NDLI: Sensory neurons and osteoblasts: close partners in a microfluidic platform

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Sensory neurons and osteoblasts: close partners in a microfluidic platform

Content Provider	Royal Society of Chemistry (RSC)
Author	Aguiar, Paulo Alves, Cecília J. Lourenço, Ana H. Almeida, Ramiro D. Leitão, Luís Ryu, Hyun R. Alencastre, Inês S. Jeon, Noo L. Sousa, Daniela M. Fernandes, Rui Neto, Estrela Lamghari, Meriem
Copyright Year	2014
Abstract	Innervation has proven to be critical in bone homeostasis/regeneration due to the effect of soluble factors, produced by nerve fibers, associated with changes in the activity of bone cells. Thus, in this study, we have established and characterized a coculture system comprising sensory neurons and osteoblasts to mimic the in vivo scenario where nerve fibers can be found in a bone microenvironment. Embryonic or adult primary dorsal root ganglion (DRG) and MC3T3-E1 osteoblastic cells were cocultured in compartmentalized microfluidic platforms and morphological and functional tests were performed. The time of adhesion and readout of axonal outgrowth were improved by the alignment of DRG with the axis of microgrooves, which showed to be a crucial step for the designed experiments. Cocultures of entire DRG from adult origin with osteoblasts were performed, showing extended DRG projections towards the axonal compartment, reaching osteoblastic cells. Immunocytochemistry showed that the neurites present within the osteoblastic compartment were immunoreactive to synapsin and calcitonin gene-related peptide suggesting the presence of specialized structures involved in this crosstalk. This evidence was further confirmed by electron microscopy where varicosities were detected as well as electron dense structures in neurite membranes. Aiming to mimic the properties of tissue extracellular matrices, MC3T3-E1 cells were seeded in the axonal side upon laminin, collagen or within 3D functionalized alginate matrices and axonal outgrowth was clearly observed. In order to analyze and quantify data with reproducible image analysis, a semi-automated algorithm was also developed. The collagen and laminin substrates displayed a higher amount of axons reaching the axonal side. Overall, the established method revealed to be a suitable tool to study the interaction between the peripheral nervous system and bone cells in different contexts mimicking the in vivo scenario.
Starting Page	586
Ending Page	595
Page Count	10
File Format	HTM / HTML PDF
ISSN	17579694
Volume Number	6
Issue Number	6
Journal	Integrative Biology
DOI	10.1039/c4ib00035h
Language	English
Publisher	Royal Society of Chemistry
Access Restriction	Open
Subject Keyword	Immunocytochemistry Bone Homeostasis Neuron Dorsal root ganglion MC3T3 Design of experiments Synapsin Calcitonin gene-related peptide Crosstalk Electron microscope Neurite Laminin Collagen Three-dimensional space Alginic acid Algorithm Peripheral nervous system
Content Type	Text
Resource Type	Article
Subject	Medicine Biochemistry Biophysics

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