Tatnall, Frances Melanie; (1991) Antiseptic cytotoxicity and the cutaneous wound: an in vitro study. Doctoral thesis (M.D), UCL (University College London).

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Abstract

Concern has been expressed about the possible harmful effects of antiseptics on cutaneous wounds. There are few clinical data to support this view but animal studies suggest that certain agents are toxic. To simplify the investigation of antiseptic toxicity investigations have been conducted in vitro using cell culture. This study set out to establish an in vitro cytotoxicity assay which is simple, accurate and reliable, so that a number of cell types and agents could be easily studied. Basal keratinocytes and fibroblasts were studied as these cells are essential to the wound healing process. A transformed keratinocyte line (SVK 14 cells) was also compared. The cells were exposed to hydrogen peroxide, chlorhexidine, sodium hypochlorite and cetrimide for 15 minutes and cell viability was assessed with a colorimetric assay which utilizes the tetrazolium salt 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT). Comparison of 50% survival levels showed that the three cell types showed similar susceptibilities to the agents tested. These findings suggest that the transformed cell line, which has the advantage of ready availibility and immortality, can replace both fibroblasts and keratinocytes in an antiseptic cytotoxicity assay. Comparison of 50% survival concentrations to the standard therapeutic concentrations showed the following ranking order of toxicity: sodium hypochlorite> cetrimide> hydrogen peroxide>chlorhexidine. These findings are In line with in vivo studies on inhibition of wound healing by antiseptic agents. The assay devised appeared to be of value in predicting the relative toxicities of antiseptic agents In vivo. Reported experimental conditions for antiseptic cytotoxicity assays vary. The effect on toxicity of following conditions was studied; Increasing exposure time; the effect of medium or serum; Increasing cell number. Increasing cell number and diluting the agents in medium and serum reduced the toxicity of all agents but had the greatest effect on sodium hypochlorite. Increasing the exposure time increased the toxicity of all agents but had the greatest effect on hydrogen peroxide. Changes in experimental variables clearly alter the toxicity of individual agents. The results of in vitro testing should be interpreted in the context of experimental conditions.

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