In-vitro evaluation of biomedical grade chitosan derivatives on primary human skin fibroblasts
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Date
2009
Authors
Muhamad Nor, Nor Asiah
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Abstract
Chitosan has been proposed for biomedical applications because of its
biocompatibility and abundance in nature. However, many barriers still exist due to its
physical and chemical limitations. Further work is needed to improve these properties,
but any changes will influence its biocompatibility. Therefore, the biosafety of this
chitosan product in terms of biocompatibility and cytotoxicity requires an in-vitro
evaluation. The chitosan film that was tested on has been locally processed as a chitosan
derivative. Primary human dermal fibroblast, which plays a major role in the process of
wound healing, was used for the evaluation. The objective of this study is to evaluate the
cytotoxicity of chitosan derivatives products and their ability to influence the
proliferation of fibroblasts. This study also aims to assess the effect of chitosan on
interleukin 8 (IL 8) and transforming growth factor- (TGF-) secretion by primary
fibroblasts in the culture system. Comparative evaluation was done on four types of
chitosan derivatives films named as NO-CMC, O-CMC, O-C and N-CMC. The results
showed that, O-C and N-CMC were found to be not cytotoxic against fibroblasts as
confirmed by 3(4, 5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide (MTT)
assay. General observation using phase contrast microscopy showed no marked
morphological changes seen on O-C and N-CMC treated cells at initial incubation
period. Cultures incubated with NO-CMC were graded as slightly changed whereby
more than 20% of the cells were rounded, loosely attached and occasional lysed cells
were present. Meanwhile, minimal cell damage was observed on O-CMC treated cells.
Similar results were obtained from quantitative approach assessment. Determination of
cell proliferation using CellTiter 96® Aqueous Non-Radioactive Cell Proliferation Assay
revealed that no fibroblasts proliferation detected in the presence of high molecular
weight (MW) of chitosan derivatives after 5 days of treatment compared to untreated
well. However, low MW has the ability to induce the proliferation of fibroblasts. In addition, stimulation of proliferation increased with time exposure. This study also
demonstrated that chitosan derivatives have an ability to influence the secretion of IL-8
by fibroblasts. In contrast, none of TGF-β secretion was detected. Although in-vivo
experiments are awaited, these findings indicate the potential use of chitosan derivatives
in wound management.
Description
Master
Keywords
Biological Science , In-vitro , Biomedical grade , Fibroblasts