In this paper, the cuticle characteristics of hair after oil treatment are discussed and analyzed. Hair lubrication mechanics were measured through atomic force microscopy (AFM), friction force microscopy (FFM), and Fourier-transform infrared (FTIR) spectrometry. From experimental results, the destruction and surface roughness of human hair treated with hair oil were evaluated. A homemade hair oil formula was applied and the treatment time was varied to observe the effects of hair care formulations on hair cuticles and surfaces. The results indicated that the extent of differences in hair cuticles varied according to different hair oil heating times. The adhesion force of human hair was approximately 17.0–36.8 nN. The results revealed high peaks associated with the increased CH2 intensity. Regions of approximately 2352–2938 cm−1 (–C–C– H–) alkyl absorption peaks and approximately 3441 cm−1 (–N–H–) amino acid absorption peaks were observed. An increasing trend of alkyl absorption peaks was also indicated. The observed 1361 cm−1 (amide, –CN–) aliphatic absorption peaks and 1626 cm−1 (>C=O) amino acid cysteine absorption peaks were typical effects of external factors on polypeptide hair fibers. The use of FTIR enabled the observation of changes in hair protein composition and surface structure.