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Central to the formation of tissue at implant surfaces are the interactions between multiple cell types including fibroblasts, endothelial cells and, in the case of bone, cells of the osteoblastic lineage. To date the importance of population dynamics and interactions have been largely neglected in the in vitro evaluation of biomaterials. To fill this gap we have developed a co-culture system using 3 cell types, primary human bone marrow stromal cells (HBMC), microvascular endothelial cells (HMVEC) and abdominal dermal fibroblasts (HDF). Proliferation of each cell type separately and differentiation of HBMC were determined by flow cytometry analysis. The medium used promoted HBMC differentiation toward osteoblasts without affecting the state of differentiation of HDF and HMVEC. Furthermore, HBMC are strongly affected by HDF and HMVEC, and vice versa. When used on a titanium coated substrate the triple cell culture system identified preferential HBMC proliferation relative to HDF if HMVEC was present. This developed culture system represents a new, optimised and potentially predictive approach to evaluate biomaterial biocompatibility early in development.

Human triple cell co-culture for evaluation of bone implant materials