Investigating the effect of β-gentiobiose, a component of Manuka Honey, on soluble ICAM production in primary human fibroblasts (HDFa)
Alkozei, Najeed Ahmad
Cardiff Metropolitan University
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1.1 Background: sICAM is a member of immunoglobulin super family that greatly contributes to wound healing, particularly in the inflammation phase. Honey has been shown to possess anti-inflammatory, anti-oxidant and antibacterial properties. Additionally, MH has been found to have strong antibacterial properties that inhibit a wide range of bacteria. Several studies have documented the efficacy of honey both in vitro and vivo, however the mechanism of action and the actual components remain to be unknown. 1.2 Aim: The aim of this study was to investigate the potential of β-gentiobiose, a component of MH to produce sICAM in HDFa. 1.3 Methods: Viability assay: 100 µL suspension containing (50000 cells/µL) HDFa cells and DMEM were exposed to various concentrations of β-gentiobiose and incubated for 24 & 48-hours at 37°c. Fluorescence was measured using an excitation wavelength of 560 (nm) and emission wavelength of 590 (nm). ELISA: A total of (38,000 cells/mL) were seeded in three 12-well plates for 48-hours at 37°c. Thereafter, settled cells were treated with the various concentrations of determinants and incubated overnight at 37°c. Determining the concentration of sICAM, EISA technique was used following the manufacturer`s standard protocol. 1.4 Results: Findings of the viability assay after 24-hours incubation suggested that β-gentiobiose has non-significant effect on the viability of HDFa cells with the (p>0.05). In contrast after 48-hours of incubation some concentrations (0.125, 0.0625 and 0.0156255%, w/v) significantly affected the viability of HDFa by (25, 15 and 14%) respectively with the (p<0.05). β-gentiobiose produced non-significant amount of sICAM, however TNF-α on the other hand had more than 6-fold non-significant increase in the level of sICAM relative to control (p=0.09). 1.5 Conclusion: The concentrations of β-gentiobiose that have been used in this study did not show important increase in expression of sICAM. Therefore, further studies are recommended in to whether increased concentration of β-gentiobiose will result in significant increase in the level of sICAM in HDFa.
BSc (Hons) Biomedical Science
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