Rho kinase inhibitor induced human dental pulp stem cells to differentiate into neurons

Srikawnawan, Wittawas; Songsaad, Anupong; Gonmanee, Thanasup; Thonabulsombat, Charoensri; Phruksaniyom, Chareerut; White, Kenneth L.; Ruangsawasdi, Nisarat

doi:10.1016/j.lfs.2022.120566

Cited by 5 publications

(6 citation statements)

References 43 publications

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“…In contrast, steady baseline patterns of low intensity were observed in the hSCAPs used as the negative control. Since these findings on intracellular calcium oscillation were consistent with previous studies that presented the functional profilings of neuronal cells derived from hDPSCs 17 , 35 , 52 and of hiPSCs derived from dentate gyrus neuronal progenitors, 53 the intra-neurospheral cells analyzed in the present study were proven to be functional neuronal cells. However, further investigation with electrophysiological tests and neurogenic maturation analyses is still necessary to thoroughly characterize the neuronal profile of these cells.…”

Section: Discussionsupporting

confidence: 92%

“… 31 Second, it was found that the Nissl body, an intensely basophilic granular consisting of a rough endoplasmic reticulum, was only present in neurons. 34 In vitro -differentiated neuronal cells derived from hSCAPs 7 and hDPSCs 35 were recently shown to present Nissl bodies, which were found through a staining process with Cresyl violet dye. Moreover, the hippocampal area of a rat model exhibited the organization of cells with Nissl bodies, which were characterized as neurons.…”

Section: Discussionmentioning

confidence: 99%

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Characterization of neural stem cells derived from human stem cells from the apical papilla undergoing three-dimensional neurosphere induction

SONGSAAD,

THAIRAT,

SEEMAUNG

et al. 2023

J. Appl. Oral Sci.

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Objectives The endogenous repairing based on the activation of neural stem cells (NSCs) is impaired by neurodegenerative diseases. The present study aims to characterize human stem cells from the apical papilla (hSCAPs) with features of mesenchymal stem cells (MSCs) and to demonstrate the neuronal differentiation of hSCAPs into NSCs through the formation of three-dimensional (3D) neurospheres, verifying the structural, immunophenotyping, self-renewal, gene expression and neuronal activities of these cells to help further improve NSCs transplantation. Methodology The hSCAPs were isolated from healthy impacted human third molar teeth and characterized as MSCs. They were then induced into 3D-neurospheres using a specific neural induction medium. Subsequently, the intra-neurospheral cells were confirmed to be NSCs by the identification of Nissl substance and the analysis of immunofluorescence staining, self-renewal ability, and gene expression of the cells. Moreover, the neuronal activity was investigated using intracellular calcium oscillation. Results The isolated cells from the human apical papilla expressed many markers of MSCs, such as self-renewal ability and multilineage differentiation. These cells were thus characterized as MSCs, specifically as hSCAPs. The neurospheres induced from hSCAPs exhibited a 3D-floating spheroidal shape and larger neurospheres, and consisted of a heterogeneous population of intra-neurospheral cells. Further investigation showed that these intra-neurospheral cells had Nissl body staining and also expressed both Nestin and SOX2. They presented a self-renewal ability as well, which was observed after their disaggregation. Their gene expression profiling also exhibited a significant amount of NSC markers ( NES , SOX1 , and PAX6 ). Lastly, a large and dynamic change of the fluorescent signal that indicated calcium ions (Ca 2+ ) was detected in the intracellular calcium oscillation, which indicated the neuronal activity of NSCs-derived hSCAPs. Conclusions The hSCAPs exhibited properties of MSCs and could differentiate into NSCs under 3D-neurosphere generation. The present findings suggest that NSCs-derived hSCAPs may be used as an alternative candidates for cell-based therapy, which uses stem cell transplantation to further treat neurodegenerative diseases.

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Section: Discussionsupporting

confidence: 92%

Section: Discussionmentioning

confidence: 99%

Characterization of neural stem cells derived from human stem cells from the apical papilla undergoing three-dimensional neurosphere induction

SONGSAAD,

THAIRAT,

SEEMAUNG

et al. 2023

J. Appl. Oral Sci.

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“…Previous studies have demonstrated DPSCs not only can express several typical neural stem/progenitor markers under the specific stimulation, but also differentiate toward phenotypes of functionally active neurons, with the higher expression of neurogenin-2, neuron-specific enolase, neurofilament-M and glial fibrillary acidic protein, and activation of voltage-gated sodium and potassium channels ( Luzuriaga et al, 2021 ). Multiple stimulations have been used to differentiate DPSCs into neuron-like cells, involving several differentiating protocols containing complex mixture of supplements ( Luzuriaga et al, 2019b ), optogenetics ( Niyazi et al, 2020 ), activation of K+ channels ( Kogo et al, 2020 ), and Rho kinase inhibitor ( Srikawnawan et al, 2022 ). Darvishi et al demonstrated human DPSCs can differentiate to functional motor neuron in organotypic culture system containing growth factors in two stages ( Darvishi et al, 2021 ).…”

Section: The Potential Mechanisms Of Dpscs In Scimentioning

confidence: 99%

The potential therapeutic roles of dental pulp stem cells in spinal cord injury

Fu,

Li,

Mao

et al. 2024

Front. Mol. Biosci.

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Spinal cord injury (SCI) can lead to serious functional disorders, which have serious impacts on patients and society. The current traditional treatments of SCI are not effective the injured spinal cord is difficult to repair and regenerate. In recent years, stem cell transplantation for the treatment of SCI has been a hot research topic. Dental pulp stem cells have strong abilities of self-renewal and multi-directional differentiation, and have been applied for tissue engineering and regenerative medicine. And dental pulp stem cells have certain advantages in neuro-regenetation, bringing new hope to biotherapy for SCI. This article reviews the characteristics of dental pulp stem cells and their research progress in the treatment of SCI.

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“…Zhao et al [ 10 ] demonstrated that mesenchymal stem cell (MSC) favoring dentin regeneration originates from periarterial cells and is regulated by Shh secreted from the neurovascular bundle. Current research on sensory nerve regeneration has focused on the potential for MSC differentiation into central neural stem or neuronal cells [ 11 ]; however, the dental pulp sensory nerves originate from peripheral nerves, namely the trigeminal nerve maxillary and mandibular branches [ 12 ]. Schwann cells (SCs) are critical for nerve regeneration, including degenerated axon, myelin debris, and inhibitory factor elimination at the distal end, and provide vital topographical cues and specific proteins to guide axon elongation in bridging the interstump gap [ 13 , 14 ].…”

Section: Introductionmentioning

confidence: 99%

The Effect of Differentiation of Human Dental Pulp Stem Cells to Glial Cells on the Sensory Nerves of the Dental Pulp

Liu,

Zou,

Liu

et al. 2024

International Journal of Dentistry

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Regeneration of sensory nerves is challenging in dental pulp regeneration. Schwann cells (SCs) are essential glial cells conducive to regenerating sensory nerve, but their source is scarce. The aim of the protocol was to investigate the regenerative potential of Schwann-like cells derived from dental pulp stem cells (SC-DPSCs) for sensory nerve regrowth. SC-DPSCs were generated from dental pulp stem cells using a three-step protocol. The expression of key markers, including myelin basic protein, S-100, and p75 neurotrophin receptor, was analyzed. Primary trigeminal neurons were cultured, and the expression of neurofilament 200, β-tubulin III, and microtubule-associated protein 2 was assessed. Simultaneous culture experiments were conducted to evaluate trigeminal neuron growth in the presence of SC-DPSCs. In addition, mRNA sequencing was performed to identify key genes involved in the differentiation process, highlighting prostaglandin-endoperoxide synthase 2 (PTGS2) as a potential candidate. The results demonstrated that SC-DPSCs expressed characteristic SCs markers and facilitated axonal growth in rat trigeminal nerves. Differentiated SC-DPSCs secreted elevated levels of nerve growth factors, including brain-derived neurotrophic factor and neurotrophin-3, promoting the growth of trigeminal nerve axons. These findings suggest the regenerative potential of SC-DPSCs in dentin–dental pulp complex; PTGS2 is considered a crucial gene in this differentiation process.

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Rho kinase inhibitor induced human dental pulp stem cells to differentiate into neurons

Cited by 5 publications

References 43 publications

Characterization of neural stem cells derived from human stem cells from the apical papilla undergoing three-dimensional neurosphere induction

Characterization of neural stem cells derived from human stem cells from the apical papilla undergoing three-dimensional neurosphere induction

The potential therapeutic roles of dental pulp stem cells in spinal cord injury

The Effect of Differentiation of Human Dental Pulp Stem Cells to Glial Cells on the Sensory Nerves of the Dental Pulp

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