Thirdhand smoke (THS) is the residual cigarette smoke that settles on indoor surface fabrics, dust and can accumulate in the environment. Therefore, it can be a risk factor for individuals who have frequent dermal contact with THS‐contaminated surfaces. In the present study, it was aimed to elucidate the toxicity of dermal THS exposure in HaCaT human keratinocytes. The THS was extracted from terrycloth exposed to 3R4F research cigarette smoke in a closed chamber and the adverse outcomes induced by THS were determined through assessment of cytotoxicity tests (MTT and NRU), intracellular GSH level, total SOD activity, matrix metalloproteinase‐1 (MMP‐1) and IL‐6 levels. The wound healing capacity of THS‐exposed keratinocytes was evaluated via scratch assay. A potent antioxidant isothiocyanate compound, sulforaphane (SFN), was used as a negative control. THS was dose‐dependently cytotoxic (12.5%–100%, v/v) to the HaCaT cells through mitochondrial cell dysfunction (p < 0.01), which was ameliorated by SFN (0.62 μM) pre‐treatment. In parallel, THS exposure significantly decreased the intracellular GSH deposits and T‐SOD activity in keratinocytes. Collagen degradation through elevated MMP‐1 expression was observed in THS‐exposed cells in parallel with the delay of wound healing and increased pro‐inflammatory response in a dose‐dependent manner (p < 0.05). The findings are expected to raise awareness about THS as an environmental pollutant for skin, particularly in the highest‐ranked countries in cigarette consumption. To conclude, these results might contribute to the studies on the importance of dermal exposure to THS in the pathogenesis of epidermal alterations and the other skin diseases.