The Effect of Manuka Honey and β-Gentiobiose on the Production of Interleukin 8 from Human Dermal Fibroblasts
Cardiff Metropolitan University
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Wound healing is an essential chemical process and the cost of wound healing in the UK is set to increase by £250 million by the end of 2019 (Dowsett et al, 2014), with the increase in the number of people with an impaired wound healing ability (Guo et al, 2010), it is essential to find alternative cost effective methods to aid this dynamic process. One way in researchers are trying to address this situation is the use of Manuka honey, the anti-bacterial effect of Manuka honey has been well documented however the inflammatory effect has been something up for debate. Human dermal fibroblasts (HDFa) are key molecules in wound healing (Martin, 1997), and will be treated with Manuka honey and one of its key components, β-Gentiobose. First a Viability assay will be carried out to measure the activity of the fibroblasts when treated with β-Gentiobiose over 24-hours and 48-hours. An enzyme-linked immunosorbent assay (ELISA) will be used to measure the concentration of interleukin (IL) 8, a pro-inflammatory cytokine that is heavily involved in the inflammatory phase of wound healing (Köhidai, 1998) the result of which will give an indication of the inflammatory effect of Manuka honey, there will be two positive controls, tumour necrosis factor α (TNFα) and IL-1β both of which have been proven to stimulate the release of IL-8 (Osawa et al, 2002). The results of the viability assay revealed that over both 24-hours and 48-hours, β- Gentiobose had a positive effect on the fibroblasts, promoting cellular activity. Making it possible to continue using β-Gentibiose at the two incubation times used. The results from the treatment of the fibroblasts with the various samples revealed that 5% Manuka honey stimulate the release of 2,335 pg/ml of IL-8 significantly more than the amount of IL-8 produced with no treatment that produced 525pg/ml. The β-Gentiobiose produced a similar amount of IL-8 to the sugar solution used both producing around 215-235pg/ml. The β-Gentiobiose produced a similar concentration of IL-8 to the sugar solution used, and reveals that the β-Gentiobose is not the key component on Manuka Honey that gives it its inflammatory properties. There are molecules within Manuka honey called Peroxisome proliferator-activated receptor (PPAR) ligands (Al-Khanbashi, Laila, 2012) that can mimic exogenous ligands and activate the Nuclear factor kappa (NF-κB) signalling pathway via toll-like receptors (TLRs) that ultimately leads to the transcription and release of IL-8 to surrounding tissues. Other research papers have similar findings using alternative cell lines such as MM6, a monocytic cell line (Nidimamidi, 2006). Thus, the results suggest that Manuka honey exhibits a proinflammatory effect when exposed to fibroblasts.
BSc (Hons) Biomedical Science
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