J Korean Neurosurg Soc.  2014 Nov;56(5):375-382. 10.3340/jkns.2014.56.5.375.

Intra-Spinal Bone Marrow Mononuclear Cells Transplantation Inhibits the Expression of Nuclear Factor-kappaB in Acute Transection Spinal Cord Injury in Rats

• Shrestha R
• Qiao JM
• Shen FG
• Bista KB
• Zhao ZN
• Yang J

• KMID: 2018094
• DOI: http://doi.org/10.3340/jkns.2014.56.5.375

Abstract

OBJECTIVE
To assess the effect of bone marrow mononuclear cells (BMMNCs) transplantation in the expression of nuclear factor-kappaB (NF-kappaB) in spinal cord injury (SCI) in rats.
METHODS
BMMNCs were isolated from tibia and femur by a density gradient centrifugation. After establishment of acute transection SCI, rats were divided into experiment (BMMNCs), experiment control (0.1 M PBS infused) and sham surgery groups (laminectomy without any SCI). Locomotor function was assessed weekly for 5 weeks post-injury using BBB locomotor score and urinary bladder function daily for 4 weeks post-injury. Activity of NF-kappaB in spinal cord was assessed by immunohistochemistry and reverse transcriptase polymerase chain reaction.
RESULTS
At each time point post-injury, sham surgery group had significantly higher Basso, Beattie, Bresnahan locomotor and urinary bladder function scores than experiment and experiment control group (p<0.05). At subsequent time interval there were gradual improvement in both experiment and experiment control group, but experiment group had higher score in comparison to experiment control group (p<0.05). Comparisons were also made for expression of activated NF-kappaB positive cells and level of NF-kappaB messenger RNA in spinal cord at various time points between the groups. Activated NF-kappaB immunoreactivity and level of NF-kappaB mRNA expression were significantly higher in control group in comparison to experiment and sham surgery group (p<0.05).
CONCLUSION
BMMNCs transplantation attenuates the expression of NF-kappaB in injured spinal cord tissue and thus helps in recovery of neurological function in rat models with SCI.

Keyword

Cell transplantation; Inflammation; Mononuclear cells; Neuroprotective agent; NF-kappaB; Spinal cord injury

MeSH Terms

Animals
Bone Marrow*
Cell Transplantation
Centrifugation, Density Gradient
Femur
Immunohistochemistry
Inflammation
Models, Animal
NF-kappa B
Rats*
Reverse Transcriptase Polymerase Chain Reaction
RNA, Messenger
Spinal Cord
Spinal Cord Injuries*
Tibia
Urinary Bladder
NF-kappa B
RNA, Messenger

Figure

• 

  Fig. 1 Total numbers of activated NF-κB positive cells in spinal cord of rats at different time post-injury. All bars represent mean values and error bars represent standard deviation. There were significantly increased activated NF-κB positive cells in experiment and experiment control group in comparison to sham surgery group (p<0.05) and significant decrease in experiment group in comparison to experimental control group (p<0.05). *Re-presents the time point of highest and significant difference in expression of NF-κB post-injury. NF-κB : nuclear factor-κB.

• 

  Fig. 2 A : Immunohistochemical analysis for the expression of NF-κB in experimental group at 6 hours post-injury. B : Immunohistochemical analysis for the expression of NF-κB in control group 6 hours post-injury. C : Immunohistochemical analysis for the expression of NF-κB in Experimental group at 1 day post-injury. D : Immunohistochemical analysis for the expression of NF-κB in control group at 1 day post-injury. E : Immunohistochemical analysis for the expression of NF-κB in Experiment group at 3 days post-injury. F : Immunohistochemical analysis for the expression of NF-κB in Control group at 3 days post-injury. G : Immunohistochemical analysis for the expression of NF-κB in experimental group at 7 days post-injury. H : Immunohistochemical analysis for the expression of NF-κB in control group at 7 days post-injury. I : Immunohistochemical analysis for the expression of NF-κB in experimental group at 14 days post-injury. J : Immunohistochemical analysis for the expression of NF-κB in control group at 14 days post-injury. K : Immunohistochemical analysis for the expression of NF-κB in Sham surgery group at 6 hours post-injury. L : Immunohistochemical analysis for the expression of NF-κB in Sham surgery group at14 days post injury. NF-κB : nuclear factor-κB.

• 

  Fig. 3 Level of NF-κB mRNA expression in spinal cord of rats at different time post-injury. All bars represent mean values and error bars represent standard deviation. There were significantly increase in NF-κB mRNA level in experiment and experiment control group in comparison to sham surgery group (p<0.05) and significant decrease in experiment group in comparison to experimental control group (p<0.05). *Represents the time point of highest and significant difference in expression of NF-κB post-injury. NF-κB : nuclear factor-κB, mRNA : messenger RNA.

• 

  Fig. 4 A : Reverse transcription polymerase chain reaction (RT PCR) analysis of nuclear factor-κB expression in spinal cord (3 day post surgery). B : RT PCR analysis of GADPH expression in spinal cord (3 day post surgery). Lane 1 : marker, Lane 2 : Sham surgery group, Lane 3 : Experimental Control group, Lane 4 : Experimental group.

• 

  Fig. 5 Open field locomotor assessment by BBB score in rats at different time post-injury. All bars represent mean values and error bars represent standard deviation. There were significantly higher BBB score in rats from experiment group in comparison to experimental control group (p<0.05). BBB : Basso, Beattie, Bresnahan.

• 

  Fig. 6 Martin Schwab's Lab urinary bladder function score in rats at different time post-injury. All bars represent mean values and error bars represent standard deviation. There were significant urinary bladder function improvement in rats from experiment group in comparison to experimental control group (p<0.05).

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