Cell‐Cell and ‐Matrix Interactions on Nanofibrous Scaffolds

In tissue engineering, it is pivotal to find the unique biological niche where cells can proliferate and differentiate in a defined configuration to regenerate functional tissues. The objective of our study was to visualize the cell matrix interactions of human gingival fibroblasts (HGFs) on the nanofibrous (NF) gelatin scaffolds. The scaffolds were prepared mimicking both the physical architecture and chemical composition of natural collagen (Liu, et. al Biomaterials 30: 4094, 2009). The HGFs were cultured on NF‐gelatin scaffolds till they were confluent. They were fixed and stained with dyes to detect the actin cytoskeleton and nucleus. Focal adhesions were identified using antibodies specific for vinculin, integrin â1 and phospho‐paxillin. An SP5 confocal microscope was used to generate 3D images of the cells on the scaffolds. The HGFs were attached and well spread throughout the scaffold with processes interacting with gelatin nanofibres. There were also focal adhesion structures as detected by the presence of vinculin, integrin and phospho‐paxillin. This research will lead to insights into how HGF cells alter their cell shape, actin cytoskeleton, and focal adhesions when cultured on these biomimetic nano‐structured scaffolds. Future experiments will determine how the cells migrate over the scaffold. The intent is to determine the cell‐scaffold interactions for tissue regeneration. Grant Funding Source : NIH/NIDCR P30 DE020742