The proapoptotic influence of AgNO3 on human keratinocytes and fibroblasts in vitro, the impact for burned patient management.

It was shown that ointments containing silver compounds delay wound healing and increase the risk of hypertrophic scarring in burned patients. The aim of the study was to establish the influence of AgNO3 on cell viability and apoptosis of keratinocytes and fibroblasts in vitro. Foreskin was used to establish primary human keratinocyte and fibroblasts cell cultures. Keratynocytes grew in DMEM/Ham's-12 at 3:1 ratio, supplemented with 10% FBS, EGF, insulin, transferin, triiodothyronine and hydrocortisone. Fibroblasts were cultured in DMEM with addition of 10% FBS. The influence of AgNO3 on keratinocyte and fibroblast cultures was evaluated by fluorescence microscopy and flow cytometry. Double staining with Annexin V-FITC and propidium iodide was performed. The AgNO3 at lower concentration (3 and 15 x 10(-4) M/dm3) than used for patient's treatment (31 x 10(-4) M/dm3) revealed to be toxic and trigger apoptosis in human keratinocytes and fibroblasts. The ointments containing nitrates should be used with caution especially in conditions where the epithelial layer is destroyed. The nitrate can negatively affect wound healing.