Page 85 - NNS 2014 Abstract Book

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TRANSPLANTED NEUROSPHERES FROM GENETICALLY
MODIFIED ADULT BONE MARROW AFTER CCI: EFFECTS
ON INJURY SIZE AND TRANSPLANT SURVIVAL
Gowans, A.1, Goshu, B.1, Koronkiewicz, A.1, Glavaski-Joksimovic,
A.2, Bohn, M.C.3 , Kozlowski, D.A.1
1DePaul University, Chicago, USA
2Medical College of Wisconsin and Zablocki Veterans Affairs Medical
Center, Milwaukee, USA
3Ann and Robert H Lurie Children’s Research Center of Chicago,
Chicago, USA
A potential treatment for traumatic brain injury is transplantation of
adult bone marrow-derived mesenchymal stem cells (BMSC). Al-
though previous studies have transplanted undifferentiated BMSC
following TBI, our lab transplanted neurospheres derived from
BMSC genetically modified with the intracellular domain of Notch1
and neo-resistance genes. These modified BMSC were transduced
with a GFP lentivirus and grown under non-adherent conditions to
promote formation of neurospheres that were transplanted one-week
post-controlled cortical impact (CCI) in the rat. Transplants were
placed in the injured cortex or striatum below the injury. Previous
data demonstrated that transplanted neurospheres decreased senso-
rimotor deficits out to one month post-transplant. Behavioral en-
hancement was better in rats with striatal vs. cortical transplants. The
current study examined whether these behavioral results could be
explained by differences in transplant survival or neuroprotection.
Brain tissue from these animals was analyzed one month post-injury
using Neurolucida. Survival of stem cells was measured qualitatively
with a rating scale for GFP positive cells. Neuroprotection or re-
placement of cells post-injury was measured via an analysis of re-
maining cortical area (lm2 ) in sections containing the forelimb
sensorimotor cortex. The volume of remaining cortex was obtained
by multiplying total area by the distance between sections,
0.24 mm2. ANOVA demonstrated that striatal neurosphere trans-
plants showed better survival (i.e. presence of more GFP positive
cells) than cortical transplants (p < 0.05). ANOVA of cortical volume
indicated that there was no significant difference between striatal
and cortical neurosphere transplants (p > 0.05 compared to CCI
only). Results indicated that striatal neurosphere transplants fol-
lowing TBI have better survival than cortical transplants, but the
neurosphere transplants in either region have no effect in reducing
contusion size through neuroprotection or replacement, suggesting
that the behavioral enhancement seen is due to other factors such as
trophic factors.
Key words
cortical volume, neurospheres, sensorimotor cortex, TBI
C2-25
INVESTIGATION OF THE EFFECTS OF CARBOXY-
FULLERENE NANOPARTICLES ON THE CEREBRAL
VASCULATURE AFTER FLUID PERCUSSION INJURY
Wynne, K.E., Prough, D.S., DeWitt, D.S.
University of Texas Medical Branch, Galveston, United States
Traumatic brain injury (TBI) causes an increase in reactive oxygen
species (ROS) that can cause cerebral vascular dysfunction. Reducing
ROS levels can preserve vascular function and improve outcome after
injury. Our preliminary results suggested that treatment with the
carboxyfullerene nanoparticle DF-1 improved Morris water maze
performance after TBI. DF-1 is an antioxidant that is capable of re-
ducing ROS levels. Here we tested the hypothesis that improved be-
havioral outcome after DF-1 treatment was due in part to a reduction
in TBI-induced cerebral vascular dysfunction.
Male Sprague-Dawley rats (n = 40) were anesthetized with iso-
flurane (1.5%), prepared for parasagittal fluid percussion TBI and
randomly assigned to receive sham, moderate TBI alone or TBI fol-
lowed by treatment with 10, 25 or 50 mg/kg DF-1 one hour post-injury
(n = 8 animals/group). Measurements of cerebral perfusion (laser
Doppler flowmetry, LDF) and mean arterial pressure (MAP) were
made from 30 minutes prior to injury and continued for two hours
following injury. Cerebral vascular resistance (CVR) was calculated
as CVR = MAP x LDF -1.
MAP was significantly increased (P < 0.05) by all three doses of
DF-1 compared to sham and injured animals. The 25 and 50 mg/kg
doses produced greater increases than the 10 mg/kg dose. Cerebral
perfusion was reduced with treatment. Lower doses (10 and 25 mg/kg)
were associated with greater reductions in perfusion compared to
injured animals and those treated with 50 mg/kg doses (P < 0.05).
CVR was significantly increased by DF-1 with higher doses producing
greater increases (P < 0.05).
These results demonstrate that DF-1 treatment increased MAP
and CVR while reducing cerebral perfusion. Larger doses seem to
return cerebral perfusion closer to baseline compared to lower
doses. Reduced cerebral perfusion would have a negative effect on
outcome after injury. This leads us to conclude that DF-1 might be
conferring benefits on the cellular or molecular level that out
weigh the effects that might be caused by a reduction in cerebral
perfusion.
Key words
nanotechnology, TBI
C2-26
THE EMERGING ROLE OF AMP-ACTIVATED PROTEIN
KINASE IN TBI AND ITS PHARMACOLOGICAL REGULA-
TION BY S-NITROSOGLUTATHIONE
Khan, M., Dhammu, T.S., Shunmugavel, A., Singh, I., Singh, A.K.
Medical University of South Carolina, Charleston, USA
A growing body of research indicates that traumatic brain injury (TBI)
patients and animal models exhibit bioenergetic and metabolic ab-
normalities, blood-brain barrier (BBB) disruption neurobehavioral
deficits. We hypothesized that AMPK, a sensor and regulator of en-
ergy balance, activation is associated with peroxynitrite-mediated
endothelial dysfunction in the acute phase whereas its activation in the
chronic phase up regulates the process of neurorepair. Therefore, we
investigated the efficacy S-nitrosoglutathione (GSNO) which invokes
functional recovery and reduces levels of peroxynitrite.
TBI was induced by controlled cortical impact (CCI) in adult male
rats. GSNO (0.05 mg/kg) and compound c (5 mg/kg) were adminis-
tered at two hours after CCI and daily thereafter. In addition to
functional recovery, the following neurovascular protection and neu-
rorepair mediators were evaluated: AMPK, peroxynitrite, eNOS, BBB
leakage, edema, HIF-1a/VEGF and vessel density.
Treatment with AMPK inhibitor compound c and BBB inducing
agent GSNO after TBI reduced peroxynitrite levels, inhibited BBB
disruption and edema formation. The treatment with GSNO also re-
duced the expression of AMPK and eNOS in the acute phase. How-
ever, a chronic treatment with compound c had adverse effect on
functional recovery. In contrast, chronic GSNO treatment of TBI not
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