A centrifugation cell adhesion assay for high-throughput screening of biomaterial surfaces.

A quantitative analysis of cell adhesion is essential in understanding physiological phenomena and designing biomaterials, implant surfaces, and tissue-engineering scaffolds. The most common cell adhesion assays used to evaluate biomaterial surfaces lack sensitivity and reproducibility and/or require specialized equipment and skill-intensive operation. We describe a modified centrifugation cell adhesion assay that uses simple and convenient techniques with standard laboratory equipment and provides reliable, quantitative measurements of cell adhesion. This centrifugation assay applies controlled and uniform detachment forces to a large population of adherent cells, providing robust statistics for quantifying cell adhesion. The applicability of this system to the design and characterization of biomaterial surfaces is shown by evaluating cell adhesion on substrates using different coating proteins, cell types, seeding times, and relative centrifugal forces (RCF). Results verify that this centrifugation cell adhesion assay represents a simple, convenient, and standard method for high-throughput characterization of a variety of biomaterial surfaces and conditions.

[1]  A. Colombatti,et al.  Centrifugal assay for fluorescence-based cell adhesion adapted to the analysis of ex vivo cells and capable of determining relative binding strengths. , 1999, BioTechniques.

[2]  Buddy D. Ratner,et al.  Some Background Concepts , 1996 .

[3]  Benjamin G Keselowsky,et al.  Surface chemistry modulates fibronectin conformation and directs integrin binding and specificity to control cell adhesion. , 2003, Journal of biomedical materials research. Part A.

[4]  G. Wessel,et al.  Intercellular recognition: quantitation of initial binding events. , 1981, Proceedings of the National Academy of Sciences of the United States of America.

[5]  C. Zhu,et al.  Measuring two-dimensional receptor-ligand binding kinetics by micropipette. , 1998, Biophysical journal.

[6]  P Ducheyne,et al.  Quantification of cell adhesion using a spinning disc device and application to surface-reactive materials. , 1997, Biomaterials.

[7]  Richard O Hynes,et al.  Integrins Bidirectional, Allosteric Signaling Machines , 2002, Cell.

[8]  J. Doroszewski,et al.  Interaction of neoplastic cells with glass surface under flow conditions. , 1977, Experimental cell research.

[9]  D. McClay,et al.  Cell adhesion to fibronectin and tenascin: quantitative measurements of initial binding and subsequent strengthening response , 1989, The Journal of cell biology.

[10]  Larry V. McIntire,et al.  Effect of flow on polymorphonuclear leukocyte/endothelial cell adhesion , 1987 .

[11]  T. Keaveny,et al.  Quantitative Assessment of Steady and Pulsatile Flow Fields in a Parallel Plate Flow Chamber , 1999, Annals of Biomedical Engineering.

[12]  K. Sung,et al.  Quantification of adhesiveness of osteoblasts to titanium surfaces in vitro by the micropipette aspiration technique. , 1996, Tissue engineering.

[13]  K. Burridge,et al.  Focal adhesions, contractility, and signaling. , 1996, Annual review of cell and developmental biology.

[14]  Nathan D. Gallant,et al.  Micropatterned surfaces to engineer focal adhesions for analysis of cell adhesion strengthening , 2002 .

[15]  A Leung,et al.  Detachment of agglutinin-bonded red blood cells. I. Forces to rupture molecular-point attachments. , 1991, Biophysical journal.

[16]  R. G. Richards,et al.  MEASUREMENT OF FIBROBLAST AND BACTERIAL DETACHMENT FROM BIOMATERIALS USING JET IMPINGEMENT , 2001, Cell biology international.

[17]  R M Hochmuth,et al.  Adhesion of red cells to foreign surfaces in the presence of flow. , 1974, Journal of biomedical materials research.

[18]  Richard O. Hynes,et al.  Integrins: Versatility, modulation, and signaling in cell adhesion , 1992, Cell.

[19]  Catherine D Reyes,et al.  Engineering integrin-specific surfaces with a triple-helical collagen-mimetic peptide. , 2003, Journal of biomedical materials research. Part A.

[20]  L. Weiss,et al.  The measurement of cell adhesion. , 1961, Experimental cell research.

[21]  B D Ratner,et al.  Cell adhesion to a series of hydrophilic-hydrophobic copolymers studied with a spinning disc apparatus. , 1988, Journal of biomedical materials research.

[22]  G. Truskey,et al.  The effect of fluid shear stress upon cell adhesion to fibronectin-treated surfaces. , 1990, Journal of biomedical materials research.