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Combining physiological relevance and throughput for in vitro cardiac contractility measurement
(2020)
Despite increasing acceptance of human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) in safety pharmacology, controversy remains about the physiological relevance of existing in vitro models for their mechanical testing. We hypothesize that existing signs of immaturity of the cell models result from an improper mechanical environment. We cultured hiPSC-CMs in a 96-well format on hyperelastic silicone membranes imitating their native mechanical environment, resulting in physiological responses to compound stimuli.We validated cell responses on the FLEXcyte 96, with a set of reference compounds covering a broad range of cellular targets, including ion channel modulators, adrenergic receptor modulators and kinase inhibitors. Acute (10 - 30 min) and chronic (up to 7 days) effects were investigated. Furthermore, the measurements were complemented with electromechanical models based on electrophysiological recordings of the used cell types.hiPSC-CMs were cultured on freely-swinging, ultra-thin and hyperelastic silicone membranes. The weight of the cell culture medium deflects the membranes downwards. Rhythmic contraction of the hiPSC-CMs resulted in dynamic deflection changes which were quantified by capacitive distance sensing. The cells were cultured for 7 days prior to compound addition. Acute measurements were conducted 10-30 minutes after compound addition in standard culture medium. For chronic treatment, compound-containing medium was replaced daily for up to 7 days. Electrophysiological properties of the employed cell types were recorded by automated patch-clamp (Patchliner) and the results were integrated into the electromechanical model of the system.Calcium channel agonist S Bay K8644 and beta-adrenergic stimulator isoproterenol induced significant positive inotropic responses without additional external stimulation. Kinase inhibitors displayed cardiotoxic effects on a functional level at low concentrations. The system-integrated analysis detected alterations in beating shape as well as frequency and arrhythmic events and we provide a quantitative measure of these.
Successful bone sawing requires a high level of skill and experience, which could be gained by the use of Virtual Reality-based simulators. A key aspect of these medical simulators is realistic force feedback. The aim of this paper is to model the bone sawing process in order to develop a valid training simulator for the bilateral sagittal split osteotomy, the most often applied corrective surgery in case of a malposition of the mandible. Bone samples from a human cadaveric mandible were tested using a designed experimental system. Image processing and statistical analysis were used for the selection of four models for the bone sawing process. The results revealed a polynomial dependency between the material removal rate and the applied force. Differences between the three segments of the osteotomy line and between the cortical and cancellous bone were highlighted.
Zusammenfassung: In der Orthopädie zählt der therapeutische Ultraschall als Mittel zur Prävention und Therapiebegleitung. Er hat mechanische, thermische und physiko-chemische Auswirkungen auf den menschlichen Körper. Um mehr Erkenntnisse über die thermischen Auswirkungen zu erlangen, wurden Versuche an einem Hydrogel-Phantom und an Probanden durchgeführt. Dabei entstand eine signifikante Erwärmung des Gewebes, welche beim Probandenversuch an der Oberfläche und beim Hydrogelversuch in der Tiefe gemessen wurde.
Summary: In orthopaedics, therapeutic ultrasound is a tool of prevention and therapy support. It has mechanical, thermal and physico-chemical effects on the human body. Tests with a hydrogel phantom and with human probands have been performed in order to obtain more knowledge about their thermal effects. Both tests measured temperature increases in cell tissue, on the surface with the human proband test and in depth with the hydrogel phantom test.
The overall objective of this study is to develop a new external fixator, which closely maps the native kinematics of the elbow to decrease the joint force resulting in reduced rehabilitation time and pain. An experimental setup was designed to determine the native kinematics of the elbow during flexion of cadaveric arms. As a preliminary study, data from literature was used to modify a published biomechanical model for the calculation of the joint and muscle forces. They were compared to the original model and the effect of the kinematic refinement was evaluated. Furthermore, the obtained muscle forces were determined in order to apply them in the experimental setup. The joint forces in the modified model differed slightly from the forces in the original model. The muscle force curves changed particularly for small flexion angles but their magnitude for larger angles was consistent.
The human arm consists of the humerus (upper arm), the medial ulna and the lateral radius (forearm). The joint between the humerus and the ulna is called humeroulnar joint and the joint between the humerus and the radius is called humeroradial joint. Lateral and medial collateral ligaments stabilize the elbow. Statistically, 2.5 out of 10,000 people suffer from radial head fractures [1]. In these fractures the cartilage is often affected. Caused by the injured cartilage, degenerative diseases like posttraumatic arthrosis may occur. The resulting pain and reduced range of motion have an impact on the patient’s quality of life. Until now, there has not been a treatment which allows typical loads in daily life activities and offers good long-term results. A new surgical approach was developed with the motivation to reduce the progress of the posttraumatic arthrosis. Here, the radius is shortened by 3 mm in the proximal part [2]. By this means, the load of the radius is intended to be reduced due to a load shift to the ulna. Since the radius is the most important stabilizer of the elbow it has to be confirmed that the stability is not affected. In the first test (Fig. 1 left), pressure distributions within the humeroulnar and humeroradial joints a native and a shortened radius were measured using resistive pressure sensors (I5076 and I5027, Tekscan, USA). The humerus was loaded axially in a tension testing machine (Z010, Zwick Roell, Germany) in 50 N steps up to 400 N. From the humerus the load is transmitted through both the radius and the ulna into the hand which is fixed on the ground. In the second test (Fig. 1 right), the joint stability was investigated using a digital image correlation system to measure the displacement of the ulna. Here, the humerus is fixed with a desired flexion angle and the unconstrained forearm lies on the ground. A rope connects the load actuator with a hook fixed in the ulna. A guide roller is used so that the rope pulls the ulna horizontally when a tensile load is applied. This creates a moment about the elbow joint with a maximum value of 7.5 Nm. Measurements were performed with varying flexion angles (0°, 30°, 60°, 90°, 120°). For both tests and each measurement, seven specimens were used. Student ́s t-test was employed to determine whether the mean values of the measurements in native specimen and operated specimens differ significantly.