Dimethyl Sulfoxide Leads to Decreased Osteogenic Differentiation of Stem Cells Derived from Gingiva via Runx2 and Collagen I Expression

Lee, Hyunjin; Park, Jun‐Beom

doi:10.1055/s-0039-1694904

Cited by 9 publications

(9 citation statements)

References 22 publications

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“…An anthraquinone dye assay were used for calcium deposit evaluation to assess osteogenic differentiation on Days 4, 7 and 14 [ 22 ]. Spheroids were washed thrice with phosphate-buffered saline and then cell spheroids were fixed in 4% paraformaldehyde in phosphate-buffered saline at room temperature for 15 min.…”

Section: Methodsmentioning

confidence: 99%

The Effects of Morinda citrifolia (Noni) on the Cellular Viability and Osteogenesis of Stem Cell Spheroids

Min

Park

2020

Medicina

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Background and objectives: Morinda citrifolia (Noni) has been widely used in herbal remedies to treat and prevent various kinds of diseases. We conducted this study to evaluate the effects of Noni extract on the maintenance of morphology, the improvement of cellular viability, and the enhancement of osteogenesis of stem cell spheroids. Materials and Methods: We cultured stem cell spheroids made with gingiva-derived stem cells in the presence of Noni extract at concentrations of 10, 100 and 200 ng/mL. We performed analysis of the cell morphology and changes in the cellular viability. We conducted alkaline phosphatase activity assays using a kit, and mineralization assays using an anthraquinone dye to evaluate the osteogenesis of stem cell spheroids with the addition of Noni extract. Results: The applied cells formed spheroids well, and the addition of Noni at 10, 100 and 200 ng/mL concentrations did not produce significant morphological changes. The quantitative values for cellular viability on Day 3 showed that the absorbance values at 450 nm were 0.314 ± 0.013, 0.318 ± 0.008, 0.304 ± 0.000 and 0.300 ± 0.011 for Noni at 0, 10, 100 and 200 ng/mL concentrations, respectively. The results of alkaline phosphatase activity with absorbance values at 405 nm were 0.189 ± 0.019, 0.174 ± 0.023, 0.192 ± 0.014 and 0.210 ± 0.062 for Noni at 0, 10, 100 and 200 ng/mL concentrations, respectively, on Day 4. There were significantly higher values of Alizarin Red S staining for Noni in the 10, 100 and 200 ng/mL groups, with the highest value at 100 ng/mL when compared with the unloaded control on Day 14. Conclusions: Based on these findings, we concluded that Noni extract might be applied for the enhanced osteogenic differentiation of stem cell spheroids.

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Section: Methodsmentioning

confidence: 99%

The Effects of Morinda citrifolia (Noni) on the Cellular Viability and Osteogenesis of Stem Cell Spheroids

Min

Park

2020

Medicina

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“…(2015) investigated the effects of different concentrations of DMSO from 0.05 to 1.0 mmol L −1 on odontoblast‐like MDPC‐23 cells and reported that cell viability was not affected by DMSO at any concentration. In a study by Lee & Park (2019), 3% and 10% DMSO caused a decrease in cell viability. In the present study, 1% DMSO was used to dissolve RSV.…”

Section: Discussionmentioning

confidence: 95%

Influence of resveratrol application with pulp‐capping materials on the genetic expression levels of stem cells

Celik

Yapar

Taghizadehghalehjoughi

et al. 2020

Int Endodontic J

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AimTo evaluate in a laboratory setting the response of human mesenchymal stem cells (MSCs) to pulp‐capping materials with and without resveratrol (RSV).MethodologyFive materials, Calcimol LC, Life, TheraCal LC, ProRoot MTA and Biodentine, were prepared according to the manufacturers’ instructions. Human MSCs were then exposed to these materials, with and without RSV, for 24 h (n = 8). Cell viability was evaluated using the MTT assay, and total cell death was quantified by annexin V‐FITC staining with flow cytometry. The expression levels of the IL‐8, IL‐10, HBD‐2 and BCL‐2 genes were investigated using real‐time polymerase chain reaction (RT‐PCR). Data obtained from MTT test were analysed using one‐way anova, and Tukey’s multiple‐comparison test. The paired Student t test was employed to compare the effects of materials on gene expression (significance level of 5%).ResultsThe group cell viabilities were Calcimol LC 53%, Life 43%, TheraCal LC 78%, ProRoot MTA 75% and Biodentine 78%. Calcimol LC and Life exhibited significant differences compared with the control groups (P < 0.05). The percentages of necrotic/late apoptotic cells associated with Calcimol LC and TheraCal LC were greater than in the other materials. However, when RSV was added to wells containing materials, cell viability increased to Calcimol LC 63%, Life 52%, TheraCal LC 82%, ProRoot MTA 91% and Biodentine 96%, and the percentages of early apoptotic and late apoptotic/necrotic cells decreased. Calcimol LC + RSV and Life + RSV differed significantly from the control group (P < 0.05). The expression of IL‐8 gene was high for all materials, ProRoot MTA caused significant overexpression, and the addition of RSV reduced the expression of IL‐8 in the Calcimol LC, TheraCal LC and ProRoot MTA groups and led to increased expression of IL‐10 in the Calcimol LC, Life and Biodentine groups. HBD‐2 and BCL‐2 exhibited increased expression in ProRoot MTA with RSV (P < 0.05).ConclusionsThe addition of RSV exerted a protective effect on MSCs and regulated the inflammatory process by altering the expression levels of pro‐ and anti‐inflammatory genes.

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“…For example, it has already been shown that DMSO can affect cell cycle [ 6 – 10 ], mitochondrial function [ 5 , 11 ], and the epigenetic landscape of various cell types [ 12 – 14 ]. More importantly, DMSO can prime the differentiation of various types of stem cells [ 7 , 15 – 18 ], and it is able to sustain mESC culture pluripotency in the absence of LIF [ 20 ]. However, in most of these studies, the range of DMSO concentrations used is not broad and is usually close to 1%, which is a much higher concentration than the amount of DMSO used in the majority of the in vitro biological studies published.…”

Section: Discussionmentioning

confidence: 99%

“…However, DMSO is also a hydrogen-bound disrupter, an intercellular electrical uncoupler, and a hydroxyl radical scavenger, among other properties [ 1 , 3 ], and could conceivably affect cell culture, depending on the cell type and the concentration used, issues that are usually overlooked. For instance, it has already been shown that apart from inducing cytotoxicity [ 4 – 6 ], DMSO can affect cell cycle [ 6 – 10 ] and mitochondrial function [ 5 , 11 ], as well as the epigenetic landscape of different cell types [ 12 – 14 ], and influence the differentiation of various types of stem cells [ 7 , 15 – 18 ]. Indeed, in 2012, Pal and colleagues reported that when exposing differentiating human embryonic stem cells (hESCs) to three different concentrations of DMSO, there was a dose-dependent downregulation of pluripotency markers and that the lowest concentration biased the differentiation process by enhancing mesodermal differentiation [ 17 ].…”

Section: Introductionmentioning

confidence: 99%

Effects of DMSO on the Pluripotency of Cultured Mouse Embryonic Stem Cells (mESCs)

Sousa

Correia

Branco

et al. 2020

Stem Cells International

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DMSO is a commonly used solvent in biological studies, as it is an amphipathic molecule soluble in both aqueous and organic media. For that reason, it is the vehicle of choice for several water-insoluble substances used in research. At the molecular and cellular level, DMSO is a hydrogen-bound disrupter, an intercellular electrical uncoupler, and a cryoprotectant, among other properties. Importantly, DMSO often has overlooked side effects. In stem cell research, the literature is scarce, but there are reports on the effect of DMSO in human embryoid body differentiation and on human pluripotent stem cell priming towards differentiation, via modulation of cell cycle. However, in mouse embryonic stem cell (mESC) culture, there is almost no available information. Taking into consideration the almost ubiquitous use of DMSO in experiments involving mESCs, we aimed to understand the effect of very low doses of DMSO (0.0001%-0.2%), usually used to introduce pharmacological inhibitors/modulators, in mESCs cultured in two different media (2i and FBS-based media). Our results show that in the E14Tg2a mESC line used in this study, even the smallest concentration of DMSO had minor effects on the total number of cells in serum-cultured mESCs. However, these effects could not be explained by alterations in cell cycle or apoptosis. Furthermore, DMSO did not affect pluripotency or differentiation potential. All things considered, and although control experiments should be carried out in each cell line that is used, it is reasonable to conclude that DMSO at the concentrations used here has a minimal effect on this particular mESC line.

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Dimethyl Sulfoxide Leads to Decreased Osteogenic Differentiation of Stem Cells Derived from Gingiva via Runx2 and Collagen I Expression

Cited by 9 publications

References 22 publications

The Effects of Morinda citrifolia (Noni) on the Cellular Viability and Osteogenesis of Stem Cell Spheroids

The Effects of Morinda citrifolia (Noni) on the Cellular Viability and Osteogenesis of Stem Cell Spheroids

Influence of resveratrol application with pulp‐capping materials on the genetic expression levels of stem cells

Effects of DMSO on the Pluripotency of Cultured Mouse Embryonic Stem Cells (mESCs)

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