Review paper: The importance of consideration of collagen cross-links in computational models of collagen-based tissues.

Collagen as the main protein in Extra Cellular Matrix (ECM) is the main load-bearing component of fibrous tissues. Nanostructure and architecture of collagen fibrils play an important role in mechanical behavior of these tissues. Extensive experimental and theoretical studies have so far been performed to capture these properties, but none of the current models realistically represent the complexity of network mechanics because still less is known about the collagen's inner structure and its effect on the mechanical properties of tissues. The goal of this review article is to emphasize the significance of cross-links in computational modeling of different collagen-based tissues, and to reveal the need for continuum models to consider cross-links properties to better reflect the mechanical behavior observed in experiments. In addition, this study outlines the limitations of current investigations and provides potential suggestions for the future work.

Computational design and evaluation of the mechanical and electrical behavior of a piezoelectric scaffold: a preclinical study.

Piezoelectric scaffolds have been recently developed to explore their potential to enhance the bone regeneration process using the concept of piezoelectricity, which also inherently occurs in bone. In addition to providing mechanical support during bone healing, with a suitable design, they are supposed to produce electrical signals that ought to favor the cell responses. In this study, using finite element analysis (FEA), a piezoelectric scaffold was designed with the aim of providing favorable ranges of mechanical and electrical signals when implanted in a large bone defect in a large animal model, so that it could inform future pre-clinical studies. A parametric analysis was then performed to evaluate the effect of the scaffold design parameters with regard to the piezoelectric behavior of the scaffold. The designed scaffold consisted of a porous strut-like structure with piezoelectric patches covering its free surfaces within the scaffold pores. The results showed that titanium or PCL for the scaffold and barium titanate (BT) for the piezoelectric patches are a promising material combination to generate favorable ranges of voltage, as reported in experimental studies. Furthermore, the analysis of variance showed the thickness of the piezoelectric patches to be the most influential geometrical parameter on the generation of electrical signals in the scaffold. This study shows the potential of computer tools for the optimization of scaffold designs and suggests that patches of piezoelectric material, attached to the scaffold surfaces, can deliver favorable ranges of electrical stimuli to the cells that might promote bone regeneration.

A review on computer modeling of bone piezoelectricity and its application to bone adaptation and regeneration.

Bone is a hierarchical, multiphasic and anisotropic structure which in addition possess piezoelectric properties. The generation of piezoelectricity in bone is a complex process which has been shown to play a key role both in bone adaptation and regeneration. In order to understand the complex biological, mechanical and electrical interactions that take place during these processes, several computer models have been developed and used to test hypothesis on potential mechanisms behind experimental observations. This paper aims to review the available literature on computer modeling of bone piezoelectricity and its application to bone adaptation and healing. We first provide a brief overview of the fundamentals of piezoelectricity and bone piezoelectric effects. We then review how these properties have been used in computational models of bone adaptation and electromechanical behaviour of bone. In addition, in the last section, we summarize current limitations and potential directions for future work.

Collagen fibril organization in chicken and porcine skeletal muscle perimysium under applied tension and compression.

The tension/compression asymmetry observed in the stress-stretch response of skeletal muscle is not well understood. The collagen network in the extracellular matrix (ECM) almost certainly plays a major role, but the details are unknown. This paper reports qualitatively and quantitatively on skeletal muscle ECM reorganization during applied deformation using confocal imaging of collagen through use of a fluorescently-tagged specific collagen binding protein (CNA35-EGFP) of porcine and chicken muscle samples under tensile and compressive deformation in both the fibre and cross-fibre directions. This reveals the overall three-dimensional structure of collagen in perimysium in planes perpendicular and parallel to the muscle fibres in both species. Furthermore, there is clear evidence of the reorganization of these structures under compression and tension applied in both the muscle fibre and cross-fibre directions. These observations improve our understanding of perimysium structure and response to three-dimensional deformations and are an important basis for constitutive models of passive skeletal muscle. Although overall behaviour was similar, some differences in perimysium structure were observed between chicken and porcine muscle tissue. Further work is required to better understand which structures are responsible for the tension and compression stress-strain asymmetry previously observed in the mechanical response of passive skeletal muscle.

A continuum model for tension-compression asymmetry in skeletal muscle.

Experiments on passive skeletal muscle on different species show a strong asymmetry in the observed tension-compression mechanical behavior. This asymmetry shows that the tension modulus is two orders of magnitude higher than the compression modulus. Until now, traditional analytical constitutive models have been unable to capture that strong asymmetry in anisotropic solids using the same material parameters. In this work we present a model which is able to accurately capture five experimental tests in chicken pectoralis muscle, including the observed tension-compression asymmetry. However, aspects of the anisotropy of the tissue are not captured by the model.

Visualisation of Collagen in fixed skeletal muscle tissue using fluorescently tagged Collagen binding protein CNA35.

Detection and visualisation of Collagen structure are important to understand the relationship between mechanical behaviour and microstructure in skeletal muscle since Collagen is the main structural protein in animal connective tissues, and is primarily responsible for their passive load-bearing properties. In the current study, the direct detection and visualization of Collagen using fluorescently tagged CNA35 binding protein (fused to EGFP or tdTomato) is reported for the first time on fixed skeletal muscle tissue. This Technical Note also establishes a working protocol by examining tissue preparation, dilution factor, exposure time etc. for sensitivity and specificity. Penetration of the binding protein into intact mature skeletal muscle was found to be very limited, but detection works well on tissue sections with higher sensitivity on wax embedded sections compared to frozen sections. CNA35 fused to tdTomato has a higher sensitivity than CNA35 fused to EGFP but both show specific detection. Best results were obtained with 15µm wax embedded sections, with blocking of non-specific binding in 1% BSA and antigen retrieval in Sodium Citrate. There was a play-off between dilution of the binding protein and time of incubation but both CNA35-tdTomato and CNA35-EGFP worked well with approximately 100µg/ml of purified protein with overnight incubation, while CNA35-tdTomato could be utilized at 5 fold less concentration. This approach can be applied to study the relationship between skeletal muscle micro-structure and to observe mechanical response to applied deformation. It can be used more broadly to detect Collagen in a variety of fixed tissues, useful for structure-functions studies, constitutive modelling, tissue engineering and assessment of muscle tissue pathologies.

The in vitro passive elastic response of chicken pectoralis muscle to applied tensile and compressive deformation.

The mechanics of passive skeletal muscle are important in impact biomechanics, surgical simulation, and rehabilitation engineering. Existing data from porcine tissue has shown a significant tension/compression asymmetry, which is not captured by current constitutive modelling approaches using a single set of material parameters, and an adequate explanation for this effect remains elusive. In this paper, the passive elastic deformation properties of chicken pectoralis muscle are assessed for the first time, to provide deformation data on a skeletal muscle which is very different to porcine tissue. Uniaxial, quasi-static compression and tensile tests were performed on fresh chicken pectoralis muscle in the fibre and cross-fibre directions, and at 45° to the fibre direction. Results show that chicken muscle elastic behaviour is nonlinear and anisotropic. The tensile stress-stretch response is two orders of magnitude larger than in compression for all directions tested, which reflects the tension/compression asymmetry previously observed in porcine tissue. In compression the tissue is stiffest in the cross-fibre direction. However, tensile deformation applied at 45° gives the stiffest response, and this is different to previous findings relating to porcine tissue. Chicken muscle tissue is most compliant in the fibre direction for both tensile and compressive applied deformation. Generally, a small percentage of fluid exudation was observed in the compressive samples. In the future these data will be combined with microstructural analysis to assess the architectural basis for the tension/compression asymmetry now observed in two different species of skeletal muscle.