Normobaric hyperoxia markedly reduces

brain damage and sensorimotor deficits

following brief ischaemia

Sohail Ejaz,1,* Julius V. Emmrich,1,2,* Sergey L. Sitnikov,1 Young T. Hong,3

Stephen J. Sawiak,3 Tim D. Fryer,3 Franklin I. Aigbirhio,3 David J. Williamson3 and
Jean-Claude Baron1,4

1 Stroke Research Group, Department of Clinical Neurosciences, University of Cambridge, UK

2 Department of Neurology, Charite´ – Universita¨tsmedizin Berlin, Germany
3 Wolfson Brain Imaging Centre, Department of Clinical Neurosciences, University of Cambridge, UK

1 08/05/2019
 Introduction
 Transient ischaemic attacks  Early spontaneous
(TIAs) are characterized both reperfusion salvages the
clinically by focal penumbra in whole or in
neurological symptoms of large part, causing full clinical
ischaemic origin lasting <24
recovery within 24 h either
h, and radiologically by a lack
of topographically congruent with no MRI sequelae
changes (Saver, 2008). (Oppenheim et al., 2006).
 Such ‘true’ TIAs account for
20–50% of all clinically-
diagnosed TIAs (Brazzelli et
al., 2014)

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  Hyperbaric oxygen as a
potential therapy for
ischaemic stroke has been
tested in 11 small-scale
randomized controlled trials
but shown in one recent
meta-analysis to have no
significant benefit on
mortality and no consistent
effect on functional scales
(Bennett et al., 2014).
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 Normobaric oxygen therapy
(NBO) has been tested both
in rodent models and in
small-scale clinical trials. The
penumbra are significantly
improved by NBO despite the
only mild increase in total
arterial O2 content (Liu et
al., 2004, 2006; Shin et al.,
2007; Baskerville et al.,
2011)
 Material dan Metode
 Using 12 subjects rodent  Microclip distal temporary
 ~3-6 month male spontaneous MCA occlusion was
hypertensive rats (~300gr body performed
weight), divided to 2 groups
 At day 14 post-MCA
(normoxia and hyperoxia)
 Anaesthesia using 4% isoflurane
occlusion, all rats underwent
11C-[R]-PK11195 PET scan
in 0.3 l/min O2 and 0.7 l/min
N2O and maintained with 2%  MRI was carried out at day
isoflurane 14 of reperfusion
 In hyperoxia group, gas mixture
switched to 1 l/min pure
oxygen on MCA occlusion.

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 Behavioral Testing

- Animal were single-housed on a 12h light/dark circle

- Neurological exam was carried out the day before surgery

and at postoperative days 1, 7, 14, 21, and 28 using Garcia’s
Neuroscore

- The modified Sticky Label Test was performed 1 day before

surgery and postoperatively Days 1, 3, 7, 11, 14, 18, 21, 25,
and 28.

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 Evaluation of ischaemic damage Statistical analysis

 Briefly, on each digitized  The modified Sticky Label

whole-brain section, two Test data  ANOVA within
independent raters blinded to each group assessing the
the subject’s experimental effects of Time.
group manually traced any
area with lack of NeuN-
immunoreactivity or
increased IB4 binding

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 Arterial PO2 measurements Result

 All 12 rats entered in the

 To determine the effect of
NBO on PAO2 level study completed the protocol
without any complication or
early death until the 28-day
termination.
MRI: Day 14
 No DWI or T2-weighted
changes were observed in any
rat (Fig. 5)

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Figure 5 Illustrative examples of coronal T2-weighted MRI in normoxic and hyperoxic rats
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10
Figure 1 Modified Sticky Label Test performance data. 08/05/2019
 Relationship between sensorimotor scores and selective neuronal loss

 There was a significant correlation between NeuN Vol-Total data and peak
modified Sticky Label Test deficit, which was relatively weak , but in the
expected biological direction, i.e. a greater behavioural deficit with greater
neuronal loss (Supplementary Fig. 1).

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 Figure 4 Total lesion volumes
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13 Figure 3 Illustrative 11C-PK11195 images. 08/05/2019
 Discussion
 This is the first study to document that, as per our hypothesis, NBO
applied during very brief MCA occlusion mimicking ‘true’ TIA is able to
almost completely prevent selective neuronal death
 NBO completely prevented the marked sensorimotor deficits present in
normoxic control rats. (Terasaki et al., 2014),
 Thus, in contrast to normoxic rats, the mild microglial activation
still present at Day 14 in NBO rats had completely resolved on
post-mortem at Day 28

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 Conclusion
 NBO administered during brief MCA occlusion in a clinically-relevant
rat model of ‘true’ TIA near-completely prevented neuronal death and
microglial inflammation as well as sensorimotor impairment.

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