We studied patterning mechanisms and cell fates during limb regeneration in the axolotl. 1) It is crucial to understand the earliest events of patterning. Since it is technically challenging to study early events, we established single cell PCR. This new tool will allow us to obtain novel insight into the initial steps of limb patterning. 2)We have examined the roles of different tissues regarding their fates and features of proximo- distal patterning. Our strategy was to transplant GFP+ skin, skeleton, muscle and Schwann cells from transgenic donors to limbs of wild type hosts, amputate through the graft and analyze fluorescent progeny combined with the use of molecular markers. Our results revealed that different subpopulations of blastema cells exist regarding two aspects. First, we found that progeny of skin and skeleton have some tissue specific memory since they did not give rise to muscle lineages. However, cells of the skin contributed to other mesenchymal tissues like cartilage or tendons, while the majority of skeleton- derived cells undergoes self- renewal. Second, we performed one cellular and two molecular assays to investigate what tissues generate cells that exhibit features of proximo- distal patterning. Both assays revealed that Schwann cell- derived progeny do not display such features while progeny of skin, skeleton and muscle did. Therefore, we conclude that the blastema is a heterogeneous mix of cells regarding tissue lineages and features of proximo- distal patterning.

Background: To ensure adequate function after orthopedic tumor reconstruction, it is important to reattach the remaining soft tissue to the implant. This study aimed at obtaining mechanical properties of textile muscle-implant and muscle-bone connections in a preliminary test. Methods: Two groups of soft-tissue attachment were mechanically tested and compared: Native bone-muscle samples obtained from human femora and muscles attached to a prosthetic implant by means of Trevira® attachment tubes. Additionally, muscle samples were tested with muscle fibers aligned parallel and perpendicular to the tension load. A uniaxial load was exerted upon all samples. Results: Failure loads of 26.7 ± 8.8 N were observed for the native bone-muscle group and of 18.1 ± 9.9 N for the Trevira® group. Elongations of 94.8 ± 36.2 % were observed for the native bone-muscle group and 79.3 ± 51.8 % for the Trevira® group. The location of failure was mainly observed in the central area of the muscle fibers. Muscle fibers with parallel fiber orientation (47.6 ± 11.5 N) yielded higher tensile strength than those with perpendicular fiber orientation (14.8 ± 4.1 N). Conclusions: Our experiments showed that higher forces were transmitted in the origin and insertion areas than in areas of flat soft tissue reconstruction using attachment tubes. The data indicate that the tested material allows reattaching muscles, but without reinforcing the insertion site. Therefore, attachment tubes with region-dependent and potentially anisotropic material behavior might be advantageous to optimize muscle-bone load transmission after surgery, which may allow lower complication rates and shorter physical recovery.

We studied patterning mechanisms and cell fates during limb regeneration in the axolotl. 1) It is crucial to understand the earliest events of patterning. Since it is technically challenging to study early events, we established single cell PCR. This new tool will allow us to obtain novel insight into the initial steps of limb patterning. 2)We have examined the roles of different tissues regarding their fates and features of proximo- distal patterning. Our strategy was to transplant GFP+ skin, skeleton, muscle and Schwann cells from transgenic donors to limbs of wild type hosts, amputate through the graft and analyze fluorescent progeny combined with the use of molecular markers. Our results revealed that different subpopulations of blastema cells exist regarding two aspects. First, we found that progeny of skin and skeleton have some tissue specific memory since they did not give rise to muscle lineages. However, cells of the skin contributed to other mesenchymal tissues like cartilage or tendons, while the majority of skeleton- derived cells undergoes self- renewal. Second, we performed one cellular and two molecular assays to investigate what tissues generate cells that exhibit features of proximo- distal patterning. Both assays revealed that Schwann cell- derived progeny do not display such features while progeny of skin, skeleton and muscle did. Therefore, we conclude that the blastema is a heterogeneous mix of cells regarding tissue lineages and features of proximo- distal patterning.

Pelvic floor disorder is an emerging research area and is highlighted in many pelvic floor studies. Assessment methods for this type of injury are lacking and new methods for prevention and diagnosis are needed. Pelvic floor disorders are common among women and can lead to suffering for the patient. Levator ani muscle injuries are the main cause for pelvic floor disorders. This muscle group is an anisotropic skeletal muscle that helps support the pelvic viscera. Assessment of this muscle is difficult due to its complex geometry and location. Therefore, two muscle phantoms were constructed to mimic different properties of the levator ani muscle. The muscle phantoms provided more availability and a more controlled setting. The muscle phantoms were examined using ultrasound-based shear wave elastography which is an elastography method that can help determine the elasticity of tissue. A PVA-graphite phantom and a water-based gelatine-graphite phantom, both with fishing lines network as fibers for anisotropy, were constructed in this project. Shear wave elastography results of the PVA phantom indicated no anisotropy but visually resembled a muscle. Although not achieving anisotropy, the shear modulus of the PVA did match the shear modulus of skeletal muscle tissue. Shear wave elastography results of the gelatine phantom indicated anisotropy but visually did not resemble a muscle due to the low shear modulus of the gelatine. A 3D model of the female bony pelvis, that was provided for this project, was measured, and compared with reference value of previous study for future construction and 3D printing of the model. Results of measurements showed similarities between the 3D model and the female pelvis except for the sagittal outlet which had a deviant value. For future work, the muscle phantom can be developed by applying the complex geometry of the levator ani muscle, assembly of the muscle phantom, and the 3D rendering of the pelvis. The combination of these two parts provides a more complete phantom where shear wave elastography can be applied in the same way as in female patients. / Bäckenbottenbesvär är ett framväxande forskningsområde och lyfts fram i många bäckenbottenstudier. Bedömningsmetoder för denna typ av skador saknas och det behövs nya metoder för förebyggande och diagnostik. Bäckenbottenbesvär är vanliga bland kvinnor och kan leda till lidande för patienten. Levator ani muskelskador är den främsta orsaken till bäckenbottensjukdomar. Denna muskelgrupp är en anisotrop skelettmuskel som hjälper till att stödja inälvorna i bäckenet. Bedömning av denna muskel är svår på grund av dess komplexa geometri och läge. Därför konstruerades två muskelfantomer för att efterlikna olika egenskaper hos levator ani-muskeln. Muskelfantomerna gav mer tillgänglighet och en mer kontrollerad inställning. Muskelfantomerna undersöktes med hjälp av ultraljudsbaserad skjuvvågselastografi som är en metod som kan hjälpa till att bestämma vävnadens elasticitet. En PVA-grafitfantom och en vattenbaserad gelatin-grafitfantom, båda med fiskelinsnätverk som fibrer för anisotropi, konstruerades i detta projekt. Resultat från skjuvvågselastografi på PVA-fantomen indikerade ingen anisotropi, men liknade visuellt en muskel. Även då anisotropi inte uppnåddes, så matchade skjuvmodulen för PVA skjuvmodulen för skelettmuskelvävnad. Resultat från skjuvvågselastografi på gelatinfantomen indikerade anisotropi dock visade resultatet ingen visuell liknelse av en skelettmuskel på grund av gelatinets låga skjuvmodul. En 3D modell av bäckenbenet, som förseddes det här projektet, mättes och jämfördes med referensvärde av tidigare studie för framtid 3D friformsframställning av modellen. Resultat av mätningar visade på liknelser mellan 3D modellen och det kvinnliga bäckenbenet förutom sagittal outlet som hade ett avvikande värde. Inför fortsättning av det här projektet kan en utveckling av muskel fantomen ske genom applicering av den komplexa geometrin hos levator ani muskeln samt sammansättning av muskel fantomen och 3D framställningen av bäckenbenet. Sammansättningen av dessa två delar ger en mer komplett fantom där skjuvvågselastografi kan appliceras på samma sätt som hos kvinnliga patienter. / Ja

Objectives: The mucosa of human detrusor strips impairs catecholamine-induced relaxation. In order to elucidate which signal transduction pathways are involved in this cross talk between the mucosa and detrusor, we have studied the effects of several pharmacological agonists and antagonists on noradrenaline-mediated relaxation in intact and mucosa-denuded detrusor strips. Patients and Methods: Strips of detrusor tissue were obtained from patients who had undergone cystectomy for bladder cancer and were set up for force measurement. KCl- or carbachol-precontracted strips were relaxed with increasing concentrations of noradrenaline in the absence and in the presence of nitric oxide synthase inhibitor, L-NAME; P2X-receptor antagonist, PPADS; ET A -receptor antagonist, BQ-123; ET B -receptor antagonist, BQ-788; cyclooxygenase inhibitor, diclofenac; AT 1 -receptor antagonist, candesartan; and NK 1 -receptor antagonist, L-703,606. Results: In intact strips, KCl-stimulated force was enhanced by all blockers; carbachol-stimulated force increased with L-703,606. In denuded strips, only L-NAME augmented the KCl-stimulated contraction. Noradrenaline relaxed the precontracted detrusor strips to a significantly larger extent and at lower concentrations in denuded than in intact strips. L-NAME, PPADS and BQ-123/BQ-788 had little effect on noradrenaline-induced relaxation, whereas diclofenac, candesartan and L-703,606 sensitized intact carbachol-stimulated detrusor strips to noradrenaline-induced relaxation. Conclusion: Inhibition of the noradrenaline-induced relaxation of precontracted human detrusor strips by the mucosa is attenuated by diclofenac, candesartan and L-703,606 suggesting the involvement of prostanoids, angiotensin and neurokinin pathways. Further experiments are required to unravel the exact mechanisms.

Aim Musculoskeletal disorders are a major public health problem in most developed countries. As a main cause of chronic pain, they have resulted in an increasing prescription of opioids worldwide. With regard to the situation in Germany, this study aimed at estimating the prevalence of musculoskeletal diseases such as chronic low back pain (CLBP) and hip/knee osteoarthritis (OA) and at depicting the applied treatment patterns. - Subject and methods German claims data from the InGef Research Database were analyzed over a 6-year period (2011–2016). The dataset contains over 4 million people, enrolled in German statutory health insurances. Inpatient and outpatient diagnoses were considered for case identification of hip/knee OA and CLBP. The World Health Organization (WHO) analgesic ladder was applied to categorize patients according to their pain management interventions. Information on demographics, comorbidities, and adjuvant medication was collected. - Results In 2016, n = 2,693,481 individuals (50.5% female, 49.5% male) were assigned to the study population; 62.5% of them were aged 18–60 years. In 2016, n = 146,443 patients (5.4%) with CLBP and n = 307,256 patients (11.4%) with hip/ knee OA were identified. Of those with pre-specified pain management interventions (CLBP: 66.3%; hip/knee OA: 65.1%), most patients received WHO I class drugs (CLBP: 73.6%; hip/knee OA: 68.7%) as the highest level. - Conclusion This study provides indications that CLBP and hip/knee OA are common chronic pain conditions in Germany, which are often subjected to pharmacological pain management. Compared to non-opioid analgesic prescriptions of the WHO I class, the dispensation of WHO class II and III opioids was markedly lower, though present to a considerable extent.

Das Dokument enthält eine fehlerhafte Darstellung der Abbildung 3.3 (Seite 38). DieOriginalabbildung ist dem entsprechenden Fachartikel im Scandinavian Journal of Medicineand Science in Sports zu entnehmen:Mersmann, F., Bohm, S., Schroll, A., Boeth, H., Duda, G., Arampatzis, A. (2015): Muscleshape consistency and muscle volume prediction of thigh muscles. Scandinavian Journal ofMedicine & Science in Sports, 25: e208–e213 doi: 10.1111/sms.12285.The document contains a corrupted rendering of figure 3.3 (page 38). The original figurecan be obtained in the respective journal article:Mersmann, F., Bohm, S., Schroll, A., Boeth, H., Duda, G., Arampatzis, A. (2015): Muscleshape consistency and muscle volume prediction of thigh muscles. Scandinavian Journal ofMedicine & Science in Sports, 25: e208–e213 doi: 10.1111/sms.12285. / Bei jugendlichen Athleten wirken sowohl die körperliche Reifung als auch die erhöhte mechanische Belastung auf die Entwicklung des Muskel- und Sehnengewebes. Die Interaktion dieser beiden adaptationswirksamen Stimuli könnte, insbesondere bei Sportlern aus Sprungdisziplinen, Dysbalancen des Kraftpotenzials der Muskulatur und der Widerstandsfähigkeit der Sehne auslösen und somit potentiell das Verletzungsrisiko der Sehne erhöhen. Vor diesem Hintergrund untersucht die vorliegende Arbeit mittels Magnetresonanztomographie, Ultraschall und Dynamometrie die Entwicklung der morphologischen und mechanischen Eigenschaften der Knieextensoren und der Patellarsehne bei jugendlichen Kader-Volleyballathleten. Durch eine Querschnitts- und zwei Längsschnittstudien konnte Evidenz dafür erbracht werden, dass sich unter dem Einfluss der sportartspezifischen Belastung Kraft und Morphologie des Muskels in Relation zu den mechano-morphologischen Eigenschaften der Patellarsehne bei den jugendlichen Athleten nicht ausgewogen entwickeln, was zu einer erhöhten chronischen Belastung und Beanspruchung des Sehnengewebes führt. Darüber hinaus können muskulotendinöse Dysbalancen im Trainingsprozess durch erhöhte Fluktuationen der Muskelkraft und einer nicht-adäquaten Modulation der Widerstandsfähigkeit der Sehne zu episodischen Spitzen der mechanischen Beanspruchung der Sehne führen. Eine derart unausgewogene Entwicklung von Muskel und Sehne könnte für die Ausprägung von Überlastungsbeschwerden in der hier untersuchten Risikogruppe für Tendinopathien prädisponieren. Im Rahmen der Forschungsbemühungen wurde darüber hinaus ein Verfahren zur Vorhersage des Volumens von Oberschenkelmuskeln auf der Basis einfach zu erhebender Parameter entwickelt. Durch das Verfahren ließe sich zukünftig der Zeit- und Kostenaufwand muskelmorphologischer Diagnostik deutlich reduzieren. / In adolescent athletes, the development of muscle and tendon tissue is influenced both by maturation and mechanical loading. The interaction of these two driving stimuli could initiate imbalances in the development of the muscle strength capacity and tendon stiffness, especially in athletes from sports that feature a high frequency of jumps. A consequence could be an increased mechanical demand placed upon the tendon by the working muscle, which might increase the risk of tendon injury. Considering the lack of information on the effects of maturation and superimposed mechanical loading on the muscle-tendon unit during adolescence, the present work examines the development of the morphological and mechanical properties of the knee extensors and patellar tendon in adolescent elite volleyball athletes by means of magnetic resonance imaging, ultrasound and dynamometry. A cross-sectional comparison and two longitudinal studies provided evidence that the two-fold stimulus of sport-specific loading and maturation results in and imbalanced development of muscle strength and morphology in relation to the mechano-morphological properties of the patellar tendon in mid-adolescent volleyball athletes and results in increased tendon stress and strain. Moreover, during a training process, fluctuations of muscle strength and an inadequate modulation of tendon stiffness initiate episodes of high-level tendon strain during maximum muscle contractions. Such an imbalanced development of muscle and tendon could predispose for the development of tendon overuse injury in adolescent volleyball athletes, which are known to be at high risk of developing tendinopathy. Moreover, in the context of the described research endeavour, a method for the volume prediction of thigh muscles on the basis of easily measurable parameters has been developed, which could greatly reduce the time and costs involved in the diagnostics of muscle morphology in the future.

Die meisten Kinder mit infantiler Zerebralparese (IZP) entwickeln eine spastische Parese, was zu Schwäche, erhöhtem Muskel-Dehnungswiderstand und Kontrakturen führt. Der Spitzfuß ist eines ihrer häufigsten Defizite. Das übergeordnete Ziel dieser Arbeit war es, nicht-invasive Behandlungsstrategien für diese Pathologie näher zu untersuchen. In der ersten Studie wurden die Effekte einer Unterschenkel-Lagerungsorthese auf die Muskelmorphometrie des Gastrocnemius unter zu Hilfenahme von Ultraschall und 3D Bewegungsanalyse untersucht. Lagerungsorthesen konnten das Gangbild verbessern, führten gleichzeitig aber zu Atrophie. Um eine alternative Therapieform zu finden, wurde in der zweiten Studie die kontraktile Aktivität des Gastrocnemius mittels Ultraschall, Bewegungsanalyse und EMG während des Gehens, Vorwärts-Bergauf, bzw. Rückwärts-Bergab, sowie in der Ebene untersucht. Das Bergaufgehen steigerte die konzentrische Exkursion der Faszikel, wohingegen das Rückwärts-Bergabgehen zu vermehrter Exzentrik führte. Da sich exzentrisches Training positiv auf Faszikellängenwachstum auswirken kann, wurde in der dritten Studie das Rückwärts-Bergabgehen mit statischem Dehnen als traditionelle Therapieform verglichen. Ultraschall, Bewegungsanalyse und handgesteuerte Dynamometrie wurden verwendet, um die Plantarflexorenkraft, die passive Sprunggelenksbeweglichkeit, die Gastrocnemius Morphometrie sowie die Steifigkeit und Dehnbarkeit auf Muskel-Sehnen und Gelenkebene zu untersuchen. Dehnen zeigte keinerlei Benefits gegenüber dem Laufbandtraining. Rückwärts-Bergabgehen war dagegen ein effektives Gangtraining und setzt vermutlich neuronale und koordinative Reize. In der Zusammenschau scheinen positive Änderungen im Gangbild bei IZP Kindern sowohl durch Unterschenkelorthesen, als auch durch Rückwärts-Bergabgehen erreichbar zu sein. Beides führte aber nicht zu Muskelwachstum. Funktionelle Verbesserungen scheinen daher auch stark von neuronal, koordinativen Aspekten abhängig zu sein. / Most children with Cerebral Palsy (CP) develop spastic paresis, which leads to muscle weakness, increased stretch-resistance and joint contractures. The gastrocnemius muscle is frequently targeted to alleviate a common deficiency known as equinus. The overall objective of this thesis was to investigate several non-invasive treatment strategies for this pathology. The first study investigated the effects of ankle foot orthotics on spastic gastrocnemius morphometrics as well as on gait by using ultrasound and motion capturing. We concluded that braces improved walking function but also lead to atrophy. During the second study, we searched for a readily available, substituting stimulus and compared the contractile activity of the gastrocnemius on treadmills, namely during flat-forward, forward-uphill and backward-downhill gait using ultrasound, motion capturing and EMG. Uphill gait promoted concentric fascicle action, while backward-downhill gait increased eccentric fascicle action. Since eccentric training had been previously shown to increase fascicle length in controls, during the third study, we compared backward-downhill walking versus static, manual stretching. Ultrasound, motion analysis and handheld dynamometry were used to test plantarflexor strength, passive ankle joint flexibility, as well as gastrocnemius morphometrics, stiffness and strain on muscle-tendon and joint level. Backward-downhill walking led to larger single stance dorsiflexion and faster achievable walking velocities while stretching aggravated knee flexion in swing. Strength, joint flexibility, as well as stiffness on muscle-tendon and joint level were not altered. Backward-downhill walking can be an effective gait treatment, probably improving coordination. Nevertheless, more intense training might be necessary to alter muscle-tendon properties. In sum, backward-downhill walking and bracing increased function without promoting or even by harming muscle growth.