Knee Contact Forces Research Articles

Machine learning based estimates of medial knee contact force are associated with cartilage loss over 2.5 years in knee osteoarthritis patients

Purpose: Mechanical loads are theorized to relate to cartilage loss, a hallmark feature of knee osteoarthritis (OA). Knee contact forces are only measurable using instrumented implants. Several studies have correlated external biomechanical gait parameters with medial contact forces (MCFs) from instrumented knees. To our knowledge, no study has related estimated MCFs with longitudinal cartilage changes. The purpose of this study was to predict MCFs from external knee loads in order to relate them to longitudinal changes in medial tibial cartilage volume over 2.5 years in knee OA patients.

Random Forest enhancement using improved Artificial Fish Swarm for the medial knee contact force prediction.

Knee contact force (KCF) is an important factor to evaluate the knee joint function for the patients with knee joint impairment. However, the KCF measurement based on the instrumented prosthetic implants or inverse dynamics analysis is limited due to the invasive, expensive price and time consumption. In this work, we propose a KCF prediction method by integrating the Artificial Fish Swarm and the Random Forest algorithm. First, we train a Random Forest to learn the nonlinear relation between gait parameters (input) and contact pressures (output) based on a dataset of three patients instrumented with knee replacement. Then, we use the improved artificial fish group algorithm to optimize the main parameters of the Random Forest based KCF prediction model. The extensive experiments verify that our method can predict the medial knee contact force both before and after the intervention of gait patterns, and the performance outperforms the classical multi-body dynamics analysis and artificial neural network model.

The Capacity of Generic Musculoskeletal Simulations to Predict Knee Joint Loading Using the CAMS-Knee Datasets

Musculoskeletal models enable non-invasive estimation of knee contact forces (KCFs) during functional movements. However, the redundant nature of the musculoskeletal system and uncertainty in model parameters necessitates that model predictions are critically evaluated. This study compared KCF and muscle activation patterns predicted using a scaled generic model and OpenSim static optimization tool against in vivo measurements from six patients in the CAMS-knee datasets during level walking and squatting. Generally, the total KCFs were under-predicted (RMS: 47.55%BW, R2: 0.92) throughout the gait cycle, but substiantially over-predicted (RMS: 105.7%BW, R2: 0.81) during squatting. To understand the underlying etiology of the errors, muscle activations were compared to electromyography (EMG) signals, and showed good agreement during level walking. For squatting, however, the muscle activations showed large descrepancies especially for the biceps femoris long head. Errors in the predicted KCF and muscle activation patterns were greatest during deep squat. Hence suggesting that the errors mainly originate from muscle represented at the hip and an associated muscle co-contraction at the knee. Furthermore, there were substaintial differences in the ranking of subjects and activities based on peak KCFs in the simulations versus measurements. Thus, future simulation study designs must account for subject-specific uncertainties in musculoskeletal predictions.

Unintended Changes in Contralateral Limb as a Result of Acute Gait Modification.

Gait modification using real-time biofeedback is a conservative intervention associated with positive outcomes. Results from systematic reviews corroborate the effectiveness of various strategies employing real-time biofeedback for reducing estimated knee joint load. The effects on the nonmodified limb, however, remain unclear. Biomechanical changes to the nonmodified limb were investigated during unilaterally implemented medial knee thrust, lateral trunk lean, and toe-in foot progression. Nineteen healthy participants were recruited. Ten trials were completed for each gait condition including baseline. Assigned magnitude for each gait modification strategy was individualized based on the mean and SD of the gait parameter during baseline. Visual real-time biofeedback was provided. During medial knee thrust, participants' nonmodified limb presented with increased: first peak medial knee contact force, internal first peak knee extensor moment, as well as knee- and hip-flexion angles at internal first peak knee extensor moment. Observed biomechanical changes are elucidative of the body's attempt to attenuate increased external loads. These findings may carry significant implications for pathological populations. Load redistribution to the nonmodified side may result in unfavorable long-term outcomes particularly in patients with bilateral diagnosis. Future studies should explore acute and chronic changes in the nonmodified limb of individuals with knee osteoarthritis.

Lower-limb joint reaction forces and moments during modified cycling in healthy controls and individuals with knee osteoarthritis

BackgroundOsteoarthritis (OA) is a clinical problem affecting an estimated 27 million adults in the United States, with the only clear treatment options being pain management. Cycling is an integral component of exercise for individuals with knee osteoarthritis, while the joint reaction forces during cycling remain unknown. MethodsThirteen subjects with medial compartment knee osteoarthritis and eleven healthy subjects performed a cycling protocol with a neutral pedal and four pedal modifications. Six hundred muscle-actuated inverse-dynamic simulations (24 subjects, 5 trials in each of 5 conditions) were performed to estimate joint reaction force differences between conditions. FindingsSubjects with knee osteoarthritis had many significant changes among them was a reduction in knee adduction-abduction moment by 45% (5° lateral wedge), 77% (10° lateral wedge), 54% (5° toe-in) and 58% (10° toe-in). Conversely the healthy subjects had no significant changes in the knee adduction-abduction moment for the lateral wedge conditions and the 5° toe-in but did decrease by 18% for the 10° toe-in condition. When comparing the cohorts across the different pedal conditions, the data showed many significant differences among the groups. InterpretationThis study showed that while cycling in different pedal modifications, the knee osteoarthritis subjects had more beneficial changes in their knee adduction-abduction moment compared to the healthy subjects with the lateral-wedge modification resulting in the greatest impact on the subjects with knee osteoarthritis. Both groups had greater contact forces at the hip and ankle across pedal modifications compared to neutral. For the knee, subjects with osteoarthritis mostly decreased their knee contact forces but the healthy subjects mostly increased these forces with all pedal modifications.

Sensitivity of the knee joint response, muscle forces and stability to variations in gait kinematics-kinetics

Sensitivity analysis of the knee joint response to variations in gait kinematics-kinetics as reported in the literature is crucial for improved understanding and more effective prevention-treatment strategies. Using our validated finite element-musculoskeletal (FE-MS) model of lower extremity, we investigate the sensitivity of knee anterior cruciate ligament (ACL), muscle, and contact forces plus stability to the reported scatter in asymptomatic gait data. Three highly loaded stance instants (25, 50 and 75%) with five levels (mean, ±0.5SD and ±SD) for each of six knee joint angles-moments are used employing Taguchi approach (25 experiments) and regression equations. ACL force drops significantly at larger flexion angles (all periods) and smaller internal moment (at 75% only) but increases with the flexion moment. Tibiofemoral (TF) medial-lateral contact force partitioning is found, contrary to the common claim, most sensitive to changes in the adduction angle and not in the adduction moment. Total TF contact force increases significantly at greater moments (but not angles), especially in the sagittal plane. Forces in lateral hamstrings are significantly influenced by changes in adduction angles-moments. Larger flexion moments (at 25 and 50%) significantly increase forces in quadriceps and on patellofemoral (PF) contact. Sagittal moment, adduction moment (at 75%) and flexion angle (at 25%) contribute most to the joint stability. A strong inverse correlation exists between the joint stability and the total TF compression force. These findings can be exploited to adapt and modify intact, injured and reconstructed knee joint responses during gait.

Muscle recruitment strategies can reduce joint loading during level walking

Joint inflammation, with consequent cartilage damage and pain, typically reduces functionality and affects activities of daily life in a variety of musculoskeletal diseases. Since mechanical loading is an important determinant of the disease process, a possible conservative treatment is the unloading of joints. In principle, a neuromuscular rehabilitation program aimed to promote alternative muscle recruitments could reduce the loads on the lower-limb joints during walking. The extent of joint load reduction one could expect from this approach remains unknown. Furthermore, assuming significant reductions of the load on the affected joint can be achieved, it is unclear whether, and to what extent, the other joints will be overloaded. Using subject-specific musculoskeletal models of four different participants, we computed the muscle recruitment strategies that minimised the hip, knee and ankle contact force, and predicted the contact forces such strategies induced at the other joints. Significant reductions of the peak force and impulse at the knee and hip were obtained, while only a minimal effect was found at the ankle joint. Adversely, the peak force and the impulse in non-targeted joints increased when aiming to minimize the load in an adjacent joint. These results confirm the potential of alternative muscle recruitment strategies to reduce the loading at the knee and the hip, but not at the ankle. Therefore, neuromuscular rehabilitation can be targeted to reduce the loading at affected joints but must be considered carefully in patients with multiple joints affected due to the potential adverse effects in non-targeted joints.

Inverse dynamic estimates of muscle recruitment and joint contact forces are more realistic when minimizing muscle activity rather than metabolic energy or contact forces

BackgroundAssessment of contact forces is essential for a better understanding of mechanical factors affecting progression of osteoarthritis. Since contact forces cannot be measured non-invasively, computer simulations are often used to assess joint loading. Contact forces are to a large extent determined by muscle forces. These muscle forces are computed using optimization techniques that solve the muscle redundancy problem by assuming that muscles are coordinated in a way that optimizes performance (e.g., minimizes muscle activity or metabolic energy). However, it is unclear which of the many proposed performance criteria best describes muscle coordination. Research questionWhich performance criterion best describes muscle recruitment patterns and knee contact forces recorded using electromyography (EMG) and load cell instrumented prostheses?. MethodsWe solved the muscle redundancy problem based on six different groups of performance criteria: muscle activations, volume-scaled activations, forces, stresses, metabolic energy, and joint contact forces. Computed muscle excitations and knee contact forces during over-ground walking were validated against recorded EMG signals and measured contact forces for four subjects with instrumented knee prostheses in the “Grand Challenge Competition to Predict in Vivo Knee Loads” dataset. ResultsPerformance criteria based on either stress or muscle activation (either unscaled or scaled by muscle volume), both to a power of 3 or 4, resulted in the best agreement between measured and simulated values. These performance criteria outperformed all other criteria in terms of agreement between simulated muscle excitations and EMG, whereas good agreement between measured and predicted contact forces was also observed for minimization of contact forces and metabolic energy. SignificanceGiven the large differences in accuracy obtained with different performance criteria (e.g., root mean square errors of contact forces differed up to 0.45 body weight), the results of our study are important to improve the validity of in silico assessment of joint loading.

Human Knee Inverse Dynamics Model of Vertical Jump Exercise

Abstract This work deals with the dynamics of the human knee during vertical jump exercise. The focus is on the joint forces necessary to produce the jump and to dissipate energy during landing. A two-dimensional (2D) sagittal plane, inverse dynamics human leg model is developed. This model uses data from a motion capture system and force plates in order to predict knee and hip joint forces during the vertical jump exercise. The model consists of three bony structures femur, tibia, and patella, ligament structures to include both cruciate and collateral ligaments, and knee joint muscles. The inverse dynamics model is solved using optimization in order to predict joint forces during this exercise. matlab software package is used for the optimization computations. Results are compared with data available in the literature. This work provides insight regarding contact forces and ligaments forces, muscle forces, and knee and hip contact forces in the vertical jump exercise.

Patients With Medial Knee Osteoarthritis Reduce Medial Knee Contact Forces by Altering Trunk Kinematics, Progression Speed, and Stepping Strategy During Stair Ascent and Descent: A Pilot Study.

Medial knee loading during stair negotiation in individuals with medial knee osteoarthritis, has only been reported in terms of joint moments, which may underestimate the knee loading. This study assessed knee contact forces (KCF) and contact pressures during different stair negotiation strategies. Motion analysis was performed in five individuals with medial knee osteoarthritis (52.8±11.0 years) and eight healthy subjects (51.0±13.4 years) while ascending and descending a staircase. KCF and contact pressures were calculated using a multi-body knee model while performing step-over-step at controlled and self-selected speed, and step-by-step strategies. At controlled speed, individuals with osteoarthritis showed decreased peak KCF during stair ascent but not during stair descent. Osteoarthritis patients showed higher trunk rotations in frontal and sagittal planes than controls. At lower self-selected speed, patients also presented reduced medial KCF during stair descent. While performing step-by-step, medial contact pressures decreased in osteoarthritis patients during stair descent. Osteoarthritis patients reduced their speed and increased trunk flexion and lean angles to reduce KCF during stair ascent. These trunk changes were less safe during stair descent where a reduced speed was more effective. Individuals should be recommended to use step-over-step during stair ascent and step-by-step during stair descent to reduce medial KCF.

Characteristics of Contact Force and Muscle Activation During Choreographed Falls With 2 Common Landing Techniques in Contemporary Dance.

This study investigated the contact forces and muscle activation during 2 choreographed fall techniques in contemporary dancers and how these were affected by the fall technique. Ten collegiate-level dancers were instructed in 2 choreographed falls: (1)an anteriorly focused fall involving ankle plantar flexion, knee flexion, and hip flexion with dispersion of forces up the anterior surface of the shank (technique 1) and (2)a laterally focused fall involving inversion at the ankle with dispersion of forces up the lateral aspect of the shank (technique 2). The knee and hip contact forces were 26.3% smaller (technique 1: 1743N vs technique 2: 1284N) and 24.1% greater (technique 1: 1334N vs technique 2: 1656N), respectively, in technique 2 (P < .03). At the time of knee contact, percentage of maximal voluntary isometric contraction (%MVIC) was 45.8% greater for rectus femoris muscle (technique 1: 7.2% vs technique 2: 10.5%) and 96.9% greater for gluteus medius muscle (technique 1: 3.2% vs technique 2: 6.3%) (P < .01) in technique 2. The results provide insight into determining safer landing strategies to avoid knee injuries in individuals who experience a fall (ie,dancers, athletes, and older adults).

Guided Growth Intervention in Children and Adolescents with Valgus Malalignment Changes the Medial and Lateral Knee Contact Force During Gait

Guided Growth Intervention in Children and Adolescents with Valgus Malalignment Changes the Medial and Lateral Knee Contact Force During Gait

Impact of variable stiffness shoes on medial knee contact forces

Impact of variable stiffness shoes on medial knee contact forces

Small Lateral Meniscus Tears Propagate and are Detrimental to Knee Kinematics and Contact Forces

Aims and Objectives: To quantify propagation of small vertical lateral meniscus tears located in the posterior horn during simulated cyclic cutting maneuvers on a robotic testing system and the resulting changes in knee kinematics and forces. Materials and Methods: 14 fresh-frozen human cadaveric knees (mean age: 74.8 ± 14.4) were mounted onto a 6-DOF robotic testing system. Knees with osteoarthritis greater than grade II were excluded. A small vertical tear in the posterior horn of the lateral meniscus medial to the popliteus tendon was created via a posterior arthrotomy. Tear length was measured with a custom-made device (accuracy 0.2 mm, inter-rater reliability 0.914). In 6 specimens, the ACL was resected arthroscopically, whereas in 8 knees the ACL was left intact. 10Nm valgus and 5Nm external rotation moment combined with 250N of axial compression were applied while the knees were continuously flexed from full extension to 90° of flexion for 500 cycles. Meniscus tear length was measured after each 100 cycles. After the measurements, the posterior capsule and skin were closed in a standardized fashion. Knee kinematics, the in situ force in the ACL, the resultant force in the lateral meniscus, and bony contact forces at the tibiofemoral joint were calculated at 30°, 60°, and 90° of knee flexion. A repeated measures ANOVA was used to compared the outcome parameters between knee states (p &lt; 0.05). Results: In intact knees, small vertical lateral meniscus tears propagated significantly from baseline to 100, 200, 300, 400, and 500 cycles (all p &lt; 0.001). This corresponds to a tear propagation of 28.7% ± 5.4%. Similarly, in ACL deficient knees meniscus tear propagation was significant throughout all sets of cycles (p = 0.001). This corresponds to a tear propagation of 26.1% ± 8.3%. While in intact knees, external tibial rotation was significantly increased by up to 45.5% after meniscus tear propagation, knee kinematics were not significantly affected in ACL deficient knees (NS). The resultant forces in the lateral meniscus increased with tear propagation in the intact knees by up to 54.1%. Likewise, the resultant forces in the lateral meniscus increased in the ACL deficient knees by up to 116.5% (p &lt; 0.05). In intact knees, bony contact forces at the tibiofemoral joint increased with meniscus tear propagation by up to 91.9% (p &lt; 0.05), which was comparable to ACL deficient knees. Conclusion: The data of this study suggest, that small vertical lateral meniscus tears propagate significantly regardless of the integrity of the ACL and after only 100 cycles of knee loading. The findings of the current robotic study are in contrast to recent clinical studies that found that small, vertical and stable tears in the posterior horn of the lateral meniscus could be left in situ. However, based on the current data, such tears do propagate over time regardless of the integrity of the ACL, which might alter knee kinematics and resultant forces in the lateral meniscus as well as bony contact forces at the tibiofemoral joint.

Foot Pronation Contributes to Altered Lower Extremity Loading After Long Distance Running.

This study presents an investigation of the changes in foot posture, joint kinematics, joint moments and joint contact forces in the lower extremity following a 5 k treadmill run. A relationship between knee and ankle joint loading and foot posture index (FPI) is developed. Twenty recreational male heel-strike runners participated in this study. All participants had a history of running exercise and were free from lower extremity injuries and foot deformities. Foot posture was assessed from a six-item FPI to quantitatively classify high supination to high pronation foot poses. The FPI is scored using a combination of observations and foot palpations. The three-dimensional marker trajectories, ground reaction force and surface electromyography (EMG) were recorded at pre and post-gait sessions conducted over-ground and 5 k running was conducted on a treadmill. Joint kinematics, joint moments and joint contact forces were computed in OpenSim. Simulated EMG activations were compared against experimental EMG to validate the model. A paired sample t-test was conducted using a 1D statistical parametric mapping method computed temporally. Hip joint moments and contact forces increased during initial foot contact following 5 k running. Knee abduction moment and superior-inferior knee contact force increased, whereas the knee extension moment decreased. Ankle plantarflexion moment and ankle contact forces increased during stance. FPI was found to be moderately correlated with peak knee and ankle moments. Recreational male runners presented increased static foot pronation after 5 k treadmill running. These findings suggest that following mid distance running foot pronation may be an early indicator of increased lower limb joint loading. Furthermore, the FPI may be used to quantify the changes in knee and ankle joint moments.

Is knee biomechanics different in uphill walking on different slopes for older adults with total knee replacement?

The purpose of this study was to investigate knee biomechanics in uphill walking on slopes of 5°, 10° and 15° for total knee replacement (TKR) patients. Twenty-five post-TKR patients and ten healthy controls performed five walking trials on level ground and different slopes on an instrumented ramp system. A 2 × 2 × 4 (limb × group × incline slope) mixed model ANOVA was used to examine selected variables. The peak knee extension moment (KEM) was greater in 15° uphill walking compared to level, 5° and 10° uphill walking. TKR patients had lower peak KEM and smaller knee extension range of motion than healthy controls in all walking conditions. The Replaced Limb showed lower peak KEM in 10° and 15° uphill walking than the Non-replaced Limb and smaller knee extension range of motion (ROM) in 10° uphill walking. Knee extension and abduction ROM increased with increased incline angles. The greater peak loading-response vertical ground reaction force was found in level walking compared to three levels of uphill walking. The peak loading-response knee abduction moment was greater in level walking compared to 10° and 15° uphill walking. However, the medial knee contact force was greater in non-replaced limb compared to replaced limb in 10° and 15° uphill walking. The results suggest 5° uphill walking may have the potential to become a safe exercise for unilateral TKR patients.

Skipping has lower knee joint contact forces and higher metabolic cost compared to running

BackgroundThe health benefits of running based exercise programs are plentiful however the high rate of injury in these programs often reduces or eliminates exercise participation. Skipping has shorter steps, reduced vertical ground reaction forces (GRFs), and lower knee extensor torques, compared to running forming the basis of the present hypothesis that skipping would have lower tibio-femoral and patello-femoral joint contact forces. Research questionThe purpose of this study was to compare knee contact forces between skipping and running at the same speed. We also compared metabolic cost of these two gaits to examine the idea that the larger vertical displacement in skipping is a primary factor in its previously reported high metabolic cost. MethodsThe study evaluated joint contact forces through musculoskeletal modeling with GRF and 3D kinematic data and metabolic cost using oxygen consumption data from 20 young, healthy, trained participants as they skipped and ran on an instrumented treadmill at 2.68 m/s. Results: Skipping, compared to running, had substantially lower tibio-femoral and patello-femoral joint contact forces and linear impulses on both per-step and per-kilometer (i.e. lower cumulative loads) bases and also 30% higher metabolic cost. The lower joint loads in skipping were directly associated with its shorter steps and the higher metabolic cost was directly associated to its larger vertical displacement through the stride. SignificanceAs joint loads may predispose individuals to running related injuries, skipping presents an attractive alternative exercise modality with additional increased aerobic benefits.

التنبؤ بقوة الاتصال عند نقطة الركبة الطبية

أجريت الدراسة الحالية لتقييم عوامل الخطر الشائعة لدى الأمهات لمرض سكري الحمل المشترك بين النساء الحوامل في مدينة الملك عبد الله الطبية بالرياض. باستخدام المنهج الوصفي التحليلي ، شملت الدراسة الحالية جميع النساء الحوامل في مدينة الملك عبد العزيز الطبية بالرياض. تم اختيار عينة مكونة من 450 امرأة حامل وفق طريقة أخذ العينات العشوائية الطبقية. تم تصميم استبيان منظم لتقييم عوامل الخطر الشائعة للأمهات لداء سكري الحمل بواسطة 449 من عينة البحث. أظهرت الدراسة أن النساء المصابات بداء سكري الحمل (GDM) يمكن أن يتعرضن لخطر الإصابة بمرض السكري في المستقبل. إلى جانب ذلك ، يختلف GDM في تناسب مباشر مع انتشار مرض السكري من النوع 2. وجدت الدراسة علاقة بين GDM والسمنة ، وتاريخ الأسرة مع مرض السكري ، والعمر ، والأنشطة البدنية. علاوة على ذلك ، أظهرت النتائج أن عينة البحث تفتقر إلى المعرفة فيما يتعلق بسكري الحمل ، أي النظام الغذائي والتمارين الرياضية. وفي هذا الصدد أوصت الباحثة بتوعية النساء بسرطان المثانة ومرض ورعاية الحوامل وخاصة المصابات بسكري الحمل.

The role of menisci in knee contact mechanics and secondary kinematics during human walking

BackgroundMeniscectomy is likely to result in an increase of joint loading on articular cartilage and initiates cartilage damages. However, the induced alterations in knee contact mechanics and secondary kinematics after the meniscal removal are still unclear during a walking gait. In this study, the role of menisci on the knee contact mechanics and secondary kinematics were investigated during a walking simulation. MethodsTwo natural knee models with or without menisci were established using a musculoskeletal multibody dynamics framework. Walking simulation were performed to qualify the knee contact forces and secondary kinematics, and ligament forces in these models. FindingsAfter the meniscal removal, the redistributed contact forces on the medial tibial cartilage increased by twofold, while the contact area remained almost the same. The subsequent increase of contact pressure suggested potential cartilage damages. In terms of the kinematic alterations, the tibia moved more posteriorly and internally with respect to the femur. And, the displacement in the medial-lateral direction reversed. In addition, the sharp force increase in the anterior cruciate ligament explained the reason why meniscal and anterior cruciate ligament injuries always happened concurrently. And, the anterior lateral ligament may act as the stabilizer in the tibial posterior displacement and varus rotation. InterpretationThis study shows that menisci served as the joint load distribution, and also as the kinematics constraints in the posterior and medial-lateral direction, which is beneficial to the rehabilitation plan-making of meniscal injuries.

Impact of antagonistic muscle co-contraction on in vivo knee contact forces

BackgroundThe onset and progression of osteoarthritis, but also the wear and loosening of the components of an artificial joint, are commonly associated with mechanical overloading of the structures. Knowledge of the mechanical forces acting at the joints, together with an understanding of the key factors that can alter them, are critical to develop effective treatments for restoring joint function. While static anatomy is usually the clinical focus, less is known about the impact of dynamic factors, such as individual muscle recruitment, on joint contact forces.MethodsIn this study, instrumented knee implants provided accurate in vivo tibio-femoral contact forces in a unique cohort of 9 patients, which were used as input for subject specific musculoskeletal models, to quantify the individual muscle forces during walking and stair negotiation.ResultsEven between patients with a very similar self-selected gait speed, the total tibio-femoral peak forces varied 1.7-fold, but had only weak correlation with static alignment (varus/valgus). In some patients, muscle co-contraction of quadriceps and gastrocnemii during walking added up to 1 bodyweight (~ 50%) to the peak tibio-femoral contact force during late stance. The greatest impact of co-contraction was observed in the late stance phase of stair ascent, with an increase of the peak tibio-femoral contact force by up to 1.7 bodyweight (66%).ConclusionsTreatment of diseased and failed joints should therefore not only be restricted to anatomical reconstruction of static limb axes alignment. The dynamic activation of muscles, as a key modifier of lower limb biomechanics, should also be taken into account and thus also represents a promising target for restoring function, patient mobility, and preventing future joint failure.Trial registrationGerman Clinical Trials Register: ID: DRKS00000606, date: 05.11.2010.