Stability of Cultured Osteoblasts on Calcium Phosphate Ceramics

2006 ◽  
Vol 309-311 ◽  
pp. 93-96 ◽  
Author(s):  
Akira Oshima ◽  
Mika Tadokoro ◽  
Motohiro Hirose ◽  
Yoshinori Takakura ◽  
Hajime Ohgushi
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Suitable Condition ◽  
The Other ◽  
Low Level ◽  
Biochemical Assays ◽  
Long Time ◽  
Mineralized Matrix ◽  
High Level ◽  
Osseous Defects

Cultured osteoblasts with mineralized matrix (regenerative cultured bone; RCB) have been used for patients having osseous defects. The RCB can be fabricated on various ceramic substrata using patient’s mesenchymal stem cells (MSCs) at our cell processing center (CPC). Since we have to transport the RCB for hospitals outside of our town, the RCB should maintain the cell viability for a long time. To determine a suitable condition for transportation of the RCB, stability of the RCB was analyzed by biochemical assays. Even outside CO2 incubator, the RCB kept high level of viability until 24 hours at 25°C and also showed low level of cytotoxicity for 24 hours at 37°C and 25°C. On the other hand, the RCB incubated for 24 hours at 4°C outside CO2 incubator resulted in extremely low level of viability with obvious cytotoxicity. These data indicated that stability of the RCB can be maintained for 24 hours at 37°C and 25°C, but not at 4°C. Therefore, the RCB derived from patient’s MSCs can be transported and utilized for the patients at hospitals far away from the CPC.

scholarly journals EFFECTS OF CAFFEINE, PARACETAMOL AND ΒNAPHTHOFLAVONE ON THE LIPID CONTENT AND DIFFERENTIATION OF HUMAN ADIPOSE DERIVED MESENCHYMAL STEM CELLS IN VITRO

2018 ◽  
Author(s):  
Mohammed H Jarrar
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Adipose Tissue ◽  
Transcription Factors ◽  
Lipid Content ◽  
Biochemical Markers ◽  
The Other ◽  
Biochemical Assays ◽  
The Common

Obesity is a complex multifactorial disorder involving the differentiation of pre-adipocytes to mature adipocytes which is achieved through the process of adipogenesis via interaction with different adipogenic transcription factors and mediators. Many drugs and their interactions has been implicated with the common feature of adipose tissue enlargement through hyperplasia and hypertrophy. Caffeine and Paracetamol are few of the most common used drugs which have been theorized by previous researches to have some influence on the adipogenic process. In our study, we investigated the effects of Caffeine, Paracetamol and β-Naphthoflavone on the differentiation of Human Adipose Derived Mesenchymal Stem Cells (HAD-MSC). Cells were cultured in vitro using differentiation inducing media with and without the presence of different combinations of the drugs. Biochemical markers of adipogenesis were evaluated using biochemical assays for triglyceride and glycerol quantification. Our results show that there is an increase of glycerol and triglyceride concentration in cells treated with caffeine suggesting its anti-lipogenesis characteristics through the enhancement of the lipolytic process.Paracetamol also appears to have anti-adipogenic effects due to its apparent role in suppressing the accumulation of triglycerides. In addition, only the use of βNaphthoflavone in combination with caffeine and Paracetamol, resulted in lower triglyceride concentrations than the latter two drugs alone which may indicate its possible enhancing anti-adipogenic properties is through its interaction with the other two drugs.

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

2020 ◽  
Vol 79 (Suppl 1) ◽  
pp. 483.2-484
Author(s):  
L. Zaripova ◽  
A. Midgley ◽  
S. Christmas ◽  
E. Baildam ◽  
R. Oldershaw
Keyword(s):  
Oxidative Stress ◽  
Cell Cycle ◽  
Stem Cells ◽  
Cell Proliferation ◽  
Mesenchymal Stem Cells ◽  
Juvenile Idiopathic Arthritis ◽  
Metabolic Activity ◽  
Healthy Controls ◽  
High Level ◽  
Pro Inflammatory Cytokines

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

scholarly journals Evaluation of Resistance Development in Bemisia tabaci Genn. (Homoptera: Aleyrodidae) in Cotton against Different Insecticides

Insects ◽  
2021 ◽  
Vol 12 (11) ◽  
pp. 996
Author(s):  
Muhammad Zaryab Khalid ◽  
Sohail Ahmed ◽  
Ibrahim Al-ashkar ◽  
Ayman EL Sabagh ◽  
Liyun Liu ◽  
...  
Keyword(s):  
Bemisia Tabaci ◽  
The Other ◽  
Resistance Development ◽  
Susceptible Population ◽  
Low Level ◽  
Development Of Resistance ◽  
Major Pest ◽  
Selection For ◽  
High Level ◽  
Very High

Cotton is a major crop of Pakistan, and Bemisia tabaci (Homoptera: Aleyrodidae) is a major pest of cotton. Due to the unwise and indiscriminate use of insecticides, resistance develops more readily in the whitefly. The present study was conducted to evaluate the resistance development in the whitefly against the different insecticides that are still in use. For this purpose, the whitefly population was selected with five concentrations of each insecticide, for five generations. At G1, compared with the laboratory susceptible population, a very low level of resistance was observed against bifenthrin, cypermethrin, acetamiprid, imidacloprid, thiamethoxam, nitenpyram, chlorfenapyr, and buprofezin with a resistance ratio of 3-fold, 2-fold, 1-fold, 4-fold, 3-fold, 3-fold, 3-fold, and 3-fold, respectively. However, the selection for five generations increased the resistance to a very high level against buprofezin (127-fold), and to a high level against imidacloprid (86-fold) compared with the laboratory susceptible population. While, a moderate level of resistance was observed against cypermethrin (34-fold), thiamethoxam (34-fold), nitenpyram (30-fold), chlorfenapyr (29-fold), and acetamiprid (21-fold). On the other hand, the resistance was low against bifenthrin (18-fold) after selection for five generations. A very low level of resistance against the field population of B. tabaci, at G1, showed that these insecticides are still effective, and thus can be used under the field conditions for the management of B. tabaci. However, the proper rotation of insecticides among different groups can help to reduce the development of resistance against insecticides.

scholarly journals Effect of lentivirus-mediated growth and differentiation factor-5 transfection on differentiation of rabbit nucleus pulposus mesenchymal stem cells

2022 ◽  
Vol 27 (1) ◽  
Author(s):  
Kun Zhu ◽  
Rui Zhao ◽  
Yuchen Ye ◽  
Gang Xu ◽  
Changchun Zhang
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Nucleus Pulposus ◽  
Lentiviral Vector ◽  
The Other ◽  
Cell Phenotype ◽  
Mrna Levels ◽  
Qrt Pcr ◽  
Differentiation Factor ◽  
Gdf5 Gene

Abstract Background Intervertebral disc degeneration (IDD) is a natural progression of age-related processes. Associated with IDD, degenerative disc disease (DDD) is a pathologic condition implicated as a major cause of chronic lower back pain, which can have a severe impact on the quality of life of patients. As degeneration progression is associated with elevated levels of inflammatory cytokines, enhanced aggrecan and collagen degradation, and changes in the disc cell phenotype. The purpose of this study was to investigate the biological and cytological characteristics of rabbit nucleus pulposus mesenchymal stem cells (NPMSCs)—a key factor in IDD—and to determine the effect of the growth and differentiation factor-5 (GDF5) on the differentiation of rabbit NPMSCs transduced with a lentivirus vector. Methods An in vitro culture model of rabbit NPMSCs was established and NPMSCs were identified by flow cytometry (FCM) and quantitative real-time PCR (qRT-PCR). Subsequently, NPMSCs were randomly divided into three groups: a transfection group (the lentiviral vector carrying GDF5 gene used to transfect NPMSCs); a control virus group (the NPMSCs transfected with an ordinary lentiviral vector); and a normal group (the NPMSCs alone). FCM, qRT-PCR, and western blot (WB) were used to detect the changes in NPMSCs. Results The GDF5-transfected NPMSCs displayed an elongated shape, with decreased cell density, and significantly increased GDF5 positivity rate in the transfected group compared to the other two groups (P < 0.01). The mRNA levels of Krt8, Krt18, and Krt19 in the transfected group were significantly higher in comparison with the other two groups (P < 0.01), and the WB results were consistent with that of qRT-PCR. Conclusions GDF5 could induce the differentiation of NPMSCs. The lentiviral vector carrying the GDF5 gene could be integrated into the chromosome genome of NPMSCs and promoted differentiation of NPMSCs into nucleus pulposus cells. Our findings advance the development of feasible and effective therapies for IDD.

Low-level visible light (LLVL) irradiation promotes proliferation of mesenchymal stem cells

2012 ◽  
Vol 28 (4) ◽  
pp. 1113-1117 ◽  
Author(s):  
Anat Lipovsky ◽  
Uri Oron ◽  
Aharon Gedanken ◽  
Rachel Lubart
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Visible Light ◽  
Low Level

Efficacy of a ciprofloxacin/amikacin combination against planktonic and biofilm cultures of susceptible and low-level resistant Pseudomonas aeruginosa

2019 ◽  
Vol 74 (11) ◽  
pp. 3252-3259 ◽  
Author(s):  
Anaïs Soares ◽  
Kévin Alexandre ◽  
Fabien Lamoureux ◽  
Ludovic Lemée ◽  
François Caron ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Bacterial Biofilm ◽  
The Other ◽  
Clinical Strain ◽  
Bacterial Reduction ◽  
Low Level ◽  
Mechanical Dispersion ◽  
Resistant Mutants ◽  
High Level

Abstract Background Eradicating bacterial biofilm without mechanical dispersion remains a challenge. Combination therapy has been suggested as a suitable strategy to eradicate biofilm. Objectives To evaluate the efficacy of a ciprofloxacin/amikacin combination in a model of in vitro Pseudomonas aeruginosa biofilm. Methods The antibacterial activity of ciprofloxacin and amikacin (alone, in combination and successively) was evaluated by planktonic and biofilm time–kill assays against five P. aeruginosa strains: PAO1, a WT clinical strain and three clinical strains overexpressing the efflux pumps MexAB-OprM (AB), MexXY-OprM (XY) and MexCD-OprJ (CD), respectively. Amikacin MIC was 16 mg/L for XY and ciprofloxacin MIC was 0.5 mg/L for CD. The other strains were fully susceptible to ciprofloxacin and amikacin. The numbers of total and resistant cells were determined. Results In planktonic cultures, regrowth of high-level resistant mutants was observed when CD was exposed to ciprofloxacin alone and XY to amikacin alone. Eradication was obtained with ciprofloxacin or amikacin in the other strains, or with the combination in XY and CD strains. In biofilm, bactericidal reduction after 8 h followed by a mean 4 log10 cfu/mL plateau in all strains and for all regimens was noticed. No regrowth of resistant mutants was observed whatever the antibiotic regimen. The bacterial reduction obtained with a second antibiotic used simultaneously or consecutively was not significant. Conclusions The ciprofloxacin/amikacin combination prevented the emergence of resistant mutants in low-level resistant strains in planktonic cultures. Biofilm persister cells were not eradicated, either with monotherapy or with the combination.

scholarly journals Jawbone microenvironment promotes periodontium regeneration by regulating the function of periodontal ligament stem cells

2017 ◽  
Vol 7 (1) ◽  
Author(s):  
Bin Zhu ◽  
Wenjia Liu ◽  
Yihan Liu ◽  
Xicong Zhao ◽  
Hao Zhang ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Tissue Regeneration ◽  
Periodontal Ligament ◽  
Tooth Development ◽  
Material Surface ◽  
Cell Aggregates ◽  
Periodontal Ligament Stem Cells ◽  
Marrow Mesenchymal Stem Cells ◽  
Mineralized Matrix

AbstractDuring tooth development, the jawbone interacts with dental germ and provides the development microenvironment. Jawbone-derived mesenchymal stem cells (JBMSCs) maintain this microenvironment for root and periodontium development. However, the effect of the jawbone microenvironment on periodontium tissue regeneration is largely elusive. Our previous study showed that cell aggregates (CAs) of bone marrow mesenchymal stem cells promoted periodontium regeneration on the treated dentin scaffold. Here, we found that JBMSCs enhanced not only the osteogenic differentiation of periodontal ligament stem cells (PDLSCs) but also their adhesion to titanium (Ti) material surface. Importantly, the compound CAs of PDLSCs and JBMSCs regenerated periodontal ligament-like fibers and mineralized matrix on the Ti scaffold surface, both in nude mice ectopic and minipig orthotopic transplantations. Our data revealed that an effective regenerative microenvironment, reconstructed by JBMSCs, promoted periodontium regeneration by regulating PDLSCs function on the Ti material.

Sign in / Sign up

Export Citation Format

Share Document