The Cyto-Genotoxicity Evaluation Of Biomedical Grade Chitosan On Primary Keratinocyte Cultures And Its Role In Scar Formation Pathways Using Skin Coculture Model
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Date
2010-07
Authors
Lim, Chin Keong
Journal Title
Journal ISSN
Volume Title
Publisher
Universiti Sains Malaysia
Abstract
Chitosan (-1, 4—D-glucosamine) is a deacetylated form of chitin with excellent
biological properties useful in wound management. However, improvement of the
physical and chemical properties of the chitosan will alter its biocompatibility.
Additionally, chitosan has been reported to be useful in scar intervention. Therefore in
this study, the in vitro biocompatibility of the modified chitosan porous skin
regenerating templates (CPSRTs) has been evaluated based on cytotoxicity analysis by
3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide (MTT) assay,
genotoxicity by comet assay, and skin keratinocyte pro-inflammatory cytokine
secretions [interleukin-1 (IL-1), tumor necrosis factor- (TNF-) and interleukin-8
(IL-8)], using primary normal human epidermal keratinocyte (pNHEK) cultures. The
most biocompatible CPSRT was then investigated for its role in keloid intervention
using keratinocyte-fibroblast cocultures. The human collagen type-I, III and V protein
expressions were assayed in Western blot analysis. Meanwhile, the secreted
transforming growth factor-β1 (TGF-β1) was analyzed by enzyme-linked
immunosorbent assay (ELISA), and the mRNA expressions of its intracellular signaling
molecules (SMAD 2, 3, 4 and 7) were quantified using real-time PCR analysis.
Description
Keywords
Cyto-Genotoxicity Evaluation Of Biomedical Grade Chitosan , Primary Keratinocyte Cultures