Effects of fibrin micromorphology on neurite growth from dorsal root ganglia cultured in three-dimensional fibrin gels.
The effect of fibrin matrix micromorphology on neurite growth was investigated by measuring the length of neurites growing in three-dimensional fibrin gels with well characterized micromorphologies. Dorsal root ganglia (DRGs) from 7-day chick embryos were entrapped and cultured in gels made from varying concentrations of fibrinogen (5-15 mg/mL) or calcium (2-10 mM). The length of growing neurites was measured with light videomicroscopy, and the number and diameter of fibrin fiber bundles were measured from scanning electron micrographs. An increase in fibrinogen concentration caused a decrease in the average fiber bundle thickness, an increase in the number of fiber bundles, and a marked decrease in neurite length. Gels made with different calcium concentrations had a similar range of variation in fibrin fiber bundle number or diameter, but these variations had little effect on neurite and associated nonneuronal cell outgrowth. These results provide insights into the process of neurite advance within fibrin and may be useful in the design of fibrin-based materials used for peripheral nerve regeneration. Furthermore, this study provides the first detailed experimental data on the micromorphology of fibrin matrices made from more than 5 mg/mL of fibrinogen and indicates that existing kinetic models of fibrin polymerization do not accurately predict fibrin structure at these higher concentrations.
Herbert
C B
CB
Department of Chemical Engineering, University of Texas, Austin 78712, USA.
Nagaswami
C
C
Bittner
G D
GD
Hubbell
J A
JA
Weisel
J W
JW
D000818:Animals; D018929:Cell Culture Techniques; D002455:Cell Division; D002478:Cells, Cultured; D002642:Chick Embryo; D005109:Extracellular Matrix / Q000502:physiology / Q000648:ultrastructure; D005337:Fibrin / Q000502:physiology* / Q000648:ultrastructure*; D005727:Ganglia, Spinal / Q000187:drug effects / Q000502:physiology*; D005782:Gels; D006801:Humans; D008855:Microscopy, Electron, Scanning; D016501:Neurites / Q000502:physiology
D005782:Gels; D005337:Fibrin