Chlorhexidine diacetate and povidone-iodine were evaluated for fibroblast toxicity on a primary line of canine embryonic fibroblasts, and for bactericidal efficacy against Staphylococcus aureus. The cultured fibroblasts or S. aureus were exposed for 30 minutes to incremental dilutions of 0.5 and 0.0005% chlorhexidine diacetate, 5.0 to 0.05% povidone-iodine, or physiologic buffered saline as a control. To determine survival, fibroblasts were trypsinized and counted; S. aureus colonies were counted on brain-heart infusion agar. Survival for both groups was expressed by calculating the number of living cells in test dilutions as a percentage of the number in control cultures. Fibroblast survival occurred at chlorhexidine concentrations less than 0.013% and at povidone-iodine concentrations less than 0.5% (p less than 0.05). Significant S. aureus survival (p less than 0.05) was noted at chlorhexidine concentrations less than 0.05% and povidone-iodine concentrations less than 1.0%. These data showed that all bactericidal concentrations of chlorhexidine diacetate and povidone-iodine were lethal to canine embryonic fibroblasts in vitro, whereas non-lethal concentrations allowed significant bacterial survival.
To correlate the results of an in vitro study with clinical response, the effects of 0.005 and 0.05% chlorhexidine diacetate and 0.1 and 1.0% povidone-iodine concentrations on wound healing were evaluated in five beagle dogs. Full-thickness skin wounds (2 X 2 cm) were irrigated once daily for 14 days with the antiseptic solutions or physiologic-buffered saline. Chlorhexidine diacetate 0.05% had significantly more bactericidal activity than povidone-iodine and saline, and both chlorhexidine diacetate concentrations had residual effects 6 hours after irrigation. Neither povidone-iodine nor saline had significant bactericidal activity. The percentages of unhealed wound area and wound contraction were calculated 7, 14, and 21 days after wounding. Healed wound area and contraction were similar in wounds treated with chlorhexidine diacetate and povidone-iodine. However, wounds treated with chlorhexidine diacetate had more healed wound area on days 7 and 14 and more contraction on days 7, 14, and 21 than saline-treated wounds. At the concentrations tested, chlorhexidine diacetate irrigations provided bactericidal activity and were more beneficial to wound healing than irrigations with saline alone. These results suggest that concentrations of chlorhexidine diacetate which are cytotoxic to tissue culture fibroblasts in vitro do not interfere with wound healing in vivo.
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