Parathyroid Hormone Secretion and Receptor Expression Determine the Age-Related Degree of Osteogenic Differentiation in Dental Pulp Stem Cells

Objective: To demonstrate the levels of parathyroid hormone secretion and genetic expressions of parathyroid hormone (PTH) and PTH1 receptor (PTH1R) genes in the dental pulp stem cells (DPSCs) from different age groups before and after induction of osteogenic differentiation. In addition, we also wanted to check their correlation with the degree of osteogenic differentiation. Methods: Human primary DPSCs from three age groups (milk tooth (SHEDs), 7–12 years old; young DPSCs (yDPSCs), 20–40 years old; old DPSCs (oDPSCs), 60+ years old) were characterized for mesenchymal stem cell (MSC) markers. DPSCs were subjected to osteogenic differentiation and functional staining. Gene expression levels were analyzed by qRT-PCR. Surface receptor analysis was done by flow cytometry. Comparative protein levels were evaluated by ELISA. Results: All SHEDs, yDPSCs, and oDPSCs were found to be expressing mesenchymal stem cell markers. SHEDs showed more mineralization than yDPSCs and oDPSCs after osteogenic induction. SHEDs exhibited higher expression of PTH and PTH1R before and after osteogenic induction, and after osteogenic induction, SHEDs showed more expression for RUNX2, ALPL, and OCN. Higher levels of PTH were observed in SHEDs and yDPSCs, and the number of PTH1R positive cells was relatively lower in yDPSCs and oDPSCs than in SHEDs. After osteogenic induction, SHEDs were superior in the secretion of OPG, and the secretions of ALPL and PTH and the number of PTH1R positive cells were relatively low in the oDPSCs. Conclusions: The therapeutic quality of dental pulp stem cells is largely based on their ability to retain their stemness characteristics. This study emphasizes the criterion of aging, which affects the secretion of PTH by these cells, which in turn attenuates their osteogenic potential.

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Exosomal circLPAR1 Promoted Osteogenic Differentiation of Homotypic Dental Pulp Stem Cells by Competitively Binding to hsa-miR-31

BioMed Research International ◽

10.1155/2020/6319395 ◽

2020 ◽

Vol 2020 ◽

pp. 1-13

Author(s):

Liangkun Xie ◽

Zheng Guan ◽

Mingzhu Zhang ◽

Sha Lyu ◽

Nattawut Thuaksuban ◽

...

Keyword(s):

Stem Cells ◽

Osteogenic Differentiation ◽

Dental Pulp ◽

Expression Profiles ◽

Circular Rna ◽

Inhibitory Effect ◽

Dental Pulp Stem Cells ◽

Great Promise ◽

Human Dental Pulp ◽

Osteogenic Induction

Human dental pulp stem cells (DPSCs) hold great promise in bone regeneration. However, the exact mechanism of osteogenic differentiation of DPSCs remains unknown, especially the role of exosomes played in. The DPSCs were cultured and received osteogenic induction; then, exosomes from osteogenic-induced DPSCs (OI-DPSC-Ex) at different time intervals were isolated and sequenced for circular RNA (circRNA) expression profiles. Gradually, increased circular lysophosphatidic acid receptor 1 (circLPAR1) expression was found in the OI-DPSC-Ex coincidentally with the degree of osteogenic differentiation. Meanwhile, results from osteogenic differentiation examinations showed that the OI-DPSC-Ex had osteogenic effect on the recipient homotypic DPSCs. To investigate the mechanism of exosomal circLPAR1 on osteogenic differentiation, we verified that circLPAR1 could competently bind to hsa-miR-31, by eliminating the inhibitory effect of hsa-miR-31 on osteogenesis, therefore promoting osteogenic differentiation of the recipient homotypic DPSCs. Our study showed that exosomal circRNA played an important role in osteogenic differentiation of DPSCs and provided a novel way of utilization of exosomes for the treatment of bone deficiencies.

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miR-217 Suppresses Osteogenic Differentiation of Human Dental Pulp Stem Cells by Down-Regulating Sirtuin 1

Journal of Biomaterials and Tissue Engineering ◽

10.1166/jbt.2020.2373 ◽

2020 ◽

Vol 10 (7) ◽

pp. 978-986

Author(s):

Haiquan Yue ◽

Yidan Guo ◽

Juan Song ◽

Ruimin Liu

Keyword(s):

Stem Cells ◽

Osteogenic Differentiation ◽

Dental Pulp ◽

Dental Pulp Stem Cells ◽

Sirtuin 1 ◽

Luciferase Reporter ◽

Induction Medium ◽

Alizarin Red ◽

Human Dental Pulp ◽

Osteogenic Induction

The paper is committed to uncovering the effect of miR-217 on osteogenic differentiation of human dental pulp stem cells (hDPSCs) and its mechanism. hDPSCs were separated from human dental pulp tissues for measurement of stemness. The osteogenic differentiation of hDPSCs was induced in an osteogenic induction medium. The hDPSCs were transfected with miR-217 mimic, miR-217 inhibitor and/or sh-SIRT1 accordingly. The expressions of miR-217 and SIRT1 were detected in hDPSCs after cell transfection and osteogenic differentiation. Calcium nodules were showed by alizarin red staining. Moreover, the expressions of osteogenic differentiation-related genes were also assessed. The binding of miR-217 to SIRT1 was predicted on starBase and further determined by dual-luciferase reporter assay. Down-regulated miR-217 and up-regulated SIRT1 were found during osteogenic differentiation of hDPSCs. The osteogenic differentiation of hDPSCs was suppressed after transfection of miR-217 mimic or sh-SIRT1 while promoted by miR-217 inhibition. Taken together, miR-217 can suppress osteogenic differentiation of hDPSCs by negatively regulating SIRT1.

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Multilineage Differentiation Potential of Human Dental Pulp Stem Cells—Impact of 3D and Hypoxic Environment on Osteogenesis In Vitro

International Journal of Molecular Sciences ◽

10.3390/ijms21176172 ◽

2020 ◽

Vol 21 (17) ◽

pp. 6172

Author(s):

Anna Labedz-Maslowska ◽

Natalia Bryniarska ◽

Andrzej Kubiak ◽

Tomasz Kaczmarzyk ◽

Malgorzata Sekula-Stryjewska ◽

...

Keyword(s):

Stem Cells ◽

Stem Cell ◽

Osteogenic Differentiation ◽

Dental Pulp ◽

Dental Pulp Stem Cells ◽

Osteogenic Potential ◽

Stem Cell Population ◽

Differentiation Potential ◽

Human Dental Pulp

Human dental pulp harbours unique stem cell population exhibiting mesenchymal stem/stromal cell (MSC) characteristics. This study aimed to analyse the differentiation potential and other essential functional and morphological features of dental pulp stem cells (DPSCs) in comparison with Wharton’s jelly-derived MSCs from the umbilical cord (UC-MSCs), and to evaluate the osteogenic differentiation of DPSCs in 3D culture with a hypoxic microenvironment resembling the stem cell niche. Human DPSCs as well as UC-MSCs were isolated from primary human tissues and were subjected to a series of experiments. We established a multiantigenic profile of DPSCs with CD45−/CD14−/CD34−/CD29+/CD44+/CD73+/CD90+/CD105+/Stro-1+/HLA-DR− (using flow cytometry) and confirmed their tri-lineage osteogenic, chondrogenic, and adipogenic differentiation potential (using qRT-PCR and histochemical staining) in comparison with the UC-MSCs. The results also demonstrated the potency of DPSCs to differentiate into osteoblasts in vitro. Moreover, we showed that the DPSCs exhibit limited cardiomyogenic and endothelial differentiation potential. Decreased proliferation and metabolic activity as well as increased osteogenic differentiation of DPSCs in vitro, attributed to 3D cell encapsulation and low oxygen concentration, were also observed. DPSCs exhibiting elevated osteogenic potential may serve as potential candidates for a cell-based product for advanced therapy, particularly for bone repair. Novel tissue engineering approaches combining DPSCs, 3D biomaterial scaffolds, and other stimulating chemical factors may represent innovative strategies for pro-regenerative therapies.

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Evaluation of the adhesion of human dental pulp stem cells to differentendodontic biomaterials before and after setting

Journal of Dental Research Dental Clinics Dental Prospects ◽

10.34172/joddd.2020.022 ◽

2020 ◽

Vol 14 (2) ◽

pp. 97-103

Author(s):

Marzie Aghazade ◽

Mohammad Samiei ◽

Marjan Imani ◽

Zahra Aghazadeh ◽

Effat Alizadeh ◽

...

Keyword(s):

Stem Cells ◽

Stem Cell ◽

Dental Pulp ◽

Culture Medium ◽

Cellular Adhesion ◽

Dental Pulp Stem Cells ◽

Treatment Modalities ◽

Root Canals ◽

Cem Cement ◽

Before And After

Background. Stem cell-based treatment modalities have been potential strategies for tissue regenerationin many conditions. Several studies have evaluated the biologic properties of DPSCs and their efficacyin the treatment of a variety of diseases. The present study was undertaken to evaluate the adhesionbehavior of DPSCs on different endodontic materials before and after setting. Methods. The crowns of the selected teeth were removed, and the root canals were prepared andobturated with gutta-percha and AH26 sealer. A retrograde cavity was prepared at root ends. Differentmaterials were placed in the cavities. Then the samples were attached to the wells with the use of achemical glue. Dental pulp stem cells were allowed to proliferate to reach a count of 2 million andtransferred to -12well plates in association with a culture medium. Finally, the samples attached to thewells were exposed to the stem cells immersed in the culture medium before and after setting. Thenadhesion of the stem cells was evaluated using SEM. Results. The SEM results showed cellular adhesion in the samples containing CEM cement both beforeand after setting. The samples containing MTA Angelus and ProRoot MTA exhibited cellular adhesionbefore setting, with no cellular adhesion after setting. The samples containing AH26 and MTA Fillapexsealers exhibited cellular adhesion after setting, with no adhesion before setting. The samples containingsimvastatin exhibited no cellular adhesion before setting; this material had dissolved in the culturemedium after setting evaluation. Conclusion. The results of the present study showed that of all the materials tested, CEM cement hadthe highest capacity for dental pulp stem cell adhesion.

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Improved osteogenic differentiation of human dental pulp stem cells in a layer-by-layer-modified gelatin scaffold

Journal of Biomaterials Applications ◽

10.1177/0885328218799162 ◽

2018 ◽

Vol 33 (4) ◽

pp. 477-487 ◽

Cited By ~ 7

Author(s):

Qiang Fu ◽

Huaijuan Ren ◽

Chen Zheng ◽

Chao Zhuang ◽

Tong Wu ◽

...

Keyword(s):

Stem Cells ◽

Stem Cell ◽

Osteogenic Differentiation ◽

Dental Pulp ◽

Dental Pulp Stem Cells ◽

Gelatin Sponge ◽

Layer By Layer ◽

Lysine Modification

Dental pulp stem cell is a new type of mesenchymal stem cell that has a potential for tissue regeneration. Gelatin sponges are often used for hemostasis in dental surgery. In this study, we aimed to evaluate the dental pulp stem cells’ proliferation and osteogenic differentiation in different layer-by-layer-modified gelatin sponge scaffolds including the G, G + P (gelatin sponge+ poly-l-lysine modification), G + M (gelatin sponge + mineralization modification), and G + M + P (gelatin sponge + mineralization modification + poly-l-lysine modification) groups in vitro and assessed them in vivo. The results showed that dental pulp stem cells had a great potential for osteogenic differentiation. In vitro, the G + M + P group not only enhanced the adhesion and proliferation of dental pulp stem cells but also facilitated their osteogenic differentiation. However, alkaline phosphatase activity was prohibited after modification. In vivo, both dental pulp stem cells and cells from nude mice grew well on the scaffold, and G + M and G + M + P groups could promote the mineralization deposit formation and the expression of osteocalcin in osteogenic differentiation of dental pulp stem cells. In conclusion, the combination of dental pulp stem cells and G + M + P scaffold has a great potential for bone tissue engineering.

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