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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References
- Roles of innervation in developing and regenerating orofacial tissues.Cell Mol Life Sci. 2014; 71: 2241-2251
- The neural stem cell microenvironment.in: StemBook. Harvard Stem Cell Institute, Cambridge, MA2008
- The wound microenvironment reprograms schwann cells to invasive mesenchymal-like cells to drive peripheral nerve regeneration.Neuron. 2017; 96: 98-114
- Dental pulp cells produce neurotrophic factors, interact with trigeminal neurons in vitro, and rescue motoneurons after spinal cord injury.Dev Biol. 2001; 238: 120-132
- Small molecules enhance neurogenic differentiation of dental-derived adult stem cells.Arch Oral Biol. 2019; 102: 26-38
- Characterization of stem cells from pulp tissue of exfoliated deciduous teeth, impacted third molars and apical papilla of impacted third molars with open apices: a comparative analysis.J Adv Oral Res. 2019; 10: 97-104
- Stem cells of the apical papilla regulate trigeminal neurite outgrowth and targeting through a BDNF-dependent mechanism.Tissue Eng A. 2014; 20: 3089-3100
- Pulp Stem Cell-Mediated Functional Pulp Regeneration.J Dent Res. 2019; 98: 27-35
- Changes in phenotype and differentiation potential of human mesenchymal stem cells aging in vitro.Stem Cell Res Ther. 2018; 9: 131
- Dental mesenchymal stromal/stem cells in different microenvironments- implications in regenerative therapy.World J Stem Cells. 2021; 13: 1863-1880
- Apical Papilla Cells Are Capable of Forming a Pulplike Tissue with Odontoblastlike Cells without the Use of Exogenous Growth Factors.J Endod. 2018; 44: 1671-1676
- Dental neuroplasticity, neuro-pulpal interactions, and nerve regeneration.Microsc Res Tech. 2003; 60: 503-515
- Pre-innervated tissue-engineered muscle promotes a pro-regenerative microenvironment following volumetric muscle loss.Commun Biol. 2020; 3: 330
- Scaffold-free prevascularized microtissue spheroids for pulp regeneration.J Dent Res. 2014; 93: 1296-1303
- EphrinB2/EphB4 Signaling Regulates DPSCs to Induce Sprouting Angiogenesis of Endothelial Cells.J Dent Res. 2019; 98: 803-812
- Different effects of implanting sensory nerve or blood vessel on the vascularization, neurotization, and osteogenesis of tissue-engineered bone in vivo.Biomed Res Int. 2014; 2014: 412570
- Conversion of stem cells from apical papilla into endothelial cells by small molecules and growth factors.Stem Cell Res Ther. 2021; 12: 266
- Dental apical papilla as therapy for spinal cord injury.J Dent Res. 2015; 94: 1575-1581
- Spheroid culture system methods and applications for mesenchymal stem cells.Cells. 2019; 8: 1620
- Natural biomaterials as instructive engineered microenvironments that direct cellular function in peripheral nerve tissue engineering.Front Bioeng Biotech. 2021; 9: 674473
- Cell migration into scaffolds under co-culture conditions in a microfluidic platform.Lab Chip. 2009; 9: 269-275
- Biomaterial and Biofilm Interactions with the Pulp-Dentin Complex-On-A-Chip.J Dent Res. 2021; 100: 1136-1143
- Mesenchymal stem cell-mediated functional tooth regeneration in swine.PLoS One. 2006; 1: e79
- Characterization of the apical papilla and its residing stem cells from human immature permanent teeth: a pilot study.J Endod. 2008; 34: 166-171
- Biomimetic organization of collagen matrices to template bone-like microstructures.Matrix Biol. 2016; 52-54: 384-396
- Hydroxyapatite nanoparticle-containing scaffolds for the study of breast cancer bone metastasis.Biomaterials. 2011; 32: 5112-5122
- Neural tissue engineering: the influence of scaffold surface topography and extracellular matrix microenvironment.J Mater Chem B. 2021; 9: 567-584
- 3D neural tissue models: from spheroids to bioprinting.Biomaterials. 2018; 154: 113-133
- Neurogenic maturation of human dental pulp stem cells following neurosphere generation induces morphological and electrophysiological characteristics of functional neurons.Stem Cells Dev. 2015; 24: 296-311
- MSC spheroids-loaded collagen hydrogels simultaneously promote neuronal differentiation and suppress inflammatory reaction through PI3K-Akt signaling pathway.Biomaterials. 2021; 265: 120448
- Three-dimensional brain-on-a-chip with an interstitial level of flow and its application as an in vitro model of Alzheimer's disease.Lab Chip. 2015; 15: 141-150
- A 3D neurovascular microfluidic model consisting of neurons, astrocytes and cerebral endothelial cells as a blood-brain barrier.Lab Chip. 2017; 17: 448-459
- Neurotrophic Proteins in Dentin and Their Effect on Trigeminal Sensory Neurons.J Endod. 2019; 45: 729-735
- Transplantation of Human Dental Pulp-Derived Stem Cells or Differentiated Neuronal Cells from Human Dental Pulp-Derived Stem Cells Identically Enhances Regeneration of the Injured Peripheral Nerve.Stem Cells Dev. 2017; 26: 1247-1257
- Impact of tissue-specific stem cells on lineage-specific differentiation: a focus on the musculoskeletal system.Stem Cell Rev Rep. 2015; 11: 119-132
- Small molecules efficiently reprogram apical papilla stem cells into neuron-like cells.Exp Ther Med. 2021; 21: 546
- Epiregulin promotes osteogenic differentiation and inhibits neurogenic trans-differentiation of adipose-derived mesenchymal stem cells via MAPKs pathway.Cell Biol Int. 2020; 44: 1046-1058
- Differentiation-dependent secretion of proangiogenic factors by mesenchymal stem cells.PLoS One. 2012; 7: e35579
Article info
Publication history
Published online: September 19, 2022
Identification
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© 2022 American Association of Endodontists.