Publications (223) View all
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Article: Novel Combination of Collagen Dynamics Analysis and Transcriptional Profiling Reveals Fibrosis-Relevant Genes and Pathways.
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ABSTRACT: Collagen deposition is a key process during idiopathic pulmonary fibrosis; however, little is known about the dynamics of collagen formation during disease development. Tissue samples of early stages of human disease are not readily available and it is difficult to identify changes in collagen content, since standard collagen analyses do not distinguish between 'old' and 'new' collagen. Therefore, the current study aimed to (i) investigate the dynamics of new collagen formation in mice using bleomycin-induced lung fibrosis in which newly synthesized collagen was labelled with deuterated water and (ii) use this information to identify genes and processes correlated to new collagen formation. Lung fibrosis was induced in female C57Bl/6 mice by bleomycin instillation. Animals were sacrificed at 1 to 5 weeks after fibrosis induction. Collagen synthesized during the week before sacrifice was labelled with deuterium by providing mice with deuterated drinking water. After sacrifice, we collected lung tissue for microarray analysis, determination of new collagen formation, and histology. Furthermore, we measured in vitro the expression of selected genes after transforming growth factor (TGF) β1-induced myofibroblast differentiation. Deuterated water labelling showed a strong increase in new collagen formation already during the first week after fibrosis induction and a complete return to baseline at five weeks. Correlation of new collagen formation data with gene expression data allowed us to create a gene expression signature of fibrosis within the lung and revealed fibrosis-specific processes, amongst which proliferation. This was confirmed by measuring cell proliferation and collagen synthesis simultaneously using deuterated water incorporation in a separate experiment. Furthermore, new collagen formation strongly correlated with gene expression of e.g. elastin, Wnt-1 inducible signalling pathway protein 1, tenascin C, lysyl oxidase, and type V collagen. Gene expression of these genes was upregulated in vitro in fibroblasts stimulated with TGFβ1. Together, these data demonstrate, using a novel combination of technologies, that the core process of fibrosis, i.e. the formation of new collagen, correlates not only with a wide range of genes involved in general extracellular matrix production and modification but also with cell proliferation. The observation that the large majority of the genes which correlated with new collagen formation also were upregulated during TGFβ1-induced myofibroblast differentiation provides further evidence for their involvement in fibrosis.Matrix biology: journal of the International Society for Matrix Biology 05/2013; · 3.56 Impact Factor -
Article: Role of Caveolin-1 in Fibrotic Diseases.
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ABSTRACT: Fibrosis underlies the pathogenesis of numerous diseases and leads to severe damage of vital body organs and, frequently, to death. Better understanding of the mechanisms resulting in fibrosis is essential for developing appropriate treatment solutions and is therefore of upmost importance. Recent evidence suggests a significant antifibrotic potential of an integral membrane protein, caveolin-1. While caveolin-1 has been widely studied for its role in the regulation of cell signaling and endocytosis, its possible implication in fibrosis remains largely unclear. In this review we survey involvement of caveolin-1 in various cellular processes and highlight different aspects of its antifibrotic activity. We hypothesize that caveolin-1 conveys a homeostatic function in the process of fibrosis by (a) regulating TGF-β1 and its downstream signaling; (b) regulating critical cellular processes involved in tissue repair, such as migration, adhesion and cellular response to mechanical stress; and (c) antagonizing profibrotic processes, such as proliferation. Finally, we consider this homeostatic function of caveolin-1 as a possible novel approach in treatment of fibroproliferative diseases.Matrix biology: journal of the International Society for Matrix Biology 04/2013; · 3.56 Impact Factor -
Article: Magnesium deficiency results in an increased formation of osteoclasts.
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ABSTRACT: Magnesium (Mg(2+)) deficiency is a frequently occurring disorder that leads to loss of bone mass, abnormal bone growth and skeletal weakness. It is not clear whether Mg(2+) deficiency affects the formation and/or activity of osteoclasts. We evaluated the effect of Mg(2+) restriction on these parameters. Bone marrow cells from long bone and jaw of mice were seeded on plastic and on bone in medium containing different concentrations of Mg(2+) (0.8 mM which is 100% of the normal value, 0.4, 0.08 and 0 mM). The effect of Mg(2+) deficiency was evaluated on osteoclast precursors for their viability after 3 days and proliferation rate after 3 and 6 days, as was mRNA expression of osteoclastogenesis-related genes and Mg(2+)-related genes. After 6 days of incubation, the number of tartrate resistant acid phosphatase-positive (TRACP(+)) multinucleated cells was determined, and the TRACP activity of the medium was measured. Osteoclastic activity was assessed at 8 days by resorption pit analysis. Mg(2+) deficiency resulted in increased numbers of osteoclast-like cells, a phenomenon found for both types of marrow. Mg(2+) deficiency had no effect on cell viability and proliferation. Increased osteoclastogenesis due to Mg(2+) deficiency was reflected in higher expression of osteoclast-related genes. However, resorption per osteoclast and TRACP activity were lower in the absence of Mg(2+). In conclusion, Mg(2+) deficiency augmented osteoclastogenesis but appeared to inhibit the activity of these cells. Together, our in vitro data suggest that altered osteoclast numbers and activity may contribute to the skeletal phenotype as seen in Mg(2+) deficient patients.The Journal of nutritional biochemistry 03/2013; · 4.29 Impact Factor -
Article: Influence of a Nanoporous Zirconia Implant Surface of on Cell Viability of Human Osteoblasts.
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ABSTRACT: Purpose: The dense nonretentive surface of zirconia implants was modified into a nanoporous surface using selective infiltration etching surface treatment. The aim of this study was to investigate the influence of such a nanoporous modified zirconia surface on the attachment of human osteoblasts. Materials and Methods: Human osteoblasts were cultured for 21 days on (i) selective infiltration etched zirconia (nanoporous surface), (ii) polished zirconia, (iii) polished titanium, or (iv) airborne particle abraded acid etched (SLA) titanium disks. After the culture period the following parameters were assessed: number of cells, the morphology of the cells, the attachment of the cells, alkaline phosphatase activity, and the level of total protein (α= 0.05). Results: Statistical analysis revealed a significantly higher cell count on the third (F = 17.4, p < 0.001) and eighth day (F = 163, p < 0.001) for nanoporous zirconia and SLA titanium surfaces compared to polished specimens. The number of cells (nanoporous zirconia 160 ± 20/mm(2) , SLA titanium 133 ± 15/mm(2) ) and cell size (nanoporous zirconia 50.7 ± 3 μm, SLA titanium 42.5 ± 4 μm) were significantly higher than polished specimens. Nanoporous zirconia specimens demonstrated comparable alkaline phosphatase activity (0.0036 ± 0.0035 ng/μl) and intracellular protein content (72.7 ± 0.9 ng/μl) compared to other tested groups. Scanning electron microscopy revealed that cells attached on the polished surface using finger-like processes, whereas on the nanoporous surface, finger-like processes were not observed, as the cell membrane appeared to be in close proximity to the underlying surface. Conclusion: The findings of this study suggest that a nanoporous zirconia surface favors cell growth and attachment compared to a polished surface. It was proposed that a nanoporous zirconia surface may improve clinical performance of zirconia implants.Journal of Prosthodontics 02/2013; · 1.01 Impact Factor -
Article: The intramembrane protease SPPL2A is critical for tooth enamel formation.
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ABSTRACT: Intramembrane proteases are critically involved in signal transduction and membrane protein turnover. Signal-peptide-peptidase-like 2a (SPPL2A), a presenilin-homologue residing in lysosomes/late endosomes, cleaves type II-oriented transmembrane proteins. We recently identified SPPL2A as the enzyme controlling turnover and functions of the invariant chain (CD74) of the MHCII complex and demonstrated critical importance of this process for B cell development. Surprisingly, we found that SPPL2A is critical for formation of dental enamel. In Sppl2a knockout mice, enamel of the erupted incisors was chalky white and rapidly eroded after eruption. SPPL2A was found to be expressed in enamel epithelium during secretory and maturation stage amelogenesis. Mineral content of enamel in Sppl2a(-/-) incisors was inhomogeneous and reduced by ∼20% compared to wild type mice with the most pronounced reduction at the mesial side. Frequently, disruption of the enamel layer and localized detachment of the most superficial enamel layer was observed in the knockout incisors leading to an uneven enamel surface. In Sppl2a null mice, morphology and function of secretory stage ameloblasts were not noticeably different from that of wild type mice. However, maturation stage ameloblasts showed reduced height and a characteristic undulation of the ameloblast layer with localized adherence of the cells to the outer enamel. This was reflected in a delayed and incomplete resorption of the proteinaceous enamel matrix. Thus, we conclude that intramembrane proteolysis by SPPL2A is essential for maintaining cellular homeostasis of ameloblasts. Since modulation of SPPL2A activity appears to be an attractive therapeutic target to deplete B cells and treat autoimmunity, interference with tooth enamel formation should be investigated as a possible adverse effect of pharmacological SPPL2A inhibitors in humans. © 2013 American Society for Bone and Mineral Research.Journal of bone and mineral research: the official journal of the American Society for Bone and Mineral Research 02/2013; · 6.04 Impact Factor
