Analysis of the glycodynamics of primary osteoblasts and bone cancer cells

Bone cells produce a variety of glycoproteins that contribute to bone health, and function in cell adhesion, stabilizing the extracellular matrix, promoting growth and differentiation, and the induction of apoptosis. Some of these processes appear to be disturbed in arthritis. In this chapter, in vitro studies aimed at an understanding of the biological effects of inflammatory stimuli in the bone of arthritis patients are described. The glycodynamics of cells can be studied using primary cultures of osteoblasts or bone cancer cell cultures, to examine the relationship between the biosynthesis of cell-surface glycoproteins and inflammatory stimuli affecting cell growth and cell death. Cell-surface carbohydrates are assessed by lectin staining of cells, and the potential of cells to synthesize glycoproteins is determined by glycosyltransferase assays. These parameters are then related to [3H]thymidine incorporation as a measure of cell proliferation, and to flow cytometry of terminal deoxynucleotidyl transferase dUTP-mediated nick end labeling (TUNEL) and annexin V-stained cells as a measure of apoptosis. These in vitro studies are aimed at an understanding of the role of glycosylation in the bone of arthritis patients, but they can also be applied to other diseases.