In this in vitro study, the biocompatibility of acrylamide-based hydrogels such as acrylamide/crotonic and acrylamide/itaconic acid, prepared by using gamma-rays, has been investigated against some biochemical parameters of human serum. The swelling kinetics of acrylamide/crotonic and acrylamide/itaconic acid hydrogels are investigated in distilled water and physiological saline solution and some swelling and diffusion parameters have been calculated. For the analysis of biocompatibility, acrylamide/crotonic and acrylamide/itaconic acid hydrogels are incubated in 10 different human sera for 24 h and their biocompatibilities with some biochemical parameters have been investigated. No significant difference in values before and after the test procedures have been found. It is therefore concluded that acrylamide/crotonic and acrylamide/itaconic acid hydrogels are biocompatible.