Human/robotic interaction: vision limits performance in simulated vitreoretinal surgery.

PURPOSE: Compare accuracy and precision in XYZ of stationary and dynamic tasks performed by surgeons with and without the use of a tele-operated robotic micromanipulator in a simulated vitreoretinal environment. The tasks were performed using a surgical microscope or while observing a video monitor. METHOD: Two experienced and two novice surgeons performed tracking and static tasks at a fixed depth with hand-held instruments on a Preceyes Surgical System R0.4. Visualization was through a standard microscope or a video display. The distances between the instrument tip and the targets (in µm) determined tracking errors in accuracy and precision. RESULTS: Using a microscope, dynamic or static accuracy and precision in XY (planar) movements were similar among test subjects. In Z (depth) movements, experience lead to more precision in both dynamic and static tasks (dynamic 35 ± 14 versus 60 ± 37 µm; static 27 ± 8 versus 36 ± 10 µm), and more accuracy in dynamic tasks (58 ± 35 versus 109 ± 79 µm). Robotic assistance improved both precision and accuracy in Z (1-3 ± 1 µm) in both groups. Using a video screen in combination with robotic assistance improved all performance measurements and reduced any differences due to experience. CONCLUSIONS: Robotics increases precision and accuracy, with greater benefit observed in less experienced surgeons. However, human control was a limiting factor in the achieved improvement. A major limitation was visualization of the target surface, in particular in depth. To maximize the benefit of robotic assistance, visualization must be optimized.

Degree of scaffold degradation influences collagen (re)orientation in engineered tissues.

Tissue engineering provides a promising tool for creating load-bearing cardiovascular tissues. Ideally, the neotissue produced by cells possesses native strength and anisotropy. By providing contact-guiding cues with microfibers, scaffold directionality can guide tissue organization. However, scaffolds transiently degrade, which may induce undesired tissue remodeling in response to applied strain. We hypothesize that in newly formed tissues, the collagen matrix does not yet provide contact guidance to the cells, and collagen orientation is altered via strain-induced remodeling. To test this hypothesis, we studied the influence of lipase-induced scaffold degradation on collagen (re)orientation at static constraint. Myofibroblasts were cultured in electrospun PCL-U4U anisotropic microfiber scaffolds, which were statically constrained perpendicular to the scaffold fibers. During 2 weeks of culture, neotissue formation aligned in the direction of the scaffold fibers, after which scaffolds were degraded to different degrees (12%, 27%, and 79% reduction in scaffold weight) and collagen (re)orientation was studied after one additional week of culturing. High degrees of scaffold degradation (79%) were associated with remodeling of the collagen toward the constraint direction, while collagen organization was maintained at low degrees of scaffold degradation. These results highlight the importance of slow scaffold degradation when aiming at maintaining collagen orientation.

Engineering fibrin-based tissue constructs from myofibroblasts and application of constraints and strain to induce cell and collagen reorganization.

Collagen content and organization in developing collagenous tissues can be influenced by local tissue strains and tissue constraint. Tissue engineers aim to use these principles to create tissues with predefined collagen architectures. A full understanding of the exact underlying processes of collagen remodeling to control the final tissue architecture, however, is lacking. In particular, little is known about the (re)orientation of collagen fibers in response to changes in tissue mechanical loading conditions. We developed an in vitro model system, consisting of biaxially-constrained myofibroblast-seeded fibrin constructs, to further elucidate collagen (re)orientation in response to i) reverting biaxial to uniaxial static loading conditions and ii) cyclic uniaxial loading of the biaxially-constrained constructs before and after a change in loading direction, with use of the Flexcell FX4000T loading device. Time-lapse confocal imaging is used to visualize collagen (re)orientation in a nondestructive manner. Cell and collagen organization in the constructs can be visualized in real-time, and an internal reference system allows us to relocate cells and collagen structures for time-lapse analysis. Various aspects of the model system can be adjusted, like cell source or use of healthy and diseased cells. Additives can be used to further elucidate mechanisms underlying collagen remodeling, by for example adding MMPs or blocking integrins. Shape and size of the construct can be easily adapted to specific needs, resulting in a highly tunable model system to study cell and collagen (re)organization.

Matrix production and organization by endothelial colony forming cells in mechanically strained engineered tissue constructs.

AIMS: Tissue engineering is an innovative method to restore cardiovascular tissue function by implanting either an in vitro cultured tissue or a degradable, mechanically functional scaffold that gradually transforms into a living neo-tissue by recruiting tissue forming cells at the site of implantation. Circulating endothelial colony forming cells (ECFCs) are capable of differentiating into endothelial cells as well as a mesenchymal ECM-producing phenotype, undergoing Endothelial-to-Mesenchymal-transition (EndoMT). We investigated the potential of ECFCs to produce and organize ECM under the influence of static and cyclic mechanical strain, as well as stimulation with transforming growth factor ß1 (TGFß1). METHODS AND RESULTS: A fibrin-based 3D tissue model was used to simulate neo-tissue formation. Extracellular matrix organization was monitored using confocal laser-scanning microscopy. ECFCs produced collagen and also elastin, but did not form an organized matrix, except when cultured with TGFß1 under static strain. Here, collagen was aligned more parallel to the strain direction, similar to Human Vena Saphena Cell-seeded controls. Priming ECFC with TGFß1 before exposing them to strain led to more homogenous matrix production. CONCLUSIONS: Biochemical and mechanical cues can induce extracellular matrix formation by ECFCs in tissue models that mimic early tissue formation. Our findings suggest that priming with bioactives may be required to optimize neo-tissue development with ECFCs and has important consequences for the timing of stimuli applied to scaffold designs for both in vitro and in situ cardiovascular tissue engineering. The results obtained with ECFCs differ from those obtained with other cell sources, such as vena saphena-derived myofibroblasts, underlining the need for experimental models like ours to test novel cell sources for cardiovascular tissue engineering.

Strain-induced collagen organization at the micro-level in fibrin-based engineered tissue constructs.

Full understanding of strain-induced collagen organization in complex tissue geometries to create tissues with predefined collagen architecture has not been achieved. This is mainly due to our limited knowledge of collagen remodeling in developing tissues. Here we investigate strain-induced collagen (re)organization in fibrin based engineered tissues using nondestructive time-lapse imaging. The tissues start from a biaxially constrained myofibroblast-populated fibrin gel and are used to study: (A) remodeling from a static equi-biaxial loading condition to static uniaxial loading; and (B) remodeling of a biaxially constrained tissue under uniaxial cyclic straining before and after a change in strain direction. Under static conditions, collagen oriented parallel to the direction of strain, whereas under cyclic conditions the orientation in the constrained tissue was perpendicular to the direction of strain. It is concluded that due to the biaxial constraints the uniaxially, cyclically strained cells can exert forces in two directions and strain shield themselves. A subsequent change in the direction of cyclic straining resulted in a rapid reorientation of collagen at the tissue surface. Reorientation was significantly slower in deeper tissue layers, where tissue remodeling was dominated by contact guidance provided by the endogenous matrix. These findings emphasize the relevance of achieving a functional collagen organization right from the start of tissue culture.

Age-related changes in material behavior of porcine mitral and aortic valves and correlation to matrix composition.

Recent studies showing significant changes in valvular matrix composition with age offer design criteria for age-specific tissue-engineered heart valves. However, knowledge regarding aging-related changes in valvular material properties is limited. Therefore, 6-week, 6-month, and 6-year-old porcine aortic valves (AV) and mitral valves (MV) were subjected to uniaxial tensile testing. In addition to standard material parameters, the radius of transition curvature (RTC) was measured to assess the acuteness of the transition region of the tension-strain curve. Radially, the MV had greater stiffness and a smaller RTC compared with the AV. Circumferentially, the center of the MV anterior leaflet (MVAC) had the highest stiffness (MVAC > AV > MV free edge [MVF]), greater stress relaxation (MVAC > MVF/AV), lowest extensibility (MVAC < AV < MVF), and smaller RTC compared with MVF (AV < MVAC < MVF). AV and MV radial strips had a larger RTC compared with circumferential strips. Aging elevated stiffness for MV and AV radial and circumferential strips, elevated stress relaxation in AV and MVF circumferential strips, and increased RTC for MV radial and MVF circumferential strips. In conclusion, there are significant age-related differences in the material properties of heart valves, which parallel differences in tissue composition and structure, likely impact valve function, and highlight the need for age-specific design goals for tissue-engineered heart valves.