We present three easily rationalized star-shaped PEGylated β-cyclodextrin (βCD) derivatives synthesized via conjugation of different molecular weight PEG chains (5000, 2000, and 550 Da) to the βCD primary face by click chemistry (βCD-PEG5000, βCD-PEG2000, βCD-PEG550 respectively). βCDPEG systems are envisioned to further carry bioactive molecules, therefore, their interactions with biological interfaces must be determined at an early stage of development. Hence, the effect of βCDPEGs chain length on cell viability was investigated. To this aim, three models were selected: Vero cells for their fibroblast-like features; HeLa cells that are commonly used for preliminary viability screening; and human peripheral monocytes which are macrophage precursors. Of the three pegylated derivatives, βCD-PEG550 was the one that significantly affected HeLa cells and human monocytes viability. Despite the popularity of PEGylation approach, our results underscore the importance of careful and systematic PEGylated materials design for their future success in drug delivery systems.