This study examines the effect of microfluidization on the physicochemical properties, stability, release behavior, and cytocompatibility of cannabidiol (CBD) nanoemulsions intended for topical application. CBD is a non-psychoactive cannabinoid characterized by anti-inflammatory and analgesic activity; however, its therapeutic use is limited by low solubility and poor bioavailability. To address these limitations, nanoemulsions were formulated using avocado oil and Tween 80 and optimized through a Box–Behnken experimental design evaluating microfluidization pressure (5000–20,000 PSI), CBD concentration (0–2%), and oil content (8–10%). Nanoemulsions were characterized over a 60-day period in terms of droplet size, dispersity index (D), and zeta potential. An increase in processing pressure led to a reduction in both droplet size and dispersity, with optimal conditions identified between 11,000 and 15,000 PSI. Higher oil and CBD concentrations were associated with an increase in the magnitude of the zeta potential, contributing to electrostatic stabilization of the system. Encapsulation efficiency reached approximately 81.4%. Cell viability assays in HaCaT keratinocytes indicated no significant cytotoxic effects. The optimized formulation exhibited a sigmoidal CBD release profile best described by Weibull and Gompertz models (R² ≈ 0.99), suggesting combined diffusion and interfacial mechanisms that support efficient topical delivery.