Effect of diode laser potentiality on proliferation of dental pulp stem cells (in vitro study)

Abstract Background The field of laser-based photochemical reactions receives a great promising for additional applications especially for targeting cells, pathogens, or molecules. Limited studies have investigated the impact of light-emitting diode on stem cell behavior. Thus, the aim of the present study was to assess the effect of diode laser irradiations on the proliferation of stem cells isolated from the human dental pulp. Isolation procedures were according to previously developed protocols for dental pulp stem cells (DPSCs). Low-level laser therapy irradiation (LLLT) was applied in two doses (0.5 J/cm2 and 1 J/cm2 for 20 s) into 96-well plates by the diode laser device (970 nm) through the fiber optic (SiroLaser fibers 320) at a distance from the opening of the wells to be accurate for equal distribution of the laser irradiation. To assess the proliferation capacity of the isolated stem cells, MTT assay was performed 24 h, 48 h, and 72 h. Results There was no significant difference among the different groups (gp 1 control, gp II dose 0.5 J/cm2, gp III dose 1 J/cm2) on day 1. While on day 2, the optical density of DPSCs subjected to dose 1 was found to be significantly higher than that of those subjected to dose 2. This was also demonstrated on day 3. It was also demonstrated that the proliferation of DPSCs subjected to dose 1 increased compared to that of DPSCs subjected to dose 2 from day 1 to day 2. There was a significant decrease in the cell number in both groups by 72 h. Conclusion In conclusion, the use of LLLT as a stimulatory factor for enhancing and proliferation of the stem cells is very promising.

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Increased VEGF-A Expression of Human Dental Pulp Stem Cells (hDPSCs) Cultured with Advanced Platelet Rich Fibrin (A-PRF)

The Open Dentistry Journal ◽

10.2174/1874210602115010569 ◽

2021 ◽

Vol 15 (1) ◽

pp. 569-574

Author(s):

Dini Asrianti Bagio ◽

Indah Julianto ◽

Anggraini Margono ◽

Endang Suprastiwi

Keyword(s):

Stem Cells ◽

Dental Pulp ◽

Dental Pulp Stem Cells ◽

Control Group ◽

Platelet Rich Fibrin ◽

Significant Difference ◽

Angiogenesis Process ◽

Human Dental Pulp ◽

Post Hoc

Background: VEGF-A expression of human dental pulp stem cells (hDPSCs) can induce the angiogenesis process of dental pulp regeneration. This in vitro study aimed to analyze the effect of various concentrations of Advanced Platelet Rich Fibrin (A-PRF) conditioned media (CM) on the increased expression of vascular endothelial growth factor-A (VEGF-A) of hDPSCs. Methods: hDPSCs were collected from ten third molars extracted from nine healthy donors, cultured, and then harvested at the end of the third passage. The hDPSCs were seeded in four different CM (control group: hDPSCs + DMEM; 1% A-PRF CM group: hDPSCs + 1% A-PRF CM; 5% A-PRF CM group: hDPSCs + 5% A-PRF CM; 10% A-PRF CM group: hDPSCs + 10% A-PRF CM). All of the groups were cultured in biological triplicates (Triplo) and observed for 5, 12, and 24 hours. The VEGF-A protein expression of hDPSCs was measured using human VEGF-A ELISA at a wavelength of 405 nm. Data was analyzed with Kruskal Wallis and post hoc Mann Whitney test with p<0.05. Results: The VEGF-A expression rate of hDPSCs among all groups was statistically significantly different at 5, 12 and 24 hours of observations (p<0.05). Post hoc analysis test showed a statistically significant difference of hDPSCs’s VEGF-A expression between 5% A-PRF groups compared to other groups at 5 and 12 hours of observation (p<0.05). However, there were no statistically significant differences observed of hDPSCs’ VEGF-A expression at 24 hours of observation between 1%, 5% and 10% A-PRF groups (p>0.05). Conclusion: 5% A-PRF CM was superior in increasing VEGF-A expression of hDPSCs at 5, 12 and 24 hours of observations.

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Low Molecular Weight Hyaluronic Acid Effect on Dental Pulp Stem Cells In Vitro

Biomolecules ◽

10.3390/biom11010022 ◽

2020 ◽

Vol 11 (1) ◽

pp. 22

Author(s):

Jan Schmidt ◽

Nela Pilbauerova ◽

Tomas Soukup ◽

Tereza Suchankova-Kleplova ◽

Jakub Suchanek

Keyword(s):

Stem Cells ◽

Molecular Weight ◽

Hyaluronic Acid ◽

Dental Pulp ◽

Degradation Kinetics ◽

Dental Pulp Stem Cells ◽

Low Molecular Weight ◽

Strong Argument ◽

The Impact

Hyaluronic acid (HA) and dental pulp stem cells (DPSCs) are attractive research topics, and their combined use in the field of tissue engineering seems to be very promising. HA is a natural extracellular biopolymer found in various tissues, including dental pulp, and due to its biocompatibility and biodegradability, it is also a suitable scaffold material. However, low molecular weight (LMW) fragments, produced by enzymatic cleavage of HA, have different bioactive properties to high molecular weight (HMW) HA. Thus, the impact of HA must be assessed separately for each molecular weight fraction. In this study, we present the effect of three LMW-HA fragments (800, 1600, and 15,000 Da) on DPSCs in vitro. Discrete biological parameters such as DPSC viability, morphology, and cell surface marker expression were determined. Following treatment with LMW-HA, DPSCs initially presented with an acute reduction in proliferation (p < 0.0016) and soon recovered in subsequent passages. They displayed significant size reduction (p = 0.0078, p = 0.0019, p = 0.0098) while maintaining high expression of DPSC markers (CD29, CD44, CD73, CD90). However, in contrast to controls, a significant phenotypic shift (p < 0.05; CD29, CD34, CD90, CD106, CD117, CD146, CD166) of surface markers was observed. These findings provide a basis for further detailed investigations and present a strong argument for the importance of HA scaffold degradation kinetics analysis.

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

Polymers ◽

10.3390/polym13142252 ◽

2021 ◽

Vol 13 (14) ◽

pp. 2252

Author(s):

Jae Eun Kim ◽

Sangbae Park ◽

Woong-Sup Lee ◽

Jinsub Han ◽

Jae Woon Lim ◽

...

Keyword(s):

Stem Cells ◽

Calcium Phosphate ◽

Zeta Potential ◽

Mechanical Strength ◽

Calcium Phosphate Cement ◽

Dental Pulp ◽

Dental Pulp Stem Cells ◽

Ion Trapping ◽

Alizarin Red

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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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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P-EP013. Superior migration propensity of dental pulp stem cells in in vitro and in vivo models of excitotoxic neurodegeneration

Clinical Neurophysiology ◽

10.1016/j.clinph.2021.02.176 ◽

2021 ◽

Vol 132 (8) ◽

pp. e82-e83

Author(s):

Sivapriya Senthilkumar ◽

Chaitra Venugopal ◽

K. Shobha ◽

Bindu M. Kutty ◽

Anandh Dhanushkodi

Keyword(s):

Stem Cells ◽

Dental Pulp ◽

Dental Pulp Stem Cells ◽

In Vivo Models ◽

Migration Propensity

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Three-dimensional bone formation including vascular networks derived from dental pulp stem cells in vitro

Human Cell ◽

10.1007/s13577-018-00228-y ◽

2018 ◽

Vol 32 (2) ◽

pp. 114-124

Author(s):

Miho Watanabe ◽

Akihiro Ohyama ◽

Hiroshi Ishikawa ◽

Akira Tanaka

Keyword(s):

Stem Cells ◽

Bone Formation ◽

Dental Pulp ◽

Three Dimensional ◽

Dental Pulp Stem Cells ◽

Vascular Networks

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Hypoxia-inducible factor 1α induces osteo/odontoblast differentiation of human dental pulp stem cells via Wnt/β-catenin transcriptional cofactor BCL9

Scientific Reports ◽

10.1038/s41598-021-04453-8 ◽

2022 ◽

Vol 12 (1) ◽

Author(s):

Shion Orikasa ◽

Nobuyuki Kawashima ◽

Kento Tazawa ◽

Kentaro Hashimoto ◽

Keisuke Sunada-Nara ◽

...

Keyword(s):

Stem Cells ◽

Dental Pulp ◽

Target Genes ◽

Hypoxia Inducible Factor ◽

Dental Pulp Stem Cells ◽

Pulp Tissue ◽

Odontoblast Differentiation ◽

Hypoxia Inducible Factor 1Α ◽

Hypoxic Conditions

AbstractAccelerated dental pulp mineralization is a common complication in avulsed/luxated teeth, although the mechanisms underlying this remain unclear. We hypothesized that hypoxia due to vascular severance may induce osteo/odontoblast differentiation of dental pulp stem cells (DPSCs). This study examined the role of B-cell CLL/lymphoma 9 (BCL9), which is downstream of hypoxia-inducible factor 1α (HIF1α) and a Wnt/β-catenin transcriptional cofactor, in the osteo/odontoblastic differentiation of human DPSCs (hDPSCs) under hypoxic conditions. hDPSCs were isolated from extracted healthy wisdom teeth. Hypoxic conditions and HIF1α overexpression induced significant upregulation of mRNAs for osteo/odontoblast markers (RUNX2, ALP, OC), BCL9, and Wnt/β-catenin signaling target genes (AXIN2, TCF1) in hDPSCs. Overexpression and suppression of BCL9 in hDPSCs up- and downregulated, respectively, the mRNAs for AXIN2, TCF1, and the osteo/odontoblast markers. Hypoxic-cultured mouse pulp tissue explants showed the promotion of HIF1α, BCL9, and β-catenin expression and BCL9-β-catenin co-localization. In addition, BCL9 formed a complex with β-catenin in hDPSCs in vitro. This study demonstrated that hypoxia/HIF1α-induced osteo/odontoblast differentiation of hDPSCs was partially dependent on Wnt/β-catenin signaling, where BCL9 acted as a key mediator between HIF1α and Wnt/β-catenin signaling. These findings may reveal part of the mechanisms of dental pulp mineralization after traumatic dental injury.

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Novel Molecule Nell-1 Promotes the Angiogenic Differentiation of Dental Pulp Stem Cells

Frontiers in Physiology ◽

10.3389/fphys.2021.703593 ◽

2021 ◽

Vol 12 ◽

Author(s):

Mengyue Li ◽

Qiang Wang ◽

Qi Han ◽

Jiameng Wu ◽

Hongfan Zhu ◽

...

Keyword(s):

Stem Cells ◽

Dental Pulp ◽

Umbilical Vein ◽

Dental Pulp Stem Cells ◽

Exposure Model ◽

Dental Tissues ◽

Normal Rat ◽

Pulp Exposure ◽

Angiogenic Markers

IntroductionThis work aimed to reveal the crucial role of Nell-1 in the angiogenic differentiation of human dental pulp stem cells (DPSCs) alone or co-cultured with human umbilical vein endothelial cell (HUVECs) in vitro and whether this molecule is involved in the pulp exposure model in vivo.MethodsImmunofluorescence was conducted to ascertain the location of Nell-1 on DPSCs, HUVECs, and normal rat dental tissues. RT-PCR, Western blot, and ELISA were performed to observe the expression levels of angiogenic markers and determine the angiogenic differentiation of Nell-1 on DPSCs alone or co-cultured with HUVECs, as well as in vitro tube formation assay. Blood vessel number for all groups was observed and compared using immunohistochemistry by establishing a rat pulp exposure model.ResultsNell-1 is highly expressed in the nucleus of DPSCs and HUVECs and is co-expressed with angiogenic markers in normal rat pulp tissues. Hence, Nell-1 can promote the angiogenic marker expression in DPSCs alone and co-cultured with other cells and can enhance angiogenesis in vitro as well as in the pulp exposure model.ConclusionNell-1 may play a positive role in the angiogenic differentiation of DPSCs.

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