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Along with providing an environment for cell attachment and proliferation, a tissue engineering scaffolds should possess physical and mechanical properties that would fit the target tissue. The present study aimed to manipulate physico-mechanical properties of polyacrylamide/gelatin hydrogels using response surface method-central composite design (RSM-CCD) to reach a scaffold with defined properties. On this demand, mixtures of gelatin and acrylamide (AAm) monomer were used to prepare semi-interpenetrating hydrogels by free radical polymerization of AAm. Selected variables for statistical modeling were chosen to be weight ratios of monomer/crosslinker, monomer/gelatin, and monomer/initiator. The desired responses were compressive modulus, compressive strength, and swelling. Results showed that desired responses could be tailored by varying these parameters with the highest impact for monomer/crosslinker ratio. The swelling ratio of hydrogels was in the range of 947–1654%, while the modulus varied between 5 and 35 kPa. The cyclic compressive test showed the durability of hydrogels under cyclic loadings. Finally, the results of cell attachment and cytocompatibility analyses indicated that the hydrogels were completely biocompatible and enhanced cell attachment. Thus, these hydrogels could potentially be used as tissue engineering scaffolds for load-bearing organs, including muscle and cartilage, or could be used for in vitro differentiation of stem cells using mechanical clues. [ABSTRACT FROM AUTHOR]