TY - JOUR A1 - Dotterweich, Julia A1 - Schlegelmilch, Katrin A1 - Keller, Alexander A1 - Geyer, Beate A1 - Schneider, Doris A1 - Zeck, Sabine A1 - Tower, Robert J. J. A1 - Ebert, Regina A1 - Jakob, Franz A1 - Schütze, Norbert T1 - Contact of myeloma cells induces a characteristic transcriptome signature in skeletal precursor cells-implications for myeloma bone disease JF - Bone N2 - Physical interaction of skeletal precursors with multiple myeloma cells has been shown to suppress their osteogenic potential while favoring their tumor-promoting features. Although several transcriptome analyses of myeloma patient-derived mesenchymal stem cells have displayed differences compared to their healthy counterparts, these analyses insufficiently reflect the signatures mediated by tumor cell contact, vary due to different methodologies, and lack results in lineage-committed precursors. To determine tumor cell contact-mediated changes on skeletal precursors, we performed transcriptome analyses of mesenchymal stem cells and osteogenic precursor cells cultured in contact with the myeloma cell line INA-6. Comparative analyses confirmed dysregulation of genes which code for known disease-relevant factors and additionally revealed upregulation of genes that are associated with plasma cell homing, adhesion, osteoclastogenesis, and angiogenesis. Osteoclast-derived coupling factors, a dysregulated adipogenic potential, and an imbalance in favor of anti-anabolic factors may play a role in the hampered osteoblast differentiation potential of mesenchymal stem cells. Angiopoietin-Like 4 (ANGPTL4) was selected from a list of differentially expressed genes as a myeloma cell contact-dependent target in skeletal precursor cells which warranted further functional analyses. Adhesion assays with full-length ANGPTL4-coated plates revealed a potential role of this protein in INA6 cell attachment. This study expands knowledge of the myeloma cell contact-induced signature in the stromal compartment of myelomatous bones and thus offers potential targets that may allow detection and treatment of myeloma bone disease at an early stage. KW - marrow stromal cells KW - Endothelial growth-factor KW - precedes multiple-myeloma KW - monoclonial gammopathy KW - in-vitro KW - mesenchymal stem-cells KW - undetermined significance KW - angiogenic cytokines KW - peripheral-blood KW - gene-expression KW - Multiple myeloma KW - Bone disease KW - Angiopoietin-like 4 KW - Gene expression profiling KW - Mesenchymal stem cells KW - Osteogenic precursor cells Y1 - 2016 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-186688 VL - 93 ER - TY - JOUR A1 - Wittmann, Katharina A1 - Sieber, Cornel A1 - von Stengel, Simon A1 - Kohl, Matthias A1 - Freiberger, Ellen A1 - Jakob, Franz A1 - Lell, Michael A1 - Engelke, Klaus A1 - Kemmler, Wolfgang T1 - Impact of whole body electromyostimulation on cardiometabolic risk factors in older women with sarcopenic obesity: the randomized controlled FORMOsA-sarcopenic obesity study JF - Clinical Interventions in Aging N2 - Background: Sarcopenic obesity (SO) is characterized by a combination of low muscle and high fat mass with an additive negative effect of both conditions on cardiometabolic risk. The aim of the study was to determine the effect of whole-body electromyostimulation (WB-EMS) on the metabolic syndrome (MetS) in community-dwelling women aged ≥70 years with SO. Methods: The study was conducted in an ambulatory university setting. Seventy-five community-dwelling women aged ≥70 years with SO living in Northern Bavaria, Germany, were randomly allocated to either 6 months of WB-EMS application with (WB-EMS&P) or without (WB-EMS) dietary supplementation (150 kcal/day, 56% protein) or a non-training control group (CG). WB-EMS included one session of 20 min (85 Hz, 350 µs, 4 s of strain–4 s of rest) per week with moderate-to-high intensity. The primary study endpoint was the MetS Z-score with the components waist circumference (WC), mean arterial pressure (MAP), triglycerides, fasting plasma glucose, and high-density lipoprotein cholesterol (HDL-C); secondary study endpoints were changes in these determining variables. Results: MetS Z-score decreased in both groups; however, changes compared with the CG were significant (P=0.001) in the WB-EMS&P group only. On analyzing the components of the MetS, significant positive effects for both WB-EMS groups (P≤0.038) were identified for MAP, while the WB-EMS group significantly differed for WC (P=0.036), and the WB-EMS&P group significantly differed for HDL-C (P=0.006) from the CG. No significant differences were observed between the WB-EMS groups. Conclusion: The study clearly confirms the favorable effect of WB-EMS application on the MetS in community-dwelling women aged ≥70 years with SO. However, protein-enriched supplements did not increase effects of WB-EMS alone. In summary, we considered this novel technology an effective and safe method to prevent cardiometabolic risk factors and diseases in older women unable or unwilling to exercise conventionally. KW - sarcopenia KW - obesity KW - whole-body electromyostimulation KW - cardiovascular KW - metabolic risk KW - metabolic syndrome KW - community-dwelling KW - older people Y1 - 2016 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-164930 VL - 11 ER - TY - JOUR A1 - Dotterweich, Julia A1 - Tower, Robert J. A1 - Brandl, Andreas A1 - Müller, Marc A1 - Hofbauer, Lorenz C. A1 - Beilhack, Andreas A1 - Ebert, Regina A1 - Glüer, Claus C. A1 - Tiwari, Sanjay A1 - Schütze, Norbert A1 - Jakob, Franz T1 - The KISS1 Receptor as an In Vivo Microenvironment Imaging Biomarker of Multiple Myeloma Bone Disease JF - PLoS One N2 - Multiple myeloma is one of the most common hematological diseases and is characterized by an aberrant proliferation of plasma cells within the bone marrow. As a result of crosstalk between cancer cells and the bone microenvironment, bone homeostasis is disrupted leading to osteolytic lesions and poor prognosis. Current diagnostic strategies for myeloma typically rely on detection of excess monoclonal immunoglobulins or light chains in the urine or serum. However, these strategies fail to localize the sites of malignancies. In this study we sought to identify novel biomarkers of myeloma bone disease which could target the malignant cells and/or the surrounding cells of the tumor microenvironment. From these studies, the KISS1 receptor (KISS1R), a G-protein-coupled receptor known to play a role in the regulation of endocrine functions, was identified as a target gene that was upregulated on mesenchymal stem cells (MSCs) and osteoprogenitor cells (OPCs) when co-cultured with myeloma cells. To determine the potential of this receptor as a biomarker, in vitro and in vivo studies were performed with the KISS1R ligand, kisspeptin, conjugated with a fluorescent dye. In vitro microscopy showed binding of fluorescently-labeled kisspeptin to both myeloma cells as well as MSCs under direct co-culture conditions. Next, conjugated kisspeptin was injected into immune-competent mice containing myeloma bone lesions. Tumor-burdened limbs showed increased peak fluorescence compared to contralateral controls. These data suggest the utility of the KISS1R as a novel biomarker for multiple myeloma, capable of targeting both tumor cells and host cells of the tumor microenvironment. KW - multiple myeloma Lesions KW - fluorescence microscopy KW - biomarkers Myelomas KW - bone imaging KW - myeloma cells KW - fluorescent dyes Y1 - 2016 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-146960 VL - 11 IS - 5 ER - TY - JOUR A1 - Wehrle, Esther A1 - Liedert, Astrid A1 - Heilmann, Aline A1 - Wehner, Tim A1 - Bindl, Ronny A1 - Fischer, Lena A1 - Haffner-Luntzer, Melanie A1 - Jakob, Franz A1 - Schinke, Thorsten A1 - Amling, Michael A1 - Ignatius, Anita T1 - The impact of low-magnitude high-frequency vibration on fracture healing is profoundly influenced by the oestrogen status in mice JF - Disease Models & Mechanisms N2 - Fracture healing is impaired in aged and osteoporotic individuals. Because adequate mechanical stimuli are able to increase bone formation, one therapeutical approach to treat poorly healing fractures could be the application of whole-body vibration, including low-magnitude high-frequency vibration (LMHFV). We investigated the effects of LMHFV on fracture healing in aged osteoporotic mice. Female C57BL/6NCrl mice (n=96) were either ovariectomised (OVX) or sham operated (non-OVX) at age 41 weeks. When aged to 49 weeks, all mice received a femur osteotomy that was stabilised using an external fixator. The mice received whole-body vibrations (20 minutes/day) with 0.3 g peak-to-peak acceleration and a frequency of 45 Hz. After 10 and 21 days, the osteotomised femurs and intact bones (contra-lateral femurs, lumbar spine) were evaluated using bending-testing, micro-computed tomography (mu CT), histology and gene expression analyses. LMHFV disturbed fracture healing in aged non-OVX mice, with significantly reduced flexural rigidity (-81%) and bone formation (-80%) in the callus. Gene expression analyses demonstrated increased oestrogen receptor β (ERβ, encoded by Esr2) and Sost expression in the callus of the vibrated animals, but decreased β-catenin, suggesting that ERβ might mediate these negative effects through inhibition of osteoanabolic Wnt/β-catenin signalling. In contrast, in OVX mice, LMHFV significantly improved callus properties, with increased flexural rigidity (+ 1398%) and bone formation (+637%), which could be abolished by subcutaneous oestrogen application (0.025 mg oestrogen administered in a 90-day-release pellet). On a molecular level, we found an upregulation of ER alpha in the callus of the vibrated OVX mice, whereas ERβ was unaffected, indicating that ERa might mediate the osteoanabolic response. Our results indicate a major role for oestrogen in the mechanostimulation of fracture healing and imply that LMHFV might only be safe and effective in confined target populations. KW - level mechanical vibrations KW - ovariectomized rats KW - bone formation KW - LMHFV KW - whole body vibration KW - receptor beta KW - replacement therapy KW - osteoblastic cells KW - early stage KW - alpha KW - Wnt KW - fracture healing KW - oestrogen receptor signalling KW - Wnt signalling Y1 - 2015 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-144700 VL - 8 ER - TY - THES A1 - Busch, Albert Franz Jakob T1 - Prälamin A und Progerie – verursachende Mutanten im Kontext nukleärer Transportprozesse, der Kernlaminaintegrität und CaaX – Prozessierung T1 - Prelamin A und truncated mutations in nuclear export, lamina integrity and CaaX processing N2 - Zur Charakterisierung nukleärer Proteinexportvorgänge wurde in dieser Arbeit zum ersten Mal ein System heterodimerisierender Fusionsproteine auf Basis des kommerziell verfügbaren ARGENT™ Regulated Heterodimerization Kit 2.0 von ARIAD verwendet. Die Expressionsvektoren wurden so verändert, dass ein CRM1 – vermittelter Proteinexport über die Zellkernhülle mittels Fluoreszenzmikroskopie in HeLa – Zellen und humanen Fibroblasten live oder nach Fixation dargestellt werden konnte. Der Export folgte in HeLa – zellen einer exponentiellen Kinetik, FN/C – Bestimmungen zwischen Wildtyp – und RD (Restriktive Dermopathie) – Fibroblasten ergaben keinen Unterschied im Proteinexport. Eine Inhibition der initialen CaaX - Prozessierung von trunkiertem Prälamin A (head/rod) durch Mevinolin ergab keine signifikante Akkumulationsveränderung des trunkierten Prälamins im Zellkern. Ergänzende subzelluläre Lokalisationsstudien unter Zuhilfenahme ausgewählter CaaX – Mutanten, um die gezeigte Unabhängigkeit der CaaX – Prozessierung zu verifizieren, stehen noch aus. FRAP – Untersuchungen in HeLa – Zellen zeigten für die episomal exprimierten trunkierten Fusionsproteine DsRed – Prälamin A Δ50 und DsRed – Prälamin A Δ90 keinen Unterschied in der lateralen Mobilität. Gegenüber dem Wildtyp – DsRed – Prälamin A ist die Beweglichkeit jedoch signifikant reduziert. Bei der Applikation von thermischem Stress (37°C – 51°C) auf Prälamin A, Prälamin A Δ50 oder Prälamin A Δ90 exprimierende HeLa – Zellen, konnte keine Veränderung hinsichtlich der subzellulären Verteilung des zusätzlich koexprimierten Markerproteins GFP – ß – Galaktosidase im Sinne nukleären Schrankenstörung festgestellt werden. Somit scheint die Kernhülle trotz der zu Zellkerndysmorphien und KPK – Fehllokalisationen führenden Prälamin A – Mutanten hinsichtlich ihrer Schrankenfunktion intakt zu bleiben. N2 - Lamin A and truncated forms were investigated adressing nuclear export, caax processing and laminar integrity. Nuclear export processes were investigated in vivo and in vivo via a modified heterdimerization assay. No difference was seen in human fibroblasts from wildtype and restrictive dermopathy patients concerning crm1-mediated nuclear export truncated prelamin A showed no enhanced nuclear localisation after inhibition of farnesylsynthesis. HGPS- and RD lamin A showed significantly decreased lateral mobility after 30 seconds in FRAP experiments. Applying heat stress to HeLa-cells showed no increased influx of the marker protein gfp-ß-galactosidase after 60 minutes. KW - Progeria infantilum KW - Progerie KW - Restriktive Dermopathie KW - Lamin A KW - Kerntransport KW - nuclear export KW - lamin A KW - progeria KW - restrictive dermopathy Y1 - 2011 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-71662 ER - TY - JOUR A1 - Seefried, Lothar A1 - Mueller-Deubert, Sigrid A1 - Schwarz, Thomas A1 - Lind, Thomas A1 - Mentrup, Birgit A1 - Kober, Melanie A1 - Docheva, Denitsa A1 - Liedert, Astrid A1 - Kassem, Moustapha A1 - Ignatius, Anita A1 - Schieker, Matthias A1 - Claes, Lutz A1 - Wilke, Winfried A1 - Jakob, Franz A1 - Ebert, Regina T1 - A small scale cell culture system to analyze mechanobiology using reporter gene constructs and polyurethane dishes N2 - Mechanical forces are translated into biochemical signals and contribute to cell differentiation and phenotype maintenance. Mesenchymal stem cells and their tissuespecific offspring, as osteoblasts and chondrocytes, cells of cardiovascular tissues and lung cells are sensitive to mechanical loading but molecules and mechanisms involved have to be unraveled. It is well established that cellular mechanotransduction is mediated e.g. by activation of the transcription factor SP1 and by kinase signaling cascades resulting in the activation of the AP1 complex. To investigate cellular mechanisms involved in mechanotransduction and to analyze substances, which modulate cellular mechanosensitivity reporter gene constructs, which can be transfected into cells of interest might be helpful. Suitable small-scale bioreactor systems and mechanosensitive reporter gene constructs are lacking. To analyze the molecular mechanisms of mechanotransduction and its crosstalk with biochemically induced signal transduction, AP1 and SP1 luciferase reporter gene constructs were cloned and transfected into various cell lines and primary cells. A newly developed bioreactor and small-scale 24-well polyurethane dishes were used to apply cyclic stretching to the transfected cells. 1 Hz cyclic stretching for 30 min in this system resulted in a significant stimulation of AP1 and SP1 mediated luciferase activity compared to unstimulated cells. In summary we describe a small-scale cell culture/bioreactor system capable of analyzing subcellular crosstalk mechanisms in mechanotransduction, mechanosensitivity of primary cells and of screening the activity of putative mechanosensitizers as new targets, e.g. for the treatment of bone loss caused by both disuse and signal transduction related alterations of mechanotransduction. KW - Bioreaktor KW - Mechanical strain KW - mechanosensitive reporter KW - gene constructs KW - bioreactor Y1 - 2010 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-68099 ER - TY - JOUR A1 - Ljunggren, Osten A1 - Barrett, Annabel A1 - Stoykov, Ivaylo A1 - Langdahl, Bente L. A1 - Lems, Willem F. A1 - Walsh, J. Bernard A1 - Fahrleitner-Pammer, Astrid A1 - Rajzbaum, Gerald A1 - Jakob, Franz A1 - Karras, Dimitrios A1 - Marin, Fernando T1 - Effective osteoporosis treatment with teriparatide is associated with enhanced quality of life in postmenopausal women with osteoporosis: the European Forsteo Observational Study JF - BMC Musculoskeletal Disorders N2 - Background: To describe changes in health-related quality of life (HRQoL) of postmenopausal women with osteoporosis treated with teriparatide for up to 18 months and followed-up for a further 18 months, and to assess the influence of recent prior and incident fractures. Methods: The European Forsteo Observational Study (EFOS) is an observational, prospective, multinational study measuring HRQoL using the EQ-5D. The primary objective was to assess changes in HRQoL during 36 months in the whole study population. A secondary post-hoc analysis examined fracture impact on HRQoL in four subgroups classified based on recent prior fracture 12 months before baseline and incident clinical fractures during the study. Changes from baseline were analysed using a repeated measures model. Results: Of the 1581 patients, 48.4% had a recent prior fracture and 15.6% of these patients had an incident fracture during follow-up. 10.9% of the 816 patients with no recent prior fracture had an incident fracture. Baseline mean EQ-VAS scores were similar across the subgroups. In the total study cohort (n = 1581), HRQoL (EQ-VAS and EQ-5D index scores) improved significantly from baseline to 18 months and this improvement was maintained over the 18-month post-teriparatide period. Improvements were seen across all five EQ-5D domains during teriparatide treatment that were maintained after teriparatide was discontinued. Subjects with incident clinical fractures had significantly less improvement in EQ-VAS than those without incident fractures. Recent prior fracture did not influence the change in EQ-VAS during treatment. Conclusions: EFOS is the first longitudinal study in women with severe postmenopausal osteoporosis in the real world setting to show a substantial improvement in HRQoL during teriparatide treatment that was sustained during subsequent treatment with other medications. The increase in HRQoL was lower in the subgroups with incident fracture but was not influenced by recent prior fracture. The results should be interpreted in the context of the design of an observational study. KW - fracture KW - osteoporosis KW - quality of life KW - teriparatide KW - EQ-5D KW - database KW - alendronate KW - persistence KW - metaanalysis KW - prevalent fractures KW - bone-mineral density KW - vertebral fractures KW - back pain KW - impact KW - responsiveness Y1 - 2013 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-122057 SN - 1471-2474 VL - 14 IS - 251 ER - TY - JOUR A1 - Steinert, Andre F. A1 - Kunz, Manuela A1 - Prager, Patrick A1 - Göbel, Sascha A1 - Klein-Hitpass, Ludger A1 - Ebert, Regina A1 - Nöth, Ulrich A1 - Jakob, Franz A1 - Gohlke, Frank T1 - Characterization of bursa subacromialis-derived mesenchymal stem cells JF - Stem Cell Research & Therapy N2 - Introduction The bursa subacromialis (BS) provides the gliding mechanism of the shoulder and regenerates itself after surgical removal. Therefore, we explored the presence of mesenchymal stem cells (MSCs) within the human adult BS tissue and characterized the BS cells compared to MSCs from bone marrow (BMSCs) on a molecular level. Methods BS cells were isolated by collagenase digest from BS tissues derived from patients with degenerative rotator cuff tears, and BMSCs were recovered by adherent culture from bone-marrow of patients with osteoarthritis of the hip. BS cells and BMSCs were compared upon their potential to proliferate and differentiate along chondrogenic, osteogenic and adipogenic lineages under specific culture conditions. Expression profiles of markers associated with mesenchymal phenotypes were comparatively evaluated by flow cytometry, immunohistochemistry, and whole genome array analyses. Results BS cells and BMSCs appeared mainly fibroblastic and revealed almost similar surface antigen expression profiles, which was \(CD44^+, CD73^+, CD90^+, CD105^+, CD106^+\),\(STRO-1^+, CD14^−, CD31^−, CD34^− , CD45^−, CD144^−\). Array analyses revealed 1969 genes upregulated and 1184 genes downregulated in BS cells vs. BMSCs, indicating a high level of transcriptome similarity. After 3 weeks of differentiation culture, BS cells and BMSCs showed a similar strong chondrogenic, adipogenic and osteogenic potential, as shown by histological, immunohistochemical and RT-PCR analyses in contrast to the respective negative controls. Conclusions Our in vitro characterizations show that BS cells fulfill all characteristics of mesenchymal stem cells, and therefore merit further attention for the development of improved therapies for various shoulder pathologies. Y1 - 2015 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-126446 VL - 6 IS - 114 ER - TY - JOUR A1 - Ebert, Regina A1 - Jakob, Franz A1 - Meissner-Weigl, Jutta A1 - Zeck, Sabine A1 - Määttä, Jorma A1 - Auriola, Seppo A1 - de Sousa, Sofia Coimbra A1 - Mentrup, Birgit A1 - Graser, Stephanie A1 - Rachner, Tilman D. A1 - Hofbauer, Lorenz C. T1 - Probenecid as a sensitizer of bisphosphonate-mediated effects in breast cancer cells N2 - Background: Anti-resorptive bisphosphonates (BP) are used for the treatment of osteoporosis and bone metastases. Clinical studies indicated a benefit in survival and tumor relapse in subpopulations of breast cancer patients receiving zoledronic acid, thus stimulating the debate about its anti-tumor activity. Amino-bisphosphonates in nM concentrations inhibit farnesyl pyrophosphate synthase leading to accumulation of isopentenyl pyrophosphate (IPP) and the ATP/ pyrophosphate adduct ApppI, which induces apoptosis in osteoclasts. For anti-tumor effects μM concentrations are needed and a sensitizer for bisphosphonate effects would be beneficial in clinical anti-tumor applications. We hypothesized that enhancing intracellular pyrophosphate accumulation via inhibition of probenecid-sensitive channels and transporters would sensitize tumor cells for bisphosphonates anti-tumor efficacy. Methods: MDA-MB-231, T47D and MCF-7 breast cancer cells were treated with BP (zoledronic acid, risedronate, ibandronate, alendronate) and the pyrophosphate channel inhibitors probenecid and novobiocin. We determined cell viability and caspase 3/7 activity (apoptosis), accumulation of IPP and ApppI, expression of ANKH, PANX1, ABCC1, SLC22A11, and the zoledronic acid target gene and tumor-suppressor KLF2. Results: Treatment of MDA-MB-231 with BP induced caspase 3/7 activity, with zoledronic acid being the most effective. In MCF-7 and T47D either BP markedly suppressed cell viability with only minor effects on apoptosis. Co-treatment with probenecid enhanced BP effects on cell viability, IPP/ApppI accumulation as measurable in MCF-7 and T47D cells, caspase 3/7 activity and target gene expression. Novobiocin co-treatment of MDA-MB-231 yielded identical results on viability and apoptosis compared to probenecid, rendering SLC22A family members as candidate modulators of BP effects, whereas no such evidence was found for ANKH, ABCC1 and PANX1. Conclusions: In summary, we demonstrate effects of various bisphosphonates on caspase 3/7 activity, cell viability and expression of tumor suppressor genes in breast cancer cells. Blocking probenecid- and novobiocin-sensitive channels and transporters enhances BP anti-tumor effects and renders SLC22A family members good candidates as BP modulators. Further studies will have to unravel if treatment with such BP-sensitizers translates into preclinical and clinical efficacy. KW - Bisphosphonates KW - Caspase 3/7 activity KW - Cell viability, KW - Probenecid KW - Novobiocin KW - Breast cancer cells Y1 - 2014 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-111174 ER - TY - JOUR A1 - Wehrle, Esther A1 - Liedert, Astrid A1 - Heilmann, Aline A1 - Wehner, Tim A1 - Bindl, Ronny A1 - Fischer, Lena A1 - Haffner-Luntzer, Melanie A1 - Jakob, Franz A1 - Schinke, Thorsten A1 - Amling, Michael A1 - Ignatius, Anita T1 - The impact of low-magnitude high-frequency vibration on fracture healing is profoundly influenced by the oestrogen status in mice JF - Disease Models & Mechanisms N2 - Fracture healing is impaired in aged and osteoporotic individuals. Because adequate mechanical stimuli are able to increase bone formation, one therapeutical approach to treat poorly healing fractures could be the application of whole-body vibration, including low-magnitude high-frequency vibration (LMHFV). We investigated the effects of LMHFV on fracture healing in aged osteoporotic mice. Female C57BL/6NCrl mice (n=96) were either ovariectomised (OVX) or sham operated (non-OVX) at age 41 weeks. When aged to 49 weeks, all mice received a femur osteotomy that was stabilised using an external fixator. The mice received whole-body vibrations (20 minutes/day) with 0.3 G: peak-to-peak acceleration and a frequency of 45 Hz. After 10 and 21 days, the osteotomised femurs and intact bones (contra-lateral femurs, lumbar spine) were evaluated using bending-testing, micro-computed tomography (μCT), histology and gene expression analyses. LMHFV disturbed fracture healing in aged non-OVX mice, with significantly reduced flexural rigidity (-81%) and bone formation (-80%) in the callus. Gene expression analyses demonstrated increased oestrogen receptor β (ERβ, encoded by Esr2) and Sost expression in the callus of the vibrated animals, but decreased β-catenin, suggesting that ERβ might mediate these negative effects through inhibition of osteoanabolic Wnt/β-catenin signalling. In contrast, in OVX mice, LMHFV significantly improved callus properties, with increased flexural rigidity (+1398%) and bone formation (+637%), which could be abolished by subcutaneous oestrogen application (0.025 mg oestrogen administered in a 90-day-release pellet). On a molecular level, we found an upregulation of ERα in the callus of the vibrated OVX mice, whereas ERβ was unaffected, indicating that ERα might mediate the osteoanabolic response. Our results indicate a major role for oestrogen in the mechanostimulation of fracture healing and imply that LMHFV might only be safe and effective in confined target populations. KW - fracture healing KW - LMHFV KW - oestrogen receptor signalling KW - whole-body vibration KW - Wnt-signalling Y1 - 2015 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-121109 VL - 8 ER - TY - JOUR A1 - Jakob, Franz A1 - Ebert, Regina A1 - Rudert, Maximilian A1 - Nöth, Ulrich A1 - Walles, Heike A1 - Docheva, Denitsa A1 - Schieker, Matthias A1 - Meinel, Lorenz A1 - Groll, Jürgen T1 - In situ guided tissue regeneration in musculoskeletal diseases and aging JF - Cell and Tissue Research N2 - In situ guided tissue regeneration, also addressed as in situ tissue engineering or endogenous regeneration, has a great potential for population-wide “minimal invasive” applications. During the last two decades, tissue engineering has been developed with remarkable in vitro and preclinical success but still the number of applications in clinical routine is extremely small. Moreover, the vision of population-wide applications of ex vivo tissue engineered constructs based on cells, growth and differentiation factors and scaffolds, must probably be deemed unrealistic for economic and regulation-related issues. Hence, the progress made in this respect will be mostly applicable to a fraction of post-traumatic or post-surgery situations such as big tissue defects due to tumor manifestation. Minimally invasive procedures would probably qualify for a broader application and ideally would only require off the shelf standardized products without cells. Such products should mimic the microenvironment of regenerating tissues and make use of the endogenous tissue regeneration capacities. Functionally, the chemotaxis of regenerative cells, their amplification as a transient amplifying pool and their concerted differentiation and remodeling should be addressed. This is especially important because the main target populations for such applications are the elderly and diseased. The quality of regenerative cells is impaired in such organisms and high levels of inhibitors also interfere with regeneration and healing. In metabolic bone diseases like osteoporosis, it is already known that antagonists for inhibitors such as activin and sclerostin enhance bone formation. Implementing such strategies into applications for in situ guided tissue regeneration should greatly enhance the efficacy of tailored procedures in the future. KW - in situ guided tissue regeneration KW - stem cells KW - scaffolds KW - regenerative medicine KW - mesenchymal tissues Y1 - 2012 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-124738 VL - 347 IS - 3 ER - TY - JOUR A1 - Walsh, J. Bernard A1 - Lems, Willem F. A1 - Karras, Dimitrios A1 - Langdahl, Bente L. A1 - Ljunggren, Osten A1 - Fahrleitner-Pammer, Astrid A1 - Barrett, Annabel A1 - Rajzbaum, Gerald A1 - Jakob, Franz A1 - Marin, Fernando T1 - Effectiveness of Teriparatide in Women Over 75 Years of Age with Severe Osteoporosis: 36-Month Results from the European Forsteo Observational Study (EFOS) JF - Calcified Tissue International N2 - This predefined analysis of the European Forsteo Observational Study (EFOS) aimed to describe clinical fracture incidence, back pain, and health-related quality of life (HRQoL) during 18 months of teriparatide treatment and 18 months post-teriparatide in the subgroup of 589 postmenopausal women with osteoporosis aged ≥75 years. Data on clinical fractures, back pain (visual analogue scale, VAS), and HRQoL (EQ-5D) were collected over 36 months. Fracture data were summarized in 6-month intervals and analyzed using logistic regression with repeated measures. A repeated-measures model analyzed changes from baseline in back pain VAS and EQ-VAS. During the 36-month observation period, 87 (14.8 %) women aged ≥75 years sustained a total of 111 new fractures: 37 (33.3 %) vertebral fractures and 74 (66.7 %) nonvertebral fractures. Adjusted odds of fracture was decreased by 80 % in the 30 to <36–month interval compared with the first 6-month interval (P < 0.009). Although the older subgroup had higher back pain scores and poorer HRQoL at baseline than the younger subgroup, both age groups showed significant reductions in back pain and improvements in HRQoL postbaseline. In conclusion, women aged ≥75 years with severe postmenopausal osteoporosis treated with teriparatide in normal clinical practice showed a reduced clinical fracture incidence by 30 months compared with baseline. An improvement in HRQoL and, possibly, an early and significant reduction in back pain were also observed, which lasted for at least 18 months after teriparatide discontinuation when patients were taking other osteoporosis medication. The results should be interpreted in the context of an uncontrolled observational study. KW - teriparatide KW - osteoporosis KW - health-related quality of life KW - fracture KW - back pain KW - age Y1 - 2012 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-124746 VL - 90 IS - 5 ER - TY - THES A1 - Busch, Albert Franz Jakob T1 - Modification of angiogenesis to abrogate abdominal aortic aneurysm growth T1 - Modifikation von Angiogenese um das Wachstum abdominaler Aortenaneurysmen zu beeinflussen N2 - Introduction: Abdominal aortic aneurysm (AAA) is a pathological saccular enlargement most often of the infrarenal aorta. Eventual rupture is fatal, making preemptive surgical therapy upon a diameter threshold of >50mm the treatment of choice. The pathophysiology, especially the initial trigger aortic remodeling is still largely unknown. However, some characteristic features involved in aneurysm growth have been established, such as medial angiogenesis, low-grade inflammation, vascular smooth muscle cell (VSMC) phenotype switch, extracellular remodeling, altered hemodynamics and an eventual humoral immune answer. Currently, no medical treatment options are available. RNA therapeutics and drug repurposing offer new possibilities to overcome this shortage. Using such to target angiogenesis in the aneurysm wall and investigate their potential mechanisms is the aim of this thesis. Material and Methods: We test our hypothesis by targeting the long non-coding RNA H19 and re-use the anti-cancer drug Lenvatinib in two murine inducible AAA models and one preclinical large animal model in the LDLR-/- pig. Furthermore, a H19-/- mouse is included to verify the results. AAA and control samples from a human biobank along with a primary human cell culture are used to verify results ex vivo by qPCR, WesternBlot, live cell imaging, histo- and immunohistochemistry along with gene array analysis, RNA knockdown, pull-down- and promotor assays. Results: H19 is significantly upregulated in AAA mice models and its knockdown limited aneurysm growth. It is well known that H19 interacts with several transcription factors. We found that cytoplasmic interaction between H19 and hypoxia-inducible factor 1-alpha (HIF1α) increased apoptosis in cultured SMCs associated with sequential p53 stabilization. In contrast, the knockdown of H19 was associated with markedly decreased apoptotic cell rates. Our data underline that HIF1α was essential in mediating the pro-apoptotic effects of H19. Secondly, Lenvatinib was applied both systemically and locally by endovascular means in mice with an established AAA. The drug significantly halted aneurysm growth and array analysis revealed myosin heavy chain 11 (MYH11) as the most differentially regulated target. This was shown to be up regulated after Lenvatinib treatment of primary AAA smooth muscle cells suggesting a salvage mechanism to obtain a contractile phenotype based on gene expression and immunohistochemistry. The same results were shown upon a local endovascular Lenvatinib-coated balloon angioplasty in the established aneurysmatic lesion of a novel atherosclerotic LDLR-/- Yucatan minipig model. Decreased phosphorylation of extracellular-signal regulated kinases 1-2 (ERK1-2) is the downstream effect of Lenvatinib-specific blockage of the vascular endothelial growth factor receptor (VEGFR2). Conclusion: Taking into account the heterogeneity of the disease, inhibition of VSMC phenotype switch, extracellular remodeling and angiogenesis seem promising targets in some if not all AAA patients. Together with surveillance and surgical therapy, these new non-invasive treatment strategies would allow for a more personalized approach to treat this disease. N2 - Einleitung: Das abdominale Aortenaneurysma (AAA) ist eine Erweiterung der infrarenalen Aorta. Die größte Gefahr ist eine Ruptur, sodass eine präemptive chirurgische Ausschaltung ab 50mm Durchmesser empfohlen wird. Insbesondere die initialen Triggermechanismen zur AAA Entstehung sind weiterhin unklar. Einige charakteristische Eigenschaften des Aneurysmawachstums sind z.B. Angiogenese in der Media, low-grade Entzündung, die vascular smooth muscle cell (VSMC) Phänotyp-Änderung, Remodelling der extrazellulären Matrix, eine veränderte Hämodynamik und eine humorale Immunantwort. Gegenwärtig sind neben den chirurgischen, keine medikamentösen Therapiealternativen vorhanden. RNA-Therapien und drug repurposing könnten dies ändern. Ziel dieser Arbeit ist die Beeinflussung der Angiogenese, um das Wachstum von AAAs zu verändern. Material und Methoden: Um diese Hypothese zu überprüfen wurden zwei verschiedene Ansätze verfolgt: Inhibition der long non-coding RNA H19 und die Verwendung von Lenvatinib in zwei Mausmodellen mit induzierbarem AAA und einem präklinischen Großtiermodell im LDLR-/- Schwein. Zusätzlich wurden Versuche in einer H19-/- Maus durchgeführt. AAA und Kontrollen aus einer humanen Biobank in Kombination mit der Verwendung einer primären Zellkultur aus AAA Patientenproben, wurden mittels qPCR, WesternBlot, live cell imaging, Histo- und Immunhistochemie sowie Microarray Analysen und RNA knockdown untersucht. Ergebnisse: Wir zeigen, dass experimenteller knockdown von H19, mittels antisense Oligonukleotiden (LNA-GapmeRs) in vivo das AAA Wachstum signifikant einschränkt. In vitro reduziert dieser knockdown deutlich die Apoptoserate von menschlichen aortalen VSMCs. Mittels array Analyse wurde hypoxia-inducible factor 1-alpha (HIF1α) als Zielgen identifiziert. Zytoplasmatische Interaktion zwischen H19 und HIF1α führt zu einer Stabilisierung von p53. Dieser Mechanismus konnte auch in H19-/- Mäusen bestätigt werden, die nach AAA Induktion kein Aneurysma entwickelten. Zweitens konnte Lenvatinib sowohl bei systemischer, wie auch bei lokaler Applikation in Mäusen mit etabliertem AAA deren Wachstum signifikant einschränken. In einer Microarray Analyse wurde hier myosin heavy chain 11 (MYH11) als am deutlichsten verändertes Gen identifiziert. In primären humanen AAA Zellen war dies nach Lenvatinib Behandlung deutlich hochreguliert und deutet damit einen Erhalt des kontraktilen VSMC Phänotyps an. Der gleiche Effekt konnte im Großtiermodell nach lokaler endovaskulärer Behandlung mit einem Lenvatinib-coated balloon in einem neuen LDLR-/- Yucatan minipig Modell gezeigt werden. Reduzierte Phophorylierung von ERK1-2 ist das Ergebnis der Lenvatinib-spezifischen Blockade von vascular endothelial growth factor receptor (VEGFR2). Schlussfolgerung: Zieht man die Heterogenität der Erkrankung AAA in Betracht, ist die Inhibition von VSMC Phänotyp Änderung, Remodelling der Matrix und Angiogenese möglicherweise ein guter Mechanismus um die Aneurysmen einiger Patienten zu behandeln. Diese neuen Ansätze werden möglicherweise in Kombination mit Überwachung und chirurgischer Therapie einen personalisierten Ansatz in der Therapie erlauben. KW - Aortenaneurysma KW - drug repurposing Y1 - 2021 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-241356 ER - TY - JOUR A1 - Rackwitz, Lars A1 - Eden, Lars A1 - Reppenhagen, Stephan A1 - Reichert, Johannes C. A1 - Jakob, Franz A1 - Walles, Heike A1 - Pullig, Oliver A1 - Tuan, Rocky S. A1 - Rudert, Maximilian A1 - Nöth, Ulrich T1 - Stem cell- and growth factor-based regenerative therapies for avascular necrosis of the femoral head JF - Stem Cell Research & Therapy N2 - Avascular necrosis (AVN) of the femoral head is a debilitating disease of multifactorial genesis, predominately affects young patients, and often leads to the development of secondary osteoarthritis. The evolving field of regenerative medicine offers promising treatment strategies using cells, biomaterial scaffolds, and bioactive factors, which might improve clinical outcome. Early stages of AVN with preserved structural integrity of the subchondral plate are accessible to retrograde surgical procedures, such as core decompression to reduce the intraosseous pressure and to induce bone remodeling. The additive application of concentrated bone marrow aspirates, ex vivo expanded mesenchymal stem cells, and osteogenic or angiogenic growth factors (or both) holds great potential to improve bone regeneration. In contrast, advanced stages of AVN with collapsed subchondral bone require an osteochondral reconstruction to preserve the physiological joint function. Analogously to strategies for osteochondral reconstruction in the knee, anterograde surgical techniques, such as osteochondral transplantation (mosaicplasty), matrix-based autologous chondrocyte implantation, or the use of acellular scaffolds alone, might preserve joint function and reduce the need for hip replacement. This review summarizes recent experimental accomplishments and initial clinical findings in the field of regenerative medicine which apply cells, growth factors, and matrices to address the clinical problem of AVN. KW - osteochondral allografts KW - autologous chondrocyte implantation KW - osteogenesis imperfecta KW - segmental collapse KW - mesenchymal cells KW - progenitor cells KW - stromal cells KW - sheep model KW - colony-stimulating factor KW - core depression Y1 - 2012 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-135413 VL - 3 IS - 7 ER - TY - JOUR A1 - Benisch, Peggy A1 - Schilling, Tatjana A1 - Klein-Hitpass, Ludger A1 - Frey, Sönke P. A1 - Seefried, Lothar A1 - Raaijmakers, Nadja A1 - Krug, Melanie A1 - Regensburger, Martina A1 - Zeck, Sabine A1 - Schinke, Thorsten A1 - Amling, Michael A1 - Ebert, Amling A1 - Jakob, Franz T1 - The Transcriptional Profile of Mesenchymal Stem Cell Populations in Primary Osteoporosis Is Distinct and Shows Overexpression of Osteogenic Inhibitors JF - PLoS One N2 - Primary osteoporosis is an age-related disease characterized by an imbalance in bone homeostasis. While the resorptive aspect of the disease has been studied intensely, less is known about the anabolic part of the syndrome or presumptive deficiencies in bone regeneration. Multipotent mesenchymal stem cells (MSC) are the primary source of osteogenic regeneration. In the present study we aimed to unravel whether MSC biology is directly involved in the pathophysiology of the disease and therefore performed microarray analyses of hMSC of elderly patients (79-94 years old) suffering from osteoporosis (hMSC-OP). In comparison to age-matched controls we detected profound changes in the transcriptome in hMSC-OP, e.g. enhanced mRNA expression of known osteoporosis-associated genes (LRP5, RUNX2, COL1A1) and of genes involved in osteoclastogenesis (CSF1, PTH1R), but most notably of genes coding for inhibitors of WNT and BMP signaling, such as Sclerostin and MAB21L2. These candidate genes indicate intrinsic deficiencies in self-renewal and differentiation potential in osteoporotic stem cells. We also compared both hMSC-OP and non-osteoporotic hMSC-old of elderly donors to hMSC of similar to 30 years younger donors and found that the transcriptional changes acquired between the sixth and the ninth decade of life differed widely between osteoporotic and non-osteoporotic stem cells. In addition, we compared the osteoporotic transcriptome to long term-cultivated, senescent hMSC and detected some signs for pre-senescence in hMSC-OP. Our results suggest that in primary osteoporosis the transcriptomes of hMSC populations show distinct signatures and little overlap with non-osteoporotic aging, although we detected some hints for senescence-associated changes. While there are remarkable inter-individual variations as expected for polygenetic diseases, we could identify many susceptibility genes for osteoporosis known from genetic studies. We also found new candidates, e.g. MAB21L2, a novel repressor of BMP-induced transcription. Such transcriptional changes may reflect epigenetic changes, which are part of a specific osteoporosis-associated aging process. KW - alkaline-phosphatase KW - in vitro KW - bone-mineral density KW - age-related osteoporosis KW - WNT signaling pathway KW - replicative senescence KW - morphogenetic protein KW - parathyroid-hormone KW - growth factor KW - skeletal overexpression Y1 - 2012 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-133379 VL - 7 IS - 9 ER - TY - JOUR A1 - Klotz, Barbara A1 - Mentrup, Birgit A1 - Regensburger, Martina A1 - Zeck, Sabine A1 - Schneidereit, Jutta A1 - Schupp, Nicole A1 - Linden, Christian A1 - Merz, Cornelia A1 - Ebert, Regina A1 - Jakob, Franz T1 - 1,25-Dihydroxyvitamin D3 Treatment Delays Cellular Aging in Human Mesenchymal Stem Cells while Maintaining Their Multipotent Capacity JF - PLoS ONE N2 - 1,25-dihydroxyvitamin D3 (1,25D3) was reported to induce premature organismal aging in fibroblast growth factor-23 (Fgf23) and klotho deficient mice, which is of main interest as 1,25D3 supplementation of its precursor cholecalciferol is used in basic osteoporosis treatment. We wanted to know if 1,25D3 is able to modulate aging processes on a cellular level in human mesenchymal stem cells (hMSC). Effects of 100 nM 1,25D3 on hMSC were analyzed by cell proliferation and apoptosis assay, beta-galactosidase staining, VDR and surface marker immunocytochemistry, RT-PCR of 1,25D3-responsive, quiescence-and replicative senescence-associated genes. 1,25D3 treatment significantly inhibited hMSC proliferation and apoptosis after 72 h and delayed the development of replicative senescence in long-term cultures according to beta-galactosidase staining and P16 expression. Cell morphology changed from a fibroblast like appearance to broad and rounded shapes. Long term treatment did not induce lineage commitment in terms of osteogenic pathways but maintained their clonogenic capacity, their surface marker characteristics (expression of CD73, CD90, CD105) and their multipotency to develop towards the chondrogenic, adipogenic and osteogenic pathways. In conclusion, 1,25D3 delays replicative senescence in primary hMSC while the pro-aging effects seen in mouse models might mainly be due to elevated systemic phosphate levels, which propagate organismal aging. KW - perspectives KW - bone marrow KW - mutant mice KW - oxidative stress KW - transcription factors KW - vitamin-D-receptor KW - differentiation KW - tissue KW - 2',7'-dichlorofluorescin KW - homeostasis Y1 - 2012 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-133392 VL - 7 IS - 1 ER - TY - JOUR A1 - Liedert, Astrid A1 - Röntgen, Viktoria A1 - Schinke, Thorsten A1 - Benisch, Peggy A1 - Ebert, Regina A1 - Jakob, Franz A1 - Klein-Hitpass, Ludger A1 - Lennerz, Jochen K. A1 - Amling, Michael A1 - Ignatius, Anita T1 - Osteoblast-Specific Krm2 Overexpression and Lrp5 Deficiency Have Different Effects on Fracture Healing in Mice JF - PLOS ONE N2 - The canonical Wnt/beta-catenin pathway plays a key role in the regulation of bone remodeling in mice and humans. Two transmembrane proteins that are involved in decreasing the activity of this pathway by binding to extracellular antagonists, such as Dickkopf 1 (Dkk1), are the low-density lipoprotein receptor related protein 5 (Lrp5) and Kremen 2 (Krm2). Lrp 5 deficiency (Lrp5(-/-)) as well as osteoblast-specific overexpression of Krm2 in mice (Col1a1-Krm2) result in severe osteoporosis occurring at young age. In this study, we analyzed the influence of Lrp5 deficiency and osteoblast-specific overexpression of Krm2 on fracture healing in mice using flexible and semi-rigid fracture fixation. We demonstrated that fracture healing was highly impaired in both mouse genotypes, but that impairment was more severe in Col1a1-Krm2 than in Lrp5(-/-) mice and particularly evident in mice in which the more flexible fixation was used. Bone formation was more reduced in Col1a1-Krm2 than in Lrp5(-/-) mice, whereas osteoclast number was similarly increased in both genotypes in comparison with wild-type mice. Using microarray analysis we identified reduced expression of genes mainly involved in osteogenesis that seemed to be responsible for the observed stronger impairment of healing in Col1a1-Krm2 mice. In line with these findings, we detected decreased expression of sphingomyelin phosphodiesterase 3 (Smpd3) and less active beta-catenin in the calli of Col1a1-Krm2 mice. Since Krm2 seems to play a significant role in regulating bone formation during fracture healing, antagonizing KRM2 might be a therapeutic option to improve fracture healing under compromised conditions, such as osteoporosis. KW - autosomal-dominant osteopetrosis KW - receptor related protein KW - high-bone-mass KW - WNT pathway KW - in-vitro KW - cells KW - gene KW - proliferation KW - osteoclasts KW - mutations Y1 - 2014 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-115782 SN - 1932-6203 VL - 9 IS - 7 ER - TY - JOUR A1 - Ebert, Regina A1 - Dotterweich, Julia A1 - Kraus, Sabrina A1 - Tower, Robert J. A1 - Jakob, Franz A1 - Schütze, Norbert T1 - Mesenchymal stem cell contact promotes CCN1 splicing and transcription in myeloma cells N2 - CCN family member 1 (CCN1), also known as cysteine-rich angiogenic inducer 61 (CYR61), belongs to the extracellular matrix-associated CCN protein family. The diverse functions of these proteins include regulation of cell migration, adhesion, proliferation, differentiation and survival/apoptosis, induction of angiogenesis and cellular senescence. Their functions are partly overlapping, largely non-redundant, cell-type specific, and depend on the local microenvironment. To elucidate the role of CCN1 in the crosstalk between stromal cells and myeloma cells, we performed co-culture experiments with primary mesenchymal stem cells (MSC) and the interleukin-6 (IL-6)-dependent myeloma cell line INA-6. Here we show that INA-6 cells display increased transcription and induction of splicing of intron-retaining CCN1 pre-mRNA when cultured in contact with MSC. Protein analyses confirmed that INA-6 cells co-cultured with MSC show increased levels of CCN1 protein consistent with the existence of a pre-mature stop codon in intron 1 that abolishes translation of unspliced mRNA. Addition of recombinant CCN1-Fc protein to INA-6 cells was also found to induce splicing of CCN1 pre-mRNA in a concentration-dependent manner. Only full length CCN1-Fc was able to induce mRNA splicing of all introns, whereas truncated recombinant isoforms lacking domain 4 failed to induce intron splicing. Blocking RGD-dependent integrins on INA-6 cells resulted in an inhibition of these splicing events. These findings expand knowledge on splicing of the proangiogenic, matricellular factor CCN1 in the tumor microenvironment. We propose that contact with MSC-derived CCN1 leads to splicing and enhanced transcription of CCN1 which further contributes to the translation of angiogenic factor CCN1 in myeloma cells, supporting tumor viability and myeloma bone disease. KW - CCN1 KW - Multiple myeloma KW - Mesenchymal stem cells KW - Splicing Y1 - 2014 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-110497 ER - TY - JOUR A1 - Steinert, Andre F. A1 - Weissenberger, Manuel A1 - Kunz, Manuela A1 - Gilbert, Fabian A1 - Ghivizzani, Steven C. A1 - Goebel, Sascha A1 - Jakob, Franz A1 - Nöth, Ulrich A1 - Rudert, Maximilian T1 - Indian hedgehog gene transfer is a chondrogenic inducer of human mesenchymal stem cells N2 - Introduction: To date, no single most-appropriate factor or delivery method has been identified for the purpose of mesenchymal stem cell (MSC)-based treatment of cartilage injury. Therefore, in this study we tested whether gene delivery of the growth factor Indian hedgehog (IHH) was able to induce chondrogenesis in human primary MSCs, and whether it was possible by such an approach to modulate the appearance of chondrogenic hypertrophy in pellet cultures in vitro. Methods: First-generation adenoviral vectors encoding the cDNA of the human IHH gene were created by cre-lox recombination and used alone or in combination with adenoviral vectors, bone morphogenetic protein-2 (Ad.BMP- 2), or transforming growth factor beta-1 (Ad.TGF-b1) to transduce human bone-marrow derived MSCs at 5 × 102 infectious particles/cell. Thereafter, 3 × 105 cells were seeded into aggregates and cultured for 3 weeks in serumfree medium, with untransduced or marker gene transduced cultures as controls. Transgene expressions were determined by ELISA, and aggregates were analysed histologically, immunohistochemically, biochemically and by RT-PCR for chondrogenesis and hypertrophy. Results: IHH, TGF-b1 and BMP-2 genes were equipotent inducers of chondrogenesis in primary MSCs, as evidenced by strong staining for proteoglycans, collagen type II, increased levels of glycosaminoglycan synthesis, and expression of mRNAs associated with chondrogenesis. IHH-modified aggregates, alone or in combination, also showed a tendency to progress towards hypertrophy, as judged by the expression of alkaline phosphatase and stainings for collagen type X and Annexin 5. Conclusion: As this study provides evidence for chondrogenic induction of MSC aggregates in vitro via IHH gene delivery, this technology may be efficiently employed for generating cartilaginous repair tissues in vivo. KW - Medizin Y1 - 2012 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-75425 ER - TY - JOUR A1 - Hochleitner, Gernot A1 - Jüngst, Tomasz A1 - Brown, Toby D A1 - Hahn, Kathrin A1 - Moseke, Claus A1 - Jakob, Franz A1 - Dalton, Paul D A1 - Groll, Jürgen T1 - Additive manufacturing of scaffolds with sub-micron filaments via melt electrospinning writing JF - Biofabrication N2 - The aim of this study was to explore the lower resolution limits of an electrohydrodynamic process combined with direct writing technology of polymer melts. Termed melt electrospinning writing, filaments are deposited layer-by-layer to produce discrete three-dimensional scaffolds for in vitro research. Through optimization of the parameters (flow rate, spinneret diameter, voltage, collector distance) for poly-ϵ-caprolactone, we could direct-write coherent scaffolds with ultrafine filaments, the smallest being 817 ± 165 nm. These low diameter filaments were deposited to form box-structures with a periodicity of 100.6 ± 5.1 μm and a height of 80 μm (50 stacked filaments; 100 overlap at intersections). We also observed oriented crystalline regions within such ultrafine filaments after annealing at 55 °C. The scaffolds were printed upon NCO-sP(EO-stat-PO)-coated glass slide surfaces and withstood frequent liquid exchanges with negligible scaffold detachment for at least 10 days in vitro. KW - additive manufacturing KW - 3D printing KW - biodegradable polymers KW - microstructures KW - nanostructures Y1 - 2015 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-254053 VL - 7 IS - 3 ER - TY - JOUR A1 - Ebert, Regina A1 - Benisch, Peggy A1 - Krug, Melanie A1 - Zeck, Sabine A1 - Meißner-Weigl, Jutta A1 - Steinert, Andre A1 - Rauner, Martina A1 - Hofbauer, Lorenz A1 - Jakob, Franz T1 - Acute phase serum amyloid A induces proinflammatory cytokines and mineralization via toll-like receptor 4 in mesenchymal stem cells JF - Stem Cell Research N2 - The role of serum amyloid A (SAA) proteins, which are ligands for toll-like receptors, was analyzed in human bone marrow-derived mesenchymal stem cells (hMSCs) and their osteogenic offspring with a focus on senescence, differentiation andmineralization. In vitro aged hMSC developed a senescence-associated secretory phenotype (SASP), resulting in enhanced SAA1/2, TLR2/4 and proinflammatory cytokine (IL6, IL8, IL1\(\beta\), CXCL1, CXCL2) expression before entering replicative senescence. Recombinant human SAA1 (rhSAA1) induced SASP-related genes and proteins in MSC, which could be abolished by cotreatment with the TLR4-inhibitor CLI-095. The same pattern of SASP-resembling genes was stimulated upon induction of osteogenic differentiation, which is accompanied by autocrine SAA1/2 expression. In this context additional rhSAA1 enhanced the SASP-like phenotype, accelerated the proinflammatory phase of osteogenic differentiation and enhanced mineralization. Autocrine/paracrine and rhSAA1 via TLR4 stimulate a proinflammatory phenotype that is both part of the early phase of osteogenic differentiation and the development of senescence. This signaling cascade is tightly involved in bone formation and mineralization, but may also propagate pathological extraosseous calcification conditions such as calcifying inflammation and atherosclerosis. KW - human atherosclerotic lesions KW - senescence KW - expression KW - toll-like receptor KW - mineralization KW - osteogenic differentiation KW - serum amyloid A KW - inflammation KW - mesenchymal stem cells KW - WNT5A KW - model KW - lines KW - stromal cells KW - RT-PCR Y1 - 2015 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-148491 VL - 15 ER - TY - JOUR A1 - Kaltdorf, Martin A1 - Breitenbach, Tim A1 - Karl, Stefan A1 - Fuchs, Maximilian A1 - Kessie, David Komla A1 - Psota, Eric A1 - Prelog, Martina A1 - Sarukhanyan, Edita A1 - Ebert, Regina A1 - Jakob, Franz A1 - Dandekar, Gudrun A1 - Naseem, Muhammad A1 - Liang, Chunguang A1 - Dandekar, Thomas T1 - Software JimenaE allows efficient dynamic simulations of Boolean networks, centrality and system state analysis JF - Scientific Reports N2 - The signal modelling framework JimenaE simulates dynamically Boolean networks. In contrast to SQUAD, there is systematic and not just heuristic calculation of all system states. These specific features are not present in CellNetAnalyzer and BoolNet. JimenaE is an expert extension of Jimena, with new optimized code, network conversion into different formats, rapid convergence both for system state calculation as well as for all three network centralities. It allows higher accuracy in determining network states and allows to dissect networks and identification of network control type and amount for each protein with high accuracy. Biological examples demonstrate this: (i) High plasticity of mesenchymal stromal cells for differentiation into chondrocytes, osteoblasts and adipocytes and differentiation-specific network control focusses on wnt-, TGF-beta and PPAR-gamma signaling. JimenaE allows to study individual proteins, removal or adding interactions (or autocrine loops) and accurately quantifies effects as well as number of system states. (ii) Dynamical modelling of cell–cell interactions of plant Arapidopsis thaliana against Pseudomonas syringae DC3000: We analyze for the first time the pathogen perspective and its interaction with the host. We next provide a detailed analysis on how plant hormonal regulation stimulates specific proteins and who and which protein has which type and amount of network control including a detailed heatmap of the A.thaliana response distinguishing between two states of the immune response. (iii) In an immune response network of dendritic cells confronted with Aspergillus fumigatus, JimenaE calculates now accurately the specific values for centralities and protein-specific network control including chemokine and pattern recognition receptors. KW - cellular signalling networks KW - computer modelling Y1 - 2023 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-313303 VL - 13 ER - TY - JOUR A1 - Ebert, Regina A1 - Weissenberger, Manuel A1 - Braun, Clemens A1 - Wagenbrenner, Mike A1 - Herrmann, Marietta A1 - Müller‐Deubert, Sigrid A1 - Krug, Melanie A1 - Jakob, Franz A1 - Rudert, Maximilian T1 - Impaired regenerative capacity and senescence‐associated secretory phenotype in mesenchymal stromal cells from samples of patients with aseptic joint arthroplasty loosening JF - Journal of Orthopaedic Research N2 - Aseptic loosening of total hip and knee joint replacements is the most common indication for revision surgery after primary hip and knee arthroplasty. Research suggests that exposure and uptake of wear by mesenchymal stromal cells (MSC) and macrophages results in the secretion of proinflammatory cytokines and local osteolysis, but also impaired cell viability and regenerative capacity of MSC. Therefore, this in vitro study compared the regenerative and differentiation capacity of MSC derived from patients undergoing primary total hip arthroplasty (MSCprim) to MSC derived from patients undergoing revision surgery after aseptic loosening of total hip and knee joint implants (MSCrev). Regenerative capacity was examined by measuring the cumulative population doubling (CPD) in addition to the number of passages until cells stopped proliferating. Osteogenesis and adipogenesis in monolayer cultures were assessed using histological stainings. Furthermore, RT‐PCR was performed to evaluate the relative expression of osteogenic and adipogenic marker genes as well as the expression of markers for a senescence‐associated secretory phenotype (SASP). MSCrev possessed a limited regenerative capacity in comparison to MSCprim. Interestingly, MSCrev also showed an impaired osteogenic and adipogenic differentiation capacity compared to MSCprim and displayed a SASP early after isolation. Whether this is the cause or the consequence of the aseptic loosening of total joint implants remains unclear. Future research should focus on the identification of specific cell markers on MSCprim, which may influence complication rates such as aseptic loosening of total joint arthroplasty to further individualize and optimize total joint arthroplasty. KW - aseptic loosening KW - mesenchymal stromal cells KW - regenerative capacity KW - senescence‐associated secretory phenotype Y1 - 2022 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-238963 VL - 40 IS - 2 SP - 513 EP - 523 ER - TY - JOUR A1 - Altmann, Stephan A1 - Mut, Jürgen A1 - Wolf, Natalia A1 - Meißner-Weigl, Jutta A1 - Rudert, Maximilian A1 - Jakob, Franz A1 - Gutmann, Marcus A1 - Lühmann, Tessa A1 - Seibel, Jürgen A1 - Ebert, Regina T1 - Metabolic glycoengineering in hMSC-TERT as a model for skeletal precursors by using modified azide/alkyne monosaccharides JF - International Journal of Molecular Sciences N2 - Metabolic glycoengineering enables a directed modification of cell surfaces by introducing target molecules to surface proteins displaying new features. Biochemical pathways involving glycans differ in dependence on the cell type; therefore, this technique should be tailored for the best results. We characterized metabolic glycoengineering in telomerase-immortalized human mesenchymal stromal cells (hMSC-TERT) as a model for primary hMSC, to investigate its applicability in TERT-modified cell lines. The metabolic incorporation of N-azidoacetylmannosamine (Ac\(_4\)ManNAz) and N-alkyneacetylmannosamine (Ac\(_4\)ManNAl) into the glycocalyx as a first step in the glycoengineering process revealed no adverse effects on cell viability or gene expression, and the in vitro multipotency (osteogenic and adipogenic differentiation potential) was maintained under these adapted culture conditions. In the second step, glycoengineered cells were modified with fluorescent dyes using Cu-mediated click chemistry. In these analyses, the two mannose derivatives showed superior incorporation efficiencies compared to glucose and galactose isomers. In time-dependent experiments, the incorporation of Ac\(_4\)ManNAz was detectable for up to six days while Ac\(_4\)ManNAl-derived metabolites were absent after two days. Taken together, these findings demonstrate the successful metabolic glycoengineering of immortalized hMSC resulting in transient cell surface modifications, and thus present a useful model to address different scientific questions regarding glycosylation processes in skeletal precursors. KW - hMSC-TERT KW - metabolic glycoengineering KW - glycocalyx KW - modified monosaccharides KW - click chemistry Y1 - 2021 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-259247 SN - 1422-0067 VL - 22 IS - 6 ER - TY - JOUR A1 - Vogt, Marius A1 - Girschick, Hermann A1 - Schweitzer, Tilmann A1 - Benoit, Clemens A1 - Holl-Wieden, Annette A1 - Seefried, Lothar A1 - Jakob, Franz A1 - Hofmann, Christine T1 - Pediatric hypophosphatasia: lessons learned from a retrospective single-center chart review of 50 children JF - Orphanet Journal of Rare Diseases N2 - Background Hypophosphatasia (HPP) is a rare, inherited metabolic disorder caused by loss-of-function mutations in the ALPL gene that encodes the tissue-nonspecific alkaline phosphatase TNAP (ORPHA 436). Its clinical presentation is highly heterogeneous with a remarkably wide-ranging severity. HPP affects patients of all ages. In children HPP-related musculoskeletal symptoms may mimic rheumatologic conditions and diagnosis is often difficult and delayed. To improve the understanding of HPP in children and in order to shorten the diagnostic time span in the future we studied the natural history of the disease in our large cohort of pediatric patients. This single centre retrospective chart review included longitudinal data from 50 patients with HPP diagnosed and followed at the University Children's Hospital Wuerzburg, Germany over the last 25 years. Results The cohort comprises 4 (8%) perinatal, 17 (34%) infantile and 29 (58%) childhood onset HPP patients. Two patients were deceased at the time of data collection. Diagnosis was based on available characteristic clinical symptoms (in 88%), low alkaline phosphatase (AP) activity (in 96%), accumulating substrates of AP (in 58%) and X-ray findings (in 48%). Genetic analysis was performed in 48 patients (31 compound heterozygous, 15 heterozygous, 2 homozygous mutations per patient), allowing investigations on genotype-phenotype correlations. Based on anamnestic data, median age at first clinical symptoms was 3.5 months (min. 0, max. 107), while median time to diagnosis was 13 months (min. 0, max. 103). Common symptoms included: impairment of motor skills (78%), impairment of mineralization (72%), premature loss of teeth (64%), musculoskeletal pain and craniosynostosis (each 64%) and failure to thrive (62%). Up to now 20 patients started medical treatment with Asfotase alfa. Conclusions Reported findings support the clinical perception of HPP being a chronic multi-systemic disease with often delayed diagnosis. Our natural history information provides detailed insights into the prevalence of different symptoms, which can help to improve and shorten diagnostics and thereby lead to an optimised medical care, especially with promising therapeutic options such as enzyme-replacement-therapy with Asfotase alfa in mind. KW - hypophosphatasia KW - alkaline phosphatase KW - asfotase alfa KW - rare bone disease KW - osteomalacia KW - rickets Y1 - 2020 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-230505 VL - 15 ER - TY - JOUR A1 - Kemmler, Wolfgang A1 - Kohl, Matthias A1 - Jakob, Franz A1 - Engelke, Klaus A1 - Stengel, Simon von T1 - Effects of high intensity dynamic resistance exercise and whey protein supplements on osteosarcopenia in older men with low bone and muscle mass. Final results of the randomized controlled FrOST study JF - Nutrients N2 - The present study aimed to evaluate the effect of high intensity dynamic resistance exercise (HIT-DRT) and whey protein supplementation (WPS) on bone mineral density (BMD) and sarcopenia parameters in osteosarcopenic men. Men ≥ 72 years with osteosarcopenia (n = 43) were randomly assigned to a HIT-RT (HIT-RT: n = 21) or a non-training control group (n = 22). Supervised HIT-RT twice/week was applied for 18 months, while the control group maintained their habitual lifestyle. Supplying WPS, total protein intake amounted to 1.5–1.6 (HIT-RT) and 1.2 g/kg/body mass/d (control). Both groups were supplied with calcium and vitamin D. Primary study outcomes were BMD and the sarcopenia Z-score. After adjusting for multiplicity, we observed significant positive effects for sarcopenia Z-score (standardized mean difference (SMD): 1.40), BMD at lumbar spine (SMD: 0.72) and total hip (SMD: 0.72). In detail, effect sizes for skeletal muscle mass changes were very pronounced (1.97, p < 0.001), while effects for functional sarcopenia parameters were moderate (0.87, p = 0.008; handgrip strength) or low (0.39, p = 0.209; gait velocity). Apart from one man who reported short periods of temporary worsening of existing joint pain, no HIT-RT/WPS-related adverse effects or injuries were reported. We consider HIT-RT supported by whey protein supplementation as a feasible, attractive, safe and highly effective option to fight osteosarcopenia in older men. KW - resistance exercise KW - osteopenia KW - sarcopenia KW - bone mineral density Y1 - 2020 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-211108 SN - 2072-6643 VL - 12 IS - 8 ER - TY - JOUR A1 - Herrmann, Marietta A1 - Engelke, Klaus A1 - Ebert, Regina A1 - Müller-Deubert, Sigrid A1 - Rudert, Maximilian A1 - Ziouti, Fani A1 - Jundt, Franziska A1 - Felsenberg, Dieter A1 - Jakob, Franz T1 - Interactions between muscle and bone — Where physics meets biology JF - Biomolecules N2 - Muscle and bone interact via physical forces and secreted osteokines and myokines. Physical forces are generated through gravity, locomotion, exercise, and external devices. Cells sense mechanical strain via adhesion molecules and translate it into biochemical responses, modulating the basic mechanisms of cellular biology such as lineage commitment, tissue formation, and maturation. This may result in the initiation of bone formation, muscle hypertrophy, and the enhanced production of extracellular matrix constituents, adhesion molecules, and cytoskeletal elements. Bone and muscle mass, resistance to strain, and the stiffness of matrix, cells, and tissues are enhanced, influencing fracture resistance and muscle power. This propagates a dynamic and continuous reciprocity of physicochemical interaction. Secreted growth and differentiation factors are important effectors of mutual interaction. The acute effects of exercise induce the secretion of exosomes with cargo molecules that are capable of mediating the endocrine effects between muscle, bone, and the organism. Long-term changes induce adaptations of the respective tissue secretome that maintain adequate homeostatic conditions. Lessons from unloading, microgravity, and disuse teach us that gratuitous tissue is removed or reorganized while immobility and inflammation trigger muscle and bone marrow fatty infiltration and propagate degenerative diseases such as sarcopenia and osteoporosis. Ongoing research will certainly find new therapeutic targets for prevention and treatment. KW - muscle KW - bone KW - mechanosensing KW - mechanotransduction KW - myokines KW - osteokines adaptation Y1 - 2020 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-203399 SN - 2218-273X VL - 10 IS - 3 ER - TY - JOUR A1 - Liedtke, Daniel A1 - Hofmann, Christine A1 - Jakob, Franz A1 - Klopocki, Eva A1 - Graser, Stephanie T1 - Tissue-Nonspecific Alkaline Phosphatase—A Gatekeeper of Physiological Conditions in Health and a Modulator of Biological Environments in Disease JF - Biomolecules N2 - Tissue-nonspecific alkaline phosphatase (TNAP) is a ubiquitously expressed enzyme that is best known for its role during mineralization processes in bones and skeleton. The enzyme metabolizes phosphate compounds like inorganic pyrophosphate and pyridoxal-5′-phosphate to provide, among others, inorganic phosphate for the mineralization and transportable vitamin B6 molecules. Patients with inherited loss of function mutations in the ALPL gene and consequently altered TNAP activity are suffering from the rare metabolic disease hypophosphatasia (HPP). This systemic disease is mainly characterized by impaired bone and dental mineralization but may also be accompanied by neurological symptoms, like anxiety disorders, seizures, and depression. HPP characteristically affects all ages and shows a wide range of clinical symptoms and disease severity, which results in the classification into different clinical subtypes. This review describes the molecular function of TNAP during the mineralization of bones and teeth, further discusses the current knowledge on the enzyme’s role in the nervous system and in sensory perception. An additional focus is set on the molecular role of TNAP in health and on functional observations reported in common laboratory vertebrate disease models, like rodents and zebrafish. KW - TNAP KW - hypophosphatasia KW - HPP KW - zebrafish KW - mineralization KW - ALPL KW - craniosynostosis KW - teeth KW - nervous system Y1 - 2020 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-220096 SN - 2218-273X VL - 10 IS - 12 PB - MDPI ER - TY - JOUR A1 - Schmalzl, Jonas A1 - Plumhoff, Piet A1 - Gilbert, Fabian A1 - Gohlke, Frank A1 - Konrads, Christian A1 - Brunner, Ulrich A1 - Jakob, Franz A1 - Ebert, Regina A1 - Steinert, Andre F. T1 - Tendon-derived stem cells from the long head of the biceps tendon JF - Bone & Joint Research N2 - Objectives The long head of the biceps (LHB) is often resected in shoulder surgery and could therefore serve as a cell source for tissue engineering approaches in the shoulder. However, whether it represents a suitable cell source for regenerative approaches, both in the inflamed and non-inflamed states, remains unclear. In the present study, inflamed and native human LHBs were comparatively characterized for features of regeneration. Methods In total, 22 resected LHB tendons were classified into inflamed samples (n = 11) and non-inflamed samples (n = 11). Proliferation potential and specific marker gene expression of primary LHB-derived cell cultures were analyzed. Multipotentiality, including osteogenic, adipogenic, chondrogenic, and tenogenic differentiation potential of both groups were compared under respective lineage-specific culture conditions. Results Inflammation does not seem to affect the proliferation rate of the isolated tendon-derived stem cells (TDSCs) and the tenogenic marker gene expression. Cells from both groups showed an equivalent osteogenic, adipogenic, chondrogenic and tenogenic differentiation potential in histology and real-time polymerase chain reaction (RT-PCR) analysis. Conclusion These results suggest that the LHB tendon might be a suitable cell source for regenerative approaches, both in inflamed and non-inflamed states. The LHB with and without tendinitis has been characterized as a novel source of TDSCs, which might facilitate treatment of degeneration and induction of regeneration in shoulder surgery. KW - biceps tendon KW - tendon-derived stem cell KW - mesenchymal stem cell KW - tissue engineering KW - shoulder Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-200370 VL - 8 IS - 9 ER - TY - JOUR A1 - Seefried, Lothar A1 - Genest, Franca A1 - Baumann, Jasmin A1 - Heidemeier, Anke A1 - Meffert, Rainer A1 - Jakob, Franz T1 - Efficacy of Zoledronic Acid in the Treatment of Nonmalignant Painful Bone Marrow Lesions: A Triple‐Blind, Randomized, Placebo‐Controlled Phase III Clinical Trial (ZoMARS) JF - Journal of Bone and Mineral Research N2 - Bone marrow lesions (BML) represent areas of deteriorated bone structure and metabolism characterized by pronounced water‐equivalent signaling within the trabecular bone on magnetic resonance imaging (MRI). BML are associated with repair mechanisms subsequent to various clinical conditions associated with inflammatory and non‐inflammatory injury to the bone. There is no approved treatment for this condition. Bisphosphonates are known to improve bone stability in osteoporosis and other bone disorders and have been used off‐label to treat BML. A randomized, triple‐blind, placebo‐controlled phase III trial was conducted to assess efficacy and safety of single‐dose zoledronic acid (ZOL) 5 mg iv with vitamin D 1000 IU/d as opposed to placebo with vitamin D 1000 IU/d in 48 patients (randomized 2:1) with BML. Primary efficacy endpoint was reduction of edema volume 6 weeks after treatment as assessed by MRI. After treatment, mean BML volume decreased by 64.53% (±41.92%) in patients receiving zoledronic acid and increased by 14.43% (±150.46%) in the placebo group (p = 0.007). A decrease in BML volume was observed in 76.5% of patients receiving ZOL and in 50% of the patients receiving placebo. Pain level (visual analogue scale [VAS]) and all categories of the pain disability index (PDI) improved with ZOL versus placebo after 6 weeks but reconciled after 6 additional weeks of follow‐up. Six serious adverse events occurred in 5 patients, none of which were classified as related to the study drug. No cases of osteonecrosis or fractures occurred. Therefore, single‐dose zoledronic acid 5 mg iv together with vitamin D may enhance resolution of bone marrow lesions over 6 weeks along with reduction of pain compared with vitamin D supplementation only. KW - bone biology KW - osteoporosis KW - bone marrow lesion/edema KW - bisphosphonates KW - zoledronic acid Y1 - 2022 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-276368 VL - 37 IS - 3 SP - 420 EP - 427 ER - TY - JOUR A1 - Kemmler, Wolfgang A1 - Kohl, Matthias A1 - Fröhlich, Michael A1 - Jakob, Franz A1 - Engelke, Klaus A1 - von Stengel, Simon A1 - Schoene, Daniel T1 - Effects of High‐Intensity Resistance Training on Osteopenia and Sarcopenia Parameters in Older Men with Osteosarcopenia—One‐Year Results of the Randomized Controlled Franconian Osteopenia and Sarcopenia Trial (FrOST) JF - Journal of Bone and Mineral Research N2 - Dynamic resistance exercise (DRT) might be the most promising agent for fighting sarcopenia in older people. However, the positive effect of DRT on osteopenia/osteoporosis in men has still to be confirmed. To evaluate the effect of low‐volume/high‐intensity (HIT)‐DRT on bone mineral density (BMD) and skeletal muscle mass index (SMI) in men with osteosarcopenia, we initiated the Franconian Osteopenia and Sarcopenia Trial (FrOST). Forty‐three sedentary community‐dwelling older men (aged 73 to 91 years) with osteopenia/osteoporosis and SMI‐based sarcopenia were randomly assigned to a HIT‐RT exercise group (EG; n = 21) or a control group (CG; n = 22). HIT‐RT provided a progressive, periodized single‐set DRT on machines with high intensity, effort, and velocity twice a week, while CG maintained their lifestyle. Both groups were adequately supplemented with whey protein, vitamin D, and calcium. Primary study endpoint was integral lumbar spine (LS) BMD as determined by quantitative computed tomography. Core secondary study endpoint was SMI as determined by dual‐energy X‐ray absorptiometry. Additional study endpoints were BMD at the total hip and maximum isokinetic hip−/leg‐extensor strength (leg press). After 12 months of exercise, LS‐BMD was maintained in the EG and decreased significantly in the CG, resulting in significant between‐group differences (p < 0.001; standardized mean difference [SMD] = 0.90). In parallel, SMI increased significantly in the EG and decreased significantly in the CG (p < 0.001; SMD = 1.95). Total hip BMD changes did not differ significantly between the groups (p = 0.064; SMD = 0.65), whereas changes in maximum hip−/leg‐extensor strength were much more prominent (p < 0.001; SMD = 1.92) in the EG. Considering dropout (n = 2), attendance rate (95%), and unintended side effects/injuries (n = 0), we believe our HIT‐RT protocol to be feasible, attractive, and safe. In summary, we conclude that our combined low‐threshold HIT‐RT/protein/vitamin D/calcium intervention was feasible, safe, and effective for tackling sarcopenia and osteopenia/osteoporosis in older men with osteosarcopenia. KW - exercise KW - osteoporosis KW - sarcopenia KW - aging KW - bone QCT Y1 - 2020 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-214609 VL - 35 IS - 9 SP - 1634 EP - 1644 ER - TY - JOUR A1 - Chaudry, Oliver A1 - Grimm, Alexandra A1 - Friedberger, Andreas A1 - Kemmler, Wolfgang A1 - Uder, Michael A1 - Jakob, Franz A1 - Quick, Harald H. A1 - von Stengel, Simon A1 - Engelke, Klaus T1 - Magnetic Resonance Imaging and Bioelectrical Impedance Analysis to Assess Visceral and Abdominal Adipose Tissue JF - Obesity N2 - Objective This study aimed to compare a state‐of‐the‐art bioelectrical impedance analysis (BIA) device with two‐point Dixon magnetic resonance imaging (MRI) for the quantification of visceral adipose tissue (VAT) as a health‐related risk factor. Methods A total of 63 male participants were measured using a 3‐T MRI scanner and a segmental, multifrequency BIA device. MRI generated fat fraction (FF) maps, in which VAT volume, total abdominal adipose tissue volume, and FF of visceral and total abdominal compartments were quantified. BIA estimated body fat mass and VAT area. Results Coefficients of determination between abdominal (r\(^{2}\) = 0.75) and visceral compartments (r\(^{2}\) = 0.78) were similar for both groups, but slopes differed by a factor of two. The ratio of visceral to total abdominal FF was increased in older men compared with younger men. This difference was not detected with BIA. MRI and BIA measurements of the total abdominal volume correlated moderately (r\(^{2}\) = 0.31‐0.56), and visceral measurements correlated poorly (r\(^{2}\) = 0.13‐0.44). Conclusions Visceral BIA measurements agreed better with MRI measurements of the total abdomen than of the visceral compartment, indicating that BIA visceral fat area assessment cannot differentiate adipose tissue between visceral and abdominal compartments in young and older participants. Y1 - 2020 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-213591 VL - 28 IS - 2 SP - 277 EP - 283 ER - TY - JOUR A1 - Mages, Michelle A1 - Shojaa, Mahdieh A1 - Kohl, Matthias A1 - Stengel, Simon von A1 - Becker, Clemens A1 - Gosch, Markus A1 - Jakob, Franz A1 - Kerschan-Schindl, Katharina A1 - Kladny, Bernd A1 - Klöckner, Nicole A1 - Lange, Uwe A1 - Middeldorf, Stefan A1 - Peters, Stefan A1 - Schoene, Daniel A1 - Sieber, Cornel C. A1 - Tholen, Reina A1 - Thomasius, Friederike E. A1 - Uder, Michael A1 - Kemmler, Wolfgang T1 - Exercise effects on Bone Mineral Density in men JF - Nutrients N2 - In contrast to postmenopausal women, evidence for a favorable effect of exercise on Bone Mineral Density (BMD) is still limited for men. This might be due to the paucity of studies, but also to the great variety of participants and study characteristics that may dilute study results. The aim of the present systematic review and meta-analysis was to evaluate the effect of exercise on BMD changes with rational eligibility criteria. A comprehensive search of six electronic databases up to 15 March 2021 was conducted. Briefly, controlled trials ≥6 months that determined changes in areal BMD in men >18 years old, with no apparent diseases or pharmacological therapy that relevantly affect bone metabolism, were included. BMD changes (standardized mean differences: SMD) of the lumbar spine (LS) and femoral neck (FN) were considered as outcomes. Twelve studies with 16 exercise and 12 control groups were identified. The pooled estimate of random-effect analysis was SMD = 0.38, 95%-CI: 0.14–0.61 and SMD = 0.25, 95%-CI: 0.00–0.49, for LS and FN, respectively. Heterogeneity between the trials was low–moderate. Funnel plots and rank and regression correlation tests indicate evidence for small study publication bias for LS but not FN-BMD. Subgroup analyses that focus on study length, type of exercise and methodologic quality revealed no significant difference between each of the three categories. In summary, we provided further evidence for a low but significant effect of exercise on BMD in men. However, we are currently unable to give even rough exercise recommendations for male cohorts. KW - Bone Mineral Density KW - exercise KW - men KW - overview Y1 - 2021 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-250247 SN - 2072-6643 VL - 13 IS - 12 ER - TY - JOUR A1 - Graser, Stephanie A1 - Liedtke, Daniel A1 - Jakob, Franz T1 - TNAP as a new player in chronic inflammatory conditions and metabolism JF - International Journal of Molecular Sciences N2 - This review summarizes important information on the ectoenzyme tissue-nonspecific alkaline phosphatase (TNAP) and gives a brief insight into the symptoms, diagnostics, and treatment of the rare disease Hypophosphatasia (HPP), which is resulting from mutations in the TNAP encoding ALPL gene. We emphasize the role of TNAP beyond its well-known contribution to mineralization processes. Therefore, above all, the impact of the enzyme on central molecular processes in the nervous system and on inflammation is presented here. KW - TNAP KW - Hypophosphatasia KW - HPP KW - mineralization KW - nervous system KW - inflammation Y1 - 2021 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-258888 SN - 1422-0067 VL - 22 IS - 2 ER - TY - JOUR A1 - Müller-Deubert, Sigrid A1 - Seefried, Lothar A1 - Krug, Melanie A1 - Jakob, Franz A1 - Ebert, Regina T1 - Epidermal growth factor as a mechanosensitizer in human bone marrow stromal cells JF - Stem Cell Research N2 - Epidermal growth factors (EGFs) e.g. EGF, heparin-binding EGF and transforming growth factor alpha and their receptors e.g. EGFR and ErbB2 control proinflammatory signaling and modulate proliferation in bone marrow stromal cells (BMSC). Interleukin-6 and interleukin-8 are EGF targets and participate in the inflammatory phase of bone regeneration via non-canonical wnt signaling. BMSC differentiation is also influenced by mechanical strain-related activation of ERK1/2 and AP-1, but the role of EGFR signaling in mechanotransduction is unclear. We investigated the effects of EGFR signaling in telomerase-immortalized BMSC, transfected with a luciferase reporter, comprising a mechanoresponsive AP1 element, using ligands, neutralizing antibodies and EGFR inhibitors on mechanotransduction and we found that EGF via EGFR increased the response to mechanical strain. Results were confirmed by qPCR analysis of mechanoresponsive genes. EGF-responsive interleukin-6 and interleukin-8 were synergistically enhanced by EGF stimulation and mechanical strain. We show here in immortalized and primary BMSC that EGFR signaling enhances mechanotransduction, indicating that the EGF system is a mechanosensitizer in BMSC. Alterations in mechanosensitivity and -adaptation are contributors to age-related diseases like osteoporosis and the identification of a suitable mechanosensitizer could be beneficial. The role of the synergism of these signaling cascades in physiology and disease remains to be unraveled. KW - mechanotransduction KW - bone marrow stromal cells KW - epidermal growth factor KW - signaling Y1 - 2017 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-170247 VL - 24 ER - TY - JOUR A1 - Liedert, Astrid A1 - Nemitz, Claudia A1 - Haffner-Luntzer, Melanie A1 - Schick, Fabian A1 - Jakob, Franz A1 - Ignatius, Anita T1 - Effects of estrogen receptor and Wnt signaling activation on mechanically induced bone formation in a mouse model of postmenopausal bone loss JF - International Journal of Molecular Sciences N2 - In the adult skeleton, bone remodeling is required to replace damaged bone and functionally adapt bone mass and structure according to the mechanical requirements. It is regulated by multiple endocrine and paracrine factors, including hormones and growth factors, which interact in a coordinated manner. Because the response of bone to mechanical signals is dependent on functional estrogen receptor (ER) and Wnt/β-catenin signaling and is impaired in postmenopausal osteoporosis by estrogen deficiency, it is of paramount importance to elucidate the underlying mechanisms as a basis for the development of new strategies in the treatment of osteoporosis. The present study aimed to investigate the effectiveness of the activation of the ligand-dependent ER and the Wnt/β-catenin signal transduction pathways on mechanically induced bone formation using ovariectomized mice as a model of postmenopausal bone loss. We demonstrated that both pathways interact in the regulation of bone mass adaption in response to mechanical loading and that the activation of Wnt/β-catenin signaling considerably increased mechanically induced bone formation, whereas the effects of estrogen treatment strictly depended on the estrogen status in the mice. KW - bone remodeling KW - mechanotransduction KW - ER signaling KW - Wnt/β-catenin signaling KW - ovariectomy Y1 - 2020 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-285487 SN - 1422-0067 VL - 21 IS - 21 ER - TY - JOUR A1 - Horas, Konstantin A1 - van Herck, Ulrike A1 - Maier, Gerrit S. A1 - Maus, Uwe A1 - Harrasser, Norbert A1 - Jakob, Franz A1 - Weissenberger, Manuel A1 - Arnholdt, Jörg A1 - Holzapfel, Boris M. A1 - Rudert, Maximilian T1 - Does vitamin D deficiency predict tumour malignancy in patients with bone tumours? Data from a multi-center cohort analysis JF - Journal of Bone Oncology N2 - Vitamin D deficiency is a global health concern that is estimated to afflict over one billion people globally. The major role of vitamin D is that of a regulator of calcium and phosphate metabolism, thus, being essential for proper bone mineralisation. Concomitantly, vitamin D is known to exert numerous extra-skeletal actions. For example, it has become evident that vitamin D has direct anti-proliferative, pro-differentiation and pro-apoptotic actions on cancer cells. Hence, vitamin D deficiency has been associated with increased cancer risk and worse prognosis in several malignancies. We have recently demonstrated that vitamin D deficiency promotes secondary cancer growth in bone. These findings were partly attributable to an increase in bone remodelling but also through direct effects of vitamin D on cancer cells. To date, very little is known about vitamin D status of patients with bone tumours in general. Thus, the objective of this study was to assess vitamin D status of patients with diverse bone tumours. Moreover, the aim was to elucidate whether or not there is an association between pre-diagnostic vitamin D status and tumour malignancy in patients with bone tumours. In a multi-center analysis, 25(OH)D, PTH and calcium levels of 225 patients that presented with various bone tumours between 2017 and 2018 were assessed. Collectively, 76% of all patients had insufficient vitamin D levels with a total mean 25(OH)D level of 21.43 ng/ml (53.58 nmol/L). In particular, 52% (117/225) of patients were identified as vitamin D deficient and further 24% of patients (55/225) were vitamin D insufficient. Notably, patients diagnosed with malignant bone tumours had significantly lower 25(OH)D levels than patients diagnosed with benign bone tumours [19.3 vs. 22.75 ng/ml (48.25 vs. 56.86 nmol/L); p = 0.04). In conclusion, we found a widespread and distressing rate of vitamin D deficiency and insufficiency in patients with bone tumours. However, especially for patients with bone tumours sufficient vitamin D levels seem to be of great importance. Thus, we believe that 25(OH)D status should routinely be monitored in these patients. Collectively, there should be an increased awareness for physicians to assess and if necessary correct vitamin D status of patients with bone tumours in general or of those at great risk of developing bone tumours. KW - bone tumour KW - vitamin D KW - hypovitaminosis D KW - vitamin D deficiency KW - malignancy KW - tumour malignancy Y1 - 2020 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-230314 VL - 25 ER - TY - JOUR A1 - Ohlebusch, Barbara A1 - Borst, Angela A1 - Frankenbach, Tina A1 - Klopocki, Eva A1 - Jakob, Franz A1 - Liedtke, Daniel A1 - Graser, Stephanie T1 - Investigation of alpl expression and Tnap-activity in zebrafish implies conserved functions during skeletal and neuronal development JF - Scientific Reports N2 - Hypophosphatasia (HPP) is a rare genetic disease with diverse symptoms and a heterogeneous severity of onset with underlying mutations in the ALPL gene encoding the ectoenzyme Tissue-nonspecific alkaline phosphatase (TNAP). Considering the establishment of zebrafish (Danio rerio) as a new model organism for HPP, the aim of the study was the spatial and temporal analysis of alpl expression in embryos and adult brains. Additionally, we determined functional consequences of Tnap inhibition on neural and skeletal development in zebrafish. We show that expression of alpl is present during embryonic stages and in adult neuronal tissues. Analyses of enzyme function reveal zones of pronounced Tnap-activity within the telencephalon and the mesencephalon. Treatment of zebrafish embryos with chemical Tnap inhibitors followed by axonal and cartilage/mineralized tissue staining imply functional consequences of Tnap deficiency on neuronal and skeletal development. Based on the results from neuronal and skeletal tissue analyses, which demonstrate an evolutionary conserved role of this enzyme, we consider zebrafish as a promising species for modeling HPP in order to discover new potential therapy strategies in the long-term. KW - nonspecific alkaline-phosphae KW - in situ hybridization KW - hypophosphatasia KW - promotes KW - model KW - neurotransmission KW - differentiation KW - mineraliztion KW - metabolism KW - vertebrate Y1 - 2020 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-230024 VL - 10 ER -