Multiphoton Imaging of Maturation in Tissue Engineering.

Donor cell-specific tissue-engineered (TE) implants are a promising therapy for personalized treatment of cardiovascular diseases, but current development protocols lack a stable longitudinal assessment of tissue development at subcellular resolution. As a first step toward such an assessment approach, in this study we establish a generalized labeling and imaging protocol to obtain quantified maturation parameters of TE constructs in three dimensions (3D) without the need of histological slicing, thus leaving the tissue intact. Focusing on intracellular matrix (ICM) and extracellular matrix (ECM) networks, multiphoton laser scanning microscopy (MPLSM) was used to investigate TE patches of different conditioning durations of up to 21 days. We show here that with a straightforward labeling procedure of whole-mount samples (so without slicing into thin histological sections), followed by an easy-to-use multiphoton imaging process, we obtained high-quality images of the tissue in 3D at various time points during development. The stacks of images could then be further analyzed to visualize and quantify the volume of cell coverage as well as the volume fraction and network of structural proteins. We showed that collagen and alpha-smooth muscle actin (α-SMA) volume fractions increased as normalized to full tissue volume and proportional to the cell count, with a converging trend to the final density of (4.0% ± 0.6%) and (7.6% ± 0.7%), respectively. The image analysis of ICM and ECM revealed a developing and widely branched interconnected matrix. We are currently working on the second step, that is, to integrate MPLSM endoscopy into a dynamic bioreactor system to monitor the maturation of intact TE constructs over time, thus without the need to take them out.

In Vivo Evaluation of a Novel Control Algorithm for Left Ventricular Assist Devices Based Upon Ventricular Stroke Work.

The physical fitness of patients with terminal heart failure and an implanted left ventricular assist device (LVAD) might be improved by load-adaptive control of the LVAD. In this study, three control strategies for LVAD were compared in eight pigs: (1) a constant stroke work (CSW) control strategy that ensures a constant ventricular load using ventricular stroke work as the control variable; (2) a work ratio (WR) controller that maintains a constant ratio of ventricular work to hydraulic pump work; and (3) a controller that maintains the pump pace at a constant speed (CS). Biventricular heart insufficiency was induced by increased isoflurane application, and preload, afterload, and contractility alterations were performed. LVAD speed changes were significantly more pronounced in all load interventions with the CSW control strategy (preload: P < 0.001 vs. CS and P = 0.004 vs. WR; afterload: P < 0.001 vs. CS and P < 0.001 vs. WR; contractility: P < 0.001 vs. CS and P < 0.001 vs. WR). However, a significant difference in systemic flow only became evident in the experiments upon afterload increase ( P < 0.001 vs. CS and P = 0.004 vs. WR). An implementation of an evolved version of the CSW control strategy that dispenses with invasively measured parameters might be feasible for clinical use.

Towards technically controlled bioreactor maturation of tissue-engineered heart valves.

Bioreactors are important tools for the pre-conditioning of tissue-engineered heart valves. The current state of the art mostly provides for timed, physical and biochemical stimulation in the bioreactor systems according to standard protocols (SOP). However, this does not meet to the individual biological variability of living tissue-engineered constructs. To achieve this, it is necessary to implement (i) sensory systems that detect the actual status of the implant and (ii) controllable bioreactor systems that allow patient-individualized pre-conditioning. During the maturation process, a pulsatile transvalvular flow of culture medium is generated within the bioreactor. For the improvement of this conditioning procedure, the relationship between the mechanical and biochemical stimuli and the corresponding tissue response has to be analyzed by performing reproducible and comparable experiments. In this work, a technological framework for maturation experiments of tissue-engineered heart valves in a pulsating bioreactor is introduced. The aim is the development of a bioreactor system that allows for continuous control and documentation of the conditioning process to increase reproducibility and comparability of experiments. This includes hardware components, a communication structure and software including online user communication and supervision. Preliminary experiments were performed with a tissue-engineered heart valve to evaluate the function of the new system. The results of the experiment proof the adequacy of the setup. Consequently, the concept is an important step for further research towards controlled maturation of tissue-engineered heart valves. The integration of molecular and histological sensor systems will be the next important step towards a fully automated, self-controlled preconditioning system.

Comparison of augmented reality and cutting guide technology in assisted harvesting of iliac crest grafts - A cadaver study.

BACKGROUND: Harvesting vascularized bone grafts with computer-assisted surgery represents the gold standard for mandibular reconstruction. However, current augmented reality (AR) approaches are limited to invasive marker fixation. This trial compared a markerless AR-guided real-time navigation with virtually planned and 3D printed cutting guides for harvesting iliac crest grafts. MATERIAL AND METHODS: Two commonly used iliac crest transplant configurations were virtually planned on 10 cadaver hips. Transplant harvest was performed with AR guidance and cutting guide technology. The harvested transplants were digitalized using cone beam CT. Deviations of angulation, distance and volume between the executed and planned osteotomies were measured. RESULTS: Both AR and cutting guides accurately rendered the virtually planned transplant volume. However, the cumulative osteotomy plane angulation differed significantly (p = 0.018) between AR (14.99 ± 11.69°) and the cutting guides (8.49 ± 5.42°). The cumulative osteotomy plane distance showed that AR-guided navigation had lower accuracy (2.65 ± 3.32 mm) than the cutting guides (1.47 ± 1.36 mm), although without significant difference. CONCLUSION: This study demonstrated the clinical usability of markerless AR-guided navigation for harvesting iliac crest grafts. Further improvement of accuracy rates might bring clinical implementation closer to reality.

Navigation of iliac crest graft harvest using markerless augmented reality and cutting guide technology: A pilot study.

BACKGROUND: Defects of the facial skeleton often require complex reconstruction with vascularized grafts. This trial elucidated the usability, visual perception and accuracy of a markerless augmented reality (AR)-guided navigation for harvesting iliac crest transplants. METHODS: Random CT scans were used to virtually plan two common transplant configurations on 10 iliac crest models, each printed four times. The transplants were harvested using projected AR and cutting guides. The duration and accuracies of the angulation, distance and volume between the planned and executed osteotomies were measured. RESULTS: AR was characterized by the efficient use of time and accurate rendition of preoperatively planned geometries. However, vertical osteotomies and complex anatomical settings displayed significant inferiority of AR guidance compared to cutting guides. CONCLUSIONS: This study demonstrated the usability of a markerless AR setup for harvesting iliac crest transplants. The visual perception and accuracy of the AR-guided osteotomies constituted remaining weaknesses against cutting guide technology.

Efficient Reject Options for Particle Filter Object Tracking in Medical Applications.

Reliable object tracking that is based on video data constitutes an important challenge in diverse areas, including, among others, assisted surgery. Particle filtering offers a state-of-the-art technology for this challenge. Becaise a particle filter is based on a probabilistic model, it provides explicit likelihood values; in theory, the question of whether an object is reliably tracked can be addressed based on these values, provided that the estimates are correct. In this contribution, we investigate the question of whether these likelihood values are suitable for deciding whether the tracked object has been lost. An immediate strategy uses a simple threshold value to reject settings with a likelihood that is too small. We show in an application from the medical domain-object tracking in assisted surgery in the domain of Robotic Osteotomies-that this simple threshold strategy does not provide a reliable reject option for object tracking, in particular if different settings are considered. However, it is possible to develop reliable and flexible machine learning models that predict a reject based on diverse quantities that are computed by the particle filter. Modeling the task in the form of a regression enables a flexible handling of different demands on the tracking accuracy; modeling the challenge as an ensemble of classification tasks yet surpasses the results, while offering the same flexibility.

Evaluation of foot position and orientation as manipulated variables to control external knee adduction moments in leg extension training.

BACKGROUND AND OBJECTIVE: Effective leg extension training at a leg press requires high forces, which need to be controlled to avoid training-induced damage. In order to avoid high external knee adduction moments, which are one reason for unphysiological loadings on knee joint structures, both training movements and the whole reaction force vector need to be observed. In this study, the applicability of lateral and medial changes in foot orientation and position as possible manipulated variables to control external knee adduction moments is investigated. As secondary parameters both the medio-lateral position of the center of pressure and the frontal-plane orientation of the reaction force vector are analyzed. METHODS: Knee adduction moments are estimated using a dynamic model of the musculoskeletal system together with the measured reaction force vector and the motion of the subject by solving the inverse kinematic and dynamic problem. Six different foot conditions with varying positions and orientations of the foot in a static leg press are evaluated and compared to a neutral foot position. RESULTS: Both lateral and medial wedges under the foot and medial and lateral shifts of the foot can influence external knee adduction moments in the presented study with six healthy subjects. Different effects are observed with the varying conditions: the pose of the leg is changed and the direction and center of pressure of the reaction force vector is influenced. Each effect results in a different direction or center of pressure of the reaction force vector. CONCLUSIONS: The results allow the conclusion that foot position and orientation can be used as manipulated variables in a control loop to actively control knee adduction moments in leg extension training.

A mock heart engineered with helical aramid fibers for in vitro cardiovascular device testing.

Mock heart circulation loops (MHCLs) serve as in-vitro platforms to investigate the physiological interaction between circulatory systems and cardiovascular devices. A mock heart (MH) engineered with silicone walls and helical aramid fibers, to mimic the complex contraction of a natural heart, has been developed to advance the MHCL previously developed in our group. A mock aorta with an anatomical shape enables the evaluation of a cannulation method for ventricular assist devices (VADs) and investigation of the usage of clinical measurement systems like pressure-volume catheters. Ventricle and aorta molds were produced based on MRI data and cast with silicone. Aramid fibers were layered in the silicone ventricle to reproduce ventricle torsion. A rotating hollow shaft was connected to the apex enabling the rotation of the MH and the connection of a VAD. Silicone wall thickness, aramid fiber angle and fiber pitch were varied to generate different MH models. All MH models were placed in a tank filled with variable amounts of water and air simulating the compliance. In this work, physiological ventricular torsion angles (15°-26°) and physiological pressure-volume loops were achieved. This MHCL can serve as a comprehensive testing platform for cardiovascular devices, such as artificial heart valves and cannulation of VADs.

Parametrization of an in-silico circulatory simulation by clinical datasets - towards prediction of ventricular function following assist device implantation.

BACKGROUND: Left ventricular assist device (LVAD) therapy has revolutionized the way end stage heart failure is treated today. Analysis of LVAD interaction with the whole cardiovascular system and its biological feedback loops is often conducted by means of computer models. Generating real time pressure volume loops (PV-loops) in patients, not using conductance catheters but routine diagnostics to feed an in-silico model could help to predict postoperative complications. METHODS: Routinely obtained hemodynamic measurements to evaluate myocardial function prior to LVAD implantation like pressure readings in the aorta, the left atrium and the left ventricle and simultaneous three-dimensional (3D) echocardiography recordings were assessed to parametrize a reduced computational model of the cardiovascular system. An automatic parameter identification procedure has been developed. RESULTS: The results constitute a patient-individual computational simulation model. An exemplary in-silico study focusing on the effect of different ventricular assist device (VAD) speeds has been conducted. Results allow for estimation of the resulting hemodynamic parameters and changes of the PV-loops. CONCLUSION: The model improves understanding and prediction of the interaction between pump and ventricles. Future modifications in exporting and merging routinely assessed real time hemodynamic patient data are necessary to investigate various clinical and pathological conditions of LVAD recipients.

Benefits of object-oriented models and ModeliChart: modern tools and methods for the interdisciplinary research on smart biomedical technology.

Computational models of biophysical systems generally constitute an essential component in the realization of smart biomedical technological applications. Typically, the development process of such models is characterized by a great extent of collaboration between different interdisciplinary parties. Furthermore, due to the fact that many underlying mechanisms and the necessary degree of abstraction of biophysical system models are unknown beforehand, the steps of the development process of the application are iteratively repeated when the model is refined. This paper presents some methods and tools to facilitate the development process. First, the principle of object-oriented (OO) modeling is presented and the advantages over classical signal-oriented modeling are emphasized. Second, our self-developed simulation tool ModeliChart is presented. ModeliChart was designed specifically for clinical users and allows independently performing in silico studies in real time including intuitive interaction with the model. Furthermore, ModeliChart is capable of interacting with hardware such as sensors and actuators. Finally, it is presented how optimal control methods in combination with OO models can be used to realize clinically motivated control applications. All methods presented are illustrated on an exemplary clinically oriented use case of the artificial perfusion of the systemic circulation.

Design of a right ventricular mock circulation loop as a test bench for right ventricular assist devices.

Right heart failure (RHF), e.g. due to pulmonary hypertension (PH), is a serious health issue with growing occurrence and high mortality rate. Limited efficacy of medication in advanced stages of the disease constitutes the need for mechanical circulatory support of the right ventricle (RV). An essential contribution to the process of developing right ventricular assist devices (RVADs) is the in vitro test bench, which simulates the hemodynamic behavior of the native circulatory system. To model healthy and diseased arterial-pulmonary hemodynamics in adults (mild and severe PH and RHF), a right heart mock circulation loop (MCL) was developed. Incorporating an anatomically shaped silicone RV and a silicone atrium, it not only enables investigations of hemodynamic values but also suction events or the handling of minimal invasive RVADs in an anatomical test environment. Ventricular pressure-volume loops of all simulated conditions as well as pressure and volume waveforms were recorded and compared to literature data. In an exemplary test, an RVAD was connected to the apex to further test the feasibility of studying such devices with the developed MCL. In conclusion, the hemodynamic behavior of the native system was well reproduced by the developed MCL, which is a useful basis for future RVAD tests.

Comparison of novel physiological load-adaptive control strategies for ventricular assist devices.

Terminal heart failure (HF) is the most prevalent cause of death in the Western world and the implantation of a left ventricular assist device (LVAD) has become the gold standard therapy today. Most of the actually implanted devices are driven at a constant speed (CS) regardless of the patient's physiological demand. A new physiological controller [power ratio (PR) controller], which keeps a constant ratio between LVAD power and left ventricular power, a previous concept [preload responsive speed (PRS) controller], which adds a variable LVAD power to reach a defined stroke work, and a CS controller were compared with an unimpaired ventricle in a full heart computer simulation model. The effects of changes in preload, afterload and left ventricular contractility are displayed by global hemodynamics and ventricular pressure-volume loops. Both physiological controllers demonstrated the desired load dependency, whereas the PR controller exceeded the PRS controller in response to an increased load and contractility. Response was inferior when preload or contractility was decreased. Thus, the PR controller might lead to an increased exercise tolerance of the patient. Additional studies are required to evaluate the controllers in vivo.

Modeling and simulation of the cardiovascular system: a review of applications, methods, and potentials.

Proper function of the cardiovascular system is indispensible to human survival. However, this system is dominated by complex interactions between different physiological processes and control mechanisms. A structured analysis and a mathematical description of this system can provide more insight, and a computer-based simulation of dynamic processes in the cardiovascular system could be applied in numerous tasks. This article gives a review of different approaches to cardio-circulatory modeling and discusses methodological aspects and fields of application for several classes of models.