Proteomic analysis and quantification of cytokines and chemokines from biomaterial surface‐adherent macrophages and foreign body giant cells

Abstract

Implantation of biomaterial devices results in the well-known foreign body reaction consisting of monocytes, macrophages, and foreign body giant cells (FBGCs) at the material/tissue interface. We continue to address the hypothesis that material surface chemistry modulates the phenotypic expression of these cells. Utilizing our human monocyte culture system, we have used surface-modified polymers displaying hydrophobic, hydrophilic, and/or ionic chemistries to determine the cytokines/chemokines released from biomaterial-adherent macrophages/FBGCs. This study broadens our approach by using proteomic analysis to identify important factors expressed by these cells and further quantifies these molecules with ELISAs. Proteomic profiles changed over time suggesting that the adherent macrophages underwent a phenotypic switch. Macrophage/FBGC-derived proinflammatory cytokines, IL-1beta and IL-6, decreased with time, while the anti-inflammatory cytokine, IL-10, gradually increased with time. Resolution of the inflammatory response was also demonstrated by a decrease in chemoattractant IL-8 and MIP-1beta production with time. Material-dependent macrophage/FBGC activation was analyzed using cytokine/chemokine production and cellular adhesion. Monocyte/macrophage adhesion was similar on all surfaces, except for the hydrophilic/neutral surfaces that showed a significant decrease in cellular density and minimal FBGC formation. Normalizing the ELISA data based on the adherent cell population provided cytokine/chemokine concentrations produced per cell. This analysis showed that although there were fewer cells on the hydrophilic/neutral surface, these adherent cells were further activated to produce significantly greater amounts of each cytokine/chemokine tested than the other surfaces. This study clearly presents evidence that material surface chemistry can differentially affect monocyte/macrophage/FBGC adhesion and cytokine/chemokine profiles derived from activated macrophages/FBGCs adherent to biomaterial surfaces.

In Vivo Host Response and Degradation of Copolymer Scaffolds Functionalized with Nanodiamonds and Bone Morphogenetic Protein 2

“…[ 31 ] Previous reports have shown that hydrophilic/neutral surfaces inhibit infl ammatory cellular adhesion. [ 15 ] This explains the reduced number of infl ammatory cells seen in the nDP modifi ed scaffolds compared to PLCL scaffolds in this study. Hydrophilic surfaces were also shown to downregulate proinfl ammatory gene expression by macrophage-like cells in vitro.…”

“…Other studies also postulate that although the number of infl ammatory cells is reduced in hydrophilic surfaces, but they are more activated thus releasing higher cytokine amounts. [ 15,34 ] However, the assessment of cytokines in vivo is challenging due to the local nature of their production and effects and the complex cellular interactions of a wide range of cytokines at any given time point.…”

“…The chemical composition of the scaffold, its surface charge, porosity, topography, size, and shape affects the intensity and duration of each of the steps of this cascade. [ 15 ] Hence, functionalizing a scaffold with nDP may affect the host's tissue response. Titania [ 16 ] and tripolyphosphate nanoparticles [ 17 ] have been shown to improve the biocompatibility of poly(lactic-co-glycolic acid) scaffolds, while nondegradable nanoparticles made of silica have been shown to induce infl ammation.…”

Adsorbed Fibrinogen Enhances Production of Bone- and Angiogenic-Related Factors by Monocytes/Macrophages

“…The surface onto which this cell population adheres has been shown to influence such secretion. 6,27,46 Our antibody array analysis revealed greater production of soluble factors by macrophages differentiated on RGD compared with those cultured on Ch-based films. Consistent with this, enhanced secretion of matrix metaloproteinases (MMPs) and TIMPs by RGD-differentiated macrophages, as well as of more cytokine production by RGD-adherent dendritic cells, has been previously described.…”

“…4,5 Importantly, the chemical properties of the material, which influence the nature and amount of proteins that rapidly adsorb to its surface after implantation, will regulate macrophage behavior. Besides directing macrophage adhesion and FBGC formation, 6 the surface chemistry of the material also impacts macrophage activation and function, namely through the production of different bioactive agents, such as cytokines, chemokines, or growth factors. 1,2 Depending on the micro-environmental stimuli, macrophages can display either pro-or anti-inflammatory features.…”