We have investigated the formation of complexes between negatively charged niosomal vesicles (hybrid niosomes), built up by dicethylphosphate [DCP], Tween 20 and Cholesterol, and three linear differently charged cationic polyions, such as alpha-polylysine, epsilon-polylysine, and polyethylvinylpyridinium bromide [PEVP], with two different substitution degrees. Our aim is to investigate the interaction mechanism between anionic-nonionic vesicles (hybrid niosomes) and linear polycations, characterizing the resulting aggregates in view of possible applications of these composite colloidal particles as vectors for multidrug delivery. In order to explore the aggregation behavior of the complexes and to gain information on the stability of the single niosomal vesicles within the aggregates, we employed dynamic light scattering (DLS), laser Doppler electrophoretic measurements, and fluorescence measurement techniques. The overall phenomenology is well described in terms of the re-entrant condensation and charge inversion behavior, observed in different colloidal systems. The aggregate size and overall charge depend on the charge ratio between vesicles and polyions, and the aggregates reach their maximum size at the point of charge inversion (re-entrant condensation). While the overall phenomenology is similar for all three polycations investigated, the stability and the integrity of the hybrid niosomal vesicles forming the aggregates strongly depend on the chemical structure of the polycations. The role of the polycations in the aggregation process is discussed by identifying specific interactions with the niosomal membrane, pointing out their importance for possible applications as drug delivery vectors.