Material surfaces affect the protein expression patterns of human macrophages: A proteomics approach

Abstract

Monocyte-derived macrophages (MDM) are key inflammatory cells and are central to the foreign body response to implant materials. MDM have been shown to exhibit changes in actin cytoskeleton, multinucleation, cell size, and function in response to small alterations in polycarbonate-urethane (PCNU) surface chemistry. Although PCNU chemistry has an influence on de novo protein synthesis, no assessments of the protein expression profiles of MDM have yet been reported. The rapid emerging field of expression proteomics facilitates the study of changes in cellular protein profiles in response to their microenvironment. The current study applied proteomic techniques, 2-dimensional electrophoresis (2-DE) combined with MALDI-ToF (matrix assisted laser desorption ionization-time of flight) mass spectrometry, to determine differences in MDM protein expression influenced by PCNU. Results indicated that MDM responded to material chemistry by modulation of structural proteins (i.e. actin, vimentin, and tubulin). Additionally, intracellular protein modulation which requires proteins responsible for trafficking (i.e. chaperone proteins) and protein structure modification (i.e. bond rearrangement and protein folding) were also altered. This study demonstrated for the first time that a proteomics approach was able to detect protein expression profile changes in MDM cultured on different material surfaces, forming the basis for utilizing further quantitative proteomics techniques that could assist in elucidation of the mechanisms involved in MDM-material interaction.

Fetal bovine serum xenoproteins modulate human monocyte adhesion and protein release on biomaterials in vitro

“…Greater cell spreading was observed for PEG and TCPS surfaces when the culture medium was supplemented with AHS instead of FBS, but not for PDMS and TCPS cultured with LPS. Adherent monocytes were stained for CD86 and CD163 at 7d (Figure 6) when a mature macrophage phenotype would be expected [35]. Adherent monocytes/macrophages on all surfaces and cultured with either AHS or FBS supplementation stained for CD86, in contrast, little to no staining was observed for CD163, a surface antigen involved in endocytosis [29].…”

Selection and validation of reference house­keeping genes in the J774A1 macrophage cell line for quantitative real-time PCR

“…In their natural environment, macrophage response to extracellular matrix (ECM) components such as laminin and fibronectin, include adhesion, proliferation, gene expression, and signaling (Förster et al, 2008;McNally et al, 2008;Sorokin, 2010). However, macrophages alter their cytokine/chemokine response profiles when cultured in hydrophobic, hydrophilic, and/or ionic surface-modified polymers (Dinnes et al, 2007;Jones et al, 2007). The natural cellular response can be maintained if the ECM components are provided on material surfaces in such a way at the micrometer and nanometer levels that they simulate the natural topography of cell growth area (Yim and Leong, 2005).…”

Fluorescence two-dimensional difference gel electrophoresis for biomaterial applications

“…In the biomaterials field, this approach has been applied to investigate the response of monocyte-derived macrophages to polycarbonate-urethane (Dinnes et al 2007) and dermal fibroblasts to titanium surfaces (Derhami et al 2001). The technique suffers from gel-to-gel variation and poor reproducibility, however, as one sample is resolved per gel in the absence of an internal standard, and therefore a large number of gels are required to perform a robust pairwise comparison.…”

“…In addition, relatively large quantities of protein are required for this approach (at least 50 mg for silver staining, the most sensitive non-fluorescent post-stain, and up to approximately 500 mg protein with colloidal Coomassie Blue staining, equivalent to two or three 75 cm 3 tissue culture flasks of confluent adherent cells). Sufficient protein was generated in the aforementioned investigations by using relatively high cell densities (Derhami et al 2001;Dinnes et al 2007) and small-format gels (less protein required, but with lower resolutionfewer distinct spots-than large-format gels) (Derhami et al 2001). These approaches are not always practicable, depending on the biomaterial system, because samples can be very scarce and high resolution is desirable.…”