Purpose: The topical application of scalp-care cosmetics, infused with functional ingredients, can offer an optimal cosmetic approach to prevent problems associated with aging. In terms of the development of anti-aging cosmetics, we studied the use of acetyl hexapeptide-8 (AH-8) for improving elasticity of the scalp.Methods: Ampoules containing 3% and 5% concentrations of AH-8 as an active ingredient were prepared and their safety and efficacy were evaluated. HaCaT cells were used to evaluate the safety of AH-8 ampoules by measurement of in vitro cytotoxicity. In in vivo experiments, we tested the AH-8 ampoules by cumulative irritation test (CIT) on healthy adults, 20–50 years of age. Relative expressions of superoxide dismutase 2 (SOD2), forkhead box O1 (FOXO1), actinin alpha 1 (ACTN1), collagen type XVII alpha 1 chain (COL17A1), and integrin subunit beta 4 (ITGB4) were assessed using quantitative real-time PCR (Q-PCR).Results: AH-8 ampoules showed significant cytotoxicity to HaCaT cells in a dose-dependent manner. CIT assessment in 30 adults revealed no skin reactions to both 3% and 5% AH-8 ampoules. After treatment of HaCaT cells with the AH-8 ampoules, mRNA levels of SOD2 and FOXO1, which are directly implicated as antioxidant factors, were increased. Of the factors that improve elasticity, ACTN1 mRNA levels had the greatest increase when treated with the 5% AH-8 ampoules (1.75 µg/mL). Also, the AH-8 ampoules increased mRNA levels of COL17A1, and ITGB4 in HaCaT cells.Conclusions: In this study, we proved that 3% and 5% AH-8 ampoules are safe for use as a scalp-care cosmetic. AH-8 ampoule had efficacy in anti-aging and improving elasticity of the scalp. This comprehensive test showed that AH-8 can be developed as a cosmetic for anti-aging and improvement of scalp elasticity.