THE mRNA EXPRESSION OF INDUCIBLE NITRIC OXIDE SYNTHASE IN KELOID USING TOCOTRIENOL-RICH FRACTION IN PRIMARY HUMAN EPIDERMAL KERATINOCYTES AND PRIMARY HUMAN DERMAL FIBROBLAST CULTURES

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dc.contributor.authorSITI MAHIRAH, YUSUF
dc.date.accessioned2017-10-30T07:38:34Z
dc.date.available2017-10-30T07:38:34Z
dc.date.issued2012
dc.description.abstractKeloid is characterized by excess collagen deposition that can damage healthy tissue. With the widespread use of Vitamin E- tocopherols based product and the discovery of tocotrienol (T3) potential to treat various skin injuries; there are possibilities of tocotrienol-rich fraction (TRF) to intercede normal, hypertrophic, or keloid scarring during the inflammatory process. However, there is a lack of scientific evidence to validate the efficacy and the therapies of TRF in scar prevention. Thus, this study was carried out to evaluate the beneficial effects of TRF in wound healing and its possible stimulation towards mRNA expression of inducible nitric oxide synthase (iNOS) in keloid human skin keratinocytes and fibroblasts. Primary human epidermal keratinocytes (pHEK) and primary human dermal fibroblasts (pHDF) were successfully established using cell dissociation method. Verification of pHEK and pHDF was conducted using Cytokeratin-6 (CK6), Involucrin, Heat Shock Protein-47 (HSP47) and Fibroblast Surface Protein (FSP) markers using imunocytochemistry analysis. The effect of TRF on pHEK and pHDF were determined by using MTT assay. The mRNA expression of iNOS in primary normal human dermal fibroblasts (pNHDF) and primary keloid human dermal fibroblasts (pKHDF) treated with TRF was evaluated using real-time PCR. Primary normal human epidermal keratinocytes (pNHEK) achieved higher cellular growth rate compared to primary keloid human epidermal keratinocytes (pKHEK). Whilst, pKHDF exhibited linear growth and sustained higher cellular growth rate compared to pNHDF. pHEK cultures were positive for the presence of CK6 and Involucrin whereas HSP47 and FSP were found in pHDF cultures. TRF ranged from 2.85 μg/ mL to180 μg/ mL. TRF (45 μg/ mL to 180 μg/ mL) was found to inhibit the growth of pHEK while for pHDF was at 90 ug/mL at 72 hours. At lower concentrations (2.8-22.5 μg/mL), TRF increased pHEK cell growth at 24 and 48 hours of incubation but have no significant effect in pHDF viability at all time intervals. TRF at 2.8 μg/mL has been found to reduce the mRNA expression of iNOS at 24 hours, which may in turn suppress the production of nitric oxide (NO) by pKHDF. The observed suppression and growth inhibition effect in pHEK and pHDF cultures may be caused by the antiproliferative and antioxidant activities of TRF. Whereas, the reduction of iNOS mRNA level in pKHDF may suggest that TRF possess antioxidant and antifibrogenic effect. This finding showed that TRF may play a role in keloid intervention by suppressing the mRNA expression of iNOS.en_US
dc.identifier.urihttp://hdl.handle.net/123456789/5172
dc.language.isoenen_US
dc.publisherPusat Pengajian Sains Perubatan Universiti Sains Malaysiaen_US
dc.subjectHuman Genomeen_US
dc.titleTHE mRNA EXPRESSION OF INDUCIBLE NITRIC OXIDE SYNTHASE IN KELOID USING TOCOTRIENOL-RICH FRACTION IN PRIMARY HUMAN EPIDERMAL KERATINOCYTES AND PRIMARY HUMAN DERMAL FIBROBLAST CULTURESen_US
dc.typeThesisen_US
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