In recent years, reed bed systems (RBSs) have been widely considered as a valid technology for sludge treatment. In this study are presented results about sludge stabilization occurring within beds in four RBSs, situated in Tuscany (Italy). The results showed that stabilization of the sludge over time occurred in all RBSs, as shown by the low content of water-soluble carbon and dehydrogenase activity, which measures indirectly the overall microbial metabolism, and by the re-synthesis of humic-like matter highlighted by the pyrolytic indices of mineralization and humification. Results about heavy metal fractionation, an appropriate technique to estimate the heavy metal bioavailability and sludge biotoxicity, showed that the process of sludge stabilization occurring in RBSs retains metals in fractions related to the stabilized organic matter, making metals less bioavailable. Moreover, the concentrations of various toxic organic compounds were below the limit of concentration suggested by the European Union's Working Document on Sludge, for land application. The effectiveness of the stabilization processes in RBs was hence clearly proven by the results that measured mineralization and humification processes, and by the low levels of bioavailable heavy metals and toxic organic compounds in stabilized sludges.