Treadmill Locomotor Training
Objective: To investigate potential interactions between neural stem cell transplantation and treadmill training in promoting locomotor recovery and structural repair in a rat contusive spinal cord injury model
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Equipment1
Not specified in provided text • Not specified • Not specified • Not mentioned
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Protocol Steps
Spinal cord injury induction
Create contusive spinal cord injury in rats
Note: Baseline timepoint for subsequent interventions
View evidence from paper
“rat contusive injury model”
Neural stem cell transplantation
Transplant neural stem cells obtained at embryonic day 14 into the injured spinal cord
Note: Timing is critical - performed at 1 week post-injury
View evidence from paper
“Combination of TMT with NSC TP at 1 week after injury synergistically improved locomotor function”
Treadmill locomotor training
Initiate treadmill training to promote locomotor recovery
Note: Combined with NSC transplantation for synergistic effects
View evidence from paper
“Combination of TMT with NSC TP at 1 week after injury synergistically improved locomotor function”
Assessment of NSC survival
Evaluate survival of grafted neural stem cells at multiple timepoints
Note: TMT increased NSC survival by >3-fold at 3 weeks and >5-fold at 9 weeks
View evidence from paper
“combining TMT increased the survival of grafted NSCs by >3-fold and >5-fold at 3 and 9 weeks after injury, respectively”
Measurement of cerebrospinal fluid IGF-1 concentration
Quantify insulin-like growth factor-1 levels in cerebrospinal fluid
Note: TMT increased IGF-1 concentration in CSF
View evidence from paper
“TMT increased the concentration of insulin-like growth factor-1 (IGF-1) in the CSF”
Intrathecal antibody infusion - IGF-1 neutralization
Administer neutralizing IGF-1 antibodies via intrathecal infusion to block IGF-1 signaling
Note: This treatment abolished the enhanced survival of NSC grafts by TMT
View evidence from paper
“Intrathecal infusion of neutralizing IGF-1 antibodies, but not antibodies against either BDNF or Neurotrophin-3 (NT-3), abolished the enhanced survival of NSC grafts by TMT”
Intrathecal antibody infusion - control treatments
Administer control antibodies (BDNF or NT-3) via intrathecal infusion as negative controls
Note: Control antibodies did not abolish enhanced NSC survival by TMT
View evidence from paper
“Intrathecal infusion of neutralizing IGF-1 antibodies, but not antibodies against either BDNF or Neurotrophin-3 (NT-3), abolished the enhanced survival of NSC grafts by TMT”
Assessment of cellular stress markers
Evaluate reactive nitrogen or oxygen species in grafted NSCs
Note: Cellular stresses were markedly attenuated by TMT
View evidence from paper
“NSCs grafted into the injured spinal cord were under cellular stresses induced by reactive nitrogen or oxygen species, which were markedly attenuated by TMT”
Assessment of locomotor recovery
Measure locomotor function recovery in treated animals
Note: Number of surviving NSCs was significantly correlated with extent of locomotor recovery
View evidence from paper
“The number of surviving NSCs was significantly correlated with the extent of locomotor recovery”
Histological assessment of tissue sparing
Evaluate tissue preservation in the injured spinal cord
Note: Combination of TP and TMT resulted in tissue sparing to larger extent than either alone
View evidence from paper
“The combination of TP and TMT also resulted in tissue sparing, increased myelination, and restoration of serotonergic fiber innervation”
Assessment of myelination
Evaluate myelination in the injured spinal cord
Note: Increased myelination observed with combination treatment
View evidence from paper
“The combination of TP and TMT also resulted in tissue sparing, increased myelination, and restoration of serotonergic fiber innervation”
Assessment of serotonergic fiber innervation
Evaluate restoration of serotonergic fiber innervation to the lumbar spinal cord
Note: Restoration was greater with combination treatment than either treatment alone
View evidence from paper
“The combination of TP and TMT also resulted in tissue sparing, increased myelination, and restoration of serotonergic fiber innervation to the lumbar spinal cord to a larger extent than that induced by either TP or TMT alone”