SDF-1 modulates periodontal ligament-Mesenchymal Stem Cells (pdl-MSCs).

BACKGROUND/OBJECTIVES: In this in vitro study, the effects of Stromal cell-derived factor-1 (SDF-1) was evaluated on the periodontal ligament-Mesenchymal Stem Cells (pdl-MSCs) functions. MATERIAL AND METHODS: Real-time cell analyzer-single plate (RTCA-SP) was employed for proliferation, and RTCA-dual purpose (DP) was utilized for pdl-MSCs migration potential treated with different SDF-1 concentrations (0, 0.1, 1, 10, 100, 200, and 400 ng/ml). Based on the dose-response findings, 10 ng/ml SDF-1 was used for further mRNA experiments. RNAs isolated at 6 and 24 h were checked using quantitative RT-PCR for mineralized tissue-associated genes including type I collagen (COL I), osteocalcin (OCN), osteopontin (OPN), and runt-related transcription factor 2 (Runx2). cRNA was synthesized for 6 h, and whole-genome array analysis was performed for over 47.000 probes. Data were subjected to quantile normalization before analysis. RESULTS: Increased proliferation and migration were observed in pdl-MSCs treated with 0.1, 1, and 10 ng/ml SDF-1. Increased COL I was observed at both time points: 6 and 24 h. While there was no significant change for OCN, OPN, and Runx2 at 6 h, SDF-1 up-regulated OCN and OPN, but down-regulated Runx2 mRNA expressions at 24 h. IL-8 and ESM1 genes were differentially expressed over twofold when the pdl-MSCs were exposed to SDF-1 at whole-genome array analysis. IL-8 induction was confirmed with RT-PCR. CONCLUSION: Findings of this study displayed that SDF-1 modulated pdl-MSCs which were important for periodontal regeneration, inducing migration and proliferation, and regulating extracellular matrix synthesis in favor of the formation of new attachment.

Sacroiliitis in systemic lupus erythematosus : The rates of involvement of the forgotten joint.

OBJECTIVE: Although musculoskeletal system involvement is a well-known manifestation in systemic lupus erythematosus (SLE), the probability of sacroiliac joint involvement and its effect on patients might be ignored. The aim of the study was to investigate the association between SLE and sacroiliitis and to evaluate the relationship between clinical parameters and sacroiliitis in SLE. METHODS: The study was designed as a case-control study. A total of 63 patients with SLE and 31 age- and sex-matched healthy controls were included in the study. The clinical and demographic variables of the study population were documented. The sacroiliac joints of patients and controls were evaluated with sacroiliac magnetic resonance imaging. Human leukocyte antigen (HLA) B27 was assessed using flow cytometry (Beckman Coulter Navios-model 3, Beckman Coulter Inc., Brea, CA, USA). Multinomial logistic regression analysis was used to determine the clinical risk factors related to sacroiliitis. RESULTS: Among the 63 patients, acute sacroiliitis was found in 25 patients (39.7%) and chronic sacroiliitis was found in 21 patients (33.3%). Sacroiliitis was higher in patients than in controls (p = 0.001). Acute sacroiliitis was more frequently observed in patients when compared with the control group (p = 0.001). Higher C­reactive protein (CRP) concentrations (odds ratio = 1.75, 95% confidence interval: 1.30-2.35; p < 0.001) were found to be a risk factor for acute sacroiliitis. CONCLUSION: The ratio of sacroiliitis was higher in patients with SLE than in controls. Increased CRP concentrations were determined as a risk factor for acute sacroiliitis. Thus, one should keep in mind that patients with SLE and higher CRP concentrations may have sacroiliitis.

Is elevated triglyceride high density lipoprotein cholesterol ratio a risk factor that causes acute coronary syndrome to appear earlier?

BACKGROUND: The purpose of this study was to assess the relationship between the triglyceride/high density lipoprotein cholesterol ratio and the risk of acute myocardial infarction in young adults. PATIENTS AND METHODS: A total of 621 patients, who underwent coronary angiography (CAG) due to Myocardial Infarction (MI) at our hospital were included in this study. Demographic characteristics, risk factor profile, laboratory test results, electrocardiographic and CAG findings were assessed in the selected groups. RESULTS: Total cholesterol, triglyceride/high density lipoprotein cholesterol (Tg/HDL) ratio, Tg levels, were higher in younger patients with MI, while glucose and high-density lipoprotein levels were lower. Using propensity score matching in the matched population comparing young patients to the older ones, serum triglyceride levels [179 (145-231) vs 148 (101-197)] and triglyceride to high density lipoprotein cholesterol ratio [5.8 (4.1-9.1) vs 3.0 (1.8-4.6)] were significantly higher, whereas high density lipoprotein levels were observed dramatically lower (32.6 ± 8.2 vs 41.7 ± 8.8). CONCLUSION: This study demonstrated that Tg/HDL ratio may be an important predictor for an acute coronary syndrome in the young adult population. Tg/HDL ratio can be used to prevent MI in young adults (Tab. 3, Fig. 1, Ref. 32.).

Comparison of Different Sources of Mesenchymal Stem Cells: Palatal versus Lipoaspirated Adipose Tissue.

OBJECTIVES: The purpose of this study was to compare the proliferation and differentiation potential of mesenchymal stem cells (MSCs) derived from palatal adipose tissue (PAT) and lipoaspirated adipose tissue (LAT). MATERIALS AND METHODS: PATs were obtained from 2 healthy female patients undergoing surgery for gingival recession, and LATs were obtained from 2 healthy female patients undergoing plastic surgery. LAT- and PAT-derived MSCs were confirmed by flow cytometry using MSC-specific surface markers. The multilineage differentiation capacity of the MSCs was analyzed. The expression of immunophenotyping, embryonic, and differentiation markers was compared between both MSC lines. The proliferation of PAT- and LAT-MSCs was evaluated using a real-time cell analyzer, and telomerase activity was determined using an ELISA-based TRAP assay. Stem cells isolated from PAT and LAT were analyzed by real-time PCR and whole genome array analysis. RESULTS: The cells isolated from PAT had MSC characteristics. In addition, PAT-MSCs had significantly higher alkaline phosphatase activity and osteogenic potential than LAT-MSCs. Although the proliferation and telomerase activities of LAT-MSCs were higher than those of PAT-MSCs, the difference was not statistically significant. The level of embryonic stem cell markers (Oct4 and Nanog) was higher in LAT-MSCs than in PAT-MSCs. The whole genome array analysis demonstrated that 255 gene sequences were differentially expressed, with more than a twofold change in expression. CONCLUSIONS: This is the first comparative analysis of the isolation and characterization of MSCs from PAT and LAT. PAT is an accessible source of MSCs, which could be used in periodontal and craniofacial tissue engineering.

Biostimulation with diode laser positively regulates cementoblast functions, in vitro.

The aim of this study was to evaluate the effects of diode laser biostimulation on cementoblasts (OCCM.30). A total of 40 root plates were obtained from healthy third molar teeth and assigned to the following two groups: (1) control group and (2) laser-treated group. Root plates were placed into the cell culture inserts, and OCCM.30 cells were seeded onto root plates. Cells were irradiated with a low level of diode laser (power: 0.3 W in continuous wave, 60 s/cm2). Proliferation and mineralized tissue-associated gene's and BMP's messenger RNA (mRNA) expressions of cementoblasts were evaluated. Total RNAs were isolated on day 3 and integrin-binding sialoprotein (Ibsp), bone gamma-carboxyglutamate protein (Bglap), Type I collagen (Col1a1), osteoblastic transcription factor, runt-related transcription factor (Runx2), and Bone Morphogenetic Protein (BMP)-2, 3, 4, 6, and 7 mRNA expressions were determined using quantitative RT-PCR. von Kossa staining was performed to evaluate biomineralization of OCCM.30 cells. In the proliferation experiment, while there was no significant difference until 96 h, laser irradiation retarded the decrease in cell proliferation trend after 96 h compared to the untreated control group. Statistically significant increase in Ibsp, Bglap, and BMP-2,3,6,7 mRNA expressions were noted in the laser groups when compared to the untreated control group (p < 0.05). Laser irradiation induced mineralized nodule formation of cementoblasts. The results of this study reveal that the biostimulation setting of diode laser modulates the behavior of cementoblasts inducing mineralized tissue-associated gene's mRNA expressions and mineralization. Therefore, biostimulation can be used during regenerative periodontal therapies to trigger cells with periodontal attachment apparatus.

Extremely high boron tolerance in Puccinellia distans (Jacq.) Parl. related to root boron exclusion and a well-regulated antioxidant system.

Recent studies indicate an extremely high level of tolerance to boron (B) toxicity in Puccinellia distans (Jacq.) Parl. but the mechanistic basis is not known. Puccinellia distans was exposed to B concentrations of up to 1000 mg B L-1 and root B uptake, growth parameters, B and N contents, H2O2 accumulation and ·OH-scavenging activity were measured. Antioxidant enzyme activities including superoxide dismutase (SOD), ascorbate peroxidase, catalase, peroxidase and glutathione reductase, and lipid peroxidation products were determined. B appears to be actively excluded from roots. Excess B supply caused structural deformations in roots and leaves, H2O2 accumulation and simultaneous up-regulation of the antioxidative system, which prevented lipid peroxidation even at the highest B concentrations. Thus, P. distans has an efficient root B-exclusion capability and, in addition, B tolerance in shoots is achieved by a well-regulated antioxidant defense system.

Neutrophil-lymphocyte ratio may predict in-hospital mortality in patients with acute type A aortic dissection.

INTRODUCTION: Inflammation has been reported to be associated with aortic dissection (AD), from the development to the prognosis of AD. In this study we aimed to find the role of the neutrophil-lymphocyte ratio (NLR) in the prediction of clinical events in patients with acute AD type A. PATIENTS AND METHODS: The study comprised 37 patients who were hospitalized at our center between 2009 and 2013 with the diagnosis of acute AD type A. RESULTS: The mean NLR was significantly higher in patients with pericardial effusion than those without effusion (15.6 ± 11.4 vs. 7.5 ± 4.8, p = 0.005). An NLR value > 8.51 yielded an area under the curve (AUC) value of 0.829 [95 % confidence interval (CI) 0.674-0.984, p = 0.004], which demonstrated a sensitivity of 77 % and specificity of 74 % for the prediction of mortality. CONCLUSIONS: The novel inflammatory marker NLR could be used to predict pericardial effusion and in-hospital mortality in patients with acute AD type A.

Zwolle risk score predicts contrast-induced acute kidney injury in STEMI patients undergoing PCI.

OBJECTIVE: Contrast-induced acute kidney injury (CI-AKI) is a common complication in patients with acute ST-elevation myocardial infarction (STEMI) undergoing primary percutaneous coronary intervention (PCI). The Mehran risk score was defined originally in elective PCI and may be predictive of CI-AKI. The aim of the present study was to investigate whether the Zwolle score predicts CI-AKI in patients with acute STEMI undergoing primary PCI. PATIENTS AND METHODS: We analyzed the data of 314 consecutive patients (mean age 56.3 ± 11.4 years) with acute STEMI undergoing primary PCI. The study population was divided into two groups according to CI-AKI development. The Mehran score, Zwolle score, baseline characteristics, and in-hospital outcomes were recorded. RESULTS: Patients with CI-AKI had higher Mehran and Zwolle scores. In a receiver operating characteristic (ROC) curve analysis, high area under the curve (AUC) values were determined for Zwolle and Mehran scores (0.85 and 0.79, respectively) for CI-AKI development. A Zwolle score greater than 2 predicted CI-AKI with a sensitivity of 76.3 % and a specificity of 75.4 %. A Mehran score greater than 5 predicted CI-AKI with a sensitivity of 71.1 % and a specificity of 73.6 %. CONCLUSION: Zwolle score predicts CI-AKI slightly better than the Mehran score in patients with STEMI undergoing primary PCI. This simple score can be used at the catheterization laboratory for risk stratification for the development of CI-AKI.

Metabolic syndrome and long-term cardiovascular outcomes in NSTEMI with unstable angina.

BACKGROUND AND AIM: Metabolic syndrome (MS) is associated with cardiovascular mortality and morbidity in patients with acute coronary syndrome. The purpose of this study was to evaluate the impact of MS on long-term clinical outcomes in patients with pure non-ST segment myocardial infarction (NSTEMI) or unstable angina pectoris (USAP). METHODS AND RESULTS: We prospectively enrolled 310 consecutive NSTEMI/USAP patients (74 females; mean age, 59.3 ± 11.9 years). The study population was divided into two groups: MS(+) and MS(-). The clinical outcomes of the patients were followed for up to 3 years. Increased 3-year cardiovascular mortality and reinfarction were observed in the MS(+) group, as compared to the MS(-) group (15 vs. 3.4%, p = 0.001, and 22.2 vs. 8.3%, p = 0.001, respectively). Hospitalization rates for heart failure and stroke were not significantly different between the two groups on follow-up. By a Cox multivariate analysis, a significant association was noted between MS and the adjusted risk of 3-year cardiovascular mortality (odds ratio 3.4, 95% confidence interval, 1.24-9.1, p = 0.02). CONCLUSION: These results suggest that MS is associated with an increased risk of 3-year cardiovascular mortality and reinfarction in patients with NSTEMI/USAP.

Two distinct types of dying back axonal degeneration in vitro.

AIMS: In many neurodegenerative diseases and following traumas, dying back degeneration is a common phenomenon that aggravates the pathology and may eventually lead to death of the affected neurone. We aimed to investigate the mechanism of dying back degeneration with an in vitro axonal injury model. METHODS: We cultured adult mouse dorsal root ganglion neurones and with a precise laser beam, cut the axons they extended. Preparations were imaged continuously and images were analysed to describe and quantify ensuing events. Potential contributions of calpains and caspases to the degeneration were explored using specific inhibitors and immunohistochemistry. In vivo implications of the results were sought in nerve sections after sciatic nerve cut. RESULTS: The proximal part of the transected axons went under basically two types of dying back degeneration, fragmentation and retraction. In fragmentation the cytoplasm became condensed and with concomitant axial collapse the axon disintegrated into small pieces. In retraction, the severed axon was pulled back to the soma in an organized manner. We demonstrated that fragmentation was associated with a high risk of cell death, while survival rate with retraction was as high as those of uninjured neurones. Regeneration of transected axon was more likely after retraction than following fragmentation. Activities of caspase-3 and calpains but not of caspase-6 were found linked with retraction and regeneration but not with the fragmentation. CONCLUSIONS: This study describes two quite distinct types of dying back degeneration that lead an injured neurone to quite different fates.

Evaluation of antiproliferative and antimetastatic effects of heparin and erythropoietin on B16f10 melanoma cell line.

A number of chemotherapeutic agents and treatment strategies have been developed or designed to treat cancer patients. However, chemotherapeutic regimens frequently cause side-effects, one of which is anemia, a severe clinical problem for cancer patients. Erythropoietin is commonly used to treat anemia and reduce blood transfusions in cancer patients. Another agent which has potential use in cancer therapy is heparin, a glycosaminoglycan with a negative charge, known to increase the clearance of tumor cells from the blood in mice and also has anti-metastatic effects. In this current study, we investigated the effects of rEpo and heparin either as single agents or in combination on B16F10 melanoma cells. Contrary to our expectations based on the previous in vitro and in vivo studies, we have not found a significant growth-promoting effect of rEpo on B16F10 cells. We have also not observed a significant cytotoxic effect of heparin on B16F10 melanoma cell as assayed by MTT test (p > 0.05). However, heparin did significantly prevent the migration/proliferation of B16F10 cell in the wound assay as compared to the control cells after 24 h of incubation (p < 0.001). In addition, this effect of heparin was not prevented when rEpo was present in the medium in the wound assay (p < 0.01 as compared to the control). These results suggest that heparin may have a therapeutic potential as an anti-metastatic drug for cancer.

Does pantoprazole protect against reperfusion injury following myocardial ischemia in rats?

BACKGROUND: Myocardial ischemia is inadequate perfusion due to reduced blood flow. Sudden onset of reperfusion could result with damage to the myocytes that have not been affected during ischemia called ischemia reperfusion (I/R) injury. Extracellular accumulation of H+ ions resulting in tissue acidosis is one of the underlying mechanisms. Inhibition of myocardial H+/K+-ATPase, namely proton pump, may lead to intracellular acidification via decreasing the extracellular H+ transport. AIM: The aim of this study is to investigate the effects of a proton pump inhibitor pantoprazole in intact rat I/R models. MATERIALS AND METHODS: A total of 30 adult male Wistar albino rats weighing 200-300 g were studied. Rats were allocated into four groups: sham (n=6), ischemia (n=8), control (n=8), and pantoprazole (n=8). Left anterior descending coronary artery was occluded for 30 minutes and then reperfused for two hours. Pantoprazole was administered via jugular vein at the dose of 9 mg/kg starting from 30 minutes before ischemia, to the first 30 minutes of reperfusion. Haemodynamic parameters were recorded and serum CK-MB levels were measured. After reperfusion, heart was removed for the measurement of myocardial infarct size. Myocardial infarct area was measured using triphenyltetrazolium chloride (TTC) staining technique. Myocardial infarction size were expressed as the percentage of the total left ventricular weight. RESULTS: Compared with other groups, plasma concentrations of CK-MB at the end of ischemia and reperfusion and myocardial infarct size were significantly lower in pantoprazole group (p < 0.008). CONCLUSIONS: Pantoprazole preconditioning induces delayed cardioprotection in intact rat I/R model, which may be triggered via H+/K+-ATPase ion channels.

Periodontal ligament cell behavior on different titanium surfaces.

AIM: The purpose of this study was to investigate proliferation, morphology, mineralization and mRNA expressions of mineralized tissue associated proteins of PDL cells on smooth (S), sandblasted small-grit (SSG), sandblasted large-grit (SLG) and sodium titanate (NaTi) coated titanium alloys, in vitro. METHODS AND MATERIALS: PDL cells were cultured with DMEM media containing 10% FBS on the S, SSG, SLG and NaTi titanium surfaces. PDL cell proliferation, mineralization and immunohistochemistry experiments for Bone Sialoprotein (BSP) were performed. The morphology of the PDL cells was examined using confocal and scanning electron microscopy (SEM). Gene expression profiles of cells were evaluated using a quantitative-polymerase chain reaction (Q-PCR) for type I collagen (COL I), Osteocalcin (OCN), osteopontin (OPN) and Runt-related transcription factor-2 (Runx2) on days 7 and 14. RESULTS: Proliferation results on days 6 and 10 were similar in groups, while those of day 13 revealed a decrease in the NaTi group when compared to the S group. NaTi surface induced BSP mRNA expression which was correlated with mineralization tests and BSP immunostaining results. Increased Runx2 mRNA expression was also noted in the NaTi surface when compared to other surfaces. CONCLUSIONS: This study considers the NaTi surface as a potential alternative to SSG and SLG surfaces. This surface might provide a promising environment for PDL ligament-anchored implants.

Relation of interatrial duration and p wave terminal force as a novel indicator of severe mitral regurgitation.

OBJECTIVES: Interatrial duration is defined as prolonged p wave on electrocardiogram. p waves with a negative terminal phase recorded in V1 enclosing an area of one small square on the electrocardiogram is significantly and strongly correlated with interatrial duration. The aim of study was to investigate whether interatrial duration with p terminal force can be used as reflection of echocardiographic severity of mitral regurgitation. MATERIALS AND METHODS: Sixty two consecutive patients with mitral regurgitation. were prospectively studied. Age/gender matched 57 subjects who had normal mitral structure and did not have mitral regurgitation. Patients with mitral regurgitation referred to a single cardiac center for echocardiography and who met the entry criteria documented moderate or severe mitral regurgitation with sinus were included. The interatrial duration was defined on the routine 12-lead electrocardiogram (50 mm/s, 10 mm/mV) using the greatest duration of p waves from D2, D3, AVF and V1. RESULTS: There was a positive correlation between interatrial duration (>/= 110 ms) and effective regurgitant orifice (r = 0.3, p < 0.001). However, left atrial diameter and brain natriuretic peptide were significantly higher in cases with mitral regurgitation. There was also strong correlation between interatrial duration (>/= 110 ms) and p terminal force and left atrial diameter. ROC analysis revealed that interatrial duration of > 110 msec. could predict of severe mitral regurgitation with 88% sensitivity and 100% specificity. CONCLUSIONS: Severe mitral regurgitation, left atrial diameter was correlated with p terminal force and interatrial duration. Significant interatrial duration (>/= 110 ms) and p terminal force might be considered as novel indicators of severe mitral regurgitation.

Protective effect of benfotiamine on methotrexate induced gastric damage in rats.

Methotrexate (MTX) is widely used for treating cancers and inflammatory diseases; it is a potential anti-metabolite and folate antagonist. We investigated potential protective effects of benfotiamine on MTX damage. We used a rat model of MTX induced gastric injury to assess changes in gastric histopathology, oxidative stress and visfatin levels due to MTX treatment. Rats were divided into four groups: an untreated control group, an MTX group treated with a single dose of MTX, a benfotiamine group treated with benfotiamine daily for two weeks, and a benfotiamine + MTX group treated with a single dose of MTX followed by benfotiamine daily for two weeks. Total tissue antioxidant status (TAS), total oxidant status (TOS) and visfatin levels were measured at the end of the experiment. At the end of the experiment, we investigated both visfatin expression and the histopathology of gastric tissues. The mean visfatin level was lower in the MTX group than in the benfotiamine group. The mean tissue TOS levels were higher in MTX group than in the control, benfotiamine or benfotiamine + MTX groups. Significant gastric gland dilation, and erosion and loss of mucosa were found on the gastric surface in the MTX group compared to the control group. The dilation, erosion and mucosal loss were decreased significantly in the benfotiamine + MTX group compared to the MTX group. Compared to the control group, visfatin immunoreactivity was reduced significantly in the MTX group. Decreased visfatin levels appear to play a role in the mechanism of gastric damage. Benfotiamine may be useful for preventing MTX induced gastric injuries.

Boron as Boric Acid Induces mRNA Expression of the Differentiation Factor Tuftelin in Pre-Osteoblastic MC3T3-E1 Cells.

The effects of boron on the formation and maintenance of mineralized structures at the molecular level are still not clearly defined. Thus, a study was conducted using MC3T3-E1 cells to determine whether boron affected mRNA expressions of genes associated with bone/alveolar bone formation around the teethMC3T3-E1 (clone 4) cells were cultured in media treated with boric acid at concentrations of 0, 0.1, 10, 100, or 1000 ng/ml. Total RNAs of each group were isolated on day 3. Gene expression profiles were determined by using RT2 Profiler PCR micro-array that included 84 genes associated with osteogenic differentiation. Tuftelin1 mRNA expression was upregulated by all boron treatments. The upregulation was confirmed by quantitative RT-PCR using the tuftelin probe. While 100 ng/ml had no effect on the integrin-α2 (Itga2) transcript and 1 ng/ml boric acid induced Itga2 mRNA expression (2.1-fold), 0.1, 10, and 1000 ng/ml boric acid downregulated the integrin-α2 gene transcript 2.2-, 1.5-, and 2.1-fold respectively. While 0.1 ng/ml boric acid induced BMP6, increased BMP1r mRNA expression (1.5 fold) was observed in 1000 ng/ml boric acid treatment. The findings suggest that boron affects the regulation of the tuftelin1 gene in osteoblastic cells. Further studies are needed to establish that the beneficial actions of boron on alveolar bone and tooth formation and maintenance include an effect on the expression of the tuftelin1 gene.