Cytotoxic effect of a new endodontic cement and mineral trioxide aggregate on L929 line culture

Introduction: The aim of this study was to compare the cytotoxicity of Mineral Trioxide Aggregate (MTA) and a New Endodontic Cement (NEC) on L929 mouse fibroblasts. Materials and Methods: Different dilutions (Neat, 1/2, 1/10, 1/100) of fresh and set materials placed adjacent flasks of L929 in DMEM medium. Cellular viability was assessed using MTT assay in three time intervals (24, 48, and 72 h after mixing). Differences in mean cell viability values between materials were assessed by using the One-way ANOVA and Bonferoni post-test. Optical microscopic analysis of morphology of the untreated control and the cement-treated cell cultures were carried out in all experimental periods. Results: It was indicated that there was not a significant difference in cytotoxicity among the materials of test and between them and the control group. However, there was a statistically significant difference between different time intervals within each group (P< 0.05) and between different concentration of test materials (P<0.05). In all samples, set materials showed better viability than fresh ones. Conclusion: According to results of this study, NEC and MTA have similar cytotoxic effect on L929 cell culture

Calcitonin Gene-Related Peptide Effects on Phenotype and IL-12 Production of Monocyte-Derived Dendritic Cells in Rheumatoid Arthritis Patients

Objective(s)Recent studies on human indicate that the introduction of therapeutic use of tolerogenic dendritic cell (DC) for chronic inflammatory conditions is imminent. For the purpose of defining CGRP potency in tolerogenic DC production, we investigated the phenotype and IL-12 production of DCs generated from the monocytes of rheumatoid arthritis (RA) patients in the presence of the calcitonin gene-related peptide (CGRP), as a multifunctional neuropeptide.Materials and MethodsDCs were generated from isolated monocytes from four resistant and two early female RA patients using IL-4, GM-CSF, and CGRP at concentrations of 0, 1, and 100 nM. Then, the phenotype of neuropeptide-treated or untreated DCs was determined using flow cytometry and the IL-12 production was measured by ELISA.ResultsOur study showed that, on the last day of the culture, at a concentration of 1 nM CGRP, the mean fluorescence intensity (MFI) for CD80 increased (14.13%) and the MFIs for CD83, CD86, and HLA-DR decreased (14.57%, 5.28%, and 6.88% respectively). Moreover, at 100 nM CGRP concentration, the MFI for CD80 increased (11.10%) and the MFIs for CD83, CD86, and HLA-DR decreased (4.27%, 18.60%, and 19.75% respectively). In addition, our results indicated that the mean concentrations of IL-12 produced at 0, 1, and 100 nm CGRP concentrations measured 13.72±2.41, 11.01±1.61, and 7±1.34 pg/ml respectively. ConclusionDecreased CD83, CD86, and HLA-DR expression and reduced IL-12 production by CGRP were found in the RA patients' monocyte-derived DCs. CD83 is a well-defined DC activation marker. HLA-DR and CD86 are appropriate molecules for inducing an immune response. IL-12 promotes cell-mediated immunity. Therefore we suggest that CGRP may be used as an inducer in the production of tolerogenic DCs

Cytotoxic Effect of Saffron Stigma Aqueous Extract on Human Transitional Cell Carcinoma and Mouse Fibroblast

<p class="MsoNormal" style="margin: 0cm 0cm 0pt; direction: ltr; unicode-bidi: embed; text-align: left;"><span style="font-size: small;"><span style="font-family: Times New Roman;"><strong>Introduction:</strong> Saffron has been suggested to have inhibitory effects on tumoral cells. We evaluated the cytotoxic effect of aqueous extract of saffron on human transitional cell carcinoma (TCC) and mouse non-neoplastic fibroblast cell lines.<strong></strong></span></span></p><p class="MsoNormal" style="margin: 0cm 0cm 0pt; direction: ltr; unicode-bidi: embed; text-align: left;"><span style="font-size: small;"><span style="font-family: Times New Roman;"><strong>Materials and Methods: </strong>Human TCC 5637 cell line and mouse fibroblast cell line (L929) were cultivated and incubated with different concentrations of aqueous extract of saffron stigma (50 </span><span style="font-family: Symbol; mso-ascii-font-family: 'Times New Roman'; mso-hansi-font-family: 'Times New Roman'; mso-char-type: symbol; mso-symbol-font-family: Symbol;"><span style="mso-char-type: symbol; mso-symbol-font-family: Symbol;">m</span></span><span style="font-family: Times New Roman;">g/mL to 4000 </span><span style="font-family: Symbol; mso-ascii-font-family: 'Times New Roman'; mso-hansi-font-family: 'Times New Roman'; mso-char-type: symbol; mso-symbol-font-family: Symbol;"><span style="mso-char-type: symbol; mso-symbol-font-family: Symbol;">m</span></span><span style="font-family: Times New Roman;">g/mL). Cytotoxic effect of saffron was evaluated by morphologic observation and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide colorimetric assay after 24, 48, 72, and 120 hours in each cell line. <strong></strong></span></span></p><p class="MsoNormal" style="margin: 0cm 0cm 0pt; direction: ltr; unicode-bidi: embed; text-align: left;"><span style="font-size: small;"><span style="font-family: Times New Roman;"><strong>Results: </strong>After 24 hours, morphological observations showed growth inhibitory effects at saffron extract concentrations higher than 200 µg/mL for L929 cells and at concentrations of 50 µg/mL to 200 µg/mL for the TCC cells. These changes became more prominent after 48 hours. However, significant growth inhibitory effects of the extract were shown at concentrations of 400 µg/mL and 800 µg/mL. Higher concentrations of saffron correlated inversely with cell population of both cell lines. Significant reduction of the survived cells was seen at concentrations of 400 µg/mL and 2000 µg/mL for TCC and L929 cell lines, respectively. After 120 hours, decrease in the percentage of survived cells at higher concentrations of saffron extract was seen in both cell lines. At a concentration of 800 </span><span style="font-family: Symbol; mso-ascii-font-family: 'Times New Roman'; mso-hansi-font-family: 'Times New Roman'; mso-char-type: symbol; mso-symbol-font-family: Symbol;"><span style="mso-char-type: symbol; mso-symbol-font-family: Symbol;">m</span></span><span style="font-family: Times New Roman;">g/mL, the survived L929 cells plummeted to less than 60% after 120 hours, while no TCC cells survived at this time. No L929 cells survived at 2000 </span><span style="font-family: Symbol; mso-ascii-font-family: 'Times New Roman'; mso-hansi-font-family: 'Times New Roman'; mso-char-type: symbol; mso-symbol-font-family: Symbol;"><span style="mso-char-type: symbol; mso-symbol-font-family: Symbol;">m</span></span><span style="font-family: Times New Roman;">g/mL.</span></span></p><strong><span style="font-size: 12pt; font-family: ";Times New Roman";; mso-bidi-language: FA; mso-ansi-language: EN-US; mso-fareast-font-family: 'Times New Roman'; mso-fareast-language: EN-US;">Conclusion: </span></strong><span style="font-size: 12pt; font-family: ";Times New Roman";; mso-bidi-language: FA; mso-ansi-language: EN-US; mso-fareast-font-family: 'Times New Roman'; mso-fareast-language: EN-US;">Saffron aqueous extract has inhibitory effects on the growth of both TCC 5637 and normal L929 cell lines. This effect is dose dependent.</span&gt

Study of Cytotoxic Effects of Saffron in MCF-7 Cells: Cytotoxicity of saffron

Current therapies for breast cancer are often limited by short-term efficacy due to the emergence of drug resistance. There has been increased interest in the use of naturally occurring compounds with chemopreventive and chemotherapeutic effects in the treatment of cancers. Saffron, dry stigmas Crocus sativusL., used in Iran as a spice, is known for its anticancer properties. In this study, the cytotoxic and apoptogenic effects of saffron in MCF-7 cells as an in vitromodel for breast cancer study were investigated. Meanwhile role of caspases were studied in its toxicity. MCF-7 and L929 cell lines were cultured in DMEM medium and incubated with different concentrations of saffron extract (100 to 2000 μg/ml). Cell viability was assessed by MTTassay. Apoptotic cells were determined using PI staining of DNA fragmentation by flow cytometry (sub-G1 peak). Role of caspase were studied using the pan-caspase inhibitor, z-VAD-fmk. Saffron extract decreased cell viability in MCF-7 cells as a concentration- and time-dependent manner. Doses of saffron inhibited 50% cell growth (IC50) against MCF-7 was 400 μg/ml after 48 h of incubation. Saffron could induce apoptosis in MCF-7 cells in which apoptosis was dependent on caspase activation. It might be concluded that saffron could cause MCF-7 cell death, in which apoptosis or programmed cell death play an important role.Saffron could be also considered as a promising chemotherapeutic agent in breast cancer treatment

Analysis of CFTR Gene Mutations in Children with Cystic Fibrosis, First Report from North-East of Iran

Objective(s):  More than 1500 registered mutations in cystic fibrosis transmembrane regulator (CFTR) gene are responsible for dysfunction of an ion channel protein and a wide spectrum of clinical manifestations in patients with cystic fibrosis (CF). This study was performed to investigate the frequency of a number of well-known CFTR mutations in North Eastern Iranian CF patients. Material and Methods: A total number of 56 documented CF patients participated in this study. Peripheral blood was obtained and DNA extraction was done by the use of routin methods. Three steps were taken for determining the target mutations: ARMS-PCR was performed for common CFTR mutations based on previous reports in Iran and neighboring countries. PCR-RFLP was done for detection of R344W and R347P, and PCR-Sequencing was performed for exon 11 in patients with unidentified mutation throughout previous steps. Samples which remained still unknown for a CFTR mutation were sequenced for exon 12.   Results: Among 112 alleles, 24 mutated alleles (21.42%) were detected: ΔF508 (10.71%), 1677delTA (3.57%), S466X (3.57%), N1303K (0.89%), G542X (0.89%), R344W (0.89%), L467F (0.89%). Eight out of 56 individuals analyzed, were confirmed as homozygous and eight samples showed heterozygous status. No mutations were detected in exon 12 of sequenced samples. Conclusion:Current findings suggest a selected package of CFTR mutations for prenatal, neonatal and carrier screening along with diagnosis and genetic counseling programs in CF patients of Khorasan