THU0495 NOVEL UNDERSTANDING OF THE PATHOGENESIS OF JUVENILE IDIOPATHIC ARTHRITIS: FOCUS ON MESENCHYMAL STEM CELLS IMPAIRMENT, SENESCENCE AND IMMUNOREGULATORY FUNCTION

Background:Juvenile idiopathic arthritis (JIA) is a well-known chronic rheumatic disease of childhood characterised by progressive joint destruction and severe systemic complications.Immune cells are known to trigger the pathophysiological cascade in JIA, but there is little information regarding the contribution made by Mesenchymal stem cells (MSCs). These cells are able to modulate the immune response and decrease the level of pro-inflammatory cytokines. With addition of regenerative property it makes MSCs potential candidates for clinical application as immunosuppressants in treatment of autoimmune diseases.Objectives:To investigate MSCs proliferation, viability and immunomodulatory function in JIA and healthy children.Methods:MSCs were separated from peripheral blood (PB) and synovial fluid (SF) of JIA patients and healthy controls. Cell proliferation rate was counted by Population doublings per day (PDD) during 9 days, in the last of which alamarBlue™ assays were performed to assess cell viability. Due to measure senescence MSCs were stained with SA-β-galactosidase. Immunofluorescence was used to examine the expression of p16, p21, p53. Oxidative stress was measured with DCFH-DA. Cell cycle analysis was evaluated with Propidium Iodide and analysed by Accuri® C6 Flow Cytometer.Commercially-available bone marrow mesenchymal stem cells (BM-MSCs) were treated with graded concentrations of pro-inflammatory cytokines (0.1-100 ng/ml) with following examination of cell viability. Mixed lymphocyte reactions (MLR) were performed to measure MSC immunomodulatory abilityin vitro.Results:The growth kinetics of JIA-MSCs were different from healthy controls. JIA-MSCs divided slowly and appeared disorganised with large cytoplasm and loads of outgrowth. They demonstrated a decrease in cell proliferation (negative PDD) and metabolic activity. Difference in growth kinetics and metabolic activity were found inside the JIA PB group with some evidence of response following biological treatment. Thus, PB-MSCs from patients treated with TNFi and anti-IL6 medications had notably higher cell proliferation and metabolic activity against JIA patients received other therapy. Considering this difference, it was hypothesised that cytokines obtained in a high amount in PB and SF of JIA patients may influence MSCs viability. To prove this BM-MSCs were treated with cytokines and demonstrated a dose-dependent decrease in metabolic activity significantly after TNFα and IL1, no significantly after treatment with IL6. Both BM-MSCs treated with cytokines and JIA-MSCs displayed high level of reactive oxygen species.Cell cycle analysis revealed that JIA-MSCs were arrested in G0/G1 phase with low number of mitotic cells. In addition, the number of senescence-associated SA-β-gal-positive cells was notably higher in JIA-MSCs. Furthermore, JIA-MSCs expressed high level of immunofluorescence for p16, p21 and p53 which played an important role in regulating the senescence progress of MSCs.Results of MLR showed the ability of BM-MSCs to decrease the percentage of activated T-helpers, T-suppressors, B-cells and natural killers proliferation, while JIA-MSCs lost this property.Conclusion:Taken together current research has demonstrated that under the influence of proinflammatory cytokines JIA-MSCs suffered from oxidative stress and disruption of metabolic activity acquire senescent morphology, shorten of telomere length, arrest in G0 phase of cell cycle and finally loss of immune regulation. We are continuing our research to determine the mechanisms that are responsible for the impaired phenotype with the aim of identifying new therapeutic strategies for the treatment of JIA.Disclosure of Interests: :None declared

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Reactive Oxygen Species Are Required for Human Mesenchymal Stem Cells to Initiate Proliferation after the Quiescence Exit

Oxidative Medicine and Cellular Longevity ◽

10.1155/2015/502105 ◽

2015 ◽

Vol 2015 ◽

pp. 1-8 ◽

Cited By ~ 27

Author(s):

O. G. Lyublinskaya ◽

Ya. G. Borisov ◽

N. A. Pugovkina ◽

I. S. Smirnova ◽

Ju. V. Obidina ◽

...

Keyword(s):

Reactive Oxygen Species ◽

Cell Cycle ◽

Stem Cells ◽

Cell Proliferation ◽

Mesenchymal Stem Cells ◽

Stem Cell ◽

Human Mesenchymal Stem Cells ◽

S Phase ◽

Oxygen Species ◽

Antioxidant Treatment

The present study focuses on the involvement of reactive oxygen species (ROS) in the process of mesenchymal stem cells “waking up” and entering the cell cycle after the quiescence. Using human endometrial mesenchymal stem cells (eMSCs), we showed that intracellular basal ROS level is positively correlated with the proliferative status of the cell cultures. Our experiments with the eMSCs synchronized in the G0phase of the cell cycle revealed a transient increase in the ROS level upon the quiescence exit after stimulation of the cell proliferation. This increase was registered before the eMSC entry to the S-phase of the cell cycle, and elimination of this increase by antioxidants (N-acetyl-L-cysteine, Tempol, and Resveratrol) blocked G1–S-phase transition. Similarly, a cell cycle arrest which resulted from the antioxidant treatment was observed in the experiments with synchronized human mesenchymal stem cells derived from the adipose tissue. Thus, we showed that physiologically relevant level of ROS is required for the initiation of human mesenchymal stem cell proliferation and that low levels of ROS due to the antioxidant treatment can block the stem cell self-renewal.

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Human bone marrow mesenchymal stem cells suppress the proliferation of hepatic stellate cells by inhibiting the ubiquitination of p27

Biochemistry and Cell Biology ◽

10.1139/bcb-2017-0127 ◽

2017 ◽

Vol 95 (6) ◽

pp. 628-633 ◽

Cited By ~ 2

Author(s):

Liang Wang ◽

Guang Bai ◽

Fei Chen

Keyword(s):

Cell Cycle ◽

Stem Cells ◽

Bone Marrow ◽

Cell Proliferation ◽

Mesenchymal Stem Cells ◽

Cell Cycle Arrest ◽

Hepatic Stellate Cells ◽

Stellate Cells ◽

Cycle Arrest ◽

Marrow Mesenchymal Stem Cells

Bone marrow mesenchymal stem cells (BMSCs) have considerable therapeutic potential for the treatment of end-stage liver disease. Previous studies have demonstrated that BMSCs secrete growth factors and cytokines that inactivate hepatic stellate cells (HSCs), which inhibited the progression of hepatic fibrosis. The aim of this study was to determine the mechanism by which BMSCs suppress the function of HSCs in fibrosis. Our results showed that co-culture of BMSCs and HSCs induced cell cycle arrest at the G10/G1 phase and cell apoptosis of HSCs, which finally inhibited the cell proliferation of HSCs. Consistent with the cell cycle arrest, co-culture of BMSCs and HSCs increased the abundance of the cell cycle protein p27. Mechanistically, we further uncovered that following the co-culture with BMSCs, the expression level of the E3 ligase S-phase kinase-associated protein 2 (SKP2) that is responsible for the ubiquitination of p27 was decreased, which attenuated the ubiquitination of p27 and increased the stability of p27 in HSCs. Collectively, our results indicated the potential involvement of the SKP2–p27 axis for the inhibitory effect of BSMCs on the cell proliferation of HSCs.

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Cell cycle-dependent expression of potassium channels and cell proliferation in rat mesenchymal stem cells from bone marrow

Cell Proliferation ◽

10.1111/j.1365-2184.2007.00458.x ◽

2007 ◽

Vol 40 (5) ◽

pp. 656-670 ◽

Cited By ~ 55

Author(s):

X. L. Deng ◽

C. P. Lau ◽

K. Lai ◽

K. F. Cheung ◽

G. K. Lau ◽

...

Keyword(s):

Cell Cycle ◽

Stem Cells ◽

Bone Marrow ◽

Cell Proliferation ◽

Mesenchymal Stem Cells ◽

Potassium Channels ◽

Cycle Dependent

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Effect of miR-124 on the Biological Characteristics of Bone Marrow Mesenchymal Stem Cells in Tumor Microenvironment

Journal of Biomaterials and Tissue Engineering ◽

10.1166/jbt.2020.2235 ◽

2020 ◽

Vol 10 (2) ◽

pp. 151-156

Author(s):

Linfu He ◽

XiaoyanHe ◽

Xiaocui Liu ◽

Wenjing Shi ◽

PeiXu ◽

...

Keyword(s):

Cell Cycle ◽

Stem Cells ◽

Bone Marrow ◽

Cell Proliferation ◽

Mesenchymal Stem Cells ◽

Tumor Microenvironment ◽

Glioma Cells ◽

Biological Characteristics ◽

Brain Glioma ◽

Marrow Mesenchymal Stem Cells

miR-124 affects migration and differentiation of mesenchymal stem cells (MSCs), but its role in bone marrow mesenchymal stem cells (BMSCs) in tumor microenvironment remain unclear. Therefore, our study aims to assess miR-124’s role in BMSCs in glioma microenvironment. BMSCs were isolated and co-cultured with glioma cells using Transwell chamber and then transfected with miR-124 mimic/NC followed by analysis of BMSCs biological characteristics (cell proliferation by CCK8, miR-124 level by Real-time PCR, cell cycle and apoptosis by flow cytometry, and cell migration by Transwell). There were significant differences of cell proliferation, migration, apoptosis and cell cycle in BMSCs group compared to co-culture and co-culture+ miR-NC group (P < 0.05), without no difference compared to co-culture + miR-124 group. In addition, co-culture + miR-124 also showed significant differences of BMSCs' biological characteristics (P < 0.05). miR-124 can alter the biological characteristics of BMSCs in the tumor microenvironment simulated by C6 brain glioma cells.

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Inhibition of hepatic stellate cell proliferation by bone marrow mesenchymal stem cells via regulation of the cell cycle in rat

Experimental and Therapeutic Medicine ◽

10.3892/etm.2012.628 ◽

2012 ◽

Vol 4 (3) ◽

pp. 375-380 ◽

Cited By ~ 8

Author(s):

SHANYU QIN ◽

HAIXING JIANG ◽

SIBIAO SU ◽

DONGXU WANG ◽

ZIYU LIANG ◽

...

Keyword(s):

Cell Cycle ◽

Stem Cells ◽

Bone Marrow ◽

Cell Proliferation ◽

Mesenchymal Stem Cells ◽

Hepatic Stellate Cell ◽

Stellate Cell ◽

Marrow Mesenchymal Stem Cells

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Peculiarities of the metabolic activity of mesenchimal stem cells under oxidative stress

Faktori eksperimental'noi evolucii organizmiv ◽

10.7124/feeo.v26.1268 ◽

2020 ◽

Vol 26 ◽

pp. 212-216

Author(s):

M. V. Kovalchuk ◽

N. S. Shuvalova ◽

V. A. Kordium

Keyword(s):

Oxidative Stress ◽

Hydrogen Peroxide ◽

Stem Cells ◽

Mesenchymal Stem Cells ◽

Metabolic Activity ◽

Therapeutic Potential ◽

Critical Concentration ◽

Test Results ◽

Standard Methods ◽

Mtt Test

Aim. Oxidative stress is considered to be one of the major damaging factors that limits the therapeutic potential of mesenchymal stem cells (MSCs). The purpose of our work was to study the metabolic activity of Wharton jelly-derived MSC of different donor origin under oxidative stress conditions induced by hydrogen peroxide. Methods. MSC were obtained by the explant method and cultured according to standard methods. Oxidative stress was caused by treating cells with different concentrations of hydrogen peroxide. The metabolic activity of MSCs was evaluated using the MTT test. Results. Analysis of the MTT test showed a biphasic dependence of the MSC response to the concentration of H2O2. Concentrations of hydrogen peroxide from 6.25 to 50 μM increased the level of metabolic activity of MSCs, and concentrations from 50 to 440 μM inhibited metabolic activity. The maximum stimulating effect was observed at concentrations of 12.5 μM and 25 μM depending on the donor. Conclusions. The response of cells to oxidative stress corresponded to the hormetic dependence, and the points of critical concentration and maximum stimulation were individual for each donor. Processes such as preconditioning MSCs with hydrogen peroxide to increase their survival rate during transplantation also require personalization of the approach depending on the points of maximum stimulation. Keywords: mesenchymal stem cells, hydrogen peroxide, oxidative stress.

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Para-nonylphenol Toxicity Induces Oxidative Stress and Arrests the Cell Cycle in Mesenchymal Stem Cells of Bone Marrow

Iranian Jornal of Toxicology ◽

10.32598/ijt.13.3.496.2 ◽

2019 ◽

Vol 13 (3) ◽

pp. 1-8

Author(s):

Mohammad Hussein Abnosi ◽

◽

Sina Masoomi ◽

Keyword(s):

Oxidative Stress ◽

Cell Cycle ◽

Stem Cells ◽

Bone Marrow ◽

Mesenchymal Stem Cells ◽

Total Protein ◽

Wistar Rat ◽

Metabolic Enzymes ◽

Electrolyte Imbalance ◽

Total Antioxidant

Background: The mechanism of para-nonylphenol (PNP) reducing the proliferation and differentiation of bone marrow mesenchymal stem cells (MSCs) is not known. The present study was designed to investigate the mechanism. Methods: MSCs were extracted under sterile condition from Wistar rat and cultured in DMEM, containing 15 % FBS and penicillin/streptomycin until the 3rd passage, then cells were treated with 0, 0.5 and 2.5 µM of PNP for 5, 10, 15 and 20 days. We studied the viability, proliferation, cell cycle and morphology of the cells. In addition, the concentration of total protein, sodium, potassium and calcium and the activity of metabolic enzymes (ALT, AST and LDH) were determined. Also, induction of oxidative stress was estimated by determining the total antioxidant and MDA levels in addition to the activity of SOD and CAT. Results: The concentrations of PNP caused a significant increase in metabolic enzymes activity and reduced the total protein dose dependently from day 5 to day 20. But only the higher PNP concentration reduced the sodium level and increased the calcium concentration during the treatment period. In addition, we observed a significant decrease in the total antioxidant level and of SOD and CAT activities whereas a significant increase in MDA was seen. Also, PNP stopped the cell cycle at “S” and “G2/M” phases. Conclusion: Para-NP, used in many industries, was able to reduce the viability and proliferation of the MSCs via metabolic and electrolyte imbalance and by induction of oxidative stress and cell cycle disruption.

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Autologous transplantation of adipose-derived mesenchymal stem cells ameliorates streptozotocin-induced diabetic nephropathy in rats by inhibiting oxidative stress, pro-inflammatory cytokines and the p38 MAPK signaling pathway

International Journal of Molecular Medicine ◽

10.3892/ijmm.2012.977 ◽

2012 ◽

Cited By ~ 12

Author(s):

Shuling Bai

Keyword(s):

Oxidative Stress ◽

Stem Cells ◽

Mesenchymal Stem Cells ◽

Diabetic Nephropathy ◽

Signaling Pathway ◽

P38 Mapk ◽

Inflammatory Cytokines ◽

Autologous Transplantation ◽

Mapk Signaling ◽

Pro Inflammatory Cytokines

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Mesenchymal Stem Cells Early Response to Low-Dose Ionizing Radiation

Frontiers in Cell and Developmental Biology ◽

10.3389/fcell.2020.584497 ◽

2020 ◽

Vol 8 ◽

Author(s):

Marina Konkova ◽

Margarita Abramova ◽

Andrey Kalianov ◽

Elizaveta Ershova ◽

Olga Dolgikh ◽

...

Keyword(s):

Oxidative Stress ◽

Cell Cycle ◽

Stem Cells ◽

Dna Damage ◽

Mesenchymal Stem Cells ◽

Ionizing Radiation ◽

Low Dose ◽

Early Response ◽

Dna Breaks ◽

Essential Components

IntroductionMesenchymal stem cells (MSCs) are applied as the therapeutic agents, e.g., in the tumor radiation therapy.Purpose of the StudyTo evaluate the human adipose MSC early response to low-dose ionizing radiation (LDIR).Materials and MethodsWe investigated different LDIR (3, 10, and 50 cGy) effects on reactive oxygen species production, DNA oxidation (marker 8-oxodG), and DNA breaks (marker ɣ H2AX) in the two lines of human adipose MSC. Using reverse transcriptase–polymerase chain reaction, fluorescence-activated cell sorting, and fluorescence microscopy, we determined expression of genes involved in the oxidative stress development (NOX4), antioxidative response (NRF2), antiapoptotic and proapoptotic response (BCL2, BCL2A1, BCL2L1, BIRC2, BIRC3, and BAX1), in the development of the nuclear DNA damage response (DDR) (BRCA1, BRCA2, ATM, and P53). Cell cycle changes were investigated by genes transcription changes (CCND1, CDKN2A, and CDKN1A) and using proliferation markers KI-67 and proliferating cell nuclear antigen (PCNA).ResultsFifteen to 120 min after exposure to LDIR in MSCs, transient oxidative stress and apoptosis of the most damaged cells against the background of the cell cycle arrest were induced. Simultaneously, DDR and an antiapoptotic response were found in other cells of the population. The 10-cGy dose causes the strongest and fastest DDR following cell nuclei DNA damage. The 3-cGy dose induces a less noticeable and prolonged response. The maximal low range dose, 50 cGy, causes a damaging effect on the MSCs.ConclusionTransient oxidative stress and the death of a small fraction of the damaged cells are essential components of the MSC population response to LDIR along with the development of DDR and antiapoptotic response. A scheme describing the early MSC response to LDIR is proposed.

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