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Development of a methodology for monitoring spinal neuroinflammation mechanisms in the neuropathic rat in vivo

Delaney, Sally-Ann (2010)
M.Phil. thesis, University of Birmingham.

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Spinal release of pro-inflammatory cytokines plays a pivotal role in the aberrant neuronal signalling that underlies neuropathic pain. This thesis sought to develop a methodology for monitoring spinal cytokine release in vivo using ‘high flux’ microdialysis sampling. In vitro studies characterising ‘high flux’ membranes Plasmaflo® (Poly-E), Prisma® (Poly-P), Terumo®(CUP-R) and commercially available probe, CMA/20 (nominal cut-off 3000,≈3000, 60 and 100 kDa respectively) revealed that Poly-E and Poly-P were significantly better in recovering IL-1β than the commercially available probe (58, 48 and 38% at 0.5 µl/min, n=4-5, respectively). Poly-E was identified as the most suitable membrane for in vivo studies. For future in vivo studies, disinfection of the membrane and blockade of non-specific peptide binding sites by siliconisation rather than conventional use of BSA, was shown not to adversely affect the dialysing capability of the Poly-E membrane. In rats with CCI-induced mechanical hypersensitivity intrathecal microdialysis was successful in measuring basal levels of IL-1β, IL-6 and TNFα (68±27, 5714±2451 and 638±175 pg/ml (mean ± SEM, n=10) respectively). Electrical stimulation capable of activating C-fibres was shown to induce IL-1β (≈ 3-fold), but not IL-6 and TNFα release in CCI animals compared to sham controls. In further studies, intrathecal microdialysis sampling was successfully performed in freely moving naïve animals. Atlanto-occipital catheterisation however, was associated with welfare concerns so a less invasive lumbar catheterisation technique was evaluated. This technique was well tolerated and may prove valuable in determining the spinal cytokine release in animal pain models.

Type of Work:M.Phil. thesis.
Supervisor(s):Whitehead, Kevin
School/Faculty:Colleges (2008 onwards) > College of Medical & Dental Sciences
Department:Department of Pharmacology
Additional Information:

Full version of thesis under embargo until: 01/07/2040.

Subjects:RM Therapeutics. Pharmacology
Institution:University of Birmingham
ID Code:925
This unpublished thesis/dissertation is copyright of the author and/or third parties. The intellectual property rights of the author or third parties in respect of this work are as defined by The Copyright Designs and Patents Act 1988 or as modified by any successor legislation. Any use made of information contained in this thesis/dissertation must be in accordance with that legislation and must be properly acknowledged. Further distribution or reproduction in any format is prohibited without the permission of the copyright holder.
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