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5. Simvastatin induces adverse effects on proliferation and mineralization of human primary osteoblasts

Author

Sabandal, Martin M. I., Schäfer, Edgar, Aed, Jonathan, Jung, Susanne, Kleinheinz, Johannes, Sielker, Sonja, and Universitäts- und Landesbibliothek Münster

Subjects
Adult, Male, Simvastatin, Osteoblasts, Mineralization, lcsh:Specialties of internal medicine, Adverse effects, 610 Medicine and health, Anticholesteremic Agents, Research, Cell Differentiation, lcsh:RC581-951, Osteogenesis, Medicine and health, Humans, Female, ddc:610, Cell Proliferation
Abstract

Background:Frequently statins were administered to reduce the LDL-concentration in circulating blood. Especially simvastatin (SV) is an often prescribed statin. Pleiotropic effects of these drugs were reported. Thus, the aim of this study was to evaluate effects of SV on osteoblastic mineralization. Methods:After informed consent primary osteoblasts were collected from tissue surplus after treatment of 14 individuals in the Department of Cranio-Maxillofacial Surgery, University Hospital Münster. The cells were passaged according to established protocols. Viability, mineralization capability and osteoblastic marker (alkaline phosphatase) were determined at day 9, 13 and 16 after adding various SV concentrations (0.05 μM, 0.1 μM, 0.5 μM, 1.0 μM). Statistical analysis was performed using the Kruskal-Wallis-test. Results:The cell cultures showed a time and dose-dependent significantly decreased viability (p < 0.01) and a significantly increased mineralization (p < 0.01) in a late mineralization stage after adding SV. The typical alteration of the alkaline phosphatase (ALP) levels during osteogenic differentiation was not recognizable. Conclusions:The pleiotropic effects found for different SV concentrations were possibly originated from other mineralization pathways beside the ALP induced one. Additionally, possible alterations of protein expression levels during mineralization and investigation of possible deviating application of SV in other treatment fields can be considered after gaining a deeper insight in the affected mechanisms., Finanziert über die DEAL-Vereinbarung mit Wiley 2019-2022.

Published
2023

8. Pleiotropic effects on proliferation and mineralization of primary human adipose tissue derived stromal cells induced by Simvastatin

Author

Sabandal, Martin M. I., Schäfer, Edgar, Petsching, Simon, Jung, Susanne, Kleinheinz, Johannes, Sielker, Sonja, and Universitäts- und Landesbibliothek Münster

Subjects
Stromazellen aus Fettgewebe, Mineralisation, pleiotrope Effekte, Simvastatin, adipose tissue derived stromal cells, mineralization, pleiotropic effects, simvastatin, Technology (Applied sciences), ddc:600, 600 Technology (Applied sciences)
Abstract

Die zirkulierende Low-Density-Lipoprotein-Konzentration im Blut kann durch die Verabreichung von Statinen gesenkt werden. Hierfür wird häufig Simvastatin (SV) verschrieben. Aufgrund der berichteten pleiotropen Wirkungen von SV war es das Ziel dieser Studie, die Mineralisierungseffekte auf Stromazellen aus menschlichem Fettgewebe nach Verabreichung von SV zu untersuchen. Nach Zustimmung der Probanden (14) wurden Stromazellen aus menschlichem Fettgewebe welches aus Gewebsüberschüssen von rouitinemäßiger Behandlungen gewonnen. Nach etablierten Protokollen wurden nach Zugabe verschiedener SV-Konzentrationen (0,01 µM, 0,1 µM, 1,0 µM, 2,0 µM) alkalische Phosphatase (Osteoblastenmarker), Mineralisierungsfähigkeit und Lebensfähigkeit an Tag 18, 21 und 28 bestimmt. Für die statistische Analyse wurde der Kruskal-Wallis-Test durchgeführt. Nach Zugabe von SV wurde eine dosisabhängige signifikante Abnahme der Lebensfähigkeit und der Werte der alkalischen Phosphatase (p, The circulating low-density lipoprotein-concentration in blood can be reduced by the administration of statins. Frequently simvastatin (SV) is prescribed. Due to reported pleiotropic effects of SV the aim of this study was to evaluate mineralization effects on human adipose tissue derived stromal cells upon administration of SV. After informed consent human adipose tissue derived stromal cells were obtained from tissue surplus of regular treatments of 14 individuals. According to established protocols after adding various SV concentrations (0.01 µM, 0.1 µM, 1.0 µM, 2.0 µM), alkaline phosphate (osteoblastic marker), mineralization capability and viability were determined at day 18, 21 and 28. Kruskal-Wallis test was performed for statistical analysis. After adding SV a dose-dependent significant decreased viability and levels of alkaline phosphatase (p

Published
2022

9. The Biologic Effect of Particulate Titanium Contaminants of Dental Implants on Human Osteoblasts and Gingival Fibroblasts.

Author

Happe, Arndt, Sielker, Sonja, Hanisch, Marcel, and Jung, Susanne

Subjects
CELL culture, ENZYME-linked immunosorbent assay, FIBROBLASTS, DENTAL implants, INTERLEUKINS, OXIDOREDUCTASES, SCANNING electron microscopy, TITANIUM, TUMOR necrosis factors, CYTOMETRY, OSTEOBLASTS, CELL survival
Abstract

Purpose: To evaluate the effects of different titanium particle concentrations on the viability of human calvarial osteoblasts and human gingival fibroblasts. Materials and Methods: Primary human calvarial osteoblasts (HCO, 3H Biomedical) and human gingival fibroblasts (HGF-1, ATCC) were cultivated and allowed to adhere for 24 hours. Titanium powder concentrations (0.01 to 1.0 mg/mL) were added, and samples were analyzed at three time points (24 hours, 7 days, 21 days). Cell viability was analyzed using living cell count, proliferation (MTT) assay, and live/dead staining. Cytotoxic effects were evaluated using lactated dehydrogenase assay. Qualitative analysis of cell viability was performed. In addition, scanning electron microscopy (SEM) analysis was performed. Release of interleukin 6 (IL-6) and tumor necrosis factor alpha (TNF-α) was estimated with human IL-6/human TNF-α ELISA. Results: Titanium concentrations of 0.1 mg/mL and 1.0 mg/mL showed medium- and long-term effects on cell growth and proliferation rates. Cytotoxic effects by release of lactate dehydrogenase were observable during the first 24 hours. Human gingival fibroblast cells showed a release factor between 2.6 and 3.4. Titanium powder seemed to be more cytotoxic to human gingival fibroblast cells than to human calvarial osteoblast cells. For human calvarial osteoblasts, only the highest concentration showed cytotoxic effects with a release factor of 2.7. Human calvarial osteoblasts secreted IL-6 only during the first 24 hours and only in the highest titanium concentration, whereas human gingival fibroblasts secreted IL-6 during the entire period. The lowest titanium concentration showed stronger secretion of IL-6 compared with the control. Incorporation of smaller and single titanium particles by cells was identified under SEM analysis. Conclusion: Cell viability is negatively correlated with titanium concentration. Further, titanium debris might lead to an inflammatory biologic response of dental peri-implant tissue. Also, cells interact with the debris, eg, with incorporation of particles. [ABSTRACT FROM AUTHOR]

Published
2019
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14. Identification of large linear plasmids in Arthrobacter spp. encoding the degradation of quinaldine to anthranilate

Author

Overhage, Jorg, Sielker, Sonja, Homburg, Stefan, Parschat, Katja, and Fetzner, Susanne

Subjects
Plasmids -- Research, Biological sciences
Abstract

Arthrobacter nitroguajacolicus Ru61a, which utilizes quinaldine as sole source of carbon and energy, was shown to contain a conjugative linear plasmid of approximately 110 kb, named pAL1. It exhibits similarities with other linear plasmids from Actinomycetales in that it has proteins covalently attached to its 5' ends. Southern hybridization with probes for the genes encoding quinaldine 4-oxidase and N-acetylanthranilate amidase indicated that pAL1 contains the gene cluster encoding the degradation of quinaldine to anthranilate. A mutant of strain R661a that had lost pAL1 indeed could not convert quinaldine, but was still able to grow on anthranilate. Conjugative transfer of pAL1 to the plasmid-less mutant of strain RiJ61a and to Arthrobacter nicotinovorans DSM 420 (pAO1) occurred at frequencies of 5.4 x 10-4 and 2-0 x 10-4 per recipient, respectively, and conferred the ability to utilize quinaldine. Five other quinaldine-degrading Gram-positive strains were isolated from soil samples; 16S rDNA sequence analysis suggested the closest relationship to different Arthrobacter species. Except for strain K2-29, all isolates contained a pAL1-like linear plasmid carrying genes encoding quinaldine conversion. A 478 bp fragment that on pAL1 represents an intergenic region showed 100% sequence identity in all isolates harbouring a pAL1-like plasmid, suggesting horizontal dissemination of the linear plasmid among the genus Arthrobacter.

Published
2005

17. Cytotoxic effects of four different root canal sealers on human osteoblasts

Author

Jung, Susanne, Sielker, Sonja, Hanisch, Marcel R., Libricht, Viktor, Schäfer, Edgar, Dammaschke, Till, and Universitäts- und Landesbibliothek Münster

Subjects
Male, Periodontal Ligament, Cytotoxicity, Cell Enumeration Techniques, lcsh:Medicine, Toxicology, Pathology and Laboratory Medicine, Research and Analysis Methods, Propidium Iodide Staining, Root Canal Filling Materials, Animal Cells, Materials Testing, Medicine and Health Sciences, Enzyme assays, Humans, ddc:610, Colorimetric assays, lcsh:Science, Bioassays and physiological analysis, Cells, Cultured, Connective Tissue Cells, Staining, Osteoblasts, MTT assay, Ligaments, lcsh:R, Biology and Life Sciences, Cell Staining, Cell Biology, Cell Cultures, Nuclear Staining, AH Plus, BioRoot RCS, cytotoxicity, MTA Fillapex, Pulp Canal Sealer, Osteoblast, Biological Tissue, Connective Tissue, Specimen Preparation and Treatment, Medicine and health, Biochemical analysis, lcsh:Q, Female, Biological Cultures, Cellular Types, Anatomy, Research Article
Abstract

The aim of this study was to evaluate the effect of an epoxy resin-based (AH-Plus), a zinc oxide eugenol containing (Pulp-Canal-Sealer) and two calcium silicate containing (MTA-Fillapex and BioRoot-RCS) sealers on primary human osteoblasts (hOB) in freshly mixed and set state. All sealers were mixed strictly according to the manufacturers´ instructions and identically samples were produced. In a pretest cytotoxic sealer concentrations were determined. Thus, for the main cell culture study, dilutions of sealer extract 1:1, 1:2, and 1:10 were used. To simulate a clinical scenario, extracts from freshly mixed sealer were added to the cells on day one. Extracts form set sealers were used for subsequent culturing for 24h, 7d, 14d, and 21d. Cell viability was analyzed by living-cell-count, MTT-assay, and living/dead-staining, cytotoxicity by LDH-assay, and changes by Richardson-staining. All data were statistically evaluated by one way ANOVA and a posthoc analysis with Bonferroni-Holm testing (p

Published
2018

23. Life stage differences in mammary gland gene expression profile in non-human primates

Author

Stute, Petra, Sielker, Sonja, Wood, Charles, Register, Thomas, Lees, Cynthia, Dewi, Fitriya, Williams, J., Wagner, Janice, Stefenelli, Ulrich, Cline, J., Stute, Petra, Sielker, Sonja, Wood, Charles, Register, Thomas, Lees, Cynthia, Dewi, Fitriya, Williams, J., Wagner, Janice, Stefenelli, Ulrich, and Cline, J.

Abstract

Breast cancer (BC) is the most common malignancy of women in the developed world. To better understand its pathogenesis, knowledge of normal breast development is crucial, as BC is the result of disregulation of physiologic processes. The aim of this study was to investigate the impact of reproductive life stages on the transcriptional profile of the mammary gland in a primate model. Comparative transcriptomic analyses were carried out using breast tissues from 28 female cynomolgus macaques (Macaca fascicularis) at the following life stages: prepubertal (n=5), adolescent (n=4), adult luteal (n=5), pregnant (n=6), lactating (n=3), and postmenopausal (n=5). Mammary gland RNA was hybridized to Affymetrix GeneChip® Rhesus Macaque Genome Arrays. Differential gene expression was analyzed using ANOVA and cluster analysis. Hierarchical cluster analysis revealed distinct separation of life stage groups. More than 2,225 differentially expressed mRNAs were identified. Gene families or pathways that changed across life stages included those related to estrogen and androgen (ESR1, PGR, TFF1, GREB1, AR, 17HSDB2, 17HSDB7, STS, HSD11B1, AKR1C4), prolactin (PRLR, ELF5, STAT5, CSN1S1), insulin-like growth factor signaling (IGF1, IGFBP1, IGFBP5), extracellular matrix (POSTN, TGFB1, COL5A2, COL12A1, FOXC1, LAMC1, PDGFRA, TGFB2), and differentiation (CD24, CD29, CD44, CD61, ALDH1, BRCA1, FOXA1, POSTN, DICER1, LIG4, KLF4, NOTCH2, RIF1, BMPR1A, TGFB2). Pregnancy and lactation displayed distinct patterns of gene expression. ESR1 and IGF1 were significantly higher in the adolescent compared to the adult animals, whereas differentiation pathways were overrepresented in adult animals and pregnancy-associated life stages. Few individual genes were distinctly different in postmenopausal animals. Our data demonstrate characteristic patterns of gene expression during breast development. Several of the pathways activated during pubertal development have been implicated in cancer development and me

Published
2018

24. Spectroscopic and biochemical studies on protein variants of quinaldine 4-oxidase: Role of E736 in catalysis and effects of serine ligands on the FeSI and FeSII clusters

Author

Kappl, Reinhard, Sielker, Sonja, Ranguelova, Kalina, Wegner, Jeannine, Parschat, Katja, Huttermann, Jurgen, and Fetzner, Susanne

Subjects
Molybdenum -- Structure, Molybdenum -- Spectra, Iron compounds -- Structure, Iron compounds -- Spectra, Electron transport -- Research, Biological sciences, Chemistry
Abstract

A study used site directed mutagenesis to construct variants of quinaldine 4-oxidase (Qox) with replacement of positions C40, C45, C48, C60, C120, and C154 of [Qox.sub.s] by serine to assess which iron ions of FeSI and FeSII are likely to be involved in intramolecular electron transfer between the molybdenum center and flavin. The properties of the Qox variants when taken collectively reveal that Fe1 of both FeSI and FeSII are the reducible iron sties whereas the Fe2 ions remain in the ferric state.

Published
2006

29. OP093

Author

Purcz, Nicolai, primary, Tiwari, Sanjay, additional, Will, Olga, additional, Heneweer, Carola, additional, Kalthoff, Holger, additional, Glüer, Claus-Christian, additional, Sielker, Sonja, additional, Jung, Susanne, additional, Sproll, Christoph, additional, Açil, Yahya, additional, and Wiltfang, Jörg, additional

Published
2013
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32. Life stage differences in mammary gland gene expression profile in non-human primates

Author

Stute, Petra, Sielker, Sonja, Wood, Charles, Register, Thomas, Lees, Cynthia, Dewi, Fitriya, Williams, J., Wagner, Janice, Stefenelli, Ulrich, Cline, J., Stute, Petra, Sielker, Sonja, Wood, Charles, Register, Thomas, Lees, Cynthia, Dewi, Fitriya, Williams, J., Wagner, Janice, Stefenelli, Ulrich, and Cline, J.

Abstract

Breast cancer (BC) is the most common malignancy of women in the developed world. To better understand its pathogenesis, knowledge of normal breast development is crucial, as BC is the result of disregulation of physiologic processes. The aim of this study was to investigate the impact of reproductive life stages on the transcriptional profile of the mammary gland in a primate model. Comparative transcriptomic analyses were carried out using breast tissues from 28 female cynomolgus macaques (Macaca fascicularis) at the following life stages: prepubertal (n=5), adolescent (n=4), adult luteal (n=5), pregnant (n=6), lactating (n=3), and postmenopausal (n=5). Mammary gland RNA was hybridized to Affymetrix GeneChip® Rhesus Macaque Genome Arrays. Differential gene expression was analyzed using ANOVA and cluster analysis. Hierarchical cluster analysis revealed distinct separation of life stage groups. More than 2,225 differentially expressed mRNAs were identified. Gene families or pathways that changed across life stages included those related to estrogen and androgen (ESR1, PGR, TFF1, GREB1, AR, 17HSDB2, 17HSDB7, STS, HSD11B1, AKR1C4), prolactin (PRLR, ELF5, STAT5, CSN1S1), insulin-like growth factor signaling (IGF1, IGFBP1, IGFBP5), extracellular matrix (POSTN, TGFB1, COL5A2, COL12A1, FOXC1, LAMC1, PDGFRA, TGFB2), and differentiation (CD24, CD29, CD44, CD61, ALDH1, BRCA1, FOXA1, POSTN, DICER1, LIG4, KLF4, NOTCH2, RIF1, BMPR1A, TGFB2). Pregnancy and lactation displayed distinct patterns of gene expression. ESR1 and IGF1 were significantly higher in the adolescent compared to the adult animals, whereas differentiation pathways were overrepresented in adult animals and pregnancy-associated life stages. Few individual genes were distinctly different in postmenopausal animals. Our data demonstrate characteristic patterns of gene expression during breast development. Several of the pathways activated during pubertal development have been implicated in cancer development and me

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