In this study, we aimed to utilize ceramide and fatty acid as a skin lipid enhancers (SLE), which increase lateral diffusivity and fluidity of the SC lipid and interact with keratin. We have prepared skin lipid enhancer-based lipid nano-particles (SLE-LNPs) composed of lipid matrix such as lecithin, ceramide, and fatty acids to enhance transdermal delivery. The measurements of particle size distribution and zeta potential of lipid nano particles with or without skin lipid enhancers have shown the effect of fatty acids to emulsion stability for 4 weeks. The morphology and structure have been investigated using SAXS, WAXS, and Cryo-TEM. In addition, in vitro skin permeation of skin carriers with skin lipid enhancers exhibited higher than that of typical skin carrier by confocal laser scanning microscopy and Franz diffusion cell experiments. In vivo human skin penetration study using Raman spectroscopy also demonstrated that the SLE-LNP has a deeper penetration depth, faster penetration rate than bare LNP. In addition, in the in vivo skin clinical trial test, the experimental group treated with SLE-LNP has a statistically significant wrinkle measurements decrease rate, melanin index decrease rate compared to the control group, resulting in a better skin efficacy enhancement effect. This results indicate that the skin lipid enhancer-corporated lipid nano particle (SLE-LNPs) formulations can effectively interact with the stratum corneum, resulting in skin delivery of the active agents such as niacinamide and adenosine, resulting improve skin efficacy.