Background/Aim: To date, no cell-based assay that focuses on the prime cause of acne initiation through activation of toll-like receptor2 and 4 and interleukin-8 (IL-8) production exists. Herein, we present an assay that evaluates acne by determining TLR2 and 4 expression and activation. Materials and Methods: Viability of keratinocytes was determined by the MTT assay. IL-8 was evaluated by ELISA. Immunocytochemistry was performed for determining receptor expression. Results: Lipoteichoic acid (LTA), peptidoglycan (PGN) and lipopolysaccharide (LPS) induced IL-8 production. Pre-treatment of cells with TLR2 and TLR4 inhibitors, before stimulation, reduced IL-8 production. Zinc gluconate was used for verification. Zinc can significantly suppress IL-8 production in the system. Treatment of cells with LTA+PGN or LPS resulted in increased TLR2 and TLR4 expression on the cell surface. This effect was prevented by zinc treatment. Conclusion: The measurement of IL-8 and TLR2 and TLR4 levels can be used for the evaluation of anti-acne treatment. Acne vulgaris is a chronic disease. It is common among adolescents and can have a huge impact on physical appearance and health both short-term and long-term as a result of the adverse effects of the medication (1). The adverse effects are induced by non-anti-acne actions such as antibiotics and steroids (2). It is known that inflammation derived from the activation of innate immunity plays a primary role in the induction of acne lesions rather than infection or an allergic reaction (3). Clinically, there are multiple types of acne lesions. However, the induction of all types of lesions is primarily initiated by colonization of Propionibacterium acnes, a Gram-positive anaerobic bacterium that plays a key role in maintaining and influencing the inflammatory phase of acne (4). The major cell wall components of P. acnes, lipoteichoic acid (LTA) and peptidoglycan (PGN) were found to stimulate immune cells to release mediators of inflammation (5). Apart from LTA and PGN, lipopolysaccharide (LPS)-a component of gram-negative bacteria-is also an immunostimulatory factor (6). The innate immune system of the skin epidermis has several defense components and mechanisms involving keratinocytes, neutrophils, mast cells and macrophages (7). The key step of acne formation is the interaction of bacterial components such as LPS, LTA or PGN with toll-like receptors (TLRs) in epidermal cells (8). TLR4 primarily mediates cellular signaling induced by Gram-negative bacteria (9), while TLR2 is required for pro-inflammatory signaling to components of Gram-positive bacteria (10). It is known that TLR2 and TLR4 are predominantly expressed on the cell surface of the infundibular keratinocytes. The activation of these TLRs results in signaling cascades that produce interleukin 8 (IL-8) (11). As a consequence, IL-8 acts as a pro-inflammatory cytokine and a powerful chemoattractant (12). This cytokine plays a major role in inflammatory events by its activity as chemokine and as an activator of crucial neutrop...