Biomechanical and biochemical factors are believed to precipitate osteoarthritis although the etiologic relationship among factors is unclear. The alpha-5 beta-1 fibronectin receptor participates in both biochemical and biomechanical signaling in chondrocytes and may integrate input from these pathways to maintain cartilage health. Changes in the cytoskeleton are affected by alpha-5 beta-1 signaling and might relate with cartilage pathologies. Fibronectin fragments (Fn-fs), which likely utilize alpha-5 beta-1 signaling, potentiate cartilage degeneration and alter actin filament organization in fibroblasts. Mechanical compression of cartilage is known to disrupt intermediate filaments. It is not known whether Fn-fs alter the chondrocyte cytoskeleton. This study examined the capacity of N-terminal 29 kDa Fn-fs to induce changes in the chondrocyte cytoskeleton and whether these changes resembled reports of Fn-f mediated changes in fibroblasts and/or compression induced changes in the chondrocyte cytoskeleton. Cytoskeleton disturbing agents colchicine, taxol, cytochalasin D and jasplakinolide were examined for their effects on Fn-f activities and interactions with chondrocytes. Agents were tested for erects on Fn-f mediated proteoglycan release and proteoglycan synthesis suppression in cartilage explant cultures. Fn-f activities were compared with native Fn in all experiments. Fluorescence cytochemistry showed no detectable differences in actin filament, vimentin filament or microtubule organization in chondrocytes incubated with or without Fn or Fn-f. Both actin filament and microtubule altering agents increased Fn and Fn-f interaction with chondrocytes but not all agent-induced increases in Fn-f binding had effects on the metabolic activities of Fn-fs. Both cytochalasin D and colchicine decreased Fn-f mediated proteoglycan release from cartilage explants, while taxol and jasplakinolide had no significant effect on proteoglycan release. Taxol but not colchicine decreased Fn-f mediated proteoglycan synthesis suppression. These data suggest that some Fn-f activities require an intact cytoskeleton while other activities are facilitated by cytoskeleton disruption. This work supports a conjectural link between biomechanical and biochemical mediators of cartilage homeostasis through common use of the cytoskeleton. The chondrocyte's capacity to re-organize the cytoskeleton alludes to possibilities that mechanical loading of cartilage might influence Fn-f activities. This provides a basis for better understanding the relationship between biomechanical and biochemical factors contributing to cartilage degeneration.