A new three-dimensional axonal outgrowth assay for central nervous system regeneration

Ishihara, Masahiro; Mochizuki-Oda, Noriko; Iwatsuki, Koichi; Kishima, Haruhiko; Iwamoto, Yumiko; Ohnishi, Yozo; Umegaki, Masao; Yoshimine, Toshiki

doi:10.1016/j.jneumeth.2011.03.020

Cited by 15 publications

(16 citation statements)

References 24 publications

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“…In our previous reports, we showed that axons from cortex in no‐cell control groups run not only on the surface of the gel but also into the gels in this coculture system (Ishihara et al, ). In contact with olfactory mucosal cells, axons from cortex run mainly on the surface of the gel, in which astrocyte‐like OECs are abundant.…”

Section: Discussionmentioning

confidence: 57%

Primary olfactory mucosal cells promote axonal outgrowth in a three-dimensional assay

Ishihara

Mochizuki-Oda

Iwatsuki

et al. 2014

Journal of Neuroscience Research

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Among the possible sources of autologous cells and tissues for use in spinal cord injury grafts, one promising source is the olfactory mucosa containing olfactory ensheathing cells and neural progenitor cells. Olfactory mucosa transplantation for spinal cord injury has been effective in animal models and in pilot clinical trials. However, the contributions of olfactory ensheathing cells and neurons in olfactory mucosa are unclear. For the present study, we prepared primary olfactory mucosal cells and used a cortex-Matrigel coculture assay system to examine the axonal outgrowth of olfactory mucosa. Axonal outgrowth from cortical slices was significantly enhanced in olfactory mucosal cells compared with noncell controls and respiratory mucosal cells, which have few olfactory ensheathing cells and neurons. Axonal outgrowth was severely reduced after treatment with an antineurotrophin cocktail. A conditioned medium in the olfactory mucosa-derived cell group contained neurotrophin-3. Some olfactory ensheathing cells and almost all neurons were immunopositive for neurotrophin-3. Axons originating from cortical slices targeted mainly the astrocyte-like olfactory ensheathing cells. Our findings demonstrate that the axonal outgrowth effect of olfactory mucosa is supported by both olfactory ensheathing cells and neurons in olfactory mucosa.

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Section: Discussionmentioning

confidence: 57%

Primary olfactory mucosal cells promote axonal outgrowth in a three-dimensional assay

Ishihara

Mochizuki-Oda

Iwatsuki

et al. 2014

Journal of Neuroscience Research

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“…In addition, the model might be a useful tool to understand the fundamental molecular mechanism underlying degeneration. Within this line, axonal regeneration (Ishihara et al, 2011) and glial response (Ziemka-Nalecz et al, 2013) after axotomy could be studied in the model presented here. Moreover, using MRI that produces detailed images of a living brain tissue over time without using radioactive ligands (Petridou et al, 2006), fiber tracts can be visualized in organotypic slices and pathway integrity assessed by calculation of FA and the study of neuronal metabolism using magnetic resonance spectroscopy (Delli Pizzi et al, 2013).…”

Section: Discussionmentioning

confidence: 99%

Modeling nigrostriatal degeneration in organotypic cultures, a new ex vivo model of Parkinson’s disease

et al. 2014

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Parkinson’s disease (PD) is the second most frequent neurodegenerative disorder afflicting 2% of the population older than 65 years worldwide. Recently, brain organotypic slices have been used to model neurodegenerative disorders, including PD. They conserve brain three-dimensional architecture, synaptic connectivity and its microenvironment. This model has allowed researchers a simple and rapid method to observe cellular interactions and mechanisms. In the present study, we developed an organotypic PD model from rat brains that includes all the areas involved in the nigrostriatal pathway in a single slice preparation, without using neurotoxins to induce the dopaminergic lesion. The mechanical transection of the nigrostriatal pathway obtained during slice preparation induced PD-like histopathology. Progressive nigrostriatal degeneration was monitored combining innovative approaches, such as diffusion tensor magnetic resonance imaging (DT-RMI) to follow fiber degeneration and mass spectrometry to quantify striatal dopamine content, together with bright-field and fluorescence microscopy imaging. A substantia nigra dopaminergic cell number decrease was observed by immunohistochemistry against rat tyrosine hydroxylase (TH) reaching 80% after 2 days in culture associated with a 30% decrease of striatal TH-positive fiber density, a 15% loss of striatal dopamine content quantified by mass spectrometry and a 70% reduction of nigrostriatal fiber fractional anisotropy quantified by DT-RMI. In addition, a significant decline of medium spiny neuron density was observed from days 7 to 16. These sagittal organotypic slices could be used to study the early stage of PD, namely dopaminergic degeneration, and the late stage of the pathology with dopaminergic and GABAergic neuron loss. This novel model might improve the understanding of PD and may represent a promising tool to refine the evaluation of new therapeutic approaches.

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“…Spinal cord and traumatic brain injuries (TBI) are sustained by millions of people each year, with almost half a million emergency department visits for TBI made annually by children 1. Research efforts toward neural tissue engineering are in the early stages of discovery, with investigations of regeneration of central nervous system (CNS) tissues,2, 3 soft biomaterials to decrease inflammation from implantable devices,4–7 and non‐invasive drug delivery to the CNS 6, 8–10. Previous efforts to develop biomaterials for neural tissue repair have focused on anti‐inflammatory coatings for implantable devices10–13 or construction of nerve guide conduits for peripheral and central nerve repair 14–22.…”

Section: Introductionmentioning

confidence: 99%

“…Although polyethylene glycol (PEG) hydrogels are frequently suggested for cell encapsulation and are non‐toxic, it has been found that neuronal cell cultures in PEG hydrogels experienced initial cell death within 1 day of culture and did not exhibit greater viability or proliferation as compared to monolayer controls, even when supplemented with soluble growth factors 45. Natural biomaterials that have been explored for neural cell culture include collagen,46 Matrigel,2 alginate,47 agarose,48 and fibrin 49. Although suitable for 3D cell culture in vitro , these hydrogels are known to exhibit fast release of loaded growth factors and degrade very quickly often not allowing adequate time for cell attachment, growth, and tissue assembly for tissue engineering applications.…”

Section: Introductionmentioning

confidence: 99%

Silk Hydrogels as Soft Substrates for Neural Tissue Engineering

Hopkins

Laporte

Tortelli

et al. 2013

Adv Funct Materials

164

156

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There is great need for soft biomaterials that match the stiffness of human tissues for tissue engineering and regeneration. Hydrogels are frequently employed for extracellular matrix functionalization and to provide appropriate mechanical cues. It is challenging, however, to achieve structural integrity and retain bioactive molecules in hydrogels for complex tissue formation that may take months to develop. This work aims to investigate mechanical and biochemical characteristics of silk hydrogels for soft tissue engineering, specifi cally for the nervous system. The stiffness of 1 to 8% silk hydrogels, measured by atomic force microscopy, is 4 to 33 kPa. The structural integrity of silk gels is maintained throughout embryonic chick dorsal root ganglion (cDRG) explant culture over 4 days whereas fi brin and collagen gels decrease in mass over time. Neurite extension of cDRGs cultured on 2 and 4% silk hydrogels exhibit greater growth than softer or stiffer gels. Silk hydrogels release < 5% of neurotrophin-3 (NT-3) over 2 weeks and 11-day old gels show maintenance of growth factor bioactivity. Finally, fi bronectin-and NT-3-functionalized silk gels elicit increased axonal bundling suggesting their use in bridging nerve injuries. These results support silk hydrogels as soft and sustainable biomaterials for neural tissue engineering.

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A new three-dimensional axonal outgrowth assay for central nervous system regeneration

Cited by 15 publications

References 24 publications

Primary olfactory mucosal cells promote axonal outgrowth in a three-dimensional assay

Primary olfactory mucosal cells promote axonal outgrowth in a three-dimensional assay

Modeling nigrostriatal degeneration in organotypic cultures, a new ex vivo model of Parkinson’s disease

Silk Hydrogels as Soft Substrates for Neural Tissue Engineering

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