Dokument: Beurteilung von altersbedingten Knorpelveränderungen an Hüftgelenken durch molekulare MRT-Bildgebung (dGEMRIC) und histologische Validierung der Methode am Schafsmodell in zwei Altersgruppen
| Titel: | Beurteilung von altersbedingten Knorpelveränderungen an Hüftgelenken durch molekulare MRT-Bildgebung (dGEMRIC) und histologische Validierung der Methode am Schafsmodell in zwei Altersgruppen | |||||||
| Weiterer Titel: | Magnetic resonance imaging (dGEMRIC) and histology of ovine hip joint cartilage in two age populations: a sheep model with assumed healthy cartilage | |||||||
| URL für Lesezeichen: | https://docserv.uni-duesseldorf.de/servlets/DocumentServlet?id=48426 | |||||||
| URN (NBN): | urn:nbn:de:hbz:061-20190211-095409-7 | |||||||
| Kollektion: | Dissertationen | |||||||
| Sprache: | Deutsch | |||||||
| Dokumententyp: | Wissenschaftliche Abschlussarbeiten » Dissertation | |||||||
| Medientyp: | Text | |||||||
| Autor: | Crumbiegel, Clemens [Autor] | |||||||
| Dateien: |
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| Beitragende: | PD Dr. Zilkens, Christoph [Gutachter] PD Dr. Schleich, Christoph [Gutachter] | |||||||
| Stichwörter: | dGEMRIC, Arthrose, biochemische MRT-Verfahren, GAG | |||||||
| Dewey Dezimal-Klassifikation: | 600 Technik, Medizin, angewandte Wissenschaften » 610 Medizin und Gesundheit | |||||||
| Beschreibungen: | Fragestellung: Das MRT-Verfahren dGEMRIC (delayed Gadolinium Enhanced MRI of Cartilage) ermöglicht eine Beurteilung von Veränderungen der Knorpel-zusammensetzung, zum Beispiel einer Verminderung und Degradation von Glykosaminoglykanen (GAG) im Rahmen degenerativer Knorpelerkrankungen. In Vorstudien konnten auch bei knorpelgesunden Probanden Varianzen in den gemessenen T1Gd-Zeiten erfasst werden. Somit war die zu Grunde liegende Arbeitshypothese der vorliegenden Studie, dass bei den Versuchsgruppen selbst in morphologisch intaktem Gelenkknorpel biochemische Unterschiede zu detektieren sind und dass das dGEMRIC-Verfahren in der Lage ist, diese Varianzen darzustellen. Es wurde aufgrund der Verfügbarkeit größerer altersgleicher Versuchsgruppen das Schaf als bereits etabliertes Arthrosemodell für die histologische Beurteilung des Gelenkknorpels herangezogen.
Methoden: Es wurden jeweils 10 Knorpelproben von Hüftköpfen gesunder Schafe aus zwei Altersgruppen (< 6 Monate; 2,5 bis 3 Jahre) entnommen; diese wurden markiert und mittels des dGEMRIC-Verfahrens sowie orientierender DESS-Sequenzen im MRT untersucht. Im Anschluss erfolgte eine Schnittbildanfertigung mittels der Trenn-Dünnschliff-Technik sowie die histologische Untersuchung durch Kollagen-ELISA und Toluidinblaufärbung. Durch zwei unabhängige Beobachter wurden die Regions of Interest (ROIs) in den MRT-Sequenzen durch eine 3D-Reformatierung den histologischen Schnittbildern zugeordnet und die entsprechenden T1Gd-Zeiten mit den Resultaten der Färbung, quantifiziert nach dem Mankin-Score, abgeglichen. Ergebnisse: Es konnten 303 ROIs untersucht werden, von denen 26 wegen zu hochgradiger Knorpelschädigung oder schlechter Bildqualität im MRT-Bild aus-geschlossen werden mussten. Insgesamt konnten somit 277 ROIs ausgewertet werden. Die Population der Lämmer zeigte weitestgehend intakte GAG, wobei 34 % der untersuchten ROIs einen Score von 0 und 66 % eine Score von 1 aufwiesen. Nur bei 2 % der Untersuchungen fanden sich Defizite in der Anfärbbarkeit. In 16 % der untersuchten ROIs bei den älteren Schafen hingegen zeigten sich Scores von 2 oder mehr. In 23 % der Untersuchungen der Versuchsgruppe Schaf fand sich eine Verminderung der Anfärbbarkeit durch Toluidinblau. Es zeigte sich, dass die T1Gd-Zeiten in der Gruppe der Lämmer im Vergleich zu denen in der Gruppe der Schafe signifikant höher waren (623,6 ms ± 72,8 ms gegenüber 540,4 ms ± 92,9 ms). Der p-Wert lag bei < 0,001. Die Kollagenfärbungen wurden aufgrund von Färbefehlern nicht ausgewertet. Diskussion: Grundlegend konnte mittels der verringerten T1Gd-Zeiten sowie der Mankin-Scores eine Verringerung des GAG-Gehaltes in der älteren Versuchsgruppe gegenüber der Versuchsgruppe der Lämmer erkannt werden. Da makroskopisch geschädigte Knorpelpräparate keinen Eingang in die Auswertung fanden, ist von einer alters- oder belastungsabhängigen Degradation des Knorpelgewebes auszugehen. Das dGEMRIC-Verfahren zeigte sich als valides Diagnostikum zur Detektion dieser Veränderungen. Die T1Gd-Zeiten glichen denen aus früheren Studien mit menschlichen Probanden. Das Schafsmodell konnte somit als sinnvolles Modell für Forschungen zur GAG-Quantifizierung herausgestellt werden. Weitere Studien zur Beurteilung von Frühveränderungen der GAG und Kollagene sind notwendig.Objective: The delayed Gadolinium Enhanced Magnetic Resonance Imaging of Cartilage (dGEMRIC) technique allows evaluation of early changes of joint cartilage like degradation of glycosaminoglycans (GAG) in case of degenerative diseases of cartilage. In previous publications significant variations in T1Gd-values within control groups with morphologically normal appearing cartilage could be noticed. Thus, the purpose of this study was to compare the cartilage status in two age groups without underlying prearthritic hip joint deformities by performing high resulotion MRI (dGEMRIC) and to reveal differences in content of GAG. Therefore, a sheep study was conducted, as sheep are an established Osteoarthritis (OA) model and different age groups of the same race are readily available for cartilage assessment, including histological validation. Materials and methods: Twenty femoral head specimen collected from ten lambs (aged < 6 months) and ten young adult sheep (aged between 2,5 and 3 years) were marked with a surgical suture and underwent dGEMRIC and morphological cartilage assessment by a Double-Echo Steady-State sequence (DESS). Slices were formed by the sawing and grinding technique. Histological analysis was performed by Collagen-ELISA and staining with conventional toluidine blue according to the Mankin scoring system. Two independent viewers matched all regions of interest (ROIs) examined by MRI with histological sections and compared T1Gd-values with corresponding Mankin-scores. Results: A total of 303 regions of interest were assessed (101 slices with 3 ROIs each). Twenty-six ROIs were excluded due to cartilage damage or insufficient MR image quality. Therefore a total of 277 ROIs were analyzed. Histological analyses revealed nearly intact GAG in samples of Group I (lambs) with 34% of ROIs scored 0 and 66% scored 1. Loss of staining intensity was found in 2% of examinations only. Distinct degenerative changes in cartilage samples of group II (young adult sheep) were found, reflected by total Mankin scores of 2 in 16% and loss of toluidine staining intensity in 23% of examinations. T1Gd-values in group I (mean T1-Gd: 623,6 ms ± 72,8 ms) were significantly (p < 0,001) higher compared with group II (mean T1-Gd: 540,4 ms ± 92,9 ms). All results of Collagen-ELISA were not accounted due to a loss of reliable staining results. Discussion: Although morphologically normal, distinct cartilage degeneration present in young adult sheep could be identified via decreased T1Gd- values. The dGEMRIC technique revealed these changes and may be a tool for the assessment of early cartilage degeneration. T1Gd- values were similar to results of human cartilage examined in previous studies. The Sheep model seemed to be useful as a model for cartilage assessment in OA. Further controlled studies including healthy human cartilage samples are required to confirm our preliminary results. | |||||||
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The Effect of Paraformaldehyde Fixation on the Delayed Gadolinium-Enhanced MRI of Cartilage (dGEMRIC) Measurement. Published online in Wiley InterScience (www.interscience.wiley.com); DOI 10.1002/jor.20767; 09/2008. | |||||||
| Lizenz: |  Urheberrechtsschutz | |||||||
| Bezug: | 2010-2019 | |||||||
| Fachbereich / Einrichtung: | Medizinische Fakultät | |||||||
| Dokument erstellt am: | 11.02.2019 | |||||||
| Dateien geändert am: | 11.02.2019 | |||||||
| Promotionsantrag am: | 16.04.2018 | |||||||
| Datum der Promotion: | 24.01.2019 |
