Prolonged Minocycline Treatment Impairs Motor Neuronal Survival and Glial Function in Organotypic Rat Spinal Cord Cultures

Pinkernelle, Josephine; Fansa, Hisham; Ebmeyer, Uwe; Keilhoff, Gerburg

doi:10.1371/journal.pone.0073422

Cited by 33 publications

(27 citation statements)

References 87 publications

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“…The OSC model appears to reproduce many of the posttraumatic events, including acute and secondary injury mechanisms (Krassioukov et al 2002;Pinkernelle et al 2013;Weightman et al 2014;Pohland et al 2015). The model serves to control the extracellular environment and has the promise of lower cost and faster discovery (Morrison et al 1998).…”

Section: Discussionmentioning

confidence: 99%

“…Cell death is one of the consequences of traumatic SCI, affecting both neurons and supporting glial cells (Beattie et al 2002). Numerous studies have reported spinal cord tissue samples isolated from embryonic or newborn rats (Krassioukov et al 2002;Cho et al 2009;Kim et al 2010;Ravikumar et al 2012;Gerardo-Nava et al 2013Pinkernelle et al 2013;Hashemian et al 2014;Weightman et al 2014;Pohland et al 2015;Sypecka et al 2015). The organotypic slice culture (OSC) model appears to reproduce much of the post-traumatic events following injury, including acute and secondary injury mechanisms.…”

Section: Introductionmentioning

confidence: 99%

“…It has been reported that there is a significant difference in function, circuitry connections and regenerative capacities between the immature spinal cord and the adult cord (Krassioukov et al 2002). The heterogeneous populations of cells found in vivo are maintained within OSCs with three-dimensional connections between neurons and supporting cells (Cho et al 2009;Kim et al 2010;Ravikumar et al 2012;Gerardo-Nava et al 2013Pinkernelle et al 2013;Hashemian et al 2014;Weightman et al 2014;Pellowska et al 2015;Pohland et al 2015) to avoid hypoxia and necrosis of the central tissue (Krassioukov et al 2002).…”

Section: Introductionmentioning

confidence: 99%

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Cell viability in three ex vivo rat models of spinal cord injury

et al. 2018

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Spinal cord injury (SCI) is a devastating disorder that has a poor prognosis of recovery. Animal models of SCI are useful to understand the pathophysiology of SCI and the potential use of therapeutic strategies for human SCI. Ex vivo models of central nervous system (CNS) trauma, particularly mechanical trauma, have become important tools to complement in vivo models of injury in order to reproduce the sequelae of human CNS injury. Ex vivo organotypic slice cultures (OSCs) provide a reliable model platform for the study of cell dynamics and therapeutic intervention following SCI. In addition, these ex vivo models support the 3R concept of animal use in SCI research – replacement, reduction and refinement. Ex vivo models cannot be used to monitor functional recovery, nor do they have the intact blood supply of the in vivo model systems. However, the ex vivo models appear to reproduce many of the post traumatic events including acute and secondary injury mechanisms. Several well‐established OSC models have been developed over the past few years for experimental spinal injuries ex vivo in order to understand the biological response to injury. In this study, we investigated cell viability in three ex vivo OSC models of SCI: stab injury, transection injury and contusion injury. Injury was inflicted in postnatal day 4 rat spinal cord slices. Stab injury was performed using a needle on transverse slices of spinal cord. Transection injury was performed on longitudinal slices of spinal cord using a double blade technique. Contusion injury was performed on longitudinal slices of spinal cord using an Infinite Horizon impactor device. At days 3 and 10 post‐injury, viability was measured using dual staining for propidium iodide and fluorescein diacetate. In all ex vivo SCI models, the slices showed more live cells than dead cells over 10 days in culture, with higher cell viability in control slices compared with injured slices. Although no change in cell viability was observed between time‐points in stab‐ and contusion‐injured OSCs, a reduction in cell viability was observed over time in transection‐injured OSCs. Taken together, ex vivo SCI models are a useful and reliable research tool that reduces the cost and time involved in carrying out animal studies. The use of OSC models provides a simple way to study the cellular consequences following SCI, and they can also be used to investigate potential therapeutics regimes for the treatment of SCI.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Cell viability in three ex vivo rat models of spinal cord injury

et al. 2018

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“…A previous study reported deleterious effects of minocycline on neurons following the administration of a high dose (90 mg/kg) to mice (5). Another study described reduced motor neuron survival following the administration of a higher concentration (100 µM) of minocycline as compared to a lower concentration (10 µM) (21).…”

Section: The Effect Of High-dose Concentrated Minocycline Through Tamentioning

confidence: 99%

The Effects of Minocycline on Spinal Root Avulsion Injury in Rat Model

Chin

Kiat²,

Faizul

et al. 2017

MJMS

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Methods: The C7 nerve roots of the animals were avulsed via an anterior extravertebral approach. Traction force was used to transect the ventral motor nerve roots at the preganglionic level. Intraperitoneal and intrathecal minocycline (50 mg/kg for the first week and 25 mg/kg for the second week) were administered to promote motor healing. The spinal cord was harvested six weeks after the injury, and structural changes following the avulsion injury and pharmacological intervention were analysed.Results: Motor neuron death and microglial proliferation were observed after the administration of minocycline via two different routes (intraperitoneal and intrathecal) following traumatic avulsion injury of the ventral nerve root. The administration of intraperitoneal minocycline reduced the microglia count but increased the motor neuron count. Intrathecal minocycline also reduced the microglial count, with a greater reduction than in the intraperitoneal group, but it decreased the motor neuron count.Conclusions: Intraperitoneal minocycline increased motor neuron survival by inhibiting microglial proliferation following traumatic avulsion injury of the nerve root. The inhibitory effect was augmented by the use of intrathecal minocycline, in which the targeted drug delivery method increased the bioavailability of the therapeutic agent. However, motor neuron survival was impaired at a higher concentration of minocycline via the intrathecal route due to the more efficient method of drug delivery. Microglial suppression via minocycline can have both beneficial and damaging effects, with a moderate dose being beneficial as regards motor neuron survival but a higher dose proving neurotoxic due to impairment of the glial response and Wallerian degeneration, which is a pre-requisite for regeneration.

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“…Although DRG explants are a relatively simple and effective model for investigating axon regeneration and myelination in vitro, their main limitation is that they only offer insights on the behaviour of sensory neurons. In order to study ex vivo the motor compartment of peripheral nerves, spinal cord explants can also be obtained, although using procedures that have a much higher degree of technical complexity (Fabbro et al, 2012;Pinkernelle et al, 2013).…”

Section: Ex Vivo Explantsmentioning

confidence: 99%

In vitro models for peripheral nerve regeneration

Geuna

Raimondo

Fregnan

et al. 2015

Eur J of Neuroscience

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The study of peripheral nerve repair and regeneration is particularly relevant in the light of the high clinical incidence of nerve lesions. However, the clinical outcome after nerve lesions is often far from satisfactory and the functional recovery is almost never complete. Therefore, a number of therapeutic approaches are being investigated, ranging from local delivery of trophic factors and other molecules to bioactive biomaterials and complex nerve prostheses. Translation of the new therapeutic approaches to the patient always requires a final pre-clinical step using in vivo animal models. The need to limit as much as possible animal use in biomedical research, however, makes the preliminary use of in vitro models mandatory from an ethical point of view. In this article, the different types of in vitro models available today for the study of peripheral nerve regeneration have been ranked by adopting a three-step stair model based on their increasing ethical impact: (i) cell line-based models, which raise no ethical concern; (ii) primary cell-based models, which have low ethical impact as animal use, although necessary, is limited; and (iii) organotypic ex vivo-based models, which raise moderate ethical concerns as the use of laboratory animals is required although with much lower impact on animal wellbeing in comparison to in vivo models of peripheral nerve regeneration. This article aims to help researchers in selecting the best experimental approach for their scientific goals driven by the 'Three Rs' (3Rs) rules (Replacement, Reduction or Refinement of animal use in research) for scientific research.

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Prolonged Minocycline Treatment Impairs Motor Neuronal Survival and Glial Function in Organotypic Rat Spinal Cord Cultures

Cited by 33 publications

References 87 publications

Cell viability in three ex vivo rat models of spinal cord injury

Cell viability in three ex vivo rat models of spinal cord injury

The Effects of Minocycline on Spinal Root Avulsion Injury in Rat Model

In vitro models for peripheral nerve regeneration

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