Aus dem Institut für Kreislaufforschung und Sportmedizin der Deutschen Sporthochschule Köln Geschäftsführender Leiter: Universitätsprofessor Dr. med. W. Bloch The role of impact loading on artery adaptations and the effect of muscle unloading on local blood supply and exercise tolerance von der Deutschen Sporthochschule Köln zur Erlangung des akademischen Grades Doktor der Sportwissenschaft genehmigte Dissertation vorgelegt von Tobias Weber Geboren in Prüm Erster Referent: Univ.-Prof. Dr. med. Wilhelm Bloch Zweiter Referent: Prof. Dr. med. Jörn Rittweger Vorsitzender des Promotionsausschusses: Univ.-Prof. Dr. med. Wilhelm Bloch Tag der Disputation: 25. Oktober 2013 Hierdurch erkläre ich, dass ich die Leitlinien guter wissenschaftlicher Praxis der Deutschen Sporthochschule Köln eingehalten habe. Köln, den 30.07.2013 ____________________ Tobias Weber Hierdurch versichere ich: Ich habe diese Arbeit selbständig und nur unter Benutzung der angegebenen Quellen und technischen Hilfen angefertigt; sie hat noch keiner anderen Stelle zur Prüfung vorgelegen. Wörtlich übernommene Textstellen, auch Einzelsätze oder Teile davon, sind als Zitate kenntlich gemacht worden. Köln, den 30.07.2013 ____________________ Tobias Weber I „Datt watt esch heij jeschriwwen hann wittmen esch minnger Mamm unn mingem Papp! Wenn et eisch net juuv wiier dat heij alles net miejellisch jewääst!“ II Contents 1 Chapter One – Abstract ..................................................................................................... 1 2 Chapter Two – Introduction .............................................................................................. 3 2.1 Structural artery adaptations ................................................................................. 6 2.1.1 Adaptations of Intima Media Thickness (IMT) ..................................................... 7 2.1.2 Arterial caliber adaptations .................................................................................... 8 2.2 Functional artery adaptations ................................................................................. 9 2.2.1 Flow mediated dilation (FMD) .............................................................................. 9 2.2.2 Skeletal muscle blood flow .................................................................................. 11 2.3 Conducted studies and applied models for exercise training and muscle unloading ............................................................................................................................. 16 2.3.1 The HEPHAISTOS study (HEP-study) ............................................................... 16 2.3.2 The molecular and functional effects of resistive vibration exercise study (EVE- study) 17 2.4 Outline of the thesis ................................................................................................ 19 3 Chapter Three – Scientific papers .................................................................................. 30 3.1 Paper 1: The HEPHAISTOS Study – Protocol and Implementation ............... 30 3.2 Paper 2: Gravitational accelerations and arterial adaptation ........................... 52 3.3 Paper 3: Muscle unloading, muscle perfusion and muscle power ..................... 74 3.4 Paper 4: Whole-body vibration and arterial adaptation .................................. 102 4 Chapter Four – Primary findings and conclusion ....................................................... 124 5 Chapter Five – Addendum ............................................................................................ 127 5.1 Zusammenfassung in deutscher Sprache ........................................................... 127 5.2 Original manuscripts ........................................................................................... 128 5.3 Curriculum Vitae ................................................................................................. 158 III Abstract 1 Chapter One – Abstract Background: Evidence suggests that muscle disuse and exercise training have direct vascular (de-) conditioning effects that can modify cardiovascular risk. The underlying mechanisms, however, are to date not entirely understood. It is clear that particularly arteries adapt to changes of mechanical stress that may predominantly act on endothelial cells (ECs) or vascular smooth muscle cells (VSMCs). Nonetheless, not all potential sources for mechanical stress have been considered yet as previous research basically investigated the conditioning effects of intrinsic hemodynamic forces. Given the gravitational environment on earth, the present thesis sought therefore to investigate the impact of gravity-induced impact loading, provoking mass accelerations, on arterial adaptations in conditions associated with exercise training and muscle disuse. Moreover, the relationship between local arterial blood flow, muscle perfusion and dynamic exercise tolerance after prolonged local muscle disuse constitutes a major topic of the present thesis, as to date no study has ever elaborated on this. Methods: Two clinical interventional studies have been carried out. Within the scope of the HEP-study, 11 healthy male subjects wore a new orthotic device (HEPHAISTOS) for 56 days to unilaterally unload the calf muscles without changing the impact of gravitational loading. The EVE-study has been conducted to study the effects of whole body vibrations (WBV) if combined with conventional resistive exercise. 26 healthy male subjects were recruited and assigned to either a resistive exercise group or a resistive vibration exercise group that performed a 6 week training intervention. Major endpoint measurements for both studies encompassed structural and functional arterial parameters as measured with sonography. In addition, soleus muscle morphology and tissue oxygenation have been assessed in the HEP study using soleus muscle biopsies and near infrared spectroscopy, respectively. Results: Muscle unloading with the HEPHAISTOS orthosis led to distinct decreases of superficial femoral artery (SFA) calibers, while wall thickness and endothelial function remained unaffected. Although muscle size and arterial calibers significantly decreased, functional exercise blood flow, tissue oxygenation and exercise tolerance did not change after the intervention. During the EVE-study, SFA calibers increased significantly and carotid artery wall thickness decreased significantly in response to resistive exercise while superposition of WBV did not reveal an additional effect. 1 Abstract Conclusion: Both studies highlight the importance of muscle work and hence the importance of intrinsic hemodynamic forces for arterial adaptations. However, gravity-induced impact loading seems to have a direct conditioning effect on arterial wall thickness and on endothelial function, thereby modulating parameters for cardiovascular risk. In addition, functional muscle perfusion seems to remain unaffected after prolonged muscle disuse and possibly as a consequence of this, dynamic exercise tolerance does not change. 2 Introduction 2 Chapter Two – Introduction The arterial system consists of a complex network of blood vessels and functions as transportation network for many cells, hormones and a multitude of other molecules within the human body. Structural as well as functional arterial properties within this system vary and are largely determined by the anatomical region they supply. For instance, dimension and function of the carotid artery (CA) strongly differs in terms of size and function compared to arterioles in calf muscles. However, the general construction of arteries is always the same, as one can basically differentiate between three main layers: a) the tunica intima, in the following referred to as ‘Intima’, is the innermost layer of an artery that is in direct contact with the circulating blood, consisting of endothelial cells (ECs); b) the tunica media, in the following referred to as ‘Media’, is the middle layer which is composed of vascular smooth muscle cells (VSMCs) and elastic tissue and c) the outer tunica adventitia, in the following referred to as ‘Adventitia’, consisting of mainly collagen fibers to anchor the artery to surrounding tissue (see Figure 1; 59). Yet, the arterial system does not merely function as a passive apparatus of rigid pipes through which blood flow is merely controlled by heart action. In fact, blood flow is fine regulated by cardiac and by vascular function in order to meet the specific metabolic demands at rest and during physical activity. Particularly peripheral blood flow supplying skeletal muscles is greatly dependent on peripheral vascular tone that in turn is orchestrated by a complex interplay of various neuro-humoral, metabolic and mechanical control mechanisms acting on VSMCs and ECs (52). Both functional as well as structural properties of arteries show the ability to acutely and chronically adjust to different functional demands and varying environmental conditions and certain parameters of arterial structure and function are thought to be directly related to physical fitness and cardiovascular risk (91). As a matter of fact, mechanical stress acting on the arterial wall is considered as the major signal to induce structural and functional artery adaptations. Numerous in- and ex-vivo studies revealed a direct conditioning effect of stresses like ‘shear’, ‘pressure’ and pressure-related ‘stretch’ upon ECs and VSMCs (15; 40; 55; 108). Thus, arterial wall shear stress would be triggered by the action of blood