Enhanced axonal regeneration following combined demyelination plus schwann cell transplantation therapy in the injured adult spinal cord.
We have treated spinal cord injured rats with demyelination plus Schwann cell transplantation and assessed neurite outgrowth in a quantifiable model of axonal regeneration. Axonal injuries of differing severity were induced in the dorsal funiculus of adult rats using a micromanipulator-controlled Scouten knife. Demyelinated regions were produced so as to overlap with the injury site by the injection of galactocerebroside antibodies plus complement one segment cranial to the axonal injury site. Schwann cells were isolated from the sciatic nerve, expanded in vitro, and transplanted into the injury site 1 day later. Animals were killed after an additional 7 days. Schwann cells were evenly distributed throughout the region of demyelination, which extended 6-7 mm cranial to the axonal injury site. The severity of axonal injury was quantified by counting degenerate axons in transverse resin sections. The degree of axonal regeneration was assessed by an electron microscopic analysis of growth cone frequency and distribution relative to the site of axonal injury. Quantification of growth cones at a distance from the site of axonal injury indicated a strong linear relationship (P < 0.001) between the number of growth cones and the number of severed axons; the ratio of growth cones to severed axons was increased by 26.5% in demyelinated plus transplanted animals compared to demyelinated animals without a transplant. Furthermore, only the demyelinated plus transplanted animals contained growth cones associated with myelin in white matter immediately outside of the region of complete demyelination. Growth cones were absent in transplanted-only animals at a distance from the site of axonal injury. These findings indicate that combined demyelination plus Schwann cell transplantation therapy enhances axonal regeneration following injury and suggests that growth cones are able to overcome myelin-associated inhibitors of neurite outgrowth in the presence of trophic support.
Keirstead
H S
HS
MRC Cambridge Centre for Brain Repair, University of Cambridge, Robinson Way, Cambridge, CB2 2PY, United Kingdom. keirstead@cord.ubc.ca
Morgan
S V
SV
Wilby
M J
MJ
Fawcett
J W
JW
D000367:Age Factors; D000818:Animals; D000906:Antibodies / Q000494:pharmacology; D001369:Axons / Q000502:physiology* / Q000648:ultrastructure; D019771:Axotomy; D003165:Complement System Proteins; D003711:Demyelinating Diseases / Q000139:chemically induced / Q000628:therapy*; D005260:Female; D005699:Galactosylceramides / Q000276:immunology; D020439:Growth Cones / Q000502:physiology / Q000648:ultrastructure; D008854:Microscopy, Electron; D009186:Myelin Sheath / Q000502:physiology; D009416:Nerve Regeneration; D009836:Oligodendroglia / Q000166:cytology / Q000502:physiology; D051381:Rats; D017207:Rats, Sprague-Dawley; D012583:Schwann Cells / Q000637:transplantation* / Q000648:ultrastructure; D013116:Spinal Cord / Q000166:cytology / Q000502:physiology; D013119:Spinal Cord Injuries / Q000628:therapy*
D000906:Antibodies; D005699:Galactosylceramides; C002454:galactocerebroside; D003165:Complement System Proteins