Abstract
Introduction
Stem cells from apical papillae (SCAPs) display potent neuroregenerative potential
by secreting neurotrophic factors and differentiating into neuronal-like cells, which
are regulated by local niches. This study aimed to explore the interactions between
SCAP-derived neuronal cell spheroids and SCAPs under various microenvironments in
a pulp-on-a-chip system.
Methods
SCAPs were induced into neuronal cells by a cocktail of chemical molecules. The expression
of neuronal markers was detected by real-time quantitative polymerase chain reaction,
Western blotting, and immunofluorescence. SCAP-derived neuronal cell spheroids were
generated with micromolded agarose gels and embedded in collagen hydrogel. After characterization,
the SCAP-derived neuronal cell spheroids were cocultured with SCAPs in a pulp-on-chip
system under various microenvironments: α-Minimum Essential Medium (MEM)/neuronal
maturation medium (NMM) (neurosphere only); undifferentiated SCAPs with α-MEM/NMM
(neurosphere + SCAP); osteogenic-induced SCAPs (Oi-SCAPs) with osteogenic medium/NMM
containing 0.02% hydroxyapatite nanoparticles (neurosphere + Oi-SCAP/hydroxyapatite
nanoparticle); and Oi-SCAPs with α-MEM/NMM (neurosphere + Oi-SCAP). Neurite outgrowth
and MAP2 and TUJ1 expression in SCAP-derived neuronal cell spheroids were assessed
by immunostaining.
Results
SCAPs were efficiently chemically induced into neuronal-like cells. The expression
levels of MAP2 and TUJ1 were upregulated in SCAP-derived neuronal cell spheroids when
cocultured with undifferentiated SCAPs, followed by Oi-SCAPs with α-MEM and neurospheres
cultured alone, while coculturing with Oi-SACPs in osteogenic medium/NMM with hydroxyapatite
nanoparticles displayed the lowest expression. Neurite spreading was more prominent
in the spheroids cocultured with undifferentiated SCAPs than in those of the other
3 groups.
Conclusion
This work revealed that local microenvironments critically regulate the neuroregenerative
potential of SCAP-derived neuronal cell spheroids.
Key Words
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Article info
Publication history
Published online: September 19, 2022
Identification
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© 2022 American Association of Endodontists.
