The preparation of smart polymeric particles in supercritical carbon dioxide (scCO(2)) presents many advantages for biomedical applications over conventional processes due to the easy elimination of trace contaminants rendering highly pure particles. Herein we report the successful optimization of poly(N-isopropylacrylamide) (PNIPAAm) synthesis strategy to obtain cell-sized hydrogel microbeads with defined and systematically varied mechanical properties. The effect of using different hydrophilic cross-linkers such as N,N-methylenebisacrylamide (MBAm), di(ethylene) glycol dimethacrylate (DEGDMA) and glycerol dimethacrylate (GDMA), on beads morphological, physico-chemical and mechanical properties was investigated. In agreement with a larger water uptake ability beads cross-linked with DEGDMA are more compliant than those containing MBAm or GDMA, having lower stiffness as accessed through oscillatory measurements on a rotational rheometer. Cytotoxicity assays showed that the obtained crosslinked PNIPAAm microbeads do not present any toxic effect on fibroblast cell cultures. Microbeads biocompatibility and adequate mechanical compliance enable their potential application on biomedical settings. (C) 2010 Elsevier B.V. All rights reserved.