The differential adhesion forces of anterior cruciate and medial collateral ligament fibroblasts: effects of tropomodulin, talin, vinculin, and alpha-actinin.
We have determined the effects of tropomodulin (Tmod), talin, vinculin, and alpha-actinin on ligament fibroblast adhesion. The anterior cruciate ligament (ACL), which lacks a functional healing response, and the medial collateral ligament (MCL), a functionally healing ligament, were selected for this study. The micropipette aspiration technique was used to determine the forces needed to separate ACL and MCL cells from a fibronectin-coated surface. Delivery of exogenous tropomodulin, an actin-filament capping protein, into MCL fibroblasts significantly increased adhesion, whereas its monoclonal antibody (mAb) significantly decreased cell adhesiveness. However, for ACL fibroblasts, Tmod significantly reduced adhesion, whereas its mAb had no effect. mAbs to talin, vinculin, and alpha-actinin significantly decreased the adhesion of both ACL and MCL cells with increasing concentrations of antibody, and also reduced stress fiber formation and cell spreading rate as revealed by immunofluorescence microscopy. Disruption of actin filament and microtubule assembly with cytochalasin D and colchicine, respectively, also significantly reduced adhesion in ACL and MCL cells. In conclusion, both ACL and MCL fibroblast adhesion depends on cytoskeletal assembly; however, this dependence differs between ACL and MCL fibroblasts in many ways, especially in the role of Tmod. These results add yet another possible factor in explaining the clinical differences in healing between the ACL and the MCL.
Sung
K L
KL
Department of Orthopaedics, University of California at San Diego, La Jolla 92093-0412, USA.
Yang
L
L
Whittemore
D E
DE
Shi
Y
Y
Jin
G
G
Hsieh
A H
AH
Akeson
W H
WH
Sung
L A
LA
D000185:Actinin / Q000276:immunology / Q000502:physiology; D000328:Adult; D016118:Anterior Cruciate Ligament / Q000166:cytology / Q000502:physiology*; D000911:Antibodies, Monoclonal / Q000494:pharmacology; D002352:Carrier Proteins / Q000276:immunology / Q000502:physiology; D002448:Cell Adhesion / Q000187:drug effects / Q000502:physiology*; D002478:Cells, Cultured; D003598:Cytoskeletal Proteins / Q000276:immunology / Q000502:physiology*; D003599:Cytoskeleton / Q000502:physiology; D005260:Female; D005347:Fibroblasts / Q000502:physiology*; D005353:Fibronectins / Q000502:physiology; D006801:Humans; D007231:Infant, Newborn; D008297:Male; D017888:Medial Collateral Ligament, Knee / Q000166:cytology / Q000502:physiology*; D008840:Microfilament Proteins; D008875:Middle Aged; D016608:Talin / Q000276:immunology / Q000502:physiology; D051348:Tropomodulin; D016596:Vinculin / Q000276:immunology / Q000502:physiology; D014945:Wound Healing / Q000502:physiology
D000911:Antibodies, Monoclonal; D002352:Carrier Proteins; D003598:Cytoskeletal Proteins; D005353:Fibronectins; D008840:Microfilament Proteins; C495829:TMOD1 protein, human; D016608:Talin; D051348:Tropomodulin; D000185:Actinin; D016596:Vinculin