A macrophage/fibroblast co-culture system using a cell migration chamber to study inflammatory effects of...

Chronic inflammatory reactions hamper the use of biomaterials after implantation. Thus, the aim of the study was to develop a novel predictive in vitro macrophage/fibroblast co-culture model based on cell migration chambers that allows a timely and locally controlled interaction of both cell types to study the inflammatory responses of biomaterials in vitro. Here, self-assembled monolayers (SAMs) with different wettability and charge properties were used as model biomaterials on which co-cultures were established by use of fence chambers having internal and external compartments. This allowed establishing separated and mixed co-cultures of both cell types before and after removal of the chamber, respectively. The key advantages of this novel co-culture model included not only to establish a timely-resolved study of cytokine release, but also the ability to assess individual macrophage migration in both macrophage mono-cultures and co-cultures. All inflammatory reactions in terms of macrophage adhesion, macrophage migration, foreign body giant cell (FBGC) formation, β1 integrin expression and pro-inflammatory cytokine production were found strongly surface property dependent. The results show that the hydrophobic CH3 surface caused the strongest inflammatory reactions, whereas the hydrophilic/anionic COOH surface caused the least inflammatory response, indicating low and high biocompatibility of the surfaces, respectively. Most importantly, we found that both macrophage motility and directional movement were increased in the presence of fibroblasts in co-cultures compared with macrophage mono-cultures. Overall, the novel co-culture system provides access to a range of parameters for studying inflammatory reactions and reveals how material surface properties affect the inflammatory responses.