Selenomethionine, a Trace Element, Increases Osteoblastic Activity of hFOB 1.19 Cells (an In Vitro Study).

Osteoporosis and resulting fractures affect a significant group of people in the world. It has been shown in many studies that selenium has positive effects on bone metabolism. Based on this information, the aim of this study is to investigate whether bone differentiation will start in a shorter time by applying selenomethionine (SeMet) to hFOB cells.First, hFOB 1.19 cells were cultured. Safe doses of SeMet were determined by MTT and LDH tests. Ossification levels were determined by alizarin red staining and measurement of alkaline phosphatase enzyme levels. The results were analyzed with statistical tests.It was observed that SeMet increased cell viability at concentrations of 10, 25, 50, 100, and 200 µM in 24 h. At these concentrations, cell viability increased above the control, the viabilities were as follows: 109.4%, 104.9%, 104.3%, 103.15%, and 100.27%. High doses of SeMet significantly reduce cell viability. According to Alizarin red staining, SeMet increases the amount of calcium deposits in hFOB cells in a dose-dependent manner. In the experimental groups, the highest ALP enzyme was determined in the 7-day SeMet application. The most effective dose was measured as 15 µM.It was determined that SeMet, which is found as a trace element in living things in nature, increases the viability of hFOB cells, which are osteoblast cell precursors, and increases osteoblastic differentiation and osteoblastic activity in these cells. Our results are at a level that sheds light on an important problem in public health.

Effects of hexagonal boron nitride nanoparticles on antimicrobial and antibiofilm activities, cell viability.

The objective of this work was to investigate the antimicrobial and antibiofilm activities of hBN nanoparticles against Streptococcus mutans 3.3, Staphylococcus pasteuri M3, Candida sp. M25 and S. mutans ATTC 25175. Minimum Inhibitory Concentration (MIC) of hBN nanoparticles were determined against Streptococcus mutans 3.3, Staphylococcus pasteuri M3, Candida sp. M25 growth. In addition, we aimed to evaluate the cytotoxic effects of hBN nanoparticles on human normal skin fibroblast (CCD-1094Sk, ATCC® CRL 2120 ™) and Madin Darby Canine Kidney (MDCK) cells by using various toxicological endpoints. Cell viability was assessed by MTT, SRB and PicoGreen assays. After experimental analyses, it was revealed that hBN nanoparticles show better MIC results. The MIC values were higher for Streptococcus mutans ATTC 25175 and Staphylococcus pasteuri M3 and lower against Streptococcus mutans 3.3, Candida sp. M25. Surprisingly, hBN nanoparticles showed a high antibiofilm activity on preformed biofilm, which inhibited biofilm growth of S. mutans 3.3, S. mutans ATTC 25175 and Candida sp.M25. These results show that hBN nanoparticles may be an option to control oral biofilms. In cell viability tests, the cells were exposed to 0.025-0.4 mg/mL concentrations of hBN nano particle suspension. The exposure time to the hBN nanoparticle suspensions were 24 h and 48 h. The results indicate that there is no cytotoxic effect on CRL 2120 and MDCK cells at the concentration range of 0.025-0.1 mg/mL. However, on both first and second day, hBN caused mild cytotoxicity on CRL-2120 cells at high hBN concentration (0.2-0.4 mg/mL). Considering all the results of this study, in appropriate concentration (0.1 mg/mL) hBN nanoparticles can be considered a potential safe oral care product.

Anti-lung Cancer and Anti-angiogenic Activities of New Designed Boronated Phenylalanine Metal Complexes.

BACKGROUND: Drug design and discovery studies still remain of great importance in the search for more convenient chemotherapeutic to avoid the drug resistance, systemic toxicity or the longterm side effects. OBJECTIVE: A series of mononuclear gold (III) and platinum (II) complexes based on 4-dihydroxyboryl- DL-phenylalanine (BPA) was designed and synthesized, for the first time, by using 2, 2'-dipyridyl (L1) and 4, 4'-diaminobibenzyl (L2) ligands. Characterization of the synthesized complexes was achieved by using 1H-NMR, IR, MS and elemental analyses. METHOD: MTT cell viability, endothelial tube formation, cancer cell colony formation and TRITCphalloidin cytoskeleton staining assays were performed on human umbilical vein endothelial (HUVEC) and human lung adenocarcinoma (A549) cells to establish the anticancer and anti-angiogenic activities of the complexes. It was determined that the organometallic complexes that include 2, 2'-dipyridyl ligand have higher antiproliferative activity than L2-based complexes in the micromolar range. Colony formation experiments showed that the anchorage-independent growth ability of A549s was significantly affected by the complexes in a concentration-dependent manner though L1-based complexes were more effective than L2-based ones. RESULTS: It was also clearly observed that the complexes have significant anti-angiogenic and cytoskeleton alterative activities. Consequently, the phenylalanine-based organometallic complexes seem to have anti-lung cancer and anti-angiogenic activities depending on the ligand type and a great potential in oncology drug development because phenylalanine amino acid has an ability to cross the cell membrane by using L-amino acid transport system. CONCLUSION: Design, synthesis and activity studies with amino acid analogs should be therefore increased to discover more efficient drugs to cure cancer diseases.

The in vitro assessment of dipyridophenazine complexes in H-ras oncogene transformed rat embryo fibroblast 5RP7 cell line.

Purpose The aim of this study is to detect apoptotic and cytotoxic/antiproliferative effects of a ligand substance and its metal derivatives. The substances were investigated by using an h-ras oncogene transformed rat embryo fibroblast cell line (5RP7). Methods The cytotoxic influences of dipyrido[3,2-a:2',3'c]phenazine ligand, dipyrido[3,2-a:2',3'c] phenazine-platinum(II) complex ([Pt(dppz)Cl2]) and dipyrido[3,2-a:2',3'c] phenazine-gold(III) complex ([Au(dppz)Cl2]Cl) were determined with MTT (3[4,5-dimetiltiyazol2-yl]-2,5-difeniltetrazolyum bromid) assay on 5RP7 cells. Results Dipyrido[3,2-a:2',3'c] phenazine, dipyrido[3,2-a:2',3'c] phenazine-platinum(II) complex ([Pt(dppz)Cl2]) and dipyrido[3,2-a:2',3'c] phenazine-gold(III) complexes ([Au(dppz)Cl2]Cl) caused significant increase in cytotoxicity in a dose and time dependent manner. The effects of dipyridophenazine ligand (dppz) and its metal derivatives on apoptosis were monitorized using cytotoxic dose (10 µM) DAPI fluorescent staining. It was shown that dppz and its compounds induced apoptosis. Conclusions These findings show that dpzz and its complexes can be studied as novel alternative chemotherapeutics in cancer treatment.

Promotion of Hair Growth by Traditionally Used Delphinium Staphisagria Seeds through Inducti on of Angiogenesis.

How the seeds of Delphinium staphisagria promote hair growth in humans is yet to be discovered. This lack of information leads us to the investigation of hair promoting effects of seeds of Delphinium staphisagria in-vitro. Extract prepared from the Seed of Delphinium staphisagria (ESDS) - traditionally used for hair loss treatment - was selected and tested for the cytotoxic and angiogenic potential in endothelial cells (HUVECs) and human keratinocytes (HaCaT) cells. The effects of extract was determined by using in-vitro colorimetric MTT [3-(4,5-dimethythiazol-2-yl)-2,5-diphenyltetrazolium bromide] cytotoxicity assay. To identify the compounds that induce angiogenesis, we applied matrigel capillary assay in-vitro using HUVECs. Vinegar and water extracts of D. staphisagria seeds significantly promoted the proliferation of human keratinocyte cells by 137, 139, 143, 149 and 147 % at the concentration of 100, 120, 200, 250 and 300 µg/mL compared with vehicle -treated control, respectively at 24 h. HUVECs viability remained the same with the control group at the concentration 1, 10, 20 and 40 µg/mL after 24 h. Results demonstrated that ESDS did not cause toxicity in human keratinocytes and endothelial cells, while inducing the angiogenic activity in-vitro. D. staphisagria seeds promote hair growth without overt cytotoxicity and through inducing angiogenesis. Based on the information from the traditional uses and our experimental results, D. staphisagria's seeds appear to be a good candidate for the promotion of hair growth.

Photoprotective Activity of Vulpinic and Gyrophoric Acids Toward Ultraviolet B-Induced Damage in Human Keratinocytes.

Vulpinic and gyrophoric acids are known as ultraviolet filters for natural lichen populations because of their chemical structures. However, to the best of our knowledge, there has been no reference to their cosmetic potential for skin protection against ultraviolet B (UVB)-induced damage and, consequently, we propose to highlight their photoprotective profiles in human keratinocytes (HaCaT). Therefore, vulpinic acid and gyrophoric acid were isolated from acetone extracts of Letharia vulpina and Xanthoparmelia pokornyi, respectively. Their photoprotective activities on irradiated HaCaT cells and destructive effects on non-irradiated HaCaT cells were compared through in vitro experimentation: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and lactate dehydrogenase assays, 4',6-diamino-2-phenylindole and tetramethylrhodamine B isothiocyanate-phalloidin staining protocols. Both of the lichen substances effectively prevented cytotoxic, apoptotic and cytoskeleton alterative activities of 2.5 J/cm(2) UVB in a dose-dependent manner. Moreover, vulpinic and gyrophoric acids showed no toxic, apoptotic or cytoskeleton alterative effects on non-irradiated HaCaT cells, except at high doses (≥400 µM) of gyrophoric acid. The findings suggest that vulpinic and gyrophoric acids can be promising cosmetic ingredients to photo-protect human skin cells and should therefore be further investigated by in vitro and in vivo multiple bioassays.

Resveratrol reduces IL-6 and VEGF secretion from co-cultured A549 lung cancer cells and adipose-derived mesenchymal stem cells.

Stem cell therapies are important treatment methodologies used in many areas of experimental or clinical medicine. In recent studies of cancer models, Mesenchymal stem cells (MSCs) suppressed the growth of cancer cells. However, also in some studies, stem cell treatments have been shown to induce cancer formation, increase tumor volume, induce the formation of new vessels, and lead to cancer invasion. The presence of MSC-secreted cytokines and their effects on cancer cells limits the reliability of MSC-based treatments. Resveratrol (trans-3,5,4'-trihydroxystilbene), an antioxidant found in red wine, has been shown to have therapeutic effects against several cancers. The aim of this study was to co-culture MSCs with A549 cancer cells to suppress the release of cancer-promoting cytokines from MSCs and to increase the applicability and reliability of stem cell therapies with resveratrol. MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) and neutral red cell viability assays were used to find safety dose of resveratrol. The MSCs secreted the cytokines IL-6 and VEGF, and the effect of resveratrol on these cytokines was analyzed by ELISA and western blot analysis of conditioned medium. One µM of resveratrol was found to be the safety dose for the A549 cancer cells and MSCs. We observed the highest release of IL-6 and VEGF from the co-cultured A549 cells and MSCs, and resveratrol was found to significantly decrease the release of these cytokines. Our study suggests that resveratrol exerts a positive effect on the release of cytokines. The safety dose of resveratrol can be administered together with stem cells during stem cell treatment.

Anti-angiogenic and antiproliferative properties of the lichen substances (-)-usnic acid and vulpinic acid.

The anti-proliferative activities of the lichen substances (-)-usnic acid and vulpinic acid on the viability of HepG2 hepatocarcinoma cells, NS20Y neuroblastoma cells and HUVEC endothelial cells were studied by the MTT assay. The anti-angiogenic potential of the substances was determined by the endothelial tube formation assay. Both lichen substances exhibited strong anti-angiogenic activity and were more cytotoxic to the cancer cell lines than to the normal cell line, but vulpinic acid has more potential as an anti-angiogenic substance because of its low cytotoxicity and stronger anti-angiogenic activity on the HUVEC cell line.

Novel ferrocenyl-containing N-acetyl-2-pyrazolines inhibit in vitro angiogenesis and human lung cancer growth by interfering with F-actin stress fiber polimeryzation.

Cancer is a significant worldwide health problem generally because of lack of widespread and comprehensive early detection methods. Lung cancer is the leading cause of cancer deaths in men worldwide and the second leading cause of cancer deaths in women. To date, the available treatment regimens are not successful. For this reason, new targets for prevention and new agents for therapy need to be identified. The biological activities of some synthesized ferrocene-containing N-acetylated-2-pyrazoline compounds were studied for this article. Their cytotoxicity (by methyl thiazol tetrazolium assay), as well as apoptotic (by 4'6-diamidino-phenylindole and F-actin staining), antitumoral (colony-forming ability assay) and antiangiogenic activities (by tube formation), were evaluated for the first time on a human non-small-cell lung cancer (A549) cell line and a human umbilicial vein endothelial cell line. All compounds were cytotoxic, antitumoral and apoptotic against tumor cells in a dose-dependent manner. Compounds 2 and 3, which were noncytotoxic, could inhibit capillary vessel formation. Especially, N-acetyl-5-ferrocenyl-3-(2-thienyl)-2-pyrazoline (2) may be used in the development of therapeutic agents for angiogenic diseases and cancer.

Cytotoxic effect of eudesmanolides isolated from flowers of Tanacetum vulgare ssp. siculum.

A phytochemical analysis of the dichloromethane extract from the flowers of a subspecies of Tanacetum vulgare growing in Sicily was carried out. Five known sesquiterpene lactones with the eudesmane skeleton have been isolated and the cytotoxic activity of these compounds was tested in vitro on A549 (human lung carcinoma epithelial-like) and V79379A (Chinese hamster lung fibroblast-like) cells using the tetrazolium salt reduction (MTT) assay. All of tested compounds induced high time- and concentration-dependent cytotoxic effects.

Induction of apoptosis and inhibition of cell proliferation by the cyclooxgenase enzyme blocker nimesulide in the Ishikawa endometrial cancer cell line.

OBJECTIVES: Endometrial cancer remains a leading cause of death in women and therefore the development of new therapies is essential. The present study evaluated the effects of nimesulide alone, cisplatin alone, and combination of cisplatin and nimesulide on an Ishikawa cell line with respect to cytotoxicity and induction of apoptosis in vitro. STUDY DESIGN: Ishikawa cells were treated with increasing doses of nimesulide alone, cisplatin alone, and a combination of cisplatin and nimesulide. Subsequently their effects on cytotoxicity were investigated by MTT assay, while apoptosis was investigated by DAPI and JC-1 staining and caspase-3 colorimetric assays. RESULTS: 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay showed that nimesulide alone and combination of cisplatin and nimesulide have growth inhibitory effect on Ishikawa cells. Nimesulide alone and the combination of cisplatin and nimesulide induced apoptosis. Apoptosis induced by nimesulide might be related to caspase-3 activation. CONCLUSIONS: These results suggest that nimesulide treatment is as effective as cisplatin treatment in Ishikawa cells. The combination of cisplatin and nimesulide treatment is more effective than cisplatin alone in Ishikawa cells.

Assessment of anti-angiogenic and anti-tumoral potentials of Origanum onites L. essential oil.

Medicinal plants and culinary herbs with anti-angiogenic and little toxicity properties have gained importance. Non-toxic anti-angiogenic phytochemicals are useful in combating cancer by preventing the formation of new blood vessels to support the tumor growth. We have investigated the essential oil of Origanum onites L. (OOEO), for a possible anti-angiogenic activity. OOEO was analyzed by gas chromatography (GC) and gas chromatography-mass spectrometry (GC/MS). The anti-proliferative activities (by MTT assay, 3-(4,5-dimethyl-2-thiazol)-2,5-diphenyl-2H-tetrazolium bromide), anti-angiogenic activities (by tube formation assay), cell migration inhibiting capability (migration assay) and apoptotic potential (DAPI staining) of OOEO were evaluated on rat adipose tissue endothelial cells (RATECs) and 5RP7 (c-H-ras transformed rat embryonic fibroblasts) cells. Our results revealed that OOEO could markedly inhibit cell viability and induced apoptosis of 5RP7 cells and also could block in vitro tube formation and migration of RATEC. These results imply that OOEO having anti-angiogenic activity might be useful in preventing angiogenesis-related diseases and in combating cancer.

Studies on the cytotoxic, apoptotic and antitumoral effects of Au(III) and Pt(II) complexes of 1, 10-phenanthroline on V79 379A and A549 cell lines.

In the present study, Au(III) and Pt(II) complexes of 1, 10-phenanthroline (phen) were synthesized and used as the test compounds. The structure elucidation of the synthesized compounds was performed by IR, (1)H-NMR and MASS spectroscopic data and the results of elemental analyses. The cytotoxic and apoptotic effects of test compounds were elucidated on V79 379A (Chinese hamster lung fibroblast like) and A549 (human lung carcinoma epithelial like) cell lines. Cytotoxicity was measured with MTT assay and antitumoral effect was determined by colony forming ability methods. In addition, nuclear fragmentation and activation of apoptotic enzyme (caspase-3) and DAPI staining were used to detect the apoptotic effect of the compounds. All the test compounds induced time and concentration-dependent cytotoxic and antitumoral effects. Significant increases in the levels of apoptosis were observed with increasing exposure concentration.

Cryptic fragment alpha4 LG4-5 derived from laminin alpha4 chain inhibits de novo adipogenesis by modulating the effect of fibroblast growth factor-2.

Cleavage of the extracellular matrix (ECM) by proteolysis unmasks cryptic sites and generates novel fragments with biological activities functionally distinct from those of the intact ECM molecule. The laminin G-like (LG)4-5 fragment has been shown to be excised from the laminin alpha4 chain in various tissues. However, the functional role of this fragment has remained unknown to date. To investigate this, we prepared alpha4 LG1-3 and alpha4 LG4-5 fragments by elastase digestion of recombinant alpha4 LG1-5, and examined their effects on de novo adipogenesis in mice at the site of injection of basement membrane extract (Matrigel) and fibroblast growth factor (FGF)-2. Although the addition of whole alpha4 LG1-5 suppressed adipogenesis to some extent, the alpha4 LG4-5 fragment could strongly suppress adipogenesis at a concentration of less than 20 nm. Addition of the alpha4 LG4 module, which contains a heparin-binding region, had a suppressive effect, but this was lost in mutants with reduced heparin-binding activity. In addition, antibodies against the extracellular domain of syndecan-2 and -4, which are known receptors for the alpha4 LG4 module, suppressed adipogenesis. Thus, these results suggest that the cryptic alpha4 LG4-5 fragment derived from the laminin alpha4 chain inhibits de novo adipogenesis by modulating the effect of FGF-2 through syndecans.