Phenotypic Modulation of Adipose-Derived Stem Cells and Fibroblasts Treated with Povidone–Iodine and Chlorhexidine in Mono and Coculture Models

Topical antiseptics are essential in wound treatment, and adipose-derived stem cells (ADSCs) have recently been proven to facilitate healing. However, the impact of antiseptics on ADSCs has not been fully elucidated, especially in relation to other relevant cell types present in the wound microenvironment, e.g., fibroblasts. This study evaluated the effects of chlorhexidine and povidone–iodine on four cellular constructs in 2D and 3D in vitro culture systems. Cell constructs were treated with two concentrations of each antiseptic, after which cell migration activity, α-SMA, and Ki67 marker expressions were assessed and compared. Both tested concentrations of povidone–iodine impaired migration and sprouting compared to chlorhexidine, which had minimal effects when used in low concentrations. The gap in the wound healing assay did not close after 24 h of povidone–iodine treatment, although, at the lower concentration, cells started to migrate in a single-cell movement pattern. Similarly, in 3D culture systems, sprouting with reduced spike formation was observed at high povidone–iodine concentrations. Both antiseptics modulated α-SMA and Ki67 marker expressions at 5 days following treatment. Although both antiseptics had cytotoxic effects dependent on drug concentration and cell type, povidone–iodine contributed more substantially to the healing process than chlorhexidine, acting especially on fibroblasts.