Endothelial-Initiated Crosstalk Regulates Dental Pulp Stem Cell Self-Renewal

Interactions with the microenvironment modulate the fate of stem cells in perivascular niches in tissues (e.g., bone) and organs (e.g., liver). However, the functional relevance of the molecular crosstalk between endothelial cells and stem cells within the perivascular niche in dental pulps is unclear. Here, we tested the hypothesis that endothelial cell–initiated signaling is necessary to maintain self-renewal of dental pulp stem cells. Confocal microscopy showed that ALDH1high and Bmi-1high stem cells are preferentially localized in close proximity to blood vessels in physiological human dental pulps. Secondary orosphere assays revealed that endothelial cell–derived factors (e.g., interleukin-6 [IL-6]) promote self-renewal of dental pulp stem cells cultured in low-attachment conditions. Mechanistic studies demonstrated that endothelial cell–derived IL-6 activates IL-6R (IL-6 Receptor) and signal transducer and activator of transcription 3 (STAT3) signaling and induces expression of Bmi-1 (master regulator of stem cell self-renewal) in dental pulp stem cells. Transplantation of dental pulp stem cells stably transduced with small hairpin RNA (shRNA)–STAT3 into immunodeficient mice revealed a decrease in the number of blood vessels surrounded by ALDH1high or Bmi-1high cells (perivascular niches) compared to tissues formed upon transplantation of vector control stem cells. And finally, in vitro capillary sprouting assays revealed that inhibition of IL-6 or STAT3 signaling decreases the vasculogenic potential of dental pulp stem cells. Collectively, these data demonstrate that endothelial cell–derived IL-6 enhances the self-renewal of dental pulp stem cells via STAT3 signaling and induction of Bmi-1. These data suggest that a crosstalk between endothelial cells and stem cells within the perivascular niche is required for the maintenance of stem cell pools in dental pulps.

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Synergy of molecularly mobile polyrotaxane surfaces with endothelial cell co-culture for mesenchymal stem cell mineralization

RSC Advances ◽

10.1039/d1ra01296g ◽

2021 ◽

Vol 11 (30) ◽

pp. 18685-18692

Author(s):

Hiroki Masuda ◽

Yoshinori Arisaka ◽

Masahiro Hakariya ◽

Takanori Iwata ◽

Tetsuya Yoda ◽

...

Keyword(s):

Stem Cells ◽

Endothelial Cells ◽

Mesenchymal Stem Cells ◽

Stem Cell ◽

Endothelial Cell ◽

Mesenchymal Stem Cell ◽

Molecular Mobility ◽

Culture System

Molecular mobility of polyrotaxane surfaces promoted mineralization in a co-culture system of mesenchymal stem cells and endothelial cells.

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The Interplay of Dental Pulp Stem Cells and Endothelial Cells in an Injectable Peptide Hydrogel on Angiogenesis and Pulp Regeneration In Vivo

Tissue Engineering Part A ◽

10.1089/ten.tea.2014.0154 ◽

2015 ◽

Vol 21 (3-4) ◽

pp. 550-563 ◽

Cited By ~ 77

Author(s):

Waruna Lakmal Dissanayaka ◽

Kenneth M. Hargreaves ◽

Lijian Jin ◽

Lakshman P. Samaranayake ◽

Chengfei Zhang

Keyword(s):

Stem Cells ◽

Endothelial Cells ◽

Dental Pulp ◽

Dental Pulp Stem Cells ◽

Pulp Regeneration ◽

Peptide Hydrogel

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Biodistribution of Intramuscularly-Transplanted Human Dental Pulp Stem Cells in Immunocompetent Healthy Rats through NIR Imaging

Cells Tissues Organs ◽

10.1159/000511569 ◽

2020 ◽

pp. 1-12

Author(s):

Pradnya Shahani ◽

Alka Kaushal ◽

Girish Waghmare ◽

Indrani Datta

Keyword(s):

Gene Expression ◽

Stem Cells ◽

Clinical Trials ◽

Blood Vessels ◽

Dental Pulp ◽

Signal Intensity ◽

Dental Pulp Stem Cells ◽

European Medicines Agency ◽

Rat Tissues ◽

Cell Based Therapy

Owing to their neural crest origin, dental pulp stem cells (DPSCs) are increasingly gaining prominence in treating nervous system disease conditions. However, as per the regulatory bodies [European-Medicines Agency (EMA), Indian-Council of Medical-Research (ICMR)], their biodistribution after transplantation needs to be evaluated for them to be considered for cell-based therapy for clinical trials. There are yet no studies describing the dynamic distribution of human origin DPSCs (hDPSCs) after transplantation in an immunocompetent, physiologically healthy animal model. Here, using near-infrared (NIR)-based whole animal and ex vivo tissue imaging, we assessed the biodistribution of intramuscularly transplanted hDPSCs in immunocompetent healthy Wistar rats. Further validation was done by quantifying gene expression of the human Alu gene in rat tissues. After 24 h of transplantation, an increase in signal intensity and area of signal was observed in the muscle of administration compared to 30 min and 6 h. At hour 24, neither increase in human Alu nor human Ki67 gene expression was seen in the rat muscle, thus confirming that the increase in signal area and intensity at hour 24 was not due to proliferation of the transplanted cells. Rather at hour 24, the NIR-signal intensity in bone marrow increased, suggesting that the NIR-tagged DPSCs have started entering into the blood vessels adjacent to the muscle, and the blood vessels being placed just beneath the subcutaneous layer might be responsible for an increase in signal intensity. Signal intensity increased distinctly in all organs at this timepoint, confirming that the cells entered the bloodstream by hour 24. Lung entrapment of DPSCs was not observed, since signal intensity was least in lungs as compared to the site of injection. Cells were retained for up to 28 days at the site of injection. These findings lay the basis to design the dosage for intramuscular delivery of hDPSCs for degenerative disease models and for future clinical trials.

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Differentiation of CD31-Positive Vascular Endothelial Cells from Organoid Culture of Dental Pulp Stem Cells

International Journal of Oral Biology ◽

10.11620/ijob.2018.43.2.077 ◽

2018 ◽

Vol 43 (2) ◽

pp. 77-82

Author(s):

Eun Jin Seo ◽

◽

Jae Kyung Park ◽

Hoim Jeong ◽

Jung Sook Kang ◽

...

Keyword(s):

Stem Cells ◽

Endothelial Cells ◽

Dental Pulp ◽

Vascular Endothelial Cells ◽

Dental Pulp Stem Cells ◽

Vascular Endothelial ◽

Organoid Culture

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Characterization of Stable Hypoxia-Preconditioned Dental Pulp Stem Cells: Implication for Pulp Regeneration

10.21203/rs.3.rs-102950/v1 ◽

2020 ◽

Author(s):

Mohammed Zayed ◽

Koichiro Iohara ◽

Hideto Watanabe ◽

Mami Ishikawa ◽

Michiyo Tominaga ◽

...

Keyword(s):

Stem Cells ◽

Stem Cell ◽

Dental Pulp ◽

Clinical Utility ◽

Dental Pulp Stem Cells ◽

Proliferation Rate ◽

Stem Cell Marker ◽

Stem Cell Markers ◽

Pulp Tissue ◽

Pulp Regeneration

Abstract Background: Dental pulp stem cells (DPSCs) have been developed as a potential source of mesenchymal stem cells (MSCs) for regeneration of dental pulp and other tissues. However, further strategies to isolate highly functional DPSCs beyond the colony-forming methods are required. Our clinical study has demonstrated safety and efficacy of DPSCs isolated by G-CSF-induced mobilization and cultured under normoxia (mobilized DPSCs, MDPSCs) for pulp regeneration. It is well known that the oxygen concentration is closely linked to the maintenance of stemness. Thus, in this investigation, hypoxia-preconditioned DPSCs (hpDPSCs) was characterized to develop and improve the clinical utility for regeneration of dental pulp in endodontics.Methods: Colony-forming DPSCs were isolated and preconditioned with hypoxia in a stable closed cultured system and compared with MDPSCs isolated from the individual dog teeth. We examined the proliferation rate, migration potential, anti-apoptotic activity and gene expression of the stem cell markers and angiogenic/neurotrophic factors. Trophic effects of the conditioned medium (CM) were also evaluated. In addition, the expression of immunomodulatory molecules upon stimulation with IFN-γ were investigated. The pulp regenerative potential and transplantation safety of hpDPSCs were further assessed in pulpectomized teeth in dogs by histological and immunohistochemical analyses and by chemistry of blood and urine. tests Results: hpDPSCs demonstrated higher proliferation rate and expression of a major regulator of oxygen homeostasis, HIF-1α, and a stem cell marker, CXCR-4. The direct migratory activity of hpDPSCs in response to G-CSF was significantly higher than MDPSCs. The CM of hpDPSCs stimulated neurite extension. However, there were no changes in angiogenic, migration and anti-apoptotic activities compared with the CM of MDPSCs. The expression of immunomodulatory gene, PTGE was significantly up-regulated by IFN gamma in hpDPSCs compared with MDPSCs. However, no difference in nitric oxide was observed. The regenerated pulp tissue was quantitatively and qualitatively similar in hpDPSC transplants compared with MDPSC transplants in dog teeth. There was no evidence of toxicity or adverse events of the hpDPSC transplantation Conclusions: These results demonstrated that hpDPSCs improved stem cell properties compared to MDPSCs, suggesting their potential clinical utility for pulp regeneration.

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Chitlac-coated Thermosets Enhance Osteogenesis and Angiogenesis in a Co-culture of Dental Pulp Stem Cells and Endothelial Cells

Nanomaterials ◽

10.3390/nano9070928 ◽

2019 ◽

Vol 9 (7) ◽

pp. 928 ◽

Cited By ~ 11

Author(s):

Rapino ◽

Di Valerio ◽

Zara ◽

Gallorini ◽

Marconi ◽

...

Keyword(s):

Stem Cells ◽

Endothelial Cells ◽

Dental Pulp ◽

Cellular Response ◽

Cell Types ◽

Culture Conditions ◽

Dental Pulp Stem Cells ◽

Alizarin Red ◽

Dental Restorations ◽

Diphenyl Tetrazolium Bromide

Dental pulp stem cells (DPSCs) represent a population of stem cells which could be useful in oral and maxillofacial reconstruction. They are part of the periendothelial niche, where their crosstalk with endothelial cells is crucial in the cellular response to biomaterials used for dental restorations. DPSCs and the endothelial cell line EA.hy926 were co-cultured in the presence of Chitlac-coated thermosets in culture conditions inducing, in turn, osteogenic or angiogenic differentiation. Cell proliferation was evaluated by 3–[4,5–dimethyl–thiazol–2–yl–]–2,5–diphenyl tetrazolium bromide (MTT) assay. DPSC differentiation was assessed by measuring Alkaline Phosphtase (ALP) activity and Alizarin Red S staining, while the formation of new vessels was monitored by optical microscopy. The IL-6 and PGE2 production was evaluated as well. When cultured together, the proliferation is increased, as is the DPSC osteogenic differentiation and EA.hy926 vessel formation. The presence of thermosets appears either not to disturb the system balance or even to improve the osteogenic and angiogenic differentiation. Chitlac-coated thermosets confirm their biocompatibility in the present co-culture model, being capable of improving the differentiation of both cell types. Furthermore, the assessed co-culture appears to be a useful tool to investigate cell response toward newly synthesized or commercially available biomaterials, as well as to evaluate their engraftment potential in restorative dentistry.

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PD-1 is required to maintain stem cell properties in human dental pulp stem cells

Cell Death and Differentiation ◽

10.1038/s41418-018-0077-8 ◽

2018 ◽

Vol 25 (7) ◽

pp. 1350-1360 ◽

Cited By ~ 10

Author(s):

Yao Liu ◽

Huan Jing ◽

Xiaoxing Kou ◽

Chider Chen ◽

Dawei Liu ◽

...

Keyword(s):

Stem Cells ◽

Stem Cell ◽

Dental Pulp ◽

Dental Pulp Stem Cells ◽

Human Dental Pulp

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Viability of Quercetin-Induced Dental Pulp Stem Cells in Forming Living Cellular Constructs for Soft Tissue Augmentation

Journal of Personalized Medicine ◽

10.3390/jpm11050430 ◽

2021 ◽

Vol 11 (5) ◽

pp. 430

Author(s):

Hytham N. Fageeh ◽

Shilpa Bhandi ◽

Mohammed Mashyakhy ◽

Ahmed Al Kahtani ◽

Zahi Badran ◽

...

Keyword(s):

Stem Cells ◽

Stem Cell ◽

Soft Tissue ◽

Dental Pulp ◽

Donor Site ◽

Dental Pulp Stem Cells ◽

Soft Tissue Augmentation ◽

Background Information ◽

Tissue Augmentation ◽

Dental Pulp Stem Cell

Autogenous gingival grafts used for root coverage or gingival augmentation procedures often result in donor site morbidity. Living cellular constructs as an exogenous alternative have been proven to be associated with lower morbidity. With the available background information, the present study aims to assess if quercetin-induced living cell constructs, derived from dental pulp stem cells, have the potential to be applied as a tool for soft tissue augmentation. The characterized dental pulp stem cells (positive for CD73, CD90, and negative for CD34, HLA-DR) were expanded in Dulbecco’s Modified Eagle’s medium (DMEM) supplemented with 10 mM quercetin. The handling properties of the quercetin-induced dental pulp stem cell constructs were assessed by visual, and tactile sensation. A microscopic characterization using hematoxylin and eosin staining, and qRT-PCR-based analysis for stemness-associated genes (OCT4, NANOG, SOX2, and cMyc) was also performed. Dental pulp stem cells without quercetin administration were used as the control. Dental pulp stem cell constructs induced by quercetin easily detached from the surface of the plate, whereas there was no formation in the control cells. It was also simple to transfer the induced cellular construct on the flattened surface. Microscopic characterization of the constructs showed cells embedded in a tissue matrix. Quercetin also increased the expression of stemness-related genes. The use of quercetin-induced DPSC living constructs for soft tissue augmentation could provide an alternative to autogenous soft tissue grafts to lower patient morbidity and improve esthetic outcomes.

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