Extracellular matrix stiffness negatively affects axon elongation, growth cone area and F-actin levels in a collagen type I 3D culture.
Col-I 3D matrices
axonal cytoskeleton
axonal growth
mechanical properties
stiffness
sympathetic axon
three-dimensional neuronal culture
Journal
Journal of tissue engineering and regenerative medicine
ISSN: 1932-7005
Titre abrégé: J Tissue Eng Regen Med
Pays: England
ID NLM: 101308490
Informations de publication
Date de publication:
02 2022
02 2022
Historique:
revised:
10
11
2021
received:
12
08
2021
accepted:
17
11
2021
pubmed:
25
11
2021
medline:
5
4
2022
entrez:
24
11
2021
Statut:
ppublish
Résumé
Three dimensional (3D) in vitro neuronal cultures can better reproduce physiologically relevant phenotypes compared to 2D-cultures, because in vivo neurons reside in a 3D microenvironment. Interest in neuronal 3D cultures is emerging, with special attention to the mechanical forces that regulate axon elongation and sprouting in three dimensions. Type I collagen (Col-I) is a native substrate since it is present in the extracellular matrix and hence emulates an in vivo environment to study axon growth. The impact of its mechanical properties needs to be further investigated. Here, we generated Col-I 3D matrices of different mechanical stiffness and evaluated axon growth in three dimensions. Superior cervical ganglion (SCG) explants from neonatal rats were cultured in soft and stiff Col-I 3D matrices and neurite outgrowth was assessed by measuring: maximum neuritic extent; neuritic halo area and fasciculation. Axonal cytoskeletal proteins were examined. Axon elongation in stiff Col-I 3D matrices was reduced (31%) following 24 h in culture compared to soft matrices. In stiff matrices, neurites fasciculated and formed less dense halos. Consistently, almost no F-actin rich growth cones were recognized, and F-actin staining was strongly reduced in the axonal compartment. This study shows that stiffness negatively affects 3D neurite outgrowth and adds insights on the cytoskeletal responses upon mechanic interactions of axons with a 3D environment. Our data will serve to facilitate the development of model systems that are mechanically well-behaved but still mimic key physiologic properties observed in vivo.
Substances chimiques
Actins
0
Collagen Type I
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
151-162Subventions
Organisme : Programa de Desarrollo de Ciencias Básicas (PEDECIBA)
Organisme : ComisiÃ3n Académica de Posgrado (CAP)
Organisme : Sociedad Uruguaya de Biociencias
Informations de copyright
© 2021 John Wiley & Sons Ltd.
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