In Vitro Ocular Cytotoxicity of BEE Venom in ARPE-19, NTM5, HCET, and HCEC-12 Cell Lines

Bee venom (BV) is a complex natural secretion of Apis mellifera with documented anti-inflammatory, antimicrobial, and anticancer properties. Despite its therapeutic potential, safety concerns remain due to its strong cytotoxic effects, particularly in sensitive tissues such as the eye. In this study, we evaluated the composition and ocular cytotoxicity of purified BV obtained from the Muğla ecotype of Apis mellifera anatoliaca. High-performance liquid chromatography with variable wavelength detection (HPLC-VWD) confirmed the presence of three principal components; melittin (71.08 ± 0.33%), phospholipase A₂ (12.98 ± 0.27%), and apamin (3.02 ± 0.25%) at retention times consistent with reference standards. Cytotoxicity was assessed in four human ocular cell lines: ARPE-19 (retinal pigment epithelium), NTM5 (trabecular meshwork), HCE-T (corneal epithelium), and HCEC-12 (corneal endothelium), using MTT viability assays and live/dead staining over 24 h and 48 h. BV exerted strong, dose and time dependent cytotoxic effects across all cell lines, with ARPE-19 and NTM5 cells exhibiting the highest sensitivity. IC₅₀ values were 22.36 µg/mL (24 h) and 12.55 µg/mL (48 h) in ARPE-19, and 20.78 µg/mL (24 h) and 13.07 µg/mL (48 h) in NTM5 cells. By contrast, HCEC-12 and HCE-T cells displayed moderate sensitivity, with IC₅₀ values between 29 and 34 µg/mL. These findings highlight tissue-specific differences in BV susceptibility. While BV may offer therapeutic promise, its narrow safety margin in ocular cells underscores the importance of controlled delivery strategies to minimize toxicity. This study provides foundational data for evaluating the ocular safety profile of BV and informs future translational applications.