Journal of Materials Science: Materials in Medicine

, Volume 21, Issue 9, pp 2637–2651 | Cite as

Functionalization of electrospun fibers of poly(ε-caprolactone) with star shaped NCO-poly(ethylene glycol)-stat-poly(propylene glycol) for neuronal cell guidance

  • Kristina Klinkhammer
  • Julia Bockelmann
  • Chariklia Simitzis
  • Gary A. Brook
  • Dirk Grafahrend
  • Jürgen Groll
  • Martin Möller
  • Jörg Mey
  • Doris Klee
Article

Abstract

Microfibers produced with electrospinning have recently been used in tissue engineering. In the development of artificial implants for nerve regeneration they are of particular interest as guidance structures for cell migration and axonal growth. Using electrospinning we produced parallel-orientated biocompatible fibers in the submicron range consisting of poly(ε-caprolactone) (PCL) and star shaped NCO-poly(ethylene glycol)-stat-poly(propylene glycol) (sPEG). Addition of the bioactive peptide sequence glycine-arginine-glycine-aspartate-serine (GRGDS) or the extracellular matrix protein fibronectin to the electrospinning solution resulted in functionalized fibers. Surface characteristics and biological properties of functionalized and non-functionalised fibers were investigated. Polymer solutions and electrospinning process parameters were varied to obtain high quality orientated fibers. A polymer mixture containing high molecular weight PCL, PCL-diol, and sPEG permitted a chemical reaction between hydroxyl groups of the diol and isocyanante groups of the sPEG. Surface analysis demonstrated that sPEG at the fiber surface minimized protein adhesion. In vitro experiments using dorsal root ganglia explants showed that the cell repellent property of pure PCL/sPEG fibers was overcome by functionalization either with GRGDS peptide or fibronectin. In this way cell migration and axonal outgrowth along fibers were significantly increased. Thus, functionalized electrospun PCL/sPEG fibers, while preventing non-specific protein adsorption, are a suitable substrate for biological and medical applications.

Abbreviations

BSA

Bovine serum albumin

DAPI

4,6-Diamidino-2-phenylindole dihydrochloride

DCM

Dichloromethane

DIV

Days in vitro

DMEM

Dulbecco’s modified eagle medium

DMSO

Dimethyl sulfoxide

DRG

Dorsal root ganglia

ECM

Extracellular matrix

FCS

Fetal calf serum

GRGDS

One letter code of the peptide sequence GlyArgGlyAspSer

Mw

Molecular weight

NF200

Neurofilament 200 kDa

NGS

Normal goat serum

NMP

N-Methylpyrrilidone

PBS

Phosphate buffer saline

PCL

Poly(ε-caprolactone)

PCL-ol

PCL diol

PEG

Poly(ethylene glycol)

PNS

Peripheral nervous system

S100

Protein antibody, marker for Schwann cells

SC

Schwann cells

SEM

Scanning electron microscopy

sPEG

Star shaped NCO-poly(ethylene glycol)-stat-poly(propylene glycol)

THF

Tetrahydrofuran

TBS-T

Tris-buffered saline with 1% Triton-X 100

wt%

Weight percent

XPS

X-ray photoelectron spectroscopy

Notes

Acknowledgments

We thank Marie Pradella for help with evaluating the cell experiments. Kristin Michael and Bernd Hoffmann kindly allowed us to use their equipment for making composite photographs of DRG explants. This work was supported by a grant from the Interdisciplinary Centre for Clinical Research “BIOMAT” within the faculty of Medicine at Aachen University (RWTH) (TV B111) and by DFG-Graduiertenkolleg 1035 “Biointerface”. Julia Bockelmann was supported by a Marie-Curie EST grant from the EU (EURON).

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Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  • Kristina Klinkhammer
    • 1
  • Julia Bockelmann
    • 2
    • 3
  • Chariklia Simitzis
    • 1
  • Gary A. Brook
    • 4
    • 5
  • Dirk Grafahrend
    • 1
  • Jürgen Groll
    • 1
  • Martin Möller
    • 1
  • Jörg Mey
    • 2
    • 3
  • Doris Klee
    • 1
  1. 1.Institute of Technical and Macromolecular Chemistry and DWI e.V.RWTH Aachen UniversityAachenGermany
  2. 2.Institute of Biology IIRWTH Aachen UniversityAachenGermany
  3. 3.EURON Graduate School of NeuroscienceMaastrichtNetherlands
  4. 4.Institute of Neuropathology, Medical FacultyRWTH Aachen UniversityAachenGermany
  5. 5.IZKF-BIOMAT, Aachen University Hospital, RWTH Aachen UniversityAachenGermany

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