Evaluating Mouse Fibroblast Interaction with Implant Surfaces in a 3D Microenvironment

Abstract

Purpose: Previous studies assessing fibroblast interactions with implants have mainly relied on measurements such as cell migration, gene expression, and cell adhesion. For these studies, testing cellular behavior at the implant surface was done by imaging the cell-implant interface using standard microscopy techniques in 2D tissue culture dishes. The true behavior of cells relative to the implant can best be assessed in a more physiologic 3D microenvironment. Materials and Methods: The embedding of the implant disks in 3D collagen gels was standardized with labeled fibroblasts to allow the imaging of fibroblast morphology and behavior when proximal to or binding to the implant disks. This allowed comparison of the behavior of laser-microgrooved and machined implant disk surfaces quantitatively in an in vitro 3D microenvironment. Results: This in vitro imaging assay revealed for the first time in a 3D microenvironment setting the statistically significant impact laser-microgrooved disk surfaces have on both cell adherence and recruitment of cells in proximity to the disk. It also allowed visualization of membrane protrusivity and cytoskeletal organization in cells adherent to the implant disk. Conclusion: This assay provides a simple and effective way of observing cell behavior on and around the implant disk surface in a more physiologic 3D setting. Within the limits of this study, it revealed that the laser-microgrooved implant surface demonstrates significant superiority in fibroblast recruitment and binding in a 3D microenvironment.