Six culture configurations were evaluated in this study. (A) Cells were cultured in mono-, co-, or tri-culture hydrogel constructs for up to 7 days and then analyzed as depicted above. After one day in culture, media was changed and half of the constructs were switched to high glucose. Shades of gray and white blocks represent hydrogel blocks of different types of cells in either mono-, co-, or tri-culture. (B) Tri-culture configurations consisted of adipocytes, MSCs, and osteoblasts (designated OMA); co-culture configurations consisted of osteoblasts and MSCs or adipocytes and MSCs (designated OMO and AMA, respectively); and mono-culture configurations consisted of osteoblasts, MSCs or adipocytes (designated OOO, MMM, and AAA, respectively).