As the global aging population continues to grow, there is an increasing societal and commercial demand for effective strategies to alleviate visible signs of skin aging—such as wrinkles, reduced elasticity, and pigmentation-which in turn is fueling rapid growth in the anti-aging cosmetics market [1], [2]. Among various aging-associated organs, the skin is the most visibly affected and often the first to exhibit signs of aging. The global facial rejuvenation market was valued at USD 24.6 billion in 2022 and is projected to reach USD 44.5 billion by 2030, with a compound annual growth rate (CAGR) of 7.7 %, driven by demographic aging, rising healthcare expenditure, and continuous technological advancements [2]. In recent years, there has been growing interest in non-invasive or minimally invasive treatments aimed at reducing wrinkles, fine lines, and skin laxity, reflecting a strong consumer demand for interventions that mitigate the visible signs of skin aging.

Aging is closely linked to skin health and appearance [3]. Skin aging, encompassing both extrinsic and intrinsic factors, is a complex process [4]. Extrinsic factors such as ultraviolet (UV) radiation and oxidative stress can accelerate skin aging through intrinsic time-dependent mechanisms. Notably, 80 % of visible skin aging symptoms are attributed to UV exposure [5]. Ultraviolet B (UVB), a type of short-wave ultraviolet light, is a significant extrinsic factor that stimulates the production of reactive oxygen species (ROS) and matrix metalloproteinases (MMPs) in the epidermis and dermis [6]. In the skin aging process, MMPs function as collagenases, gelatinases, and stromelysins, breaking down skin collagen within the dermis [7]. This activity of MMPs contributes to wrinkle formation by degrading, fragmenting, and disorganizing the extracellular matrix components of the skin [7].

Retinol, a subtype of vitamin A and a well-established anti-aging compound, is widely incorporated into cosmetic formulations [8]. Compared to retinoic acid, retinol has a lower incidence of adverse effects when applied topically [9]. Retinol exerts anti-aging effects by addressing multiple clinical signs, such as wrinkles, fine lines, and uneven pigmentation. These effects are achieved through the inhibition of MMP activity and gene expression, reduction of oxidative stress, and stimulation of collagen synthesis [10]. Studies have shown that topical application of retinol significantly mitigates aging signs by increasing epidermal thickness and enhancing the production of extracellular matrix components, including type I collagen, fibronectin, and elastin, in aged human skin [11].

Probiotics and prebiotics, along with their metabolites, have been identified as promising agents for modulating microbial balance, reducing cellular senescence, and mitigating inflammation caused by senescence-associated secretory phenotype factors [12]. In parallel, microbiota-derived enzymes, such as Arazyme, have also emerged as potential therapeutic agents for managing skin aging and inflammation through different mechanisms [13]. Microbial proteases have been increasingly explored for their potential applications in dermatology and cosmetology, due to their roles in extracellular matrix remodeling, anti-inflammatory effects, and skin regeneration [14]. Arazyme is a 51.5 kDa metalloprotease produced by Serratia proteamaculans, originally isolated from the gut of the spider Nephila clavata. It is an approved food additive and has been reported to exert hepatoprotective and glucose-lowering effects in a non-alcoholic fatty liver disease model [15], [16]. In skin keratinocytes, Arazyme has been shown to inhibit TNFα- and IFNγ-induced secretion of inflammatory cytokines, including MCP-1, IL-6, and IL-8, while enhancing the expression of skin barrier-associated proteins [17]. In an atopic dermatitis model, Arazyme treatment significantly reduced the infiltration of inflammatory cells into the dermal layer and alleviated atopic dermatitis-like skin lesions [18]. Moreover, Arazyme has been reported to exert anti-inflammatory effects, in part, by modulating neutrophil activity—a mechanism that is particularly relevant given the critical role of neutrophils in UVB-induced skin inflammation [19]. Despite these promising effects, experimental evidence regarding the anti-wrinkle potential of Arazyme has not yet been reported. This study aimed to investigate the anti-wrinkle effects of Arazyme in an in vitro model of UVB- and oxidative stress-induced skin aging in keratinocytes and fibroblasts. We hypothesized that Arazyme would (1) suppress the expression of MMPs, (2) promote collagen synthesis, and (3) attenuate cellular senescence by modulating oxidative stress and associated signaling pathways.