Nylon as an in vitro scaffold for three‐dimensional study of neural cells

Abstract

The requirement for improved modeling of cells in culture for study of cell-to-cell interactions has led to an increased focus on three-dimensional (3D) in vitro models. In this study, NG108-15 neural cells and rat sciatic nerve Schwann cells were successfully grown as monocultures and as a coculture on the biocompatible nylon mesh substrates. This has allowed proliferation and interactions between the two cell types to be monitored on the mesh over time. Electrophysiological recordings validated the NG108-15 neural cell differentiation which confirmed the similarity of recorded action potentials in two-dimensional culture environments. A variety of assays have been used, demonstrating the use of nylon as an appropriate substrate for neural cell in vitro studies, and nylon can easily be imaged using inverted microscopy. These studies confirm the suitability of the nylon 3D culture presented for modeling neural cell behavior. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 1575-1584, 2018.

In Vitro, In Vivo and Ex Vivo Models for Peripheral Nerve Injury and Regeneration

“…Blasiak et al developed a 3D-printed surface to model peripheral nerve growth using Veroclear, a material similar to acrylic, and designed channels into the surface ranging from 2500 µm to 500 µm wide, and 500 µm deep [ 139 ]. Batth et al have described a 3-dimensional substrate constructed from nylon mesh to coculture a neuronal cell line and primary rat sciatic nerve Schwann cells and investigate their cellular interactions including recording action potentials [ 140 ].…”

“…There have also been models that have used a combination of the aforementioned materials, such as a PDMS based platform and collagen membrane overlay connected to a peripheral nerve graft, described by Siddique et al [ 141 ]. Their design allowed for access to perform microsurgery and image spinal cord sections as they grew ventral root axons on the collagen surface [ 140 ]. The growth of axons from the collagen membrane into the peripheral nerve graft simulated a repair of a motor nerve using a peripheral nerve graft [ 142 ].…”

“…While a 2D ex vivo model involves directed axonal growth along a biocompatible surface such as a microchannel made from PDMS, a 3D ex vivo model involves directed axonal growth usually within a biocompatible gel, such as collagen. The 3-dimensional orientation of extracellular matrix components and architectural orientation of peripheral nerve is very difficult to model in vitro [ 140 ]. Three-dimensional organization is important because release of cellular growth factors and appropriate interaction of these factors with neighboring cellular elements following nerve injury occurs within a three-dimensional structure in vivo .…”

“…An interesting approach to modeling a mixed motor-sensory nerve in vitro was demonstrated by Brushart et al [ 152 ]. They used spinal cord slices taken from mice expressing Yellow Fluorescent Protein (YFP) to create an in vitro peripheral nerve regeneration model with motor axons that were able to be visualized clearly as they grew from a the ventral root [ 140 ]. The group has expanded this model by coculturing these YFP expressing motor neurons with Red Fluorescent Protein (RFP) expressing sensory neurons taken from mice DRG [ 151 ].…”