Abstract
Event Abstract Back to Event The effect of Centella asiatica (L.) urban on the organotypic model of spinal cord injury Ahmad P. Nabilah1, 2, 3, Yogeswaran Lokanathan1, A Mohd Ilham4 and Ruszymah Hj Idrus1, 2* 1 Universiti Kebangsaan Malaysia Medical Center, Tissue Engineering Center, Malaysia 2 Universiti Kebangsaan Malaysia, Department of Physiology, Faculty of Medicine, Malaysia 3 Universiti Kebangsaan Malaysia, Faculty of Dentistry, Malaysia 4 Universiti Teknologi MARA, Faculty of Pharmacy, Malaysia Neurotrophic effect of Centella asiatica (L.) urban (CA) has been studied in vitro and in vivo on neuroblastoma cell lines and rats’ hippocampus, respectively with reports of increased neurite outgrowth. However, its effect on the spinal cord has not been studied widely. Here we evaluated the effect of CA on the spinal cord using an in vitro organotypic spinal cord injury (OSCI) model. Spinal cords from postnatal day 8 Wistar rat pups were embedded in 10% gelatin and sliced into 300µm transverse slices, then, cultured organotypically for 24 hours. CA cytotoxicity was studied on spinal cord primary culture using Presto Blue® Cell Viability Reagent. CA is added to the media at 0, 100, 200, 400 and 800 µg/ml for 7 days, then slices were fixed and immunostained for neurite outgrowths with anti-beta III Tubulin antibody [clone: TUJ-1] and anti-glial fibrillary acidic protein (GFAP) antibodies. Average number of neurite outgrowth and mean axonal length were then measured. The IC50 for CA on spinal cord primary culture was determined to be 1102.5µg/ml. The organotypic model was successfully developed where the intact, adherent and viable transverse slices of spinal cord were acquired using 7 and 8 days old rat pups, 10% gelatin embedding, and a slice thickness of 300µm. The tissue cultured on transwell showed 100% viability at least up to 8 days. We found that, CA up to 800 µg/ml does not show any cytotoxic or inhibitory effect to the tissue. Neurite outgrowth stained positive for beta III Tubulin and astrocytes stained positive for GFAP. Keywords: Spinal Cord, in vivo, in vitro, Neurite outgrowth, Neurotrophic effect Conference: 14th Meeting of the Asian-Pacific Society for Neurochemistry, Kuala Lumpur, Malaysia, 27 Aug - 30 Aug, 2016. Presentation Type: Poster Presentation Session Topic: 14th Meeting of the Asian-Pacific Society for Neurochemistry Citation: Nabilah AP, Lokanathan Y, Mohd Ilham A and Hj Idrus R (2016). The effect of Centella asiatica (L.) urban on the organotypic model of spinal cord injury. Conference Abstract: 14th Meeting of the Asian-Pacific Society for Neurochemistry. doi: 10.3389/conf.fncel.2016.36.00126 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 04 Aug 2016; Published Online: 11 Aug 2016. * Correspondence: Prof. Ruszymah Hj Idrus, Universiti Kebangsaan Malaysia Medical Center, Tissue Engineering Center, Bandar Tun Razak, Kuala Lumpur, Malaysia, ruszyidrus@gmail.com Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract The Authors in Frontiers Ahmad P Nabilah Yogeswaran Lokanathan A Mohd Ilham Ruszymah Hj Idrus Google Ahmad P Nabilah Yogeswaran Lokanathan A Mohd Ilham Ruszymah Hj Idrus Google Scholar Ahmad P Nabilah Yogeswaran Lokanathan A Mohd Ilham Ruszymah Hj Idrus PubMed Ahmad P Nabilah Yogeswaran Lokanathan A Mohd Ilham Ruszymah Hj Idrus Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. Please enable Javascript in your browser settings in order to see all the content on this page.
Highlights
Spinal cord (SC) injury (SCI) occurs due to trauma sustained by the victims of motor vehicle accidents (66%), falls (28%), violence (4%), sports (2%) and other accidents in homes or work places (Ibrahim et al 2013)
Analysis was done using images taken with Nikon Eclipse Ti Fluorescence microscope (Nikon, Japan) and the NISelement software. 5 regions of interest (ROI) were marked on each SC slice at the edge of the SC slices where the neurite outgrowths sprouted out
For the mean longest neurite, the longest neurite measured in each ROI are added up and divided by the number of ROI to find the mean longest neurite for the slice
Summary
Spinal cord (SC) injury (SCI) occurs due to trauma sustained by the victims of motor vehicle accidents (66%), falls (28%), violence (4%), sports (2%) and other accidents in homes or work places (Ibrahim et al 2013). The trauma itself results in primary injury to the SC when pieces of vertebral bones puncture the tissue causing disruption of descending and ascending nerve tracts connecting the brain to the peripheral motor and visceral organs (Sadowsky et al 2002). Steroids are the first medication given to patient to halt excessive inflammatory reactions to prevent further damage. Surgical treatment such as decompression laminectomy is done to alleviate pressure from the SC. No conclusive cure has been found (Ramer et al 2014)
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