TY  - THES
A1  - Melms, Hannah
T1  - Charakterisierung und Analyse mesenchymaler Stammzellen dentalen Ursprungs mit Fokus auf die dentalen Aspekte der Hypophosphatasie - Etablierung eines in vitro Modells -
T1  - Characterization and analysis of mesenchymal stem cells of dental origin with focus on the dental aspects of hypophosphatasia - Establishment of an in vitro model -
N2  - Im seltenen Krankheitsbild der Hypophosphatasie (HPP) treten aufgrund der Fehlfunktion der Gewebe-unspezifischen Alkalischen Phosphatase (tissue-nonspecific alkaline phosphatase, TNAP) skelettale und dentale Symptome in sehr variabler Ausprägung auf. Der vorzeitige Verlust von Milchzähnen ist das zahnmedizinische Leitsymptom und in vielen Fällen ein erstes Anzeichen dieser Erkrankung. In dieser Arbeit wurde ein in vitro Modell der HPP etabliert und der Fokus auf die dentalen Aspekte dieser Erkrankung gelegt. Hierzu wurden mesenchymale Stammzellen (MSCs) aus Bereichen analysiert, die bei einer Erkrankung von dieser Mineralisierungsstörung betroffen sind. Es wurden dentale Stammzellen aus der Pulpa (dental pulp stem cells, DPSCs) und dem parodontalen Ligament (periodontal ligament stem cells, PDLSCs) isoliert und im Vergleich zu Stammzellen aus dem Knochenmark (bone marrow mesenchymal stem cells, BMSCs) charakterisiert. Um den Einfluss der Spendervariabilität zu reduzieren, wurden aus dem gesamten dentalen Probenmaterial nur vollständige Probenpaare aus DPSCs und PDLSCs von 5 Spendern für die vergleichenden Analysen verwendet. Die dentalen MSCs konnten somit paarweise direkt miteinander verglichen werden. 
DPSCs gelten seit ihrer Entdeckung von Gronthos et al. im Jahr 2000 als geeignete Quelle für die Stammzellgewinnung mit vielversprechenden Anwendungsmöglichkeiten im Bereich des Tissue Engineering und der regenerativen muskuloskelettalen Medizin. PDLSCs sind aufgrund der parodontalen Problematik der HPP von besonderem Interesse in dieser Arbeit. Die Isolation von Stammzellen aus Pulpa und PDL konnte mit dem Nachweis der sogenannten Minimalkriterien für MSCs bestätigt werden. 
In diesem durch Enzyminhibition mit Levamisol induzierten in vitro Modell der HPP wurde die TNAP-abhängige Genexpression, die Enzym-Aktivität und das osteogene Differenzierungspotenzial an diesen drei Mineralisierungs-assoziierten MSCs untersucht. Die erweiterte Genexpressionsanalyse in Kooperation mit der Core Unit Systemmedizin der Universität Würzburg mit einer RNA-Sequenzierung der PDLSCs ergab interessante Einblicke in die differentielle Genexpression nach der TNAP-Inhibition während der osteogenen Differenzierung und Ansatzpunkte für weitere Analysen. Die beobachteten Genregulationen waren nach dem derzeitigen Verständnis pathologischer Zusammenhänge nachvollziehbar und simulierten in vitro HPP-relevante Signalwege repräsentativ. Insbesondere die signifikante Genregulation von P2X7 und DMP1, sowie Zusammenhänge aus dem Wnt-Signalweg zeigen hinsichtlich der dentalen Aspekte der HPP neue Ansatzpunkte auf. Die erhöhte P2X7-Expression in diesem in vitro HPP-Modell scheint mit der Parodontitis-Problematik der HPP zu korrelieren und verdeutlicht unter anderem die multifaktorielle Ätiologie und Pathogenese der Parodontitis. Die Tatsache, dass die experimentellen Beobachtungen in Einklang mit dem klinischen Bild der HPP gebracht werden können, bestätigt die Relevanz des hier etablierten in vitro Modells. Zusammenfassend konnten anhand dieses in vitro Modells der HPP neue Aspekte aufgedeckt werden, die nicht nur im Hinblick auf die dentale Problematik der HPP aufschlussreich sind.
N2  - In the rare clinical picture of hypophosphatasia (HPP) skeletal and dental symptoms occur in very variable severity due to a dysfunction of the enzyme tissue-nonspecific alkaline phosphatase (TNAP). The premature loss of deciduous teeth is the main dental symptom and in many cases the first sign of this disease. In this thesis, an in vitro model for HPP was established and the research focus was put on the dental aspects of this disease. Mesenchymal stem cells (MSCs) from areas affected by this mineralization disorder were analyzed. Dental pulp stem cells (DPSCs) and periodontal ligament stem cells (PDLSCs) were isolated and characterized in comparison to bone marrow mesenchymal stem cells (BMSCs). In order to reduce the influence of donor variability, only complete pairs of DPSCs and PDLSCs from five donors were used for comparative analysis. The mesenchymal stem cell character of the isolated cells from the dental pulp and the PDL was confirmed by the verification of the so-called minimal criteria for MSCs. 
In this in vitro model for HPP, which was induced by inhibition of TNAP enzyme with levamisole, the TNAP-dependent gene expression, the enzyme activity and the osteogenic differentiation potential of these three mineralization-associated MSCs were investigated. Additionally, an extended gene expression analysis was performed in cooperation with the Core Unit System Medicine of the University of Würzburg using the RNA sequencing technique. Analysis of RNAs isolated from PDLSCs provided interesting insights into the differential gene expression due to TNAP inhibition during osteogenic differentiation and consequently starting points for further analyses. The observed gene regulation was in line with the current understanding of pathological relationships and simulated HPP-relevant signaling pathways in vitro. In particular, the significant gene regulation of P2X7 and DMP1, as well as correlations from the Wnt signaling pathway, reveal new considerable approaches for the dental aspects of HPP. The increased P2X7 expression in this in vitro HPP model seems to correlate with the periodontal disease of HPP and illustrates, among other things, the multifactorial etiology and pathogenesis of periodontitis. The fact that the experimental observations can be reconciled with the clinical picture of HPP confirms the relevance of the in vitro model established here. In summary, this in vitro model for HPP has revealed a number of new interesting aspects - not only with regard to the dental problems of HPP.
KW  - Hypophosphatasie
KW  - Stammzellen
KW  - Hypophosphatasia
KW  - Mesenchymale Stammzellen
KW  - mesenchymal stem cells
KW  - dentale Stammzellen
KW  - dental stem cells
KW  - osteogene Differenzierung
KW  - osteogenic differentiation
Y1  - 2019
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-189844
ER  - 
TY  - JOUR
A1  - Jarius, Sven
A1  - Ruprecht, Klemens
A1  - Wildemann, Brigitte
A1  - Kuempfel, Tania
A1  - Ringelstein, Marius
A1  - Geis, Christian
A1  - Kleiter, Ingo
A1  - Kleinschnitz, Christoph
A1  - Berthele, Achim
A1  - Brettschneider, Johannes
A1  - Hellwig, Kerstin
A1  - Hemmer, Bernhard
A1  - Linker, Ralf A.
A1  - Lauda, Florian
A1  - Hayrettin, Christoph A.
A1  - Tumani, Hayrettin
A1  - Melms, Arthur
A1  - Trebst, Corinna
A1  - Stangel, Martin
A1  - Marziniak, Martin
A1  - Hoffmann, Frank
A1  - Schippling, Sven
A1  - Faiss, Jürgen H.
A1  - Neuhaus, Oliver
A1  - Ettrich, Barbara
A1  - Zentner, Christian
A1  - Guthke, Kersten
A1  - Hofstadt-van Oy, Ulrich
A1  - Reuss, Reinhard
A1  - Pellkofer, Hannah
A1  - Ziemann, Ulf
A1  - Kern, Peter
A1  - Wandinger, Klaus P.
A1  - Bergh, Florian Then
A1  - Boettcher, Tobias
A1  - Langel, Stefan
A1  - Liebetrau, Martin
A1  - Rommer, Paulus S.
A1  - Niehaus, Sabine
A1  - Münch, Christoph
A1  - Winkelmann, Alexander
A1  - Zettl, Uwe K
A1  - Metz, Imke
A1  - Veauthier, Christian
A1  - Sieb, Jörn P.
A1  - Wilke, Christian
A1  - Hartung, Hans P.
A1  - Aktas, Orhan
A1  - Paul, Friedemann
T1  - Contrasting disease patterns in seropositive and seronegative neuromyelitis optica: A multicentre study of 175 patients
JF  - Journal of Neuroinflammation
N2  - Background: The diagnostic and pathophysiological relevance of antibodies to aquaporin-4 (AQP4-Ab) in patients with neuromyelitis optica spectrum disorders (NMOSD) has been intensively studied. However, little is known so far about the clinical impact of AQP4-Ab seropositivity. 
Objective: To analyse systematically the clinical and paraclinical features associated with NMO spectrum disorders in Caucasians in a stratified fashion according to the patients' AQP4-Ab serostatus. 
Methods: Retrospective study of 175 Caucasian patients (AQP4-Ab positive in 78.3%). 
Results: Seropositive patients were found to be predominantly female (p < 0.0003), to more often have signs of co-existing autoimmunity (p < 0.00001), and to experience more severe clinical attacks. A visual acuity of <= 0.1 during acute optic neuritis (ON) attacks was more frequent among seropositives (p < 0.002). Similarly, motor symptoms were more common in seropositive patients, the median Medical Research Council scale (MRC) grade worse, and MRC grades <= 2 more frequent, in particular if patients met the 2006 revised criteria (p < 0.005, p < 0.006 and p < 0.01, respectively), the total spinal cord lesion load was higher (p < 0.006), and lesions >= 6 vertebral segments as well as entire spinal cord involvement more frequent (p < 0.003 and p < 0.043). By contrast, bilateral ON at onset was more common in seronegatives (p < 0.007), as was simultaneous ON and myelitis (p < 0.001); accordingly, the time to diagnosis of NMO was shorter in the seronegative group (p < 0.029). The course of disease was more often monophasic in seronegatives (p < 0.008). Seropositives and seronegatives did not differ significantly with regard to age at onset, time to relapse, annualized relapse rates, outcome from relapse (complete, partial, no recovery), annualized EDSS increase, mortality rate, supratentorial brain lesions, brainstem lesions, history of carcinoma, frequency of preceding infections, oligoclonal bands, or CSF pleocytosis. Both the time to relapse and the time to diagnosis was longer if the disease started with ON (p < 0.002 and p < 0.013). Motor symptoms or tetraparesis at first myelitis and > 1 myelitis attacks in the first year were identified as possible predictors of a worse outcome.
KW  - cerebrospinal-fluid
KW  - intractable hiccup
KW  - extensiv transverse myelitis
KW  - multiple sclerosis
KW  - anti-aquaporin-4 antibody
KW  - NMO-IGG
KW  - aquaporin-4 autoantibodies
KW  - immune-response
KW  - myasthenia gravis
KW  - immunoglobulin-G
Y1  - 2012
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-133636
VL  - 9
IS  - 14
ER  -