJ-MSCs secretome. Wharton’s jelly was extracted from umbilical cord, with the enzymatic-explant mixed method used to isolate the MSCs which were stained with CD 105, CD 73, CD 90, and CD 45 FITC conjugated antibodies. Furthermore, WJ-MSCs were treated with culture media supplemented with resveratrol at doses of 0.1 µM, 0.2 µM, 0.4 µM, 0.5 µM, 0.8 µM, and 1 µM, with 2% and 10% FBS used in the control group. The cell viability was measured using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphyniltetrazolium bromide (MTT) assay., while the level of EGF, HGF, and vascular endothelial growth factor (VEGF)-A were measured using Enzyme Linked Immunosorbent Assay (ELISA). Data analysis, were carried out using Analysis of variance (ANOVA) and posthoc Tukey using SPSS 19.0 software. The viability of WJ-MSCs was maintained in all resveratrol doses compared to the control. This lead to a decrease in cell viability in resveratrol at 1 µM dose and at 0.1 µM it increased the HGF (130 ± 11.2 pg/mL) and EGF (76.67 ± 15.67 pg/mL) levels, and decreased the VEGF (65 ± 5.02 pg/mL) level compared to other groups. These cells secretome composition tend to have beneficial effect on ocular surface regeneration in DED. 0.1 µM resveratrol was the optimal dose used to increase EGF and HGF secretion from WJ-MSCs, while decreasing the VEGF level. In addition, the various compositions of growth factors within WJ-MSCs leads to a cell-free based regenerative therapy in DED.

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