Biochemical Characterization of the Cell-Biomaterial Interface by Quantitative Proteomics

Abstract

The growth and differentiation of cells in multicellular organisms are regulated by the complex interplay of biochemical and mechanical signals. In the past decades, a plethora of data on the roles of mechanical and structural cues in modulating cellular behaviors has emerged (1-5). It is increasingly evident that cell fates can be changed by engineering the physical properties of the microenvironment to which the cells are exposed (6 -8). These observations have inspired the development of functionalized biomaterials that can directly and specifically interact with tissue components, and support or even direct the appropriate cellular activities (9, 10). Although promising progress has been observed in the past few years, several gaps in knowledge in this field have hindered the development of such "intelligent" biomaterials. In particular, the understanding of the mechanism in which the cell orchestrates physiological and morphological changes by translating mechanical and structural information into biochemical signals is still very limited.As a standard experimental model, cell lines cultured in vitro as a monolayer over solid substrata are usually used to study the effects of biomaterial surfaces on cellular phenotypes. With this simple model system, ingenious experiments have shown that physical forces applied through the extracellular matrix (ECM) 1 1 can induce changes in cell adhesion molecules and stress-induced ion channels, which then lead to changes in the cytoskeleton and gene expressions (11-13). We term the biochemical structure present at the interface between the substratum and the cellular interior the adherence surface (AS), which is composed of the basal plasma membrane with associated structures such as the ECM on one side and the focal adherence complexes on the other. In monolayer cell culture systems, the AS is the only part of the cells in direct contact with the substratum, and is therefore responsible for the first line of communication between the cells and the biomaterial. It is likely that the AS is the organelle that mediates the communication of mechanical and tectonic signals from the substratum to biochemical transducers in the cells. Given the complexity of this process, it is clear that the understanding of this phenomenon cannot be achieved merely by studying individual biological parts in isolation. It is necessary, therefore, to systematically characterize the biochemical factors that mediate the interactions between cells and materials to yield insights into intracellular signaling processes that are responsible for such cellular responses. Toward this goal, we seek to investigate the biochemical basis of how different biomaterials may impose changes in the composition of the AS of adherent cells.

ARAP2 Signals through Arf6 and Rac1 to Control Focal Adhesion Morphology*

“…Preparation of Focal Adhesion Fraction-FA fraction was prepared following methods described previously with some modifications (40,41). Briefly, cells were plated in 12-well plates at 120,000 cells/well.…”

Development of a Human Extracellular Matrix for Applications Related with Stem Cells and Tissue Engineering

“…Proteomic studies confirmed that the majority of proteins which comprise the ECM [ 34 , 35 ] were present in hffECM after the decellularization induced by osmotic shock of HFF, demonstrating that the procedure prevents enzymatic degradation of the ECM. To date, proteomics has not been applied to define the ECM required for the maintenance and differentiation of hESC.…”

Haralick's texture analysis to predict cellular proliferation on randomly oriented electrospun nanomaterials

“…To properly design and develop functional and responsive biomaterials that successfully interact with tissue components, a deeper understanding of the behavior of cells at the biomaterial interface is very important. 21,22 It is widely known that integrin receptors, a family of cell surface adhesion receptors, play a key role in cell–biomaterial interaction and cell mechanotransduction. 22–25 The physical ( e.g.…”