Chitosan thin films, processed by solvent casting, were functionalized by incorporating nanoliposomes based on natural plant and marine lecithin. The physicochemical properties were characterized by water contact angle, kinetic swelling test, and torsion harmonic atomic force microscopy (TH-AFM) analysis. The surface wettability, swelling ratio, roughness, and local stiffness of chitosan thin films can be modified and controlled by the addition of nanoliposomes. The water contact angle decreases from 75° (±2°) to 50° (±1°) and 38° (±1°) as the amount of soybean and salmon nanoliposomes increases in the chitosan films, respectively. the films reach the equilibrium state after seven days in the presence of phosphate buffer saline. The surface topography of the films is identical in terms of asymmetry and width distribution of roughness measurements, but showed a significant increase in asperity height when incorporating soy nanoliposomes. This variation induces a decrease in the contact stiffness (0.6 Gpa) and in the value of the adhesive force (11.7 nN). A major goal is to understand the underlying cell-matrix interactions. For this purpose, human mesenchymal stem cells (hMSCs) were seeded on the films, the results of the in vitro biocompatibility analysis showed that the thin films of the nanoliposomes/chitosan mixture are not toxic after seeding the cells for 24 hours. The films promote cell adhesion and proliferation when the concentration of soy and salmon nanoliposomes is less than 2 mg/mL and 4 mg/mL, respectively. Salmon nanoliposomes/chitosan blend films especially showed improved cell proliferation in hMSC culture with the concentration of 1 mg/mL. Nanoliposomes functionalized chitosan thin films at certain concentrations could offer...... middle of paper...... w surface energy like polymers. The Owens-Wendt theory divides the surface energy into two components: one due to dispersive interactions and one due to polar interactions 46. Owens and Wendt demonstrated that the total surface energy of a solid can be expressed as the sum of dispersive contributions and components of the polar force. These can be determined from contact angle data, θ, of polar and non-polar liquids with known dispersive and polar parts of their surface energy, via the following relationships: Two liquids (water and diiodomethane) were used for these measurements. The same Digidrop meter was used for measuring the surface energy of chitosan and blended films. The total surface energy, polar component and dispersion component of chitosan and nanoliposomes mix thin films were previously determined from the contact angle of water and diiodomethane.