Modulation of the contractile and biosynthetic activity of chondrocytes seeded in collagen-glycosaminoglycan matrices.
Studies have demonstrated that articular chondrocytes can express the gene for alpha-smooth muscle actin (SMA) and can contract porous polymeric matrices employed for tissue engineering, thereby altering the pore structure and distorting the shape of the scaffold. The objectives of this study were to determine whether an agent known to disrupt microfilament organization in chondrocytes could reduce this contractility and to assess whether there was an association between the contractile behavior of chondrocytes and their biosynthetic activity. Staurosporine, an antibiotic known to inhibit protein kinase C and disrupt cytoskeletal structure, was used as the agent to modulate the chondrocytic phenotype and contractile and biosynthetic activity of serially passaged adult canine chondrocytes seeded in type 1 collagen-glycosaminoglycan scaffolds. Cells in monolayer culture treated with as little as 3 nM staurosporine for 4 days contained type II procollagen, whereas few cells in the untreated control cultures demonstrated type II procollagen synthesis. Treatment with staurosporine also led to a decrease in the amount of SMA synthesized by the cells. Consistent with this decreased expression of the contractile actin isoform, cells cultured in the collagen-glycosaminoglycan scaffolds and treated with 5 nM staurosporine contracted the scaffold significantly less than untreated cells (15% diameter contraction by treated cells, compared with more than 50% contraction by untreated cells). The staurosporine-treated cells were biosynthetically active, displaying higher rates of protein and glycosaminoglycan synthesis, as indicated by rates of incorporation of [(3)H]proline and [(35)S]sulfate, respectively, compared with untreated cells. The long-held notion that changes in cytoskeletal structure influence phenotypic characteristics of cultured chondrocytes may now be extended to relate expression of a specific muscle actin isoform to certain cell processes. Moreover, the finding that chondrocytes with a lower level of expression of SMA and reduced contractility display higher rates of biosynthesis warrants further study.
Lee
C R
CR
Department of Mechanical Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
Grodzinsky
A J
AJ
Spector
M
M
D000199:Actins / Q000378:metabolism; D000818:Animals; D015153:Blotting, Western; D019902:Chondrocytes / Q000187:drug effects / Q000378:metabolism*; D003094:Collagen / Q000378:metabolism*; D004247:DNA / Q000378:metabolism; D004285:Dogs; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006025:Glycosaminoglycans / Q000378:metabolism*; D007150:Immunohistochemistry; D019311:Staurosporine / Q000494:pharmacology; D023822:Tissue Engineering
D000199:Actins; D004791:Enzyme Inhibitors; D006025:Glycosaminoglycans; D003094:Collagen; D004247:DNA; D019311:Staurosporine