M Mehdi Khoshgoftar

Mechanical stimulation to stimulate formation of a physiological collagen architecture in tissue-engineered cartilage; a numerical study

The load-bearing capacity of todays tissue-engineered (TE) cartilage is insufficient. The arcade-like collagen network in native cartilage plays an important role in its load-bearing properties. Inducing the formation of such collagen architecture in engineered cartilage can, therefore, enhance mechanical properties of TE cartilage. Considering the well-defined relationship between tensile strains and collagen alignment in the literature, we assume that cues for inducing this orientation should come from mechanical loading. In this study, strain fields prescribed by loading conditions of unconfined compression, sliding indentation and a novel loading regime of compression–sliding indentation are numerically evaluated to assess the probability that these would trigger a physiological collagen architecture. Results suggest that sliding indentation is likely to stimulate the formation of an appropriate superficial zone with parallel fibres. Adding lateral compression may stimulate the formation of a deep zone with perpendicularly aligned fibres. These insights may be used to improve loading conditions for cartilage tissue engineering.

The effect of tissue-engineered cartilage biomechanical and biochemical properties on its post-implantation mechanical behavior

The insufficient load-bearing capacity of today’s tissue-engineered (TE) cartilage limits its clinical application. Focus has been on engineering cartilage with enhanced mechanical stiffness by reproducing native biochemical compositions. More recently, depth dependency of the biochemical content and the collagen network architecture has gained interest. However, it is unknown whether the mechanical performance of TE cartilage would benefit more from higher content of biochemical compositions or from achieving an appropriate collagen organization. Furthermore, the relative synthesis rate of collagen and proteoglycans during the TE process may affect implant performance. Such insights would assist tissue engineers to focus on those aspects that are most important. The aim of the present study is therefore to elucidate the relative importance of implant ground substance stiffness, collagen content, and collagen architecture of the implant, as well as the synthesis rate of the biochemical constituents for the post-implantation mechanical behavior of the implant. We approach this by computing the post-implantation mechanical conditions using a composition-based fibril-reinforced poro-viscoelastic swelling model of the medial tibia plateau. Results show that adverse implant composition and ultrastructure may lead to post-implantation excessive mechanical loads, with collagen orientation being the most critical variable. In addition, we predict that a faster synthesis rate of proteoglycans compared to that of collagen during TE culture may result in excessive loads on collagen fibers post-implantation. This indicates that even with similar final contents, constructs may behave differently depending on their development. Considering these aspects may help to engineer TE cartilage implants with improved survival rates.

Influence of tissue- and cell-scale extracellular matrix distribution on the mechanical properties of tissue-engineered cartilage

The insufficient load-bearing capacity of today’s tissue- engineered (TE) cartilage limits its clinical application. Generally, cartilage TE studies aim to increase the extracellular matrix (ECM) content, as this is thought to determine the load-bearing properties of the cartilage. However, there are apparent inconsistencies in the literature regarding the correlation between ECM content and mechanical properties of TE constructs. In addition to the amount of ECM, the spatial inhomogeneities in ECM distribution at the tissue scale as well as at the cell scale may affect the mechanical properties of TE cartilage. The relative importance of such structural inhomogeneities on mechanical behavior of TE cartilage is unknown. The aim of the present study was, therefore, to theoretically elucidate the influence of these inhomogeneities on the mechanical behavior of chondrocyte-agarose TE constructs. A validated non-linear fiber-reinforced poro-elastic swelling cartilage model that can accommodate for effects of collagen reinforcement and swelling by proteoglycans was used. At the tissue scale, ECM was gradually varied from predominantly localized in the periphery of the TE construct toward an ECM-rich inner core. The effect of these inhomogeneities in relation to the total amount of ECM was also evaluated. At the cell scale, ECM was gradually varied from localized in the pericellular area, toward equally distributed throughout the interterritorial area. Results from the tissue-scale model indicated that localization of ECM in either the construct periphery or in the inner core may reduce construct stiffness compared with that of constructs with homogeneous ECM. Such effects are more significant at high ECM amounts. At the cell scale, localization of ECM around the cells significantly reduced the overall stiffness, even at low ECM amounts. The compressive stiffness gradually increased when ECM distribution became more homogeneous and the osmotic swelling pressure in the interterritorial area increased. We conclude that for the same amount of ECM content in TE cartilage constructs, superior mechanical properties can be achieved with more homogeneous ECM distribution at both tissue and cell scale. Inhomogeneities at the cell scale are more important than those at the tissue scale.

Direct Noninvasive Measurement and Numerical Modeling of Depth-Dependent Strains in Layered Agarose Constructs

Biomechanical factors play an important role in the growth, regulation, and maintenance of engineered biomaterials and tissues. While physical factors (e.g. applied mechanical strain) can accelerate regeneration, and knowledge of tissue properties often guide the design of custom materials with tailored functionality, the distribution of mechanical quantities (e.g. strain) throughout native and repair tissues is largely unknown. Here, we directly quantify distributions of strain using noninvasive magnetic resonance imaging (MRI) throughout layered agarose constructs, a model system for articular cartilage regeneration. Bulk mechanical testing, giving both instantaneous and equilibrium moduli, was incapable of differentiating between the layered constructs with defined amounts of 2% and 4% agarose. In contrast, MRI revealed complex distributions of strain, with strain transfer to softer (2%) agarose regions, resulting in amplified magnitudes. Comparative studies using finite element simulations and mixture (biphasic) theory confirmed strain distributions in the layered agarose. The results indicate that strain transfer to soft regions is possible in vivo as the biomaterial and tissue changes during regeneration and maturity. It is also possible to modulate locally the strain field that is applied to construct-embedded cells (e.g. chondrocytes) using stratified agarose constructs.

Tissue- and Cell-Level Inhomogeneities Significantly Alter the Mechanical Behavior of Tissue-Engineered Cartilage

The insufficient load-bearing capacity of today’s tissue engineered (TE) cartilage is an important limiting factor for its clinical application. It is believed that the mechanical quality of TE cartilage constructs would be optimal if it had both a structure and composition resembling native cartilage. Cartilage TE studies therefore aim to reach extracellular matrix (ECM) content that resembles that of native tissue. However, the correlation between ECM content and mechanical properties of TE constructs is not unique and the correlation between matrix content and mechanical properties vary considerably.© 2012 ASME

Role of Collagen Content and Architecture in the Load Bearing Capabilities of Tissue-Engineered Cartilage

A promising treatment for damaged cartilage is to replace it with tissue-engineered (TE) cartilage. However, the insufficient load-bearing capacity of today’s TE cartilage is an important limiting factor in its clinical application. In native cartilage, collagen fibers resist tension and proteoglycans (PG’s) attract water through osmotic pressure and resist its flow, which allows cartilage to withstand high compressive forces. One of the main challenges for tissue engineering of mechanically stable cartilage is therefore to find the cues to create an engineered tissue with an ultrastructure similar to that of native tissue. Currently, it is possible to tissue engineer cartilage with almost native PG content but collagen reaches only 1/4 of the native content [1]. Furthermore, the specific depth dependent arcade-like organization of collagen in native cartilage (i.e. vertical fibers in the deep zone and horizontal fibers in the superficial zone), which is optimized for distributing loads, has not been addressed in TE’d cartilage. However, the relative importance of matrix component content and collagen network architecture to the mechanical performance of TE cartilage is poorly understood, perhaps because this would require substantial effort on time consuming and labor-intensive experimental studies. The aim of this study is to explore if it is sufficient to produce a tissue with abundant proteoglycans and/or collagen, or whether reproducing the specific arcade-like collagen network in the implant is essential to develop sufficient load-bearing capacity, using a numerical approach.Copyright

Approach to Design Loading Protocols for Cartilage Tissue Engineering: Hypothesis, Experiment and Model

Tissue engineered cartilage has reached the level of maturity where the cells, either chondrocytes, BMSC’s or other cells are stimulated to produce a tissue of which the biochemical content qualitatively resembles that of native cartilage. Quantitatively, the proteoglycan content approaches that of native content in long term cultures, but to obtain native collagen fractions is still challenging. Engineered cartilage matrix is either homogeneously distributed, or shows gradual variation from the periphery to the center, caused by nutritional effects.Copyright