Contractile forces generated by articular chondrocytes in collagen-glycosaminoglycan matrices.
The objective of the study was to directly measure the force of contraction of adult articular chondrocytes and to examine their contractile behavior in collagen-glycosaminoglycan analogs of extracellular matrix by live cell imaging in vitro. The contractile forces generated by passages 2 and 3 adult canine articular chondrocytes were measured using a cell force monitor. The contractile behavior of the cells was also directly imaged as they were cultured in collagen-glycosaminoglycan scaffolds. Passage 2 cells seeded in a collagen-glycosaminoglycan scaffold were capable of generating a force of 0.3 nN/cell. Chondrocytes subcultured through a third passage generated a force twice that level, paralleling the increase in the alpha-smooth muscle actin (SMA) content of the cells as demonstrated by Western blot analysis. Treatment of passage 3 cells with staurosporine reduced the force of contraction by approximately one-half, reflecting the effects of this agent in reducing the SMA content of the cells and disrupting the microfilaments. These values compare with 1 nN previously reported for lapine dermal fibroblasts of passage 5-7, using the same apparatus. Direct live cell imaging documented the contractile behavior of the articular chondrocytes in the collagen-glycosaminoglycan matrix in the time frame in which the force was directly measured in the cell force monitor. This imaging demonstrated how the cells acted individually and in unison to buckle the collagen struts making up the matrix. Adult articular chondrocytes are capable of generating a SMA-enabled force of contraction sufficient to model extracellular matrix molecules.
Zaleskas
Janice M
JM
Department of Orthopaedic Surgery, Brigham and Women's Hospital, Harvard Medical School, 75 Francis Street, Boston, MA 02115, USA.
Kinner
Bernd
B
Freyman
Toby M
TM
Yannas
Ioannis V
IV
Gibson
Lorna J
LJ
Spector
Myron
M
D000818:Animals; D002358:Cartilage, Articular / Q000166:cytology / Q000502:physiology; D002478:Cells, Cultured; D019902:Chondrocytes / Q000166:cytology / Q000502:physiology*; D003094:Collagen / Q000502:physiology* / Q000648:ultrastructure; D004285:Dogs; D005109:Extracellular Matrix / Q000502:physiology / Q000648:ultrastructure; D006025:Glycosaminoglycans / Q000502:physiology* / Q000648:ultrastructure; D008422:Materials Testing; D040542:Mechanotransduction, Cellular / Q000502:physiology; D020409:Molecular Motor Proteins / Q000502:physiology*; D013314:Stress, Mechanical; D023822:Tissue Engineering / Q000379:methods*
D006025:Glycosaminoglycans; D020409:Molecular Motor Proteins; D003094:Collagen