Quantification of Varying Adhesion Levels in Chondrocytes Using the Cytodetacher

Abstract

The potential to promote cell adhesion must be evaluated in the development of tissue-engineered implants and scaffolds. One measure of cell adherence is the force necessary to detach the cell. The objective of this study was to evaluate the cytodetacher (Athanasiou, K. A., et al. Development of the cytodetachment technique to quantify cellular adhesiveness. Biomaterials 20: 2405-2415, 1999), modified to test cells grown on substrata, as a means of calculating cell adhesion forces. Live and formalin-fixed bovine and rabbit chondrocytes underwent cytodetachment to verify that the cytodetacher provides satisfactory resolution to differentiate between live and fixed cells. Fixed cells had significantly greater mechanical adhesiveness than those prepared live: the values for the fixed rabbit and bovine chondrocytes were 1.01 and 1.56 microN, respectively, versus 0.14 and 0.17 microN for the live cells (p<0.05). The sensitivity of the cytodetacher was also gauged by detaching live rabbit chondrocytes seeded for varying amounts of time (40, 80, and 120 min). For the 40, 80, and 120 min time points the maximum detachment forces were found to be significantly different: 2.87 x 10(-2), 6.75 x 10(-2), and 14.30 x 10(-)2 microN, respectively. This study validates the use of the modified cytodetacher as an effective means of evaluating the strength of adhesion of cells attached to a substratum.

The effect of remodelling and contractility of the actin cytoskeleton on the shear resistance of single cells: a computational and experimental investigation

“…The absence of bond rupture was confirmed by fluorescent imaging of the focal adhesion protein vinculin, which was observed at the periphery of the cell. Previous in vitro experiments have investigated cell adhesion and detachment during shear deformation [13,41]. A computational study [17], based on the experimental findings of Huang et al [13], illustrates that a strengthening of the cell-substrate interface in tandem with cytoplasm stiffening during cell spreading results in increased cell detachment forces.…”

Nanotribological Perspectives in Tissue Engineering

“…The role of shear strength on cell-material adhesion has been investigated also by Yamamoto et al [93] and Athanasioiu and coworkers [94,95]. Such a method was based on the design of a cytodetachment instrument able to quantify the force required to displace and to detach cells attached to a substratum, Fig.…”

“…The averages of the cell detachment shear strengths measured on murine fibroblasts after 24 h of incubation of glass by Yamamoto et al fall in the range 500 -750 kPa, and shear force in the range 350 -580 nN. Athanasiou and coworkers used the cytodetachment to measure single-cell adhesiveness and the effect of protein on cell adhesiveness [94,95]. They found that chondrocytes present an adhesion force around 20 nN and that the fibronectin strongly increases the adhesion of the cell to the substrate, see Fig.…”

Biomechanics of single chondrocytes under direct shear

“…Direct shearing of single chondrocytes was performed using a modified cytodetachment apparatus (Athanasiou et al 1999;Hoben et al 2002). This device was adapted from its original setup to allow for the videocapture of cells experiencing mechanical shear.…”