The current study aimed at in vitro investigating the kinetic study and thermodynamic analysis of mebendazole drug released from electrospun cellulose nanofiber in which guar gum is used as a release controller. The nanofibers were fabricated by electrospinning technique. The fibers were boosted by different controller guar gum 10 at 50, 250, and 500 ppm concentrations. The drug release was investigated on each fiber at 25 °C, 31 °C, 37 °C, and 43 °C for 72 h. The results showed that guar gum can be used as a drug controlling agent in nanofiber. The drug release becomes more difficult where the concentration of guar gum in the nanofiber is higher. Various models for kinetic modeling were investigated, among which the Sahlin-Peppas model fitted the experimental data efficiently. Kinetic studies have shown that both diffusion and swelling mechanisms contribute to the drug release process. This is due to the hydrophilic nature of guar gum. If the value of the controller is greater, the diffusion mechanism dominates the process. Thermodynamic analysis showed that drug release at all controlling concentrations is not spontaneous (ΔG>0) and is an endothermic process (ΔH>0), leading to increased disorder (ΔS<0). Activation energy increases with the increase in the amount of guar gum controller, which means that more energy is needed to release the drug.