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.
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Master
Keywords
Biological Science , In-vitro , Biomedical grade , Fibroblasts
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