In this study, L‐Arginine (L‐Arg) molecules are self‐assembled on HA coatings for potential orthopedic applications. By immersing HA coatings in L‐Arg solution in phosphate buffer saline (PBS) with a pH of 7.4 at room temperature for 24 hours, L‐Arg assembled on HA. The samples were characterized by using Scanning Electron Microscopy (SEM), X‐ray Photoelectron Spectroscopy (XPS), Attenuated Total Reflection‐Fourier Transform Infrared Spectroscopy (ATR‐FTIR), X‐ray Diffraction (XRD), Atomic Force Microscopy (AFM) and ninhydrin assay. The response of human bone marrow derived mesenchymal stem cells (hBMSCs) to HA and L‐Arg/HA coatings both in growth and osteogenic differentiation medium are reported. In the growth medium, cells are found to be more spread out on L‐Arg/HA coatings than HA coatings. Moreover, cell aggregates which are indicators of differentiation are observed on HA coatings spontaneously in the growth medium and cell aggregates are only observed on L‐Arg/HA coatings in the osteogenic differentiation medium. Overall, this study showed that L‐Arg self‐assembles on HA and it hinders osteogenic differentiation of hBMSCs in the growth medium.