A long-term usage investigation of a scalp lotion containing Eucalyptus extract, which increases the amount of ceramide in the skin, was carried out to explore the change in physical properties of the hair fiber. Half-head or whole-head usage studies of a scalp lotion with Eucalyptus extract were carried out for the following groups: Japanese female, Japanese senior female, Japanese male, and Caucasian female panelists. As a result, the improvement in hair luster and bounce in the root part of the hair were recognized by the panelists after the long-term application of the scalp lotion with Eucalyptus extract. Measurement of hair gloss intensity and bending stress at the root suggests that this improvement is based on changes in these physical properties. These results indicate that the recognition of panelists is based on an actual change in the hair fiber properties. The efficacy of Eucalyptus extract is expressed regardless of race, age, or gender, since similar results were confirmed in all panelist groups. In order to investigate the cause of these phenomena, we measured the elasticity (Young's modulus) of the new-growth part of the cortex in Eucalyptus extract-treated hair and placebo hair by the nano-indentation method of atomic force microscopy (AFM). These results suggest that the Young's modulus of the new-growth part of the cortex in Eucalyptus extract treated-hair increases in comparison with placebo hair. The IR spectra of treated samples of hair show changes that appear to confirm a decrease in the alpha-helix structure and an increase in the beta-sheet structure.