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Journal of Molecular Neuroscience

, Volume 49, Issue 2, pp 334–346 | Cite as

Angioneural Crosstalk in Scaffolds with Oriented Microchannels for Regenerative Spinal Cord Injury Repair

  • Aybike Saglam
  • Anat Perets
  • Adam Charles Canver
  • Ho-Lung Li
  • Katherine Kollins
  • Gadi Cohen
  • Itzhak Fischer
  • Philip LazaroviciEmail author
  • Peter I. LelkesEmail author
Article

Abstract

The aim of our work is to utilize the crosstalk between the vascular and the neuronal system to enhance directed neuritogenesis in uniaxial guidance scaffolds for the repair of spinal cord injury. In this study, we describe a method for angioneural regenerative engineering, i.e., for generating biodegradable scaffolds, produced by a combination of controlled freezing (freeze-casting) and lyophilization, which contain longitudinally oriented channels, and provide uniaxial directionality to support and guide neuritogenesis from neuronal cells in the presence of endothelial cells. The optimized scaffolds, composed of 2.5 % gelatin and 1 % genipin crosslinked, were characterized by an elastic modulus of ~51 kPa and longitudinal channels of ~50 μm diameter. The scaffolds support the growth of endothelial cells, undifferentiated or NGF-differentiated PC12 cells, and primary cultures of fetal chick forebrain neurons. The angioneural crosstalk, as generated by first forming endothelial cell monolayers in the scaffolds followed by injection of neuronal cells, leads to the outgrowth of long aligned neurites in the PC12/endothelial cell co-cultures also in the absence of exogenously added nerve growth factor. Neuritogenesis was not observed in the scaffolds in the absence of the endothelial cells. This methodology is a promising approach for neural tissue engineering and may be applicable for regenerative spinal cord injury repair.

Keywords

Genipin Gelatin Controlled freeze-drying Angioneural tissue engineering NGF Longitudinal channels Endothelial cells Neuronal cells Differentiation Neuritogenesis 

Abbreviations

2-D

Two dimensional

3-D

Three dimensional

CFS

Controlled freeze-dried scaffold

DMEM

Dubelco's modified eagle medium

EA.hy926

Hybrid endothelial cell line

ECs

Endothelial cells

ECM

Extracellular matrix

EPS

Endothelial progenitor cells

FBN

Chicken forebrain primary neurons

FBS

Fetal bovine serum

GFP-PC12

Green fluorescence protein transduced PC12 cells

Medium A

DMEM supplemented with 7.5 % fetal bovine serum FBS 7.5 % horse serum

Medium B

Dulbecco's modified Eagle's medium supplemented with 10 % FBS

NGF

Nerve growth factor

PC12

Pheochromocytoma cells

RFS

Regular freeze-dried scaffold

SCI

Spinal cord injury

SEM

Scanning electron microscope

Notes

Acknowledgments

This study was supported by grants (to PIL) from the Craig H. Neilson Foundation, the Louis and Bessie Stein Foundation (to PIL and PL), and Israel Ministry of Science and Technology (PL). PL holds the Jacob Gitlin Chair in Physiology and is affiliated with and supported by the David R. Bloom Center for Pharmacy and the Dr. Adolf and Klara Brettler Center for Research in Molecular Pharmacology and Therapeutics at the Hebrew University of Jerusalem, Israel. We would like to thank Dr. Cora Edgell for her generous gift of the EA.hy926 cells, Dr. Ulrike G.K. Wegst and her team at the Department of Materials Sciences, Drexel University for introducing us to the controlled freezing technique, Dr. J. Yasha Kresh (Department of Cariothoracic Surgery, Drexel University College of Medicine) for the use of the rheometer, and Dr. Gianluca Gallo (Department of Anatomy and Neurobiology, Drexel University College of Medicine) for teaching us how to isolate embryonic chick forebrain neurons. We are grateful to Dr. Qingwei Zhang and Ms. Gozde Senel, Drexel University, School of Biomedical Engineering, Science and Health Systems for their assistance with the SEM. We also would like to thank Ms. Pimchanok Pimton, Ms. Tantri Syawalwulanti, and Ms. Zehava Cohen for their help with the figures.

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Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Aybike Saglam
    • 1
  • Anat Perets
    • 1
  • Adam Charles Canver
    • 1
  • Ho-Lung Li
    • 2
  • Katherine Kollins
    • 3
  • Gadi Cohen
    • 4
  • Itzhak Fischer
    • 3
  • Philip Lazarovici
    • 1
    • 4
    Email author
  • Peter I. Lelkes
    • 1
    • 5
    Email author
  1. 1.School of Biomedical Engineering, Science and Health SystemsDrexel UniversityPhiladelphiaUSA
  2. 2.Department of Mechanical Engineering and MechanicsDrexel UniversityPhiladelphiaUSA
  3. 3.Department of Neurobiology and AnatomyDrexel University College of MedicinePhiladelphiaUSA
  4. 4.School of Pharmacy Institute for Drug Research, Faculty of MedicineThe Hebrew University of JerusalemJerusalemIsrael
  5. 5.Department of Bioengineering, College of EngineeringTemple UniversityPhiladelphiaUSA

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