Connie Wang

The proper alignment of cells when engineering extracellular matrix (ECM) scaffolds is imperative in constructing functional tissue engineered grafts. However, the phenomenon that drives co-alignment of cells with ECM fibers, also known as contact guidance, remains poorly defined in terms of its underlying mechanisms. Therefore, this study aims to determine whether fibrinolysis inhibition could affect the cell’s ability to exhibit contact guidance in an aligned 3D fibrin matrix by determining whether it affects the cell’s ability to exhibit motility in an isotropic 3D fibrin matrix. To explore this effect, fibrin gels seeded with Neonatal Human Dermal Fibroblasts (nHDFs) were prepared and treated with aminocaproic acid (ACA), a known fibrinolysis inhibitor. The ACA-treated cells’ morphology was compared with the untreated control group through fluorescent staining and imaging. Specialized, automated software was used to perform morphometric analysis on the imaged cells. The results found no significant statistical difference between the control and treatment group in terms of two-dimensional cell area, number of protrusions per cell, and protrusion orientation. However, the protrusion lengths within the treatment group (Mdn = 13.49) were found to be shorter than the control group (Mdn = 16.03) when performing a Mann-Whitney U test, U = 42600, P < .01: a novel finding. The results thus suggest that nHDFs utilize proteolytic pathways to an extent to exhibit pseudopodia extension. These findings would prove useful in future studies of various topics such as cancer metastasis and tissue graft manufacturing.