Abstract
Variations in substrate chemistry and the micro-structure were shown to have a significant effect on the biology of human mesenchymal stromal cells (hMSCs). This occurs when differences in the surface properties indirectly modulate pathways within numerous signaling networks that control cell fate. To understand how the surface features affect hMSC gene expression, we performed RNA-sequencing analysis of bone marrow-derived hMSCs cultured on tissue culture-treated polystyrene (TCP) and poly(l-lactide) (PLLA) based substrates of differing topography (Fl: flat and Fs: fibrous) and chemistry (Pr: pristine and Am: aminated). Whilst 80% of gene expression remained similar for cells cultured on test substrates, the analysis of differentially expressed genes (DEGs) revealed that surface topography significantly altered gene expression more than surface chemistry. The Fl and Fs topologies introduced opposite directional alternations in gene expression when compared to TCP control. In addition, the effect of chemical treatment interacted with that of topography in a synergistic manner with the Pr samples promoting more DEGs than Am samples in all gene ontology function groups. These findings not only highlight the significance of the culture surface on regulating the overall gene expression profile but also provide novel insights into cell-material interactions that could help further design the next-generation biomaterials to facilitate hMSC applications. At the same time, further studies are required to investigate whether or not the observations noted correlate with subsequent protein expression and functionality of cells.
Highlights
Human mesenchymal stromal cells are multi-potent, plastic-adherent, and fibroblast-like cells found in the human body
The scope of this study focuses on the impact of culture surfaces on the transcriptome profile of hbm- MSCs
While the effects of substrate topography and chemistry on Human mesenchymal stromal cells (hMSCs) behavior are reported in the literature, we present a global perspective of the cell-material interactions
Summary
Human mesenchymal stromal cells (hMSCs) are multi-potent, plastic-adherent, and fibroblast-like cells found in the human body. The fundamental questions that need to be addressed are (i) whether surface topography and chemistry mediate gradual changes or directional alternations and (ii) what kind of interactive effects they would impose on each other (Figure 1) To answer these questions, in the current study, we performed RNA-seq analysis on human bone marrow-derived MSCs (hbm-MSCs) cultured on flat (Fl) and fibrous (Fs) poly(L-lactide) (PLLA) substrates. 277 gene expressions were either detected uniquely for a particular sample or shown to be differentially up-regulated or down-regulated by different culture conditions Such a differential response of MSCs, in terms of proliferative and differentiation potential, towards surface topography or surface chemistry have been reported in the literature [25,49,50,51]. FigurFeig7u.reG7e. nGeenoentoonltoogloygyanaanlaylsyissisfoforrDDEEGGss:: DDEEGGssaasssoscoicaitaedtedwiwthitbhiobloioglicoaglicparlocpersosecse,scseelslu, lcaerllular compcoomnepnotnse, natns,damndomleocluelcaurlafrufnucntciotinonwweerreeggrroouuppeedd aannddccoouunntetde.d
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