Spinal cord organotypic slice cultures for the study of regenerating motor axon interactions with 3D scaffolds

Gerardo‐Nava, José; Hodde, Dorothée; Katona, István; Bozkurt, Ahmet; Grehl, Torsten; Steinbusch, Harry W.M.; Weis, Joachim; Brook, Gary

doi:10.1016/j.biomaterials.2014.02.007

Cited by 40 publications

(32 citation statements)

References 31 publications

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“…[ 37 ] Even now, there was not any applicable technique or functional materials for the fast and persistent acceleration of the motor neurons growth and development. [ 38,39 ] Here, in order to detect the actual application effect of the G-NFs as multifunctional soft cell modulation scaffold for cellular electrical stimulation, a 100 mV pulse electrical signal stimulation was applied by the G-NFs to the primary motor neurons in vitro ( Figure S7, Supporting Information). Figure 3 showed the neuritogenesis and maturation of the motor neurons in a period of 3 d in culture.…”

Section: Doi: 101002/adma201503319mentioning

confidence: 99%

Soft Graphene Nanofibers Designed for the Acceleration of Nerve Growth and Development

et al. 2015

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Soft graphene nanofibers with recoverable electrical conductivity and excellent physicochemical stability are prepared by a controlled assembly technique. By using the soft graphene nanofibers for cellular electrical stimulation, the common inhibitory effect of long-term electrical stimulation on nerve growth and development is avoided, which usually happens with traditional 2D conductive materials.

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Section: Doi: 101002/adma201503319mentioning

confidence: 99%

Soft Graphene Nanofibers Designed for the Acceleration of Nerve Growth and Development

et al. 2015

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“…; Gerardo‐Nava et al. ). We have also investigated secondary injury mechanisms such as scarring in these model systems (data submitted for publication).…”

Section: Discussionmentioning

confidence: 99%

“…A slightly longer period of time was used by Gerardo‐Nava et al. ()who kept the slices in culture for 7–14 days to investigate the interaction between axons in postnatal spinal cord. Slice thickness of the spinal cord tissue is recommended at 350–400 μm as this has been reported to avoid hypoxia and necrosis of the central tissue (Krassioukov et al.…”

Section: Introductionmentioning

confidence: 99%

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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“…Vyas et al . () have developed a more advanced in vitro organotypic co‐culture system, later also used by other authors (Gerardo‐Nava et al ., ), that accurately models peripheral nerve repair in the adult mammal. This model is based on entire spinal cord explants with reconstructed ventral nerve roots.…”

Section: Organotypic Ex Vivo‐based Modelsmentioning

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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Spinal cord organotypic slice cultures for the study of regenerating motor axon interactions with 3D scaffolds

Cited by 40 publications

References 31 publications

Soft Graphene Nanofibers Designed for the Acceleration of Nerve Growth and Development

Soft Graphene Nanofibers Designed for the Acceleration of Nerve Growth and Development

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

In vitro models for peripheral nerve regeneration

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