Page 87 - NNS 2014 Program & Abstract Book

2Institute of Medical Science, Faculty of Medicine, University of
Toronto, Toronto, Canada
3Division of Neurosurgery and Spinal Program, University of Tor-
onto, Toronto, Canada
Spinal cord injury (SCI)-induced vascular disruption (VD) is a trigger
for secondary injury. Thus, targeting VD may reduce tissue loss and
impairment. Mesenchymal Stem Cells (MSCs), including those from
the human umbilical cord matrix (HUCMCs), have pericytic attri-
butes. They are well suited to addressing VD and the multi-factorial
dynamic SCI pathophysiology. However, SCI treatment with live cells
is severely hampered by logistical and practical considerations.
A novel, viable, safe and effective alternative to acute cell therapy
for SCI.
R&D ELISA arrays were employed to profile serum-free media con-
ditioned by age- and passage-matched cells cultured for 2 weeks in
DMEM/F12 + 1% Glutamax. Concentrated HUCMC secretome ( >9kDa)
was systemically infused immediately after traumatic 1-minute C7 clip-
compression SCI in 250–300g female Wistar rats, which were sacrificed
48 hours later. Vascular permeability was evaluated by spectrophoto-
metric quantification of 2% Evans Blue dye infused 30 minutes pre-
sacrifice in snap-frozen homogenates of lesional spinal cord tissue. Pre-
sacrifice very-high-resolution ultrasound (VHRUS) measurements of
haemorrhagic lesional volume were made.
HUCMCs secrete more and greater concentrations of pro-angiogenic
factors (activin, angiogenin, angiopoietin-1, amphiregulin and coagu-
lation factor III), anti-inflammatory cytokines (G-CSF, GM-CSF, IL-6,
IL-8, ENA-78, MCP-3, midkine, MIP-1alpha/beta and MIP-3alpha) and
neurotrophic factors (FGF2, FGF7 and GDNF) than adult bone marrow
stromal cells (BMSCs) and (adult and newborn dermal) fibroblasts. At
48 hours post-SCI, lesion-induced vascular permeability (p = 0.0067,
n = 5) and lesion volume size (p = 0.001, n = 4) were both reduced by
concentrated HUCMC-CM relative to controls (concentrated Alpha-
MEM). Inflammation (measured by MPO activity) and haemorrhage
(measured using Drabkin’s reagent) were only slightly reduced intra-
and peri-lesionally.
HUCMCs have a more potent secretome than BMSCs and fibro-
blasts, and can reduce SCI-induced VD.
Key words
cervical, conditioned medium, Drabkin’s, Evans blue, mesenchymal,
secretome, spinal cord injury, ultrasound, umbilical cord matrix,
vascular permeability
B2-10
FUNCTIONAL IMPROVEMENT WITH INTRANASALLY-
DELIVERED HUMAN OLFACTORY STEM CELLS AND EX-
ERCISE AND ENRICHMENT IN SPINAL CORD INJURY
Peduzzi, J.1, Jackson, C.H.1, Kronner, J.1, Nelson, J.O.1, Jain, T.1,
Morck, K.1, Drzyzga, V.1, Girgla, T.1, Kasprzak, M.2, Meythaler, J.M.2
1Department of Anatomy, Detroit, USA
2Department of Physical Medicine and Rehabilitation at Oakwood,
Wayne State School of Medicine, Detroit, USA
We hypothesized that exercise and environmental enrichment may
produce greater functional improvement when combined with intra-
nasally-delivered progenitor cells after spinal cord injury. Thirty male,
athymic Nude rats (3 groups, 10 rats each) were injured at T9 spinal
level using the MASCIS device. Two weeks later, Group 1 received
intranasal saline, Groups 2 & 3 received intranasally-delivered human
olfactory-derived progenitor cells and Group 3 additionally received
enrichment and exercise. Exercise consisted of passive cycling, low-
and high-level swimming, perturbation training and voluntary ex-
ercise in exercise balls. Environmental enrichment involved expo-
sure to a social environment with novel objects. Outcome measures
included the BBB, the Louisville Swim, Inclined Plane and Beam
tests. A statistical difference (p = 0.027) using repeated measures
ANOVA was obtained with the Beam test where the progenitor,
exercise/enrichment group improved more than the progenitor cells
alone group. Poor health in the Nude rats may be responsible for lack
of even greater functional improvement. Immunohistochemistry
using anti-human antibody revealed that the intranasally-delivered
progenitor cells homed to the region of damage in the spinal cord.
The greater functional improvement in rats receiving the olfactory
progenitor cells, exercise and enrichment may suggest that progen-
itor cells require input in order to form appropriate circuitry nec-
essary for functional improvement. This therapeutic approach has
great potential for clinical translation because a person’s own ol-
factory progenitor cells with a normal neural fate could be used. The
cells are obtainable and deliverable with minimally invasive tech-
niques. The recently discovered method of intranasal delivery (no
injections) would mean that even patients with severe spinal injuries
may be able to be treated subacutely.
Key words
enriched environment, exercise, intranasal delivery, progenitor cell
B3-01
ACUTE STRESS COMPLICATING MILD TRAUMATIC
BRAIN INJURY IN RODENTS
Garcia, R.L., Estevez Castillo, C.R., Robertson, C.S.
Baylor College of Medicine, Department of Neurosurgery, Houston,
USA
Mild traumatic brain injury (mTBI) in humans often occurs in the
setting of acute stress (AS), especially with military injuries. It is not
clear if AS significantly contributes to the chronic symptoms that
occur after mTBI. Rodent models of mTBI do not share this charac-
teristic because the animals are anesthetized at the time of injury. The
purpose of this research was to study the effect of AS either imme-
diately before or after a mild cortical impact injury (mCCI) on be-
havioral consequences of the injury.
Forty Long Evans rats, weighing 300–350 grams, were enrolled
and randomly assigned to five groups: sham (n = 8), AS (n = 8),
mCCI (n = 8), AS induced 1 hr after mCCI (n = 8), and mCCI in-
duced 1 hr after AS (n = 8). To induce AS, rats were placed in a 1¢ by
1¢ enclosure with a cotton ball scented with trimethylthiazoline and
subjected to white noise at random intervals for 15 minutes. Rats
undergoing mCCI were anesthetized and subjected to a mCCI [3 m/
sec, 2.5 mm deformation]. Outcome measures were beam walking
and balance tests, novel object recognition test, open field test, and
two Morris water maze variations for spatial navigation and working
memory.
The mCCI animals had impaired performance on beam balance
testing compared to sham (p = .029), and a trend for impaired working
memory on the Morris water maze testing (p = .086). The AS animals
had significant differences on the open field test, with greater distance
traveled (p < .001) and greater velocity of movement (p < .001) com-
pared to sham, but no impairments on the motor or cognitive tasks.
When mCCI was complicated by AS, there was no greater impairment
on the behavioral tasks than with mCCI alone. These animals with the
combined injury did not have the increased activity on the open field
testing as those with AS alone.
A-55