Software-supported image quantification of angiogenesis in an in vitro culture system: Application to studies of biocompatibility

Healing of soft tissue trauma and bone discontinuities following implantation involves acute inflammatory reactions and the formation of blood vessels (angiogenesis). During angiogenesis new capillary vessels arise from the existing vasculature. Endothelial cells (EC) are the major cell type involved in angiogenesis. Corrosion of orthopaedic metallic implant materials (e.g. CoCr alloys) can cause locally high concentrations of heavy metal ions in the peri-implant tissues. Some divalent metal ions (Co²⁺, Ni²⁺, Zn²⁺) lead to the activation of EC in vitro. Upon exposure to these ions, EC release cytokines and chemokines and increase the expression of cell surface adhesion molecules, which represents the pro-inflammatory phenotype. In this study we have examined whether metal ions influence the other endothelial aspect of wound healing, the angiogenic response. Therefore, we utilized an in vitro model of angiogenesis and examined the effects of divalent cobalt ions on the in vitro vessel formation. The quantification of the cobalt/ion-exerted effects on angiogenesis in vitro was performed using a contrast-rich vital staining and analysed by software-supported image quantification.