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Physically crosslinked hydrogels based on poly(2-hydroxyethyl methacrylate) (PHEMA) and polyurethane macroiniferter (PUMI) were prepared. The synthesis of polyurethane-block-poly(2-hydroxyethyl methacrylate) (PU-b-PHEMA) was verified by spectroscopic analyses. Due to the low fibronectin adsorption from culture media, cell attachment on PU-b-PHEMA surface was poor compared with the PUMI control. To improve the cell affinity of PU-b-PHEMA, fibronectin was conjugated via surface hydroxyl groups. These biomimetic PU-b-PHEMA hydrogel surfaces were tested for tissue engineering applications. A short-term cell culture study revealed that, compared with the unmodified PU-b-PHEMA, fibronectin-conjugated PU-b-PHEMA hydrogel showed more uniform and dense cell attachment and spreading, indicating a potential use for tissue engineering applications.