Niels J. B. Driessen

Tissue Engineering of Human Heart Valve Leaflets: A Novel Bioreactor for a Strain-Based Conditioning Approach

Current mechanical conditioning approaches for heart valve tissue engineering concentrate on mimicking the opening and closing behavior of the leaflets, either or not in combination with tissue straining. This study describes a novel approach by mimicking only the diastolic phase of the cardiac cycle, resulting in tissue straining. A novel, yet simplified, bioreactor system was developed for this purpose by applying a dynamic pressure difference over a closed tissue engineered valve, thereby inducing dynamic strains within the leaflets. Besides the use of dynamic strains, the developing leaflet tissues were exposed to prestrain induced by the use of a stented geometry. To demonstrate the feasibility of this strain-based conditioning approach, human heart valve leaflets were engineered and their mechanial behavior evaluated. The actual dynamic strain magnitude in the leaflets over time was estimated using numerical analyses. Preliminary results showed superior tissue formation and non-linear tissue-like mechanical properties in the strained valves when compared to non-loaded tissue strips. In conclusion, the strain-based conditioning approach, using both prestrain and dynamic strains, offers new possibilities for bioreactor design and optimization of tissue properties towards a tissue-engineered aortic human heart valve replacement.

Autologous Human Tissue-Engineered Heart Valves Prospects for Systemic Application

Background— Tissue engineering represents a promising approach for the development of living heart valve replacements. In vivo animal studies of tissue-engineered autologous heart valves have focused on pulmonary valve replacements, leaving the challenge to tissue engineer heart valves suitable for systemic application using human cells. Methods and Results— Tissue-engineered human heart valves were analyzed up to 4 weeks and conditioning using bioreactors was compared with static culturing. Tissue formation and mechanical properties increased with time and when using conditioning. Organization of the tissue, in terms of anisotropic properties, increased when conditioning was dynamic in nature. Exposure of the valves to physiological aortic valve flow demonstrated proper opening motion. Closure dynamics were suboptimal, most likely caused by the lower degree of anisotropy when compared with native aortic valve leaflets. Conclusions— This study presents autologous tissue-engineered heart valves based on human saphenous vein cells and a rapid degrading synthetic scaffold. Tissue properties and mechanical behavior might allow for use as living aortic valve replacements.

A structural constitutive model for collagenous cardiovascular tissues incorporating the angular fiber distribution

Accurate constitutive models are required to gain further insight into the mechanical behavior of cardiovascular tissues. In this study, a structural constitutive framework for cardiovascular tissues is introduced that accounts for the angular distribution of collagen fibers. To demonstrate its capabilities, the model is applied to study the biaxial behavior of the arterial wall and the aortic valve. The pressure-radius relationships of the arterial wall accurately describe experimentally observed sigma-shaped curves. In addition, the nonlinear and anisotropic mechanical properties of the aortic valve can be analyzed with the proposed model. We expect that the current model offers strong possibilities to further investigate the complex mechanical behavior of cardiovascular tissues, including their response to mechanical stimuli.

Remodelling of the angular collagen fiber distribution in cardiovascular tissues

Understanding collagen fiber remodelling is desired to optimize the mechanical conditioning protocols in tissue-engineering of load-bearing cardiovascular structures. Mathematical models offer strong possibilities to gain insight into the mechanisms and mechanical stimuli involved in these remodelling processes. In this study, a framework is proposed to investigate remodelling of angular collagen fiber distribution in cardiovascular tissues. A structurally based model for collagenous cardiovascular tissues is extended with remodelling laws for the collagen architecture, and the model is subsequently applied to the arterial wall and aortic valve. For the arterial wall, the model predicts the presence of two helically arranged families of collagen fibers. A branching, diverging hammock-type fiber architecture is predicted for the aortic valve. It is expected that the proposed model may be of great potential for the design of improved tissue engineering protocols and may give further insight into the pathophysiology of cardiovascular diseases.

Improved Prediction of the Collagen Fiber Architecture in the Aortic Heart Valve

Living tissues show an adaptive response to mechanical loading by changing their internal structure and morphology. Understanding this response is essential for successful tissue engineering of load-bearing structures, such as the aortic valve. In this study, mechanically induced remodeling of the collagen architecture in the aortic valve was investigated. It was hypothesized that, in uniaxially loaded regions, the fibers aligned with the tensile principal stretch direction. For biaxial loading conditions, on the other hand, it was assumed that the collagen fibers aligned with directions situated between the principal stretch directions. This hypothesis has already been applied successfully to study collagen remodeling in arteries. The predicted fiber architecture represented a branching network and resembled the macroscopically visible collagen bundles in the native leaflet. In addition, the complex biaxial mechanical behavior of the native valve could be simulated qualitatively with the predicted fiber directions. The results of the present model might be used to gain further insight into the response of tissue engineered constructs during mechanical conditioning.

Stress related collagen ultrastructure in human aortic valves : implications for tissue engineering

Understanding the response of tissue structures to mechanical stress is crucial for optimization of mechanical conditioning protocols in the field of heart valve tissue engineering. In heart valve tissue, it is unclear to what extent mechanical loading affects the collagen fibril morphology. To determine if local stress affects the collagen fibril morphology, in terms of fibril diameter, its distribution, and the fibril density, this was investigated in adult native human aortic valve leaflets. Transmission electron microscopy images of collagen fibrils were analyzed at three locations: the commissures, the belly, and the fixed edge of the leaflets. Subsequently, the mechanical behavior of human aortic valves was used in a computational model to predict the stress distribution in the valve leaflet during the diastolic phase of the cardiac cycle. The local stresses at the three locations were related to the collagen fibril morphology. The fibril diameter and density varied significantly between the measured locations, and appeared inversely related. The average fibril diameter increased from the fixed edge, to the belly, and to the commissures of the leaflets, while fibril density decreased. Interestingly, these differences corresponded well with the level of stress at the locations. The presented data showed that large tissue stress is associated with greater average fibril diameter, lower fibril density, and wider fibril size distribution compared with low stress locations in the leaflets. The findings here provide insight in the effect of mechanical loading on the collagen ultrastructure, and are valuable to improve conditioning protocols for tissue engineering.

A microstructurally motivated model of the mechanical behavior of tissue engineered blood vessels.

Mechanical models have potential to guide the development and use of engineered blood vessels as well as other engineered tissues. This paper presents a microstructurally motivated, pseudoelastic, mechanical model of the biaxial mechanics of engineered vessels in the physiologic pressure range. The model incorporates experimentally measured densities and alignments of engineered collagen. Specifically, these microstructural and associated mechanical inputs were measured directly from engineered blood vessels that were cultured over periods of 5–7.5xa0weeks. To the best of our knowledge, this is the first successful application of either a phenomenological or a microstructurally motivated mechanical model to engineered vascular tissues. Model development revealed the need to use novel theoretical configurations to describe the strain history of engineered vessels. The constitutive equations developed herein suggested that collagen remodeled between 5 and 7.5xa0weeks during a 7.5-week culture period. This remodeling led to strain energies for collagen that differed with alignment, which likely resulted from undulations that varied with alignment. Finally, biaxial data emphasized that axial extensions increase stresses in engineered vessels in the physiologic pressure range, thereby providing a guideline for surgical use: engineered vessels should be implanted at appropriate axial extension to minimize adverse stress responses.

Non-Destructive and Non-Invasive Assessment of Mechanical Properties in Heart Valve Tissue Engineering

Despite recent progress, mechanical properties of tissue engineered heart valves still lack mechanical strength compared to native aortic valves [1]. Although cyclic tissue straining in bioreactor systems is known to enhance tissue formation [2], specific optimal loading protocols have not yet been defined. To get a better insight in the effects of mechanical loading on tissue development, mechanical behavior of tissue constructs should be monitored and controlled during culture. However, currently used methods for mechanical characterization (e.g. tensile tests, indentation tests) are destructive and can therefore only be performed at the end stage of tissue culture. An experimental-numerical approach was previously proposed by which leaflet deformation was assessed during culture in a bioreactor system, real-time and non-invasively [3]. Further development of this approach now enables a non-invasive and non-destructive assessment of mechanical properties of engineered heart valve leaflets.Copyright

Inverse Mechanical Characterization of Tissue Engineered Heart Valves

Over the last few years, research interest in tissue engineering as an alternative for current treatment and replacement strategies for cardiovascular and heart valve diseases has significantly increased. In vitro mechanical conditioning is an essential tool for engineering strong implantable tissues [1]. Detailed knowledge of the mechanical properties of the native tissue as well as the properties of the developing engineered constructs is vital for a better understanding and control of the mechanical conditioning process. The nonlinear and anisotropic behavior of soft tissues puts high demands on their mechanical characterization. Current standards in mechanical testing of soft tissues include (multiaxial) tensile testing and indentation tests. Uniaxial tensile tests do not provide sufficient information for characterizing the full anisotropic material behavior, while biaxial tensile tests are difficult to perform, and boundary effects limit the test region to a small central portion of the tissue. In addition, characterization of the local tissue properties from a tensile test is non-trivial. Indentation tests may be used to overcome some of these limitations. Indentation tests are easy to perform and when indenter size is small relative to the tissue dimensions, local characterization is possible. We have demonstrated that by recording deformation gradients and indentation force during a spherical indentation test the anisotropic mechanical behavior of engineered cardiovascular constructs can be characterized [2]. In the current study this combined numerical-experimental approach is used on Tissue Engineered Heart Valves (TEHV).Copyright

Stress Dependent Collagen Fibril Diameter Distribution in Human Aortic Valves

The mechanical properties of collagenous tissues are known to depend on a wide variety of factors, such as the type of tissue and the composition of its extracellular matrix. Relating mechanical roles to individual matrix components in such a complex system is difficult, if not impossible. However, as collagen is the main load bearing component in connective tissues, the relation between collagen and tissue biomechanics has been studied extensively in various types of tissues. The type of collagen, the amount and type of inter- and intramolecular covalent cross-links and collagen fibril morphology are involved in the tissues mechanical behavior (Beekman et al., 1997; Parry et al., 1978; Avery and Bailey, 2005). From literature it is known that the the collagen fibril diameter distribution can be directly related to the mechanical properties of the tissue. In particular, the diameter distribution of collagen fibrils is largely determined by the tissues requirement for tensile strength and elasticity (Parry et al., 1978).Copyright