In Vitro Cytotoxicity Evaluation Of Processed Natural Coral On Human Fibroblast And Osteoblast Cell Lines
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Date
2010-08
Authors
Omar, Nor Shamsuria
Journal Title
Journal ISSN
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Publisher
Universiti Sains Malaysia
Abstract
The aim of this study was to evaluate the in vitro cytotoxicity of the locally produced processed natural coral (Health Campus, Universiti Sains Malaysia) from the Porites species in terms of proliferation, apoptosis study and cell attachment by using human osteoblast (HOS) and fibroblast (MRC-5) cell lines (ATCC, USA). The in vitro cytotoxicity of the processed natural coral (PNC) was evaluated using test on extract and direct contact as per ISO 10993-5 (1999). HOS cells were used to study the magnitude of proliferation when exposed to the extraction medium of two different particle sizes of PNC, granules (0.5-1mm) and powder (1-50μm). The proliferation of HOS cells was analyzed using MTT and NR assays. The material is considered toxic if the proliferation rate was less than 50%. For the apoptosis study, HOS cells were exposed to the extraction medium of PNC powder at a concentration of 200mg/ml for 48 and 72 hours. Then the cell pellet (1x 106 cells) was resuspended in Annexin-V-FLOUS labelling solution and subjected to flow cytometric (FCM) analysis. Proliferation study via direct contact of PNC disc was carried out using HOS and MRC-5 cell lines. Those cells were exposed to the PNC material at 1, 24, 72 and 168 hours and analyzed using NR assay and the medium was never changed through out the incubation periods. The material is considered toxic if the proliferation rate was less than 50%. For the cell attachment study, MRC-5 cells and HOS cells were cultured on the PNC discs and rubber latex (positive control) and observed under inverted microscope after 72 hours of incubation period. For the Scanning Electron Microscopic (SEM) analysis, HOS cells were cultured on the PNC disc and thermanox plastic (negative control) and observed after 72 hours of incubation period. Proliferation study of HOS cells on the extraction of PNC granules and powder showed that both particles were not cytotoxic. Also, both PNC granules and powder did not induce any cytotoxicity as was evident from their proliferation rate, which was above 50%. The flow cytometry analysis showed that the viable cell percentage for the HOS cells was high and the apoptotic cell percentage was low, indicating that PNC did not cause a remarkable damage to the cells.
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Keywords
In vitro cytotoxicity evaluation of processed natural coral , on human fibroblast and osteoblast cell lines