Enhanced Osteogenesis of Dental Pulp Stem Cells In Vitro Induced by Chitosan–PEG-Incorporated Calcium Phosphate Cement

The use of bone graft materials is required for the treatment of bone defects damaged beyond the critical defect; therefore, injectable calcium phosphate cement (CPC) is actively used after surgery. The application of various polymers to improve injectability, mechanical strength, and biological function of injection-type CPC is encouraged. We previously developed a chitosan–PEG conjugate (CS/PEG) by a sulfur (VI) fluoride exchange reaction, and the resulting chitosan derivative showed high solubility at a neutral pH. We have demonstrated the CPC incorporated with a poly (ethylene glycol) (PEG)-grafted chitosan (CS/PEG) and developed CS/PEG CPC. The characterization of CS/PEG CPC was conducted using Fourier transform infrared spectroscopy (FT-IR) and X-ray diffraction (XRD). The initial properties of CS/PEG CPCs, such as the pH, porosity, mechanical strength, zeta potential, and in vitro biocompatibility using the WST-1 assay, were also investigated. Moreover, osteocompatibility of CS/PEG CPCs was carried out via Alizarin Red S staining, immunocytochemistry, and Western blot analysis. CS/PEG CPC has enhanced mechanical strength compared to CPC, and the cohesion test also demonstrated in vivo stability. Furthermore, we determined whether CS/PEG CPC is a suitable candidate for promoting the osteogenic ability of Dental Pulp Stem Cells (DPSC). The elution of CS/PEG CPC entraps more calcium ion than CPC, as confirmed through the zeta potential test. Accordingly, the ion trapping effect of CS/PEG is considered to have played a role in promoting osteogenic differentiation of DPSCs. The results strongly suggested that CS/PEG could be used as suitable additives for improving osteogenic induction of bone substitute materials.

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Effect of Vitamins D and E on the Proliferation, Viability, and Differentiation of Human Dental Pulp Stem Cells: An In Vitro Study

International Journal of Dentistry ◽

10.1155/2020/8860840 ◽

2020 ◽

Vol 2020 ◽

pp. 1-10

Author(s):

Lina M. Escobar ◽

Zita Bendahan ◽

Andrea Bayona ◽

Jaime E. Castellanos ◽

María-Clara González

Keyword(s):

Stem Cells ◽

Cell Proliferation ◽

Dental Pulp ◽

Morphological Changes ◽

In Vitro Study ◽

Osteoblastic Differentiation ◽

Dental Pulp Stem Cells ◽

Alizarin Red ◽

Human Dental Pulp

Introduction. The aim of the present study was to determine the effects of vitamins D and E on the proliferation, morphology, and differentiation of human dental pulp stem cells (hDPSCs). Methods. In this in vitro experimental study, hDPSCs were isolated, characterized, and treated with vitamins D and E, individually and in combination, utilizing different doses and treatment periods. Changes in morphology and cell proliferation were evaluated using light microscopy and the resazurin assay, respectively. Osteoblast differentiation was evaluated with alizarin red S staining and expression of RUNX2, Osterix, and Osteocalcin genes using real-time RT-PCR. Results. Compared with untreated cells, the number of cells significantly reduced following treatment with vitamin D (49%), vitamin E (35%), and vitamins D + E (61%) after 144 h. Compared with cell cultures treated with individual vitamins, cells treated with vitamins D + E demonstrated decreased cell confluence, with more extensive and flatter cytoplasm that initiated the formation of a significantly large number of calcified nodules after 7 days of treatment. After 14 days, treatment with vitamins D, E, and D + E increased the transcription of RUNX2, Osterix, and Osteocalcin genes. Conclusions. Vitamins D and E induced osteoblastic differentiation of hDPSCs, as evidenced by the decrease in cell proliferation, morphological changes, and the formation of calcified nodules, increasing the expression of differentiation genes. Concurrent treatment with vitamins D + E induces a synergistic effect in differentiation toward an osteoblastic lineage.

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Extracellular IL-37 Enhances Osteogenic and Odontogenic Differentiation of Human Dental Pulp Stem Cells via Autophagy Pathway

10.21203/rs.3.rs-291517/v1 ◽

2021 ◽

Author(s):

Na Li ◽

Yan Chen ◽

Ming Yan ◽

Yanqiu Wang ◽

Jintao Wu ◽

...

Keyword(s):

Stem Cells ◽

Dental Pulp ◽

Dental Pulp Stem Cells ◽

Inflammatory Factor ◽

Alizarin Red ◽

Dental Tissues ◽

Alp Activity ◽

Odontogenic Differentiation ◽

Marrow Mesenchymal Stem Cells

Abstract BackgroundThe osteogenic and odontogenic differentiation of dental pulp stem cells (DPSCs) contributes to the restoration and regeneration of dental tissues. Previous study indicated that IL-37 has often been identified as an anti-inflammatory factor that affects other pro-inflammatory signals. It is known to be a factor capable of inducing in vitro osteogenic differentiation of bone marrow mesenchymal stem cells (BMMSCs). The aims of this study were to explore the effects of IL-37 on the differentiation of DPSCs.MethodsDPSCs were cultured in growth medium with different concentration of IL-37, ALP activity was done to detect the optimal concentration for the following experiments. CCK-8 were conducted to assess the effect of IL-37 on proliferation of DPSCs. To assess differentiation, alkaline phosphatase activity, ALP staining, alizarin red S staining and real‐time RT‐PCR of DSPP, Runx2, ALP, and OSX were measured. Western blot was conducted to examine the levels of autophagy related markers (Beclin1, P62, LC3). ResultsCells cultured with 1 ng/mL IL-37 owned the highest ALP activity. IL-37 enhanced the osteogenic and odontogenic differentiation of DPSCs following upregulated the expression of Beclin1, downregulated the expression of P62, and reduced the ratio of LC3II/I, whereas depletion of autophagy suppressed DPSCs osteogenic and odontogenic differentiation. ConclusionIL-37 increased osteogenic and odontogenic differentiation via autophagy.

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Covalent Decoration of Cortical Membranes with Graphene Oxide as a Substrate for Dental Pulp Stem Cells

Nanomaterials ◽

10.3390/nano9040604 ◽

2019 ◽

Vol 9 (4) ◽

pp. 604 ◽

Cited By ~ 2

Author(s):

Roberta Di Carlo ◽

Susi Zara ◽

Alessia Ventrella ◽

Gabriella Siani ◽

Tatiana Da Ros ◽

...

Keyword(s):

Stem Cells ◽

Graphene Oxide ◽

Calcium Phosphate ◽

Dental Pulp ◽

Membrane Integrity ◽

Dental Pulp Stem Cells ◽

Covalent Functionalization ◽

Force Microscopy ◽

Cortical Membranes

(1) Background: The aim of this study was to optimize, through a cheap and facile protocol, the covalent functionalization of graphene oxide (GO)-decorated cortical membrane (Lamina®) in order to promote the adhesion, the growth and the osteogenic differentiation of DPSCs (Dental Pulp Stem Cells); (2) Methods: GO-coated Laminas were fully characterized by Scannsion Electron Microscopy (SEM) and Atomic Force Microscopy (AFM) analyses. In vitro analyses of viability, membrane integrity and calcium phosphate deposition were performed; (3) Results: The GO-decorated Laminas demonstrated an increase in the roughness of Laminas, a reduction in toxicity and did not affect membrane integrity of DPSCs; and (4) Conclusions: The GO covalent functionalization of Laminas was effective and relatively easy to obtain. The homogeneous GO coating obtained favored the proliferation rate of DPSCs and the deposition of calcium phosphate.

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Gold nanoparticles in injectable calcium phosphate cement enhance osteogenic differentiation of human dental pulp stem cells

Nanomedicine Nanotechnology Biology and Medicine ◽

10.1016/j.nano.2017.08.014 ◽

2018 ◽

Vol 14 (1) ◽

pp. 35-45 ◽

Cited By ~ 26

Author(s):

Yang Xia ◽

Huimin Chen ◽

Feimin Zhang ◽

Chongyun Bao ◽

Michael D. Weir ◽

...

Keyword(s):

Stem Cells ◽

Gold Nanoparticles ◽

Calcium Phosphate ◽

Osteogenic Differentiation ◽

Calcium Phosphate Cement ◽

Dental Pulp ◽

Dental Pulp Stem Cells ◽

Human Dental Pulp

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Osteogenic stimulation of human dental pulp stem cells with self‐setting biphasic calcium phosphate cement

Journal of Biomedical Materials Research Part B Applied Biomaterials ◽

10.1002/jbm.b.34512 ◽

2020 ◽

Vol 108 (4) ◽

pp. 1669-1678

Author(s):

Yingzhi Gu ◽

Rui Zhuang ◽

Xianju Xie ◽

Yuxing Bai

Keyword(s):

Stem Cells ◽

Calcium Phosphate ◽

Calcium Phosphate Cement ◽

Dental Pulp ◽

Biphasic Calcium Phosphate ◽

Dental Pulp Stem Cells ◽

Human Dental Pulp ◽

Stimulation Of

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Assessment of a PCL-3D Printing-Dental Pulp Stem Cells Triplet for Bone Engineering: An In Vitro Study

Polymers ◽

10.3390/polym13071154 ◽

2021 ◽

Vol 13 (7) ◽

pp. 1154

Author(s):

Raúl Rosales-Ibáñez ◽

Nieves Cubo-Mateo ◽

Amairany Rodríguez-Navarrete ◽

Arely M. González-González ◽

Tomás E. Villamar-Duque ◽

...

Keyword(s):

Stem Cells ◽

3D Printing ◽

Dental Pulp ◽

In Vitro Study ◽

Axial Direction ◽

Dental Pulp Stem Cells ◽

Bone Engineering ◽

Lateral Direction ◽

Alizarin Red

The search of suitable combinations of stem cells, biomaterials and scaffolds manufacturing methods have become a major focus of research for bone engineering. The aim of this study was to test the potential of dental pulp stem cells to attach, proliferate, mineralize and differentiate on 3D printed polycaprolactone (PCL) scaffolds. A 100% pure Mw: 84,500 ± 1000 PCL was selected. 5 × 10 × 5 mm3 parallelepiped scaffolds were designed as a wood-pilled structure composed of 20 layers of 250 μm in height, in a non-alternate order ([0,0,0,90,90,90°]). 3D printing was made at 170 °C. Swine dental pulp stem cells (DPSCs) were extracted from lower lateral incisors of swine and cultivated until the cells reached 80% confluence. The third passage was used for seeding on the scaffolds. Phenotype of cells was determined by flow Cytometry. Live and dead, Alamar blue™, von Kossa and alizarin red staining assays were performed. Scaffolds with 290 + 30 μm strand diameter, 938 ± 80 μm pores in the axial direction and 689 ± 13 μm pores in the lateral direction were manufactured. Together, cell viability tests, von Kossa and Alizarin red staining indicate the ability of the printed scaffolds to support DPSCs attachment, proliferation and enable differentiation followed by mineralization. The selected material-processing technique-cell line (PCL-3D printing-DPSCs) triplet can be though to be used for further modelling and preclinical experiments in bone engineering studies.

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The effect of fibroblast growth factor 7 on human dental pulp stem cells for differentiation to AQP5 ‐positive and α SMA ‐positive cells in vitro and in vivo

Clinical and Experimental Dental Research ◽

10.1002/cre2.423 ◽

2021 ◽

Author(s):

Yoshihiko Akashi ◽

Atsushi Nemoto ◽

Kei Nakajima ◽

Katsutoshi Kokubun ◽

Satoshi Murakami ◽

...

Keyword(s):

Stem Cells ◽

Growth Factor ◽

Fibroblast Growth Factor ◽

Dental Pulp ◽

Dental Pulp Stem Cells ◽

Fibroblast Growth ◽

Human Dental Pulp

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Odontoblast-Like Cells Differentiated from Dental Pulp Stem Cells Retain Their Phenotype after Subcultivation

International Journal of Cell Biology ◽

10.1155/2018/6853189 ◽

2018 ◽

Vol 2018 ◽

pp. 1-12 ◽

Cited By ~ 12

Author(s):

Paula A. Baldión ◽

Myriam L. Velandia-Romero ◽

Jaime E. Castellanos

Keyword(s):

Stem Cells ◽

Dental Pulp ◽

Matrix Protein ◽

Dental Materials ◽

Dental Pulp Stem Cells ◽

Cell Physiology ◽

Pulp Tissue ◽

Alizarin Red ◽

Dentin Matrix Protein ◽

Mineral Deposition

Odontoblasts, the main cell type in teeth pulp tissue, are not cultivable and they are responsible for the first line of response after dental restauration. Studies on dental materials cytotoxicity and odontoblast cells physiology require large quantity of homogenous cells retaining most of the phenotype characteristics. Odontoblast-like cells (OLC) were differentiated from human dental pulp stem cells using differentiation medium (containing TGF-β1), and OLC expanded after trypsinization (EXP-21) were evaluated and compared. Despite a slower cell growth curve, EXP-21 cells express similarly the odontoblast markers dentinal sialophosphoprotein and dentin matrix protein-1 concomitantly with RUNX2 transcripts and low alkaline phosphatase activity as expected. Both OLC and EXP-21 cells showed similar mineral deposition activity evidenced by alizarin red and von Kossa staining. These results pointed out minor changes in phenotype of subcultured EXP-21 regarding the primarily differentiated OLC, making the subcultivation of these cells a useful strategy to obtain odontoblasts for biocompatibility or cell physiology studies in dentistry.

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