To address implant-related infections, antibacterial solutions specific to biomaterials are required to prevent bacterial proliferation. Traditional antibiotic usage has been found insufficient, motivating researchers to investigate alternative strategies such as surface modification and the application of antifouling or infection-resistant properties. A developing interest lies in designing surfaces that mimic natural antibacterial nanotopographies. In this study, we conducted a quantitative analysis of the outcomes from plasma nanotexturing, with particular emphasis on how the organization of topography influences antibacterial efficacy and the regulation of cell alignment. Plasma nanotexturing was applied to chitosan surfaces, which gradually transformed from nanopores to pillars and eventually into tilted pillars, as the plasma parameters (fluence and angle) increased. We used directed plasma nanosynthesis, a plasma-based technique that primarily induces topographical alterations on the surfaces. The surfaces were systematically characterized, incorporating methods such as scanning electron microscopy (SEM), atomic force microscopy (AFM), and X-ray photoelectron spectroscopy (XPS). A comprehensive comparison of the nanotextures was executed by utilizing a trapezoidal method to calculate aspect ratios and assess texture orientation by examining the gaps in the nanostructures. We evaluated antibacterial properties against E. coli and S. aureus strains and assessed the survival and alignment of human bone marrow mesenchymal stem cells. Our findings reveal a significant reduction in bacterial adhesion (>80%) and growth on nanotextured surfaces, underscoring their potential for clinical applications. Moreover, we measured cell alignment, presenting the results in both a color-coded and numerical format to demonstrate the preferential alignment orientation induced specially by the tilted nanotexture. These insights highlight the profound impacts of plasma nanotexturing, indicating its potential for innovative biomedical applications such as advanced wound healing and tissue engineering.