Confocal laser scanning microscopy (CLSM) was used in conjunction with in vitro cell culture to investigate cellular interactions with orthopaedic biomaterials. Transfected rat and human osteoblasts were seeded on two potential isoelastic hip prosthesis materials, carbon fibre reinforced polyetheretherketone (PEEK) and epoxy. Titanium 318 alloy was employed as a control. Determination of the material surface contour, an important factor influencing cellular adhesion, proliferation and function, was performed using the industry standard Talysurf® and compared to analogous results obtained using the CLSM. The latter technique consistently gave higher values of material roughness but offers the advantage that it can be used to correlate roughness with cell distribution on the same samples, whereas Talysurf® measurement of roughness requires clean rigid samples. Image analysis and processing, performed on cells after attachment and culture on the materials for 48 hours, provided cell morphology data. Cells cultured on titanium were larger, with a higher percentage of cytoplasm, than those grown on either of the other materials. The macroscopic surface of epoxy resulted in smaller cells with altered morphology, which orientated themselves along carbon fibres. In conclusion, we believe CLSM offers great potential for investigating the cellular interactions of biomaterials involving minimal sample preparation, non-invasive optical sectioning of samples and minimal opportunity for generation of cellular deformation and sample preparation artefacts.
Macnair, R.; Wilkinson, R.; MacDonald, C.; Goldie, I.; Jones, D. B.; and Grant, M. H.
"Application of Confocal Laser Scanning Microscopy to Cytocompatibility Testing of Potential Orthopaedic Materials in Immortalised Osteoblast-Like Cell Lines,"
Cells and Materials: Vol. 6
, Article 8.
Available at: http://digitalcommons.usu.edu/cellsandmaterials/vol6/iss1/8