Mesenchymal stem cells (MSCs) are used extensively in cartilage tissue engineering. We have developed a photopolymerizable poly(ethylene glycol diacrylate) (PEGDA) and poly(ethylene glycol) (PEG) semi-interpenetrating network that facilitates the in vitro chondrogenesis of human MSCs (hMSCs). Network parameters were altered and tested for their effects on subsequent matrix elaboration. The mesh size, calculated for each network based on equilibrium swelling ratios, was larger with lower PEGDA:PEG ratios and with higher PEGDA molecular weight. Changes in xi correlated with changes in extracellular matrix content and deposition in hMSC-seeded networks cultured in vitro for 6 weeks in defined chondrogenic medium. Networks constructed with PEGDA (6 kDa) and PEG (88 kDa) at 1:2 displayed intercellular deposition of proteoglycan. Furthermore, their proteoglycan contents were significantly higher than with PEGDA (6 kDa) hydrogels constructed without the PEG component and those constructed at a PEGDA:PEG ratio of 2:1, which both exhibited pericellular proteoglycan deposition. However, networks constructed with PEGDA (12 and 20 kDa) and PEG (88 kDa) exhibited intercellular deposition of proteoglycan regardless of the ratio employed. Collagen content was lower in networks constructed with PEGDA (12 and 20 kDa) and PEG (88 kDa) at a ratio of 1:2 than in those fabricated at the same PEGDA molecular weights at a ratio of 2:1. This study demonstrated that semi-interpenetrating network parameters influence not only extracellular matrix content, but also the deposition of the matrix molecules by hMSCs undergoing chondrogenesis. It is important that these parameters be considered carefully when creating scaffolds for tissue-engineered cartilage.