Hot spring (HS) waters manifest diverse positive effects on the skin due to their unique chemical compositions. Sodium hyaluronate acid (HA) comprises N-acetylglucosamine and D-glucuronic acid, and distinguishes itself with superior qualities in skin regeneration, providing moisturizing and anti-aging benefits. The combination of HA with HS water is widely applied across ophthalmology, pneumology, nutrition, and cosmetics. This study delved into the application of HA in cosmetology, with a focus on its interaction with HS water and its effects on moisture retention and promoting wound healing. In particular, with the alkaline pH levels of the Yumoto HS, HA molecules may undergo dissociation to be ionized resulting in a negatively charged polymer and interacting with positively charged ions in the HS water through electrostatic interactions. The shifted peaks in the FTIR result and zeta potential shifts to a less negative region in the case of HA-HS compared to HA-DI indicate an ionic interaction between HS water and HA. Moisture tests confirmed the sustained hydration when HA is dissolved in HS water, underscoring its potential to improve skin hydration at certain concentrations, specifically at 0.5% and 1%. Additionally, MTT assay results demonstrated that HS water stimulates the growth of fibroblast cells compared to distilled water, implying its potential beneficial effect in wound healing. These findings suggested the multifaceted benefits of HAHS in skincare, highlighting its role in enhancing skin hydration and potentially accelerating wound healing processes, thus presenting avenues for the development of advanced cosmeceutical formulations.