Abstract
To examine the influence of substrate topology on the behavior of fibroblasts, tissue engineering scaffolds were electrospun from polycaprolactone (PCL) and a blend of PCL and gelatin (PCL+Gel) to produce matrices with both random and aligned nanofibrous orientations. The addition of gelatin to the scaffold was shown to increase the hydrophilicity of the PCL matrix and to increase the proliferation of NIH3T3 cells compared to scaffolds of PCL alone. The orientation of nanofibers within the matrix did not have an effect on the proliferation of adherent cells, but cells on aligned substrates were shown to elongate and align parallel to the direction of substrate fiber alignment. A microarray of cyotoskeleton regulators was probed to examine differences in gene expression between cells grown on an aligned and randomly oriented substrates. It was found that transcriptional expression of eight genes was statistically different between the two conditions, with all of them being upregulated in the aligned condition. The proteins encoded by these genes are linked to production and polymerization of actin microfilaments, as well as focal adhesion assembly. Taken together, the data indicates NIH3T3 fibroblasts on aligned substrates align themselves parallel with their substrate and increase production of actin and focal adhesion related genes.
Highlights
Using electrospun materials as scaffolds for engineering tissue replacements remains a promising research area
For the PCL and gelatin (PCL+Gel) sample, a smaller peak is present indicating the presence of gelatin within the sample, the height of the smaller peak is 11% of the 100% gelatin peak, which confirms the ratio of PCL to gelatin in the PCL+Gel sample
We examined the influence of substrate nanofiber orientation on the expression of cytoskeleton regulators in fibroblasts
Summary
Using electrospun materials as scaffolds for engineering tissue replacements remains a promising research area. Electrospun scaffolds can be fabricated from numerous biodegradable materials and their nanofibrous structure can possess features which mimic the architecture of the native extracellular matrix (ECM) of many tissues. The electrospinning apparatus only requires a few components and is highly customizable to produce tailored nanofibrous matrices. Electrospun scaffolds composed of highly aligned fibers are of particular interest due to their ability to modulate many cellular behaviors. Cells cultured on substrates with an oriented microtopology have been shown to behave differently than cells on randomly oriented or smooth.
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