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The use of bee venom (Apis mellifera L., BV) occasionally causes side effects such as inflammation and allergic reactions in the recipients. Several case reports also suggested the treatment of BV has some limitations in its clinical uses, due to the occurrence of dermal necrosis and anaphylatic reactions. It is generally understood that bee venom allergy is mainly the result of its allergic component, phospholipase A2 (PLA2). The present study was aimed to generate PLA2-free bee venom (PBV) and evaluate its efficacy as skin care and cosmetic preparation, comparing with original bee venom (BV). Our results showed that both BV and PBV exhibited significant protective effects in UVB-irradiated human keratinocyte (HaCaT) and human dermal fibroblast (HDF) cells and they also induced type I collagen synthesis in UVB-irradiated HDF cells except BV at 3 μg/ml. Furthermore, BV and PBV showed the inhibition of UVB-stimulated matrix metalloproteinase-1 (MMP-1), a major collagen degrading enzyme in skin. However, BV, unlike PBV, exhibited strong cytotoxicities in skin cells (both HaCaT and HDF) at its working concentrations of anti-wrinkle effect. The underlying cell signaling mechanisms of anti-wrinkle effects of BV and PBV were demonstrated by the activation of ERK1/2, and p38. Conclusively, PBV appears to be the bee venom of choice with less cytotoxicity and higher efficacy on UVB-irradiated skin cells in comparison with original bee venom (BV). Therefore, PBV can better be used as a cosmetic ingredient exhibiting excellent anti-wrinkle effect against photoaging than original BV.