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Tissue-Engineered Strategies to Repair Chondrogenic and Osteogenic Defects

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Tissue-Engineered Strategies to Repair Chondrogenic and Osteogenic Defects DISSERTATION Tissue-engineered strategies to repair chondrogenic and osteogenic defects. ausgeführt zum Zwecke der Erlangung des akademischen Grades eines Doktors der technischen Wissenschaften unter der Leitung von a.o. Univ.Prof. Dipl.-Ing. Dr.techn. Heinz Redl Institut für Verfahrenstechnik, Umwelttechnik und Technische Biowissenschaften (E166) eingereicht an der Technischen Universität Wien Fakultät für Technische Chemie von Bernhard Rieder, MSc 01329280 Krummgasse 14/1 1030 Wien Wien, am …………………………….. EIDESSTAATLICHE ERKLÄRUNG Hiermit erkläre ich an Eides statt, dass ich die vorliegende Dissertation „Tissue-engineered strategies to repair chondrogenic and osteogenic defects“ selbstständig verfasst und keine anderen als die angegebenen Quellen und Hilfsmittel verwendet habe. Wien, am ………………………….. Bernhard Rieder 2 Table of Contents ACKNOWLEDGMENTS ....................................................................................................................... 4 KURZFASSUNG .................................................................................................................................... 5 ABSTRACT ............................................................................................................................................ 7 GENERAL INTRODUCTION ............................................................................................................... 9 1. Problem and Possible Solution .................................................................................................... 9 2. Bone and Cartilage – The Developmental Siblings................................................................... 10 3. The Structure and Function of Articular Cartilage and Bone .................................................... 11 3.1. Structure and Composition of Articular Cartilage ............................................................. 11 3.2. Mechanism of Articular Cartilage and Bone Development .............................................. 16 3.3. Acute and Chronic Articular Cartilage Pathologies .......................................................... 21 3.4. Surgical Treatment Strategies in Clinics ........................................................................... 21 4. Tissue Engineering .................................................................................................................... 23 4.1. Cells ................................................................................................................................... 23 4.2. Growth Factors .................................................................................................................. 24 4.3. Biomaterials and Scaffolds ................................................................................................ 25 4.4. Mechanical Stimulation ..................................................................................................... 27 AIM OF THE THESIS .......................................................................................................................... 30 CHAPTER I .......................................................................................................................................... 32 Tissue-engineered hypertrophic chondrocyte grafts enhanced long bone repair .............................. 33 Perfusion enhances hypertrophic chondrocyte matrix deposition, but not the bone formation ........ 57 CHAPTER II ......................................................................................................................................... 76 Repopulation of an auricular cartilage scaffold, AuriScaff, perforated with an enzyme combination ........................................................................................................................................................... 77 Chondroinductive properties of the decellularized auricular cartilage scaffold is further enhanced by mechanical loading .......................................................................................................................... 103 CHAPTER III ...................................................................................................................................... 122 Hydrostatic pressure-generated reactive oxygen species induce osteoarthritic conditions in cartilage pellet cultures .................................................................................................................................. 123 REFERENCES .................................................................................................................................... 148 LIST OF ABBREVIATIONS ............................................................................................................. 171 CURRICULUM VITAE ..................................................................................................................... 175 3 ACKNOWLEDGMENTS First and foremost I want to thank my thesis supervisor Prof. Dr. Heinz Redl for giving me the opportunity to conduct this thesis, provide the necessary funding and embedding me in an environment with exceptional scientists. Furthermore, I want to thank FH-Prof. Mag. Dr. Dominik Rünzler and FH- Prof. DI Dr. Carina Huber-Gries for the opportunity to work and learn in their lab and the financial support aid, especially in the first years of the thesis. Moreover, I want to thank Andi for his constant support. Without a doubt I can clearly state that without his guidance this thesis would be non-existent. He is not only a brilliant scientist but has also became a good friend over the years, who showed me what a passion for science and teaching can achieve. In addition I would like to thank: • Nane and Anna, for always being there for me since my early beginning in the lab. Their knowledge and common sense were impressive and refreshing. • Raphael, for being one of my best friends during my studies and beyond. Casually talking about similar problems helped to dissuade from evil paths and get me back on track. • Jon, for being a typical Yankee whose constant positive mood was infectious and brightened up my days. His passion for science and knowledge in his field of expertise were unmet. • Babette, Daniel, Michi, Elias, Carina, Janine, Sabrina, Bettina, Safet, Sophie, Babsi, Eli, Dani, and all the other guys and girls in the lab for the great time and support. • Conny, Marian, and Sylvia for being a part of the CartiScaff team. My genuine thanks to my best friends Dora and Xavi, the infamous double. Seeing you evolve from curious students into scientists who outperform their former teachers not only with knowledge but also with enthusiasm warms my heart and encourages me to strive for my better self. I am sure that without the two of you, coming to the lab would have been less exciting and a lot tougher - I will miss every minute of it. At last I would like to express my sincere gratitude to my parents and my siblings. To my older brother Franz, whose enthusiasm for biology brought me into science in the first place. To Waltraud, who shows that having a perfect relationship with (soon) two little kids is achievable. To Constanze, who constantly proves that being an awesome mother and a busy teacher with various side projects is manageable. To Antonia, who persistently follows her dreams without hesitation (although they regularly change). To Martin, who engaged in a relationship at a young age but shows enormous level of maturity ever since. To Viktoria, who balances party, student life, and relationship in an exceptional manner hardly seen before. To my father and my mother, for always supporting me financially and morally and never questioning the direction of my decisions. 4 KURZFASSUNG Erkrankungen des Bewegungsapparates (MSD, engl. musculoskeletal disorders) betreffen hunderte Millionen Menschen auf der ganzen Welt, die an einer körperlichen Beeinträchtigung, welche mit starken langfristigen Schmerzen einhergeht, leiden. Die erhöhte Lebenserwartung in Kombination mit einem Wechsel von unverarbeiteten Lebensmitteln zu einer Ernährung mit hohem Glukose- und Fettgehalt sowie eingeschränkte körperliche Aktivität bei der Arbeit und zu Hause erhöht die weltweite Verbreitung von MSD kontinuierlich. Diese Krankheiten und Syndrome haben Dimensionen erreicht, die eine hohe wirtschaftliche Belastung für die Gesellschaft zur Folge haben. Schätzungen der Gesamtkosten reichen von 240 Mrd. EUR in der EU bis zu 213 Mrd. USD in den USA und erfordern daher ausgeklügelte Lösungen, um ein defektes Gesundheitssystem zu verhindern. Daher bedarf es einer Überarbeitung der derzeitigen Behandlungsoptionen und den Ersatz dieser durch überlegene Ansätze, um Kosten zu reduzieren. Diese Behandlungsverfahren beinhalten immer noch die Transplantation von autologem oder allogenem Gewebe, die zu einer Morbidität an der Spenderstelle, sowie Immunreaktionen oder Erkrankungen bedingt durch Transfer führen kann. Das aufstrebende Gebiet „Tissue Engineering und Regenerative Medizin“ versucht daher, dem Mangel aktueller Therapien entgegenzuwirken. Dabei werden neue Technologien und Techniken entwickelt, um ein funktionelles Gewebe herzustellen, welches in den Patienten implantiert werden kann und sofort seine vorgesehene Aufgabe erfüllt. In dieser Arbeit werden verschiedene Konzepte zur Knochen- und Knorpelreparatur untersucht. Obwohl Knochen eine angeborene Regenerationsfähigkeit besitzen, übertreffen große komplexe Frakturen oft die natürliche Heilkraft des Knochens und führen zu nicht zusammengewachsenen
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