Estragole is considered as a promising candidate for food preservation due to its antioxidant and antimicrobial properties. However, its sensitivity to light and oxygen, its volatility and hydrophobicity prevent its large application. In order to increase the stability of estragole, conventional liposomes (CL) and drug-in-cyclodextrin-inliposomes (DCL) were prepared by the ethanol-injection method using Phospholipon 90H in combination with cholesterol and then freeze-dried using hydroxypropyl-ß-cyclodextrin (HP-ß-CD) as a cryoprotectant. Fresh and reconstituted vesicles demonstrated nanometric vesicles size, spherical shape and negative surface charge. Moreover, all suspensions were homogeneous. Compared to CL, DCL improved estragole encapsulation efficiency, and conserved the same loading rate during freeze-drying. Freeze-dried CL and DCL retained estragole more efficiently compared to the reconstituted ones, as evidenced by release study performed by multiple headspace extraction. Finally, we demonstrated, for the first time, that HP-ß-CD did not affect the membrane fluidity of fresh and reconstituted Phospholipon 90H/cholesterol CL and DCL.