Cytotoxicity of chlorhexidine and neem extract on cultured human gingival fibroblasts through fluorescence-activated cell sorting analysis : An in-vitro study

ABSTRACT

Objective: To assess the influence of chlorhexidine (CHX), neem vehicle control (NVC), and neem extract (NE) on cultured human gingival fibroblasts (hGFs) using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and fluorescence-activated cell sorting (FACS) analysis. Materials and Methods: Fibroblasts were derived from healthy gingival biopsy specimens harvested aseptically. The effects of CHX, NVC, and NE were evaluated on cultured hGFs through FACS and MTT assay. Results: MTT assay with hGFs indicated altered morphology with maximum cell death at 10% CHX, while NVC and NE showed similar results at a concentration of 75% and above. On FACS analysis, beyond 1%, CHX adversely affected the cell cycle phase distribution whereas NE exerted a detrimental effect only at 100%. Moreover, both with NVC and NE cells were well differentiated in all the three phases of the cell cycle, with distinction getting lost at 50% to finally causing cell death at 100%. Conclusions: CHX beyond 1% concentration exhibited a toxic effect on hGFs at 1, 5, and 15 min time exposure. However, NE did not adversely affect the fibroblasts even up to 50% concentration showing a less toxic effect in comparison with CHX on these cells. The cytoprotective and oral friendly quality of NE emphaisze the superiority of NE over CHX.

Key words:

3-(4 - 5-dimethylthiazol-2-yl)-2 - 5-diphenyltetrazolium bromide assay - chlorhexidine - cytotoxicity - fluorescence-activated cell sorting analysis - human gingival fibroblasts - neem extract

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