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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Date
2012
Authors
Yusuf, Siti Mahirah
Journal Title
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Publisher
Universiti Sains Malaysia
Abstract
Keloid 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).
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Keywords
Keloid is characterized by excess collagen deposition , that can damage healthy tissue