Effect of collagen gel stiffness on neurite extension

Although collagen is commonly used as components of tissue-engineered nerve-guidance channels, little is known about the effect of the mechanical properties of commonly used gel concentrations on the extension of neurites. This study focused on neurite extension of dissociated chick dorsal root ganglia in vitro over a range of collagen concentrations (0.4–2.0 mg/ml). Neurite length increased in all gels between day 1 and day 4, except at the highest collagen concentration, where a 9% decrease was noted at day 4. Although maximum neurite extension was seen in lower concentration gels (0.6–0.8 mg/ml), mechanical stiffness of each gel significantly increased with increasing concentration, from 2.2 Pa at 0.4 mg/ml to 17.0 Pa at 2.0 mg/ml. A previous model of mechanical stiffness versus neurite outgrowth did not fit this data well, likely because of interactions between the growth cone and the collagen fibers. Overall, these results provided insight regarding factors that influence neurite elongation and may be utilized to further optimize tissue-engineered scaffolds.

[1]  David A Stenger,et al.  Survival and neurite outgrowth of rat cortical neurons in three-dimensional agarose and collagen gel matrices , 2001, Neuroscience Letters.

[2]  E. Glandt,et al.  Partitioning of spherical particles into fibrous matrices: 2. Monte Carlo simulation , 1990 .

[3]  R. Buxbaum,et al.  A cytomechanical investigation of neurite growth on different culture surfaces , 1992, The Journal of cell biology.

[4]  F. Lanni,et al.  Cell traction forces on soft biomaterials. I. Microrheology of type I collagen gels. , 2001, Biophysical journal.

[5]  S. Ichinose,et al.  A study of induction of nerve regeneration using bioabsorbable tubes. , 2001, Journal of reconstructive microsurgery.

[6]  M. Spector,et al.  Connective tissue response to tubular implants for peripheral nerve regeneration: The role of myofibroblasts , 2000, The Journal of comparative neurology.

[7]  V. Lemmon,et al.  Neurite growth on different substrates: permissive versus instructive influences and the role of adhesive strength , 1992, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[8]  D A Lauffenburger,et al.  Maximal migration of human smooth muscle cells on fibronectin and type IV collagen occurs at an intermediate attachment strength , 1993, The Journal of cell biology.

[9]  James W. Fawcett,et al.  Building a Bridge: Engineering Spinal Cord Repair , 2002, Experimental Neurology.

[10]  L. Breckenridge,et al.  Effects of target tissue on growth of snail neurones in collagen gel culture , 1998, Neuroreport.

[11]  C. Heath,et al.  The development of bioartificial nerve grafts for peripheral-nerve regeneration. , 1998, Trends in biotechnology.

[12]  W. Saltzman,et al.  PC12 CELL AGGREGATION AND NEURITE GROWTH IN GELS OF COLLAGEN, LAMININ AND FIBRONECTIN , 1996, International Journal of Developmental Neuroscience.

[13]  V. Martins,et al.  Thermal and Rheological Behavior of Collagen. Chitosan blends , 2002 .

[14]  M. Schachner,et al.  Control of neuronal morphology in vitro: Interplay between adhesive substrate forces and molecular instruction , 1995, Journal of neuroscience research.

[15]  T. Gómez,et al.  Chick sensory neuronal growth cones distinguish fibronectin from laminin by making substratum contacts that resemble focal contacts. , 1996, Journal of neurobiology.

[16]  M. Brocco,et al.  Survival and process regrowth of purified chick retinal ganglion cells cultured in a growth factor lacking medium at low density. Modulation by extracellular matrix proteins. , 1999, Brain research. Developmental brain research.

[17]  E. Sackmann,et al.  Assembly of collagen matrices as a phase transition revealed by structural and rheologic studies. , 2003, Biophysical journal.

[18]  J. Hubbell,et al.  Three-dimensional Migration of Neurites Is Mediated by Adhesion Site Density and Affinity* , 2000, The Journal of Biological Chemistry.

[19]  R T Tranquillo,et al.  The fibroblast-populated collagen microsphere assay of cell traction force--Part 2: Measurement of the cell traction parameter. , 1995, Journal of biomechanical engineering.

[20]  C. Krarup,et al.  Factors that influence peripheral nerve regeneration: An electrophysiological study of the monkey median nerve , 2002, Annals of neurology.

[21]  Richard B. Dickinson,et al.  Quantitative Analysis of Adhesion-Mediated Cell Migration in Three-Dimensional Gels of RGD-Grafted Collagen , 2004, Annals of Biomedical Engineering.

[22]  Tatsuo Nakamura,et al.  Peripheral nerve regeneration across an 80-mm gap bridged by a polyglycolic acid (PGA)–collagen tube filled with laminin-coated collagen fibers: a histological and electrophysiological evaluation of regenerated nerves , 2000, Brain Research.

[23]  M. Tadié,et al.  Axonal regrowth through collagen tubes bridging the spinal cord to nerve roots , 1997, Journal of neuroscience research.

[24]  W. Saltzman,et al.  Quantification of human neutrophil motility in three-dimensional collagen gels. Effect of collagen concentration. , 1992, Biophysical journal.

[25]  Tatsuo Nakamura,et al.  Regeneration of canine peroneal nerve with the use of a polyglycolic acid-collagen tube filled with laminin-soaked collagen sponge: a comparative study of collagen sponge and collagen fibers as filling materials for nerve conduits. , 2001, Journal of biomedical materials research.

[26]  W. Saltzman,et al.  Neutrophil motility in extracellular matrix gels: mesh size and adhesion affect speed of migration. , 1997, Biophysical journal.

[27]  R T Tranquillo,et al.  Neuronal contact guidance in magnetically aligned fibrin gels: effect of variation in gel mechano-structural properties. , 2001, Biomaterials.

[28]  W. Friess,et al.  Effects of processing conditions on the rheological behavior of collagen dispersions. , 2001, European journal of pharmaceutics and biopharmaceutics : official journal of Arbeitsgemeinschaft fur Pharmazeutische Verfahrenstechnik e.V.

[29]  Cheryl Miller,et al.  Synergistic effects of physical and chemical guidance cues on neurite alignment and outgrowth on biodegradable polymer substrates. , 2002, Tissue engineering.

[30]  R. Buxbaum,et al.  Tensile regulation of axonal elongation and initiation , 1991, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[31]  S. Mackinnon,et al.  The Role of Conduits in Nerve Repair: A Review , 1996, Reviews in the neurosciences.

[32]  O. Guntinas-Lichius,et al.  Local application of extracellular matrix proteins fails to reduce the number of axonal branches after varying reconstructive surgery on rat facial nerve. , 2000, Restorative neurology and neuroscience.

[33]  W. Gispen,et al.  Collagen Containing Neurotrophin-3 (NT-3) Attracts Regrowing Injured Corticospinal Axons in the Adult Rat Spinal Cord and Promotes Partial Functional Recovery , 1998, Experimental Neurology.

[34]  Xavier Navarro,et al.  Magnetically Aligned Collagen Gel Filling a Collagen Nerve Guide Improves Peripheral Nerve Regeneration , 1999, Experimental Neurology.

[35]  Guo-Hua Li,et al.  Computer model of growth cone behavior and neuronal morphogenesis , 1995 .

[36]  R V Bellamkonda,et al.  Agarose gel stiffness determines rate of DRG neurite extension in 3D cultures. , 2001, Biomaterials.