Accumulation of PrPSc, an insoluble and protease-resistant pathogenic isoform of the cellular prion protein (PrPC), is a hallmark in prion diseases. Branched polyamines, including PPI (poly(propylene mime)) dendrimers, are able to remove protease resistant PrPSc and abolish infectivity, offering possible applications for therapy. These dendrimer types are thought to act through their positively charged amino surface groups. In the present study, the molecular basis of the antiprion activity of dendrimers was further investigated, employing modified PPI dendrimers in which the positively charged amino surface groups were substituted with neutral carbohydrate units of maltose (mPPI) or maltotriose (m3PPI). Modification of surface groups greatly reduced the toxicity associated with unmodified PPI but did not abolish its antiprion activity, suggesting that the presence of cationic surface groups is not essential for dendrimer action. PPI and mPPI dendrimers of generation 5 were equally effective in reducing levels of protease-resistant PrPSc (PrPres) in a dose- and time-dependent manner in ScN2a cells and in pre-existing aggregates in homogenates from infected brain. Solubility assays revealed that total levels of PrPSc in scrapie-infected mouse neuroblastoma (ScN2a) cells were reduced by mPPI. Coupled with the known ability of polyamino dendrimers to render protease-resistant PrPSc in pre-existing aggregates of PrPSc susceptible to proteolysis, these findings strongly suggest that within infected cells dendrimers reduce total amounts of PrPSc by mediating its denaturation and subsequent elimination.