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A nerve guidance conduit with topographical and biochemical cues: potential application using human neural stem cells.

Jenkins PM, Laughter MR, Lee DJ, Lee YM, Freed CR, Park D - Nanoscale Res Lett (2015)

Bottom Line: Biochemical cues were integrated into the conduit by functionalizing the polyurea with RGD to improve cell attachment.We determined that electrospinning the polymer solution across a two electrode system with dissolvable sucrose fibers produced a polymer conduit with the appropriate biomimetic properties.Human neural stem cells were cultured on the conduit to evaluate its ability to promote neuronal growth and axonal extension.

View Article: PubMed Central - PubMed

Affiliation: Department of Bioengineering, University of Colorado Denver Anschutz Medical Campus, 12800 E. 19th Avenue, Aurora, CO, 80045, USA.

ABSTRACT
Despite major advances in the pathophysiological understanding of peripheral nerve damage, the treatment of nerve injuries still remains an unmet medical need. Nerve guidance conduits present a promising treatment option by providing a growth-permissive environment that 1) promotes neuronal cell survival and axon growth and 2) directs axonal extension. To this end, we designed an electrospun nerve guidance conduit using a blend of polyurea and poly-caprolactone with both biochemical and topographical cues. Biochemical cues were integrated into the conduit by functionalizing the polyurea with RGD to improve cell attachment. Topographical cues that resemble natural nerve tissue were incorporated by introducing intraluminal microchannels aligned with nanofibers. We determined that electrospinning the polymer solution across a two electrode system with dissolvable sucrose fibers produced a polymer conduit with the appropriate biomimetic properties. Human neural stem cells were cultured on the conduit to evaluate its ability to promote neuronal growth and axonal extension. The nerve guidance conduit was shown to enhance cell survival, migration, and guide neurite extension.

No MeSH data available.


Related in: MedlinePlus

A schematic illustration of two-electrode electrospinning setup. An electric field perpendicular to the two electrodes is formed resulting in parallel nanofiber deposition across the gap. As more layers of aligned nanofibers are deposited across the gap, the electric charge distribution begins to shift, which leads to random nanofiber deposition on top of the layers of aligned nanofibers
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Fig1: A schematic illustration of two-electrode electrospinning setup. An electric field perpendicular to the two electrodes is formed resulting in parallel nanofiber deposition across the gap. As more layers of aligned nanofibers are deposited across the gap, the electric charge distribution begins to shift, which leads to random nanofiber deposition on top of the layers of aligned nanofibers

Mentions: Prior to electrospinning, sucrose fibers with diameters between 200 and 500 μm were formed using a fiber drawing method. Sucrose was heated to 75 °C until melted with a thick consistency. Then, the end of a microscope slide was dipped into the melted sucrose and fibers were drawn. The collector was constructed using two copper wire electrodes spaced 3.5 cm apart, and the sucrose fibers were fit to span the gap between the copper wire electrodes. Eight percent (w/w) polymer solutions in HFP were prepared for PSHU-RGD/PCL (30:70) blend, PSHU/PCL (30:70) blend, and pure PCL. The dual two-electrode electrospinning setup is depicted in Fig. 1. The collector was placed between the two needles distanced 10 cm from each needle, and the polymer solution was ejected at 1 ml/h through a 21-gauge stainless steel flat-tip needle at room temperature and relative humidity at 30 %. A positive 7.5-kV electrostatic potential was applied to both needles for the two blended solutions and a 9-kV potential for the pure PCL solution. For PSHU-RGD/PCL conduits, the PSHU-RGD/PCL blend was electrospun for the initial 15–20 min, and then the PSHU/PCL blend was used to deposit the remainder of the nanofibers. The pure PCL conduits were electrospun using its respective solution during the entire electrospinning process. The flat scaffold sheet was then allowed to air dry for 2 h before being removed from the collector, hand-rolled into a 1.2-mm-diameter tube, and cut into 1.5-cm-long conduits. Both ends of the conduits were loosely tied using nylon fishing line and then soaked in water for 48 h to dissolve out the sucrose fibers.Fig. 1


A nerve guidance conduit with topographical and biochemical cues: potential application using human neural stem cells.

Jenkins PM, Laughter MR, Lee DJ, Lee YM, Freed CR, Park D - Nanoscale Res Lett (2015)

A schematic illustration of two-electrode electrospinning setup. An electric field perpendicular to the two electrodes is formed resulting in parallel nanofiber deposition across the gap. As more layers of aligned nanofibers are deposited across the gap, the electric charge distribution begins to shift, which leads to random nanofiber deposition on top of the layers of aligned nanofibers
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4469602&req=5

Fig1: A schematic illustration of two-electrode electrospinning setup. An electric field perpendicular to the two electrodes is formed resulting in parallel nanofiber deposition across the gap. As more layers of aligned nanofibers are deposited across the gap, the electric charge distribution begins to shift, which leads to random nanofiber deposition on top of the layers of aligned nanofibers
Mentions: Prior to electrospinning, sucrose fibers with diameters between 200 and 500 μm were formed using a fiber drawing method. Sucrose was heated to 75 °C until melted with a thick consistency. Then, the end of a microscope slide was dipped into the melted sucrose and fibers were drawn. The collector was constructed using two copper wire electrodes spaced 3.5 cm apart, and the sucrose fibers were fit to span the gap between the copper wire electrodes. Eight percent (w/w) polymer solutions in HFP were prepared for PSHU-RGD/PCL (30:70) blend, PSHU/PCL (30:70) blend, and pure PCL. The dual two-electrode electrospinning setup is depicted in Fig. 1. The collector was placed between the two needles distanced 10 cm from each needle, and the polymer solution was ejected at 1 ml/h through a 21-gauge stainless steel flat-tip needle at room temperature and relative humidity at 30 %. A positive 7.5-kV electrostatic potential was applied to both needles for the two blended solutions and a 9-kV potential for the pure PCL solution. For PSHU-RGD/PCL conduits, the PSHU-RGD/PCL blend was electrospun for the initial 15–20 min, and then the PSHU/PCL blend was used to deposit the remainder of the nanofibers. The pure PCL conduits were electrospun using its respective solution during the entire electrospinning process. The flat scaffold sheet was then allowed to air dry for 2 h before being removed from the collector, hand-rolled into a 1.2-mm-diameter tube, and cut into 1.5-cm-long conduits. Both ends of the conduits were loosely tied using nylon fishing line and then soaked in water for 48 h to dissolve out the sucrose fibers.Fig. 1

Bottom Line: Biochemical cues were integrated into the conduit by functionalizing the polyurea with RGD to improve cell attachment.We determined that electrospinning the polymer solution across a two electrode system with dissolvable sucrose fibers produced a polymer conduit with the appropriate biomimetic properties.Human neural stem cells were cultured on the conduit to evaluate its ability to promote neuronal growth and axonal extension.

View Article: PubMed Central - PubMed

Affiliation: Department of Bioengineering, University of Colorado Denver Anschutz Medical Campus, 12800 E. 19th Avenue, Aurora, CO, 80045, USA.

ABSTRACT
Despite major advances in the pathophysiological understanding of peripheral nerve damage, the treatment of nerve injuries still remains an unmet medical need. Nerve guidance conduits present a promising treatment option by providing a growth-permissive environment that 1) promotes neuronal cell survival and axon growth and 2) directs axonal extension. To this end, we designed an electrospun nerve guidance conduit using a blend of polyurea and poly-caprolactone with both biochemical and topographical cues. Biochemical cues were integrated into the conduit by functionalizing the polyurea with RGD to improve cell attachment. Topographical cues that resemble natural nerve tissue were incorporated by introducing intraluminal microchannels aligned with nanofibers. We determined that electrospinning the polymer solution across a two electrode system with dissolvable sucrose fibers produced a polymer conduit with the appropriate biomimetic properties. Human neural stem cells were cultured on the conduit to evaluate its ability to promote neuronal growth and axonal extension. The nerve guidance conduit was shown to enhance cell survival, migration, and guide neurite extension.

No MeSH data available.


Related in: MedlinePlus