Collagen mimetic peptide (CMP) with a specific amino acid sequence, -(Pro-Hyp-Gly)(x)-, forms a triple helix conformation that resembles the native protein structure of natural collagens. CMP previously has been shown to associate with type I collagen molecules and fibers via a strand invasion process. We hypothesized that when poly(ethylene glycol) (PEG) hydrogel, a non-adhesive tissue engineering scaffold, is conjugated with CMP, it may retain cell-secreted collagens and also form physical crosslinks that can be manipulated by cells. A photopolymerizable CMP derivative was synthesized and copolymerized with poly(ethylene oxide) diacrylate to create a novel PEG hydrogel. In a model retention experiment, diffusional loss of type I collagen that was added to the hydrogel was limited. Chondrocytes were encapsulated in the hydrogel to examine its use as a tissue engineering scaffold. After 2 weeks, the biochemical analysis of the CMP-conjugated PEG gel revealed an 87% increase in glycosaminoglycan content and a 103% increase in collagen content compared to that of control PEG hydrogels. The histology and immunohistochemistry analyses also showed increased staining of extracellular matrix. These results indicate that the CMP enhances the tissue production of cells encapsulated in the PEG hydrogel by providing cell-manipulated crosslinks and collagen binding sites that simulate natural extracellular matrix.