TY - JOUR

T1 - A rapid, quantitative method for assessing axonal extension on biomaterial platforms

AU - Cregg, Jared M

AU - Wiseman, Sherri L

AU - Pietrzak-Goetze, Nicole M

AU - Smith, Martyn R

AU - Jaroch, David B

AU - Clupper, Daniel C

AU - Gilbert, Ryan J

PY - 2010/4

Y1 - 2010/4

N2 - Measuring outgrowth of neuronal explants is critical in evaluating the ability of a biomaterial to act as a permissive substrate for neuronal adhesion and growth. Previous methods lack the ability to quantify robust outgrowth, or lack the capacity to quantify growth on opaque substrates because they exploit the transparent nature of culture dishes to segregate neuronal processes from an image background based on color intensity. In this study, we sought to investigate the ability of opaque silica sol-gel materials to facilitate axonal outgrowth; therefore, a method was developed for quantifying outgrowth of neurites from dorsal root ganglion explants on these unique surfaces. Dorsal root ganglia were isolated from stage-nine chick embryos and cultured for 48 h on sol-gel materials presenting agarose and chitosan polysaccharides individually or in combination. Explants were then imaged, and basic image analysis software was used by three independent observers to obtain axonal length and axonal area measurements. Robust axon length and axonal spread measurements for ganglia cultured on agarose-chitosan sol-gel matrices yield an estimate of strong neural compatibility for these substrates over silica matrices presenting no polysaccharides, or silica matrices presenting chitosan or agarose individually. We suggest that this simple protocol for quantifying material biocompatibility offers an analysis strategy that can be used universally to the same end.

AB - Measuring outgrowth of neuronal explants is critical in evaluating the ability of a biomaterial to act as a permissive substrate for neuronal adhesion and growth. Previous methods lack the ability to quantify robust outgrowth, or lack the capacity to quantify growth on opaque substrates because they exploit the transparent nature of culture dishes to segregate neuronal processes from an image background based on color intensity. In this study, we sought to investigate the ability of opaque silica sol-gel materials to facilitate axonal outgrowth; therefore, a method was developed for quantifying outgrowth of neurites from dorsal root ganglion explants on these unique surfaces. Dorsal root ganglia were isolated from stage-nine chick embryos and cultured for 48 h on sol-gel materials presenting agarose and chitosan polysaccharides individually or in combination. Explants were then imaged, and basic image analysis software was used by three independent observers to obtain axonal length and axonal area measurements. Robust axon length and axonal spread measurements for ganglia cultured on agarose-chitosan sol-gel matrices yield an estimate of strong neural compatibility for these substrates over silica matrices presenting no polysaccharides, or silica matrices presenting chitosan or agarose individually. We suggest that this simple protocol for quantifying material biocompatibility offers an analysis strategy that can be used universally to the same end.

KW - Animals

KW - Axons/physiology

KW - Biocompatible Materials/chemistry

KW - Cell Adhesion/drug effects

KW - Cell Count

KW - Cell Proliferation/drug effects

KW - Cells, Cultured

KW - Chick Embryo

KW - Chitosan/chemistry

KW - Ganglia, Spinal/drug effects

KW - Materials Testing/methods

KW - Nerve Regeneration/physiology

KW - Neurogenesis/drug effects

KW - Neurons/cytology

KW - Sepharose/chemistry

KW - Time Factors

KW - Tissue Engineering/methods

KW - Tissue Scaffolds/chemistry

U2 - 10.1089/ten.TEC.2009.0108

DO - 10.1089/ten.TEC.2009.0108

M3 - Journal article

C2 - 19409034

VL - 16

SP - 167

EP - 172

JO - Tissue Engineering - Part C: Methods

JF - Tissue Engineering - Part C: Methods

SN - 1937-3384

IS - 2

ER -