Interaction of leech neurons with topographical gratings: comparison with rodent and human neuronal lines and primary cells

Abstract

Controlling and improving neuronal cell migration and neurite outgrowth are critical elements of tissue engineering applications and development of artificial neuronal interfaces. To this end, a promising approach exploits nano/microstructured surfaces, which have been demonstrated to be capable of tuning neuronal differentiation, polarity, migration and neurite orientation. Here, we investigate the neurite contact guidance of leech neurons on plastic gratings (GRs; anisotropic topographies composed of alternating lines of grooves and ridges). By high-resolution microscopy, we quantitatively evaluate the changes in tubulin cytoskeleton organization and cell morphology and in the neurite and growth cone development. The topography-reading process of leech neurons on GRs is mediated by filopodia and is more responsive to 4-µm-period GRs than to smaller period GRs. Leech neuron behaviour on GRs is finally compared and validated with several other neuronal cells, from murine differentiated embryonic stem cells and primary hippocampal neurons to differentiated human neuroblastoma cells.

Nanoimprinted Anisotropic Topography Preferentially Guides Axons and Enhances Nerve Regeneration

“…To generate the anisotropic topography resembling a linear grooved pattern, we used nanoimprinting lithography with metallic stampers of three different spacings (Blu‐ray Disc: BD, Digital Video Disc: DVD, and Compact Disc: CD) on EVA copolymer. Similar approach has been utilized by various groups to generate surface micropattern to study neuron‐topography interaction . We chose to work with EVA copolymer because of its biocompatible nature, optical clarity, and tunable stiffness, with Young's moduli between 3 and 42 MPa ( Table 1 ).…”

“…Similar approach has been utilized by various groups to generate surface micropattern to study neuron-topography interaction. [32,33] We chose to work with EVA copolymer because of its biocompatible nature, optical clarity, and tunable stiffness, with Young's moduli between 3 and 42 MPa ( Table 1). BD, DVD, and CD stampers were selected for two reasons.…”

“…Based on the physical dimension of the cell body, neurite, and growth cone, numerous studies have been conducted to examine the interaction of neurons and surface micropatterns with comparable dimensions. These surface micropatterns include beads, electrospun fibers, nanotubes, pillars, ridges/grooves/gratings, or nanorough surfaces . However, most of these in vitro studies focus on the interaction of surface topography with uninjured neurons.…”

“…Among all patients admitted to level I trauma centers, PNS injury is estimated to occur in ≈5% of patients, [1] this results conducted to examine the interaction of neurons and surface micropatterns with comparable dimensions. These surface micropatterns include beads, [13,14] electrospun fibers, [15][16][17][18] nanotubes, [19,20] pillars, [21][22][23][24][25] ridges/grooves/gratings, [26][27][28][29][30][31][32][33][34][35] or nanorough surfaces. [36][37][38] However, most of these in vitro studies focus on the interaction of surface topography with uninjured neurons.…”

Human Neuronal SHSY5Y Cells on PVDF:PTrFE Copolymer Thin Films

“…Biocompatibility is one of the most important characteristics of biomedical polymer materials whose surface is required to interact with a biological system. It is recognized that both chemical and physical surface features, from several microns down to nanometer length scales, are crucial for influencing cell responses such as their adhesion, growth, and functionality . The biological performance of a material is dependent on its chemical characteristics, morphological features (expressed through roughness, fractal dimension, anisotropy) and mechanical compliance.…”