Sensitivity of cartilage mechanical behaviour to spatial variations in material properties.

Articular cartilage tissue exhibits a spatial dependence in material properties that govern mechanical behaviour. A mathematical model of cartilage tissue under one dimensional confined compression testing is developed for normal tissue that takes account of these variations in material properties. Modifications to the model representative of a selection of mechanisms driving osteoarthritic cartilage are proposed, allowing application of the model to both physiological and pathophysiological, osteoarthritic tissue. Incorporating spatial variations into the model requires the specification of more parameters than are required in the absence of these variations. A global sensitivity analysis of these parameters is implemented to identify the dominant mechanisms of mechanical response, in normal and osteoarthritic cartilage tissue, to both static and dynamic loading. The most sensitive parameters differ between dynamic and static mechanics of the cartilage, and also differ between physiological and osteoarthritic pathophysiological cartilage. As a consequence changes in cartilage mechanics in response to alterations in cartilage structure are predicted to be contingent on the nature of loading and the health, or otherwise, of the cartilage. In particular the mechanical response of cartilage, especially deformation, is predicted to be much more sensitive to cartilage stiffness in the superficial zone given the onset of osteoarthritic changes to material properties, such as superficial zone increases in permeability and reductions in fixed charge. In turn this indicates that any degenerative changes in the stiffness associated with the superficial cartilage collagen mesh are amplified if other elements of osteoarthritic disease are present, which provides a suggested mechanism-based explanation for observations that the range of mechanical parameters representative of normal and osteoarthritic tissue can overlap substantially.

Anatomy of the Facial Glideplanes, Deep Plane Spaces and Ligaments: Implications for Surgical and Non-Surgical Lifting Procedures.

BACKGROUND: The soft tissue glideplanes of the face are functionally important and have a role in facial rejuvenation surgery. The aim of this study was to improve our understanding of soft tissue mobility of the face and its impact on the redraping of tissues involved in facelifting. The consequences of "no-release" and "extensive-release" lifting were analyzed to explain the difference in efficacy and potential longevity between these two contrasting philosophies. MATERIALS METHODS: Preliminary dissections and macro sectioning were followed by a definitive series of standardized layered dissections on fifty cadaver heads, along with histology, sheet plastination, and mechanical testing. RESULTS: The previously described spaces are potential surgical dissection planes deep to the superficial fascia layer. The classically described retaining ligaments are local reinforcements of a system of small retaining fibers (retinacula cutis and deep retinacula fibers) which provide support of the soft tissues of the face and neck against gravitational sagging while allowing certain mobility. This mobility is utilized when mobile tissues are lifted without surgical release. However, the process of dragging up these fibers results in a loss of their previous, anti-gravitational, supportive orientation. CONCLUSION: No-release lifting techniques, such as thread lifts and minimal-invasive facelifts, tighten "tissue laxity" with a change of the gravity-opposing tissue architecture, placing the weight of the flap solely on the fixation, which limits longevity of the lift. The alternative, to perform a full release with redraping, enables reattachment of the flap to a higher position, with preservation of the original deep fascial architecture with its antigravity orientation and natural mobility, conceivably improving the longevity of the lift.

Lifting the Anterior Midcheek and Nasolabial Fold: Introduction to the Melo Fat Pad Anatomy and Its Role in Longevity and Recurrence.

BACKGROUND: A limitation of current facelift techniques is the early postoperative reappearance of anterior midcheek laxity associated with recurrence of the nasolabial fold (NLF). OBJECTIVES: This study was undertaken to examine the regional anatomy of the anterior midcheek and NLF with a focus on explaining the early recurrence phenomenon and to explore the possibility of alternative surgical methods that prolong NLF correction. METHODS: Fifty cadaver heads were studied (16 embalmed, 34 fresh; mean age, 75 years). Following preliminary dissections and macrosectioning, a series of standardized layered dissections were performed, complemented by histology, sheet plastination, and microcomputed tomography. Mechanical testing of the melo fat pad (MFP) and skin was performed to gain insight on which structure is responsible for transmission of the lifting tension in a composite facelift procedure. RESULTS: Anatomic dissections, sheet plastination, and microcomputed tomography demonstrated the 3-dimensional architecture and borders of the MFP. Histology of a lifted midcheek demonstrated that a composite MFP lift causes a change in connective tissue organization from a hanging-down pattern into a pulled-upward pattern, suggesting traction on the skin. Mechanical testing confirmed that, in a composite lift, despite the sutures being placed directly into the deep aspect of the MFP, the lifting tension distal to the suture is transmitted through the skin and not through the MFP. CONCLUSIONS: The usual method of performing a composite midcheek lift results in the skin, and not the MFP itself, bearing the load of the nondissected tissues distal to the lifting suture. For this reason, early recurrence of the NLF occurs following skin relaxation in the postoperative period. Accordingly, specific surgical procedures for remodeling the MFP should be explored, possibly in combination with volume restoration of the fat and bone, for more lasting improvement of the NLF.

Modelling articular cartilage: the relative motion of two adjacent poroviscoelastic layers.

In skeletal joints two layers of adjacent cartilage are often in relative motion. The individual cartilage layers are often modelled as a poroviscoelastic material. To model the relative motion, noting the separation of scales between the pore level and the macroscale, a homogenization based on multiple scale asymptotic analysis has been used in this study to derive a macroscale model for the relative translation of two poroviscoelastic layers separated by a very thin layer of fluid. In particular the fluid layer thickness is essentially zero at the macroscale so that the two poroviscoelastic layers are effectively in contact and their interaction is captured in the derived model via a set of interfacial conditions, including a generalization of the Beavers-Joseph condition at the interface between a viscous fluid and a porous medium. In the simplifying context of a uniform geometry, constant fixed charge density, a Newtonian interstitial fluid and a viscoelastic scaffold, modelled via finite deformation theory, we present preliminary simulations that may be used to highlight predictions for how oscillatory relative movement of cartilage under load influences the peak force the cartilage experiences and the extent of the associated deformations. In addition to highlighting such cartilage mechanics, the systematic derivation of the macroscale models will enable the study of how nanoscale cartilage physics, such as the swelling pressure induced by fixed charges, manifests in cartilage mechanics at much higher lengthscales.

Embrittlement of collagen in early-stage human osteoarthritis.

Articular cartilage is a remarkable material with mechanical performance that surpasses engineering standards. Collagen, the most abundant protein in cartilage, plays an important role in this performance, and also in disease. Building on observations of network-level collagen changes at the earliest stages of osteoarthritis, this study explores the physical role of the collagen fibril in the disease process. Specifically, we focus on the material properties of collagen fibrils in the cartilage surface. Ten human tibial plateaus were characterised by atomic force microscopy (AFM) and Raman spectroscopy, with histological scoring used to define disease state. Measures of tropocollagen remained stable with disease progression, yet a marked mechanical change was observed. A slight stiffening coupled with a substantial decrease in loss tangent suggests a physical embrittlement caused by increased inter-molecular interactions.

The combined impact of tissue heterogeneity and fixed charge for models of cartilage: the one-dimensional biphasic swelling model revisited.

Articular cartilage is a complex, anisotropic, stratified tissue with remarkable resilience and mechanical properties. It has been subject to extensive modelling as a multiphase medium, with many recent studies examining the impact of increasing detail in the representation of this tissue's fine scale structure. However, further investigation of simple models with minimal constitutive relations can nonetheless inform our understanding at the foundations of soft tissue simulation. Here, we focus on the impact of heterogeneity with regard to the volume fractions of solid and fluid within the cartilage. Once swelling pressure due to cartilage fixed charge is also present, we demonstrate that the multiphase modelling framework is substantially more complicated, and thus investigate this complexity, especially in the simple setting of a confined compression experiment. Our findings highlight the importance of locally, and thus heterogeneously, approaching pore compaction for load bearing in cartilage models, while emphasising that such effects can be represented by simple constitutive relations. In addition, simulation predictions are observed for the sensitivity of stress and displacement in the cartilage to variations in the initial state of the cartilage and thus the details of experimental protocol, once the tissue is heterogeneous. These findings are for the simplest models given only heterogeneity in volume fractions and swelling pressure, further emphasising that the complex behaviours associated with the interaction of volume fraction heterogeneity and swelling pressure are likely to persist for simulations of cartilage representations with more fine-grained structural detail of the tissue.

A preliminary modeling investigation into the safe correction zone for high tibial osteotomy.

BACKGROUND: High tibial osteotomy (HTO) re-aligns the weight-bearing axis (WBA) of the lower limb. The surgery reduces medial load (reducing pain and slowing progression of cartilage damage) while avoiding overloading the lateral compartment. The optimal correction has not been established. This study investigated how different WBA re-alignments affected load distribution in the knee, to consider the optimal post-surgery re-alignment. METHODS: We collected motion analysis and seven Tesla MRI data from three healthy subjects, and combined this data to create sets of subject-specific finite element models (total=45 models). Each set of models simulated a range of potential post-HTO knee re-alignments. We shifted the WBA from its native alignment to between 40% and 80% medial-lateral tibial width (corresponding to 2.8°-3.1° varus and 8.5°-9.3° valgus), in three percent increments. We then compared stress/pressure distributions in the models. RESULTS: Correcting the WBA to 50% tibial width (0° varus-valgus) approximately halved medial compartment stresses, with minimal changes to lateral stress levels, but provided little margin for error in undercorrection. Correcting the WBA to a more commonly-used 62%-65% tibial width (3.4°-4.6° valgus) further reduced medial stresses but introduced the danger of damaging lateral compartment tissues. To balance optimal loading environment with that of the historical risk of under-correction, we propose a new target: WBA correction to 55% tibial width (1.7°-1.9° valgus), which anatomically represented the apex of the lateral tibial spine. CONCLUSIONS: Finite element models can successfully simulate a variety of HTO re-alignments. Correcting the WBA to 55% tibial width (1.7°-1.9° valgus) optimally distributes medial and lateral stresses/pressures.

Characterizing the macro and micro mechanical properties of scaffolds for rotator cuff repair.

BACKGROUND: Retearing after rotator cuff surgery is a major clinical problem. Numerous scaffolds are being used to try to reduce retear rates. However, few have demonstrated clinical efficacy. We hypothesize that this lack of efficacy is due to insufficient mechanical properties. Therefore, we compared the macro and nano/micro mechanical properties of 7 commercially available scaffolds to those of the human supraspinatus tendons, whose function they seek to restore. METHODS: The clinically approved scaffolds tested were X-Repair, LARS ligament, Poly-Tape, BioFiber, GraftJacket, Permacol, and Conexa. Fresh frozen cadaveric human supraspinatus tendon samples were used. Macro mechanical properties were determined through tensile testing and rheometry. Scanning probe microscopy and scanning electron microscopy were performed to assess properties of materials at the nano/microscale (morphology, Young modulus, loss tangent). RESULTS: None of the scaffolds tested adequately approximated both the macro and micro mechanical properties of human supraspinatus tendon. Macroscale mechanical properties were insufficient to restore load-bearing function. The best-performing scaffolds on the macroscale (X-Repair, LARS ligament) had poor nano/microscale properties. Scaffolds approximating tendon properties on the nano/microscale (BioFiber, biologic scaffolds) had poor macroscale properties. CONCLUSION: Existing scaffolds failed to adequately approximate the mechanical properties of human supraspinatus tendons. Combining the macroscopic mechanical properties of a synthetic scaffold with the micro mechanical properties of biologic scaffold could better achieve this goal. Future work should focus on advancing techniques to create new scaffolds with more desirable mechanical properties. This may help improve outcomes for rotator cuff surgery patients.

Effect of crosslinking in cartilage-like collagen microstructures.

The mechanical performance of biological tissues is underpinned by a complex and finely balanced structure. Central to this is collagen, the most abundant protein in our bodies, which plays a dominant role in the functioning of tissues, and also in disease. Based on the collagen meshwork of articular cartilage, we have developed a bottom-up spring-node model of collagen and examined the effect of fibril connectivity, implemented by crosslinking, on mechanical behaviour. Although changing individual crosslink stiffness within an order of magnitude had no significant effect on modelling predictions, the density of crosslinks in a meshwork had a substantial impact on its behaviour. Highly crosslinked meshworks maintained a 'normal' configuration under loading, with stronger resistance to deformation and improved recovery relative to sparsely crosslinked meshwork. Stress on individual fibrils, however, was higher in highly crosslinked meshworks. Meshworks with low numbers of crosslinks reconfigured to disease-like states upon deformation and recovery. The importance of collagen interconnectivity may provide insight into the role of ultrastructure and its mechanics in the initiation, and early stages, of diseases such as osteoarthritis.

Vitamin D receptor expression in human bone tissue and dose-dependent activation in resorbing osteoclasts.

The effects of vitamin D on osteoblast mineralization are well documented. Reports of the effects of vitamin D on osteoclasts, however, are conflicting, showing both inhibition and stimulation. Finding that resorbing osteoclasts in human bone express vitamin D receptor (VDR), we examined their response to different concentrations of 25-hydroxy vitamin D3 [25(OH)D3] (100 or 500 nmol·L-1) and 1,25-dihydroxy vitamin D3 [1,25(OH)2D3] (0.1 or 0.5 nmol·L-1) metabolites in cell cultures. Specifically, CD14+ monocytes were cultured in charcoal-stripped serum in the presence of receptor activator of nuclear factor kappa-B ligand (RANKL) and macrophage colony-stimulating factor (M-CSF). Tartrate-resistant acid phosphatase (TRAP) histochemical staining assays and dentine resorption analysis were used to identify the size and number of osteoclast cells, number of nuclei per cell and resorption activity. The expression of VDR was detected in human bone tissue (ex vivo) by immunohistochemistry and in vitro cell cultures by western blotting. Quantitative reverse transcription-PCR (qRT-PCR) was used to determine the level of expression of vitamin D-related genes in response to vitamin D metabolites. VDR-related genes during osteoclastogenesis, shown by qRT-PCR, was stimulated in response to 500 nmol·L-1 of 25(OH)D3 and 0.1-0.5 nmol·L-1 of 1,25(OH)2D3, upregulating cytochrome P450 family 27 subfamily B member 1 (CYP27B1) and cytochrome P450 family 24 subfamily A member 1 (CYP24A1). Osteoclast fusion transcripts transmembrane 7 subfamily member 4 (tm7sf4) and nuclear factor of activated T-cell cytoplasmic 1 (nfatc1) where downregulated in response to vitamin D metabolites. Osteoclast number and resorption activity were also increased. Both 25(OH)D3 and 1,25(OH)2D3 reduced osteoclast size and number when co-treated with RANKL and M-CSF. The evidence for VDR expression in resorbing osteoclasts in vivo and low-dose effects of 1,25(OH)2D3 on osteoclasts in vitro may therefore provide insight into the effects of clinical vitamin D treatments, further providing a counterpoint to the high-dose effects reported from in vitro experiments.

An overview of multiphase cartilage mechanical modelling and its role in understanding function and pathology.

There is a long history of mathematical and computational modelling with the objective of understanding the mechanisms governing cartilage׳s remarkable mechanical performance. Nonetheless, despite sophisticated modelling development, simulations of cartilage have consistently lagged behind structural knowledge and thus the relationship between structure and function in cartilage is not fully understood. However, in the most recent generation of studies, there is an emerging confluence between our structural knowledge and the structure represented in cartilage modelling. This raises the prospect of further refinement in our understanding of cartilage function and also the initiation of an engineering-level understanding for how structural degradation and ageing relates to cartilage dysfunction and pathology, as well as informing the potential design of prospective interventions. Aimed at researchers entering the field of cartilage modelling, we thus review the basic principles of cartilage models, discussing the underlying physics and assumptions in relatively simple settings, whilst presenting the derivation of relatively parsimonious multiphase cartilage models consistent with our discussions. We proceed to consider modern developments that start aligning the structure captured in the models with observed complexities. This emphasises the challenges associated with constitutive relations, boundary conditions, parameter estimation and validation in cartilage modelling programmes. Consequently, we further detail how both experimental interrogations and modelling developments can be utilised to investigate and reduce such difficulties before summarising how cartilage modelling initiatives may improve our understanding of cartilage ageing, pathology and intervention.

A constituent-based preprocessing approach for characterising cartilage using NIR absorbance measurements.

Near-infrared spectroscopy is a widely adopted technique for characterising biological tissues. The high dimensionality of spectral data, however, presents a major challenge for analysis. Here, we present a second-derivative Beer's law-based technique aimed at projecting spectral data onto a lower dimension feature space characterised by the constituents of the target tissue type. This is intended as a preprocessing step to provide a physically-based, low dimensionality input to predictive models. Testing the proposed technique on an experimental set of 145 bovine cartilage samples before and after enzymatic degradation, produced a clear visual separation between the normal and degraded groups. Reduced proteoglycan and collagen concentrations, and increased water concentrations were predicted by simple linear fitting following degradation (all [Formula: see text]). Classification accuracy using the Mahalanobis distance was [Formula: see text] between these groups.

Advancing musculoskeletal research with nanoscience.

Nanoscience has arrived. Biological applications of nanoscience are particularly prominent and can be useful in a range of disciplines. Advances in nanoscience are underpinning breakthroughs in biomedical research and are beginning to be adopted by the rheumatology and musculoskeletal science communities. Within these fields, nanoscience can be applied to imaging, drug delivery, implant development, regenerative medicine, and the characterization of nanoscale features of cells, matrices and biomaterials. Nanoscience and nanotechnology also provide means by which the interaction of cells with their environment can be studied, thereby increasing the understanding of disease and regenerative processes. Although its potential is clear, nanoscience research tends to be highly technical, generally targeting an audience of physicists, chemists, materials scientists and engineers, and is difficult for a general audience to follow. This Review aims to step back from the most technical aspects of nanoscience and provide a widely accessible view of how it can be applied to advance the field of rheumatology, with an emphasis on technologies that can have an immediate impact on rheumatology and musculoskeletal research.

Imaging and modeling collagen architecture from the nano to micro scale.

The collagen meshwork plays a central role in the functioning of a range of tissues including cartilage, tendon, arteries, skin, bone and ligament. Because of its importance in function, it is of considerable interest for studying development, disease and regeneration processes. Here, we have used second harmonic generation (SHG) to image human tissues on the hundreds of micron scale, and developed a numerical model to quantitatively interpret the images in terms of the underlying collagen structure on the tens to hundreds of nanometer scale. Focusing on osteoarthritic changes in cartilage, we have demonstrated that this combination of polarized SHG imaging and numerical modeling can estimate fibril diameter, filling fraction, orientation and bundling. This extends SHG microscopy from a qualitative to quantitative imaging technique, providing a label-free and non-destructive platform for characterizing the extracellular matrix that can expand our understanding of the structural mechanisms in disease.

Rough fibrils provide a toughening mechanism in biological fibers.

Spider silk is a fascinating natural composite material. Its combination of strength and toughness is unrivalled in nature, and as a result, it has gained considerable interest from the medical, physics, and materials communities. Most of this attention has focused on the one to tens of nanometer scale: predominantly the primary (peptide sequences) and secondary (ß sheets, helices, and amorphous domains) structure, with some insights into tertiary structure (the arrangement of these secondary structures) to describe the origins of the mechanical and biological performance. Starting with spider silk, and relating our findings to collagen fibrils, we describe toughening mechanisms at the hundreds of nanometer scale, namely, the fibril morphology and its consequences for mechanical behavior and the dissipation of energy. Under normal conditions, this morphology creates a nonslip fibril kinematics, restricting shearing between fibrils, yet allowing controlled local slipping under high shear stress, dissipating energy without bulk fracturing. This mechanism provides a relatively simple target for biomimicry and, thus, can potentially be used to increase fracture resistance in synthetic materials.

The critical role of water in spider silk and its consequence for protein mechanics.

Due to its remarkable mechanical and biological properties, there is considerable interest in understanding, and replicating, spider silk's stress-processing mechanisms and structure-function relationships. Here, we investigate the role of water in the nanoscale mechanics of the different regions in the spider silk fibre, and their relative contributions to stress processing. We propose that the inner core region, rich in spidroin II, retains water due to its inherent disorder, thereby providing a mechanism to dissipate energy as it breaks a sacrificial amide-water bond and gains order under strain, forming a stronger amide-amide bond. The spidroin I-rich outer core is more ordered under ambient conditions and is inherently stiffer and stronger, yet does not on its own provide high toughness. The markedly different interactions of the two proteins with water, and their distribution across the fibre, produce a stiffness differential and provide a balance between stiffness, strength and toughness under ambient conditions. Under wet conditions, this balance is destroyed as the stiff outer core material reverts to the behaviour of the inner core.

Spider silk as a load bearing biomaterial: tailoring mechanical properties via structural modifications.

Spider silk shows great potential as a biomaterial: in addition to biocompatibility and biodegradability, its strength and toughness are greater than native biological fibres (e.g. collagen), with toughness exceeding that of synthetic fibres (e.g. nylon). Although the ultimate tensile strength and toughness at failure are unlikely to be limiting factors, its yield strain of 2% is insufficient, particularly for biomedical application because of the inability to mimic the complex ultrastructure of natural tissues with current tissue engineering approaches. To harness the full potential of spider silk as a biomaterial, it is therefore necessary to increase its yield strain. In this paper, we discuss the means by which the mechanical properties of spider silk, particularly the yield strain, can be optimized through structural modifications.

Ultrasound assessment of articular cartilage: analysis of the frequency profile of reflected signals from naturally and artificially degraded samples.

This article investigates in vitro the hypothesis that the frequency profile of ultrasound reflections may be used to characterize degradation and osteoarthritic progression in articular cartilage, irrespective of the effects of transducer orientation. To this end, ultrasound echoes were taken in the time domain from the articular surface and osteochondral junction of normal, collagen meshwork-disrupted, proteoglycan-depleted, and osteoarthritic samples, converted to the frequency domain by fast Fourier transform and analyzed. Our results show the significant effects of specific enzymatic degradation programs on the ultrasound frequency profile of reflections from the cartilage surface and osteochondral junction, and their manifestation in the tissue surrounding a focal osteoarthritic defect. Collagen meshwork disruption was most apparent in the profile of reflections from the articular surface, while proteoglycan depletion was most clearly observed in the reflections from the osteochondral junction. The reflected signals from the osteochondral junction may further contain information about the subchondral bone. From these results we proposed that the analysis of specific frequencies of reflected ultrasound signals has the potential to differentiate normal from degraded articular cartilage-on-bone, when the angle of incidence can be controlled within a +/-1.2 degrees limit. This encourages further research into the effects of progressive artificial degradation of the cartilage matrix and subchondral bone on the spectral profile to quantify the relationship between the frequency profile and the level of specific degradation in naturally degraded joints.

Indentation stiffness does not discriminate between normal and degraded articular cartilage.

BACKGROUND: Relative indentation characteristics are commonly used for distinguishing between normal healthy and degraded cartilage. The application of this parameter in surgical decision making and an appreciation of articular cartilage biomechanics has prompted us to hypothesise that it is difficult to define a reference stiffness to characterise normal articular cartilage. METHODS: This hypothesis is tested for validity by carrying out biomechanical indentation of articular cartilage samples that are characterised as visually normal and degraded relative to proteoglycan depletion and collagen disruption. Compressive loading was applied at known strain rates to visually normal, artificially degraded and naturally osteoarthritic articular cartilage and observing the trends of their stress-strain and stiffness characteristics. FINDINGS: While our results demonstrated a 25% depreciation in the stiffness of individual samples after proteoglycan depletion, they also showed that when compared to the stiffness of normal samples only 17% lie outside the range of the stress-strain behaviour of normal samples. INTERPRETATION: We conclude that the extent of the variability in the properties of normal samples, and the degree of overlap (81%) of the biomechanical properties of normal and degraded matrices demonstrate that indentation data cannot form an accurate basis for distinguishing normal from abnormal articular cartilage samples with consequences for the application of this mechanical process in the clinical environment.

A novel approach to the development of benchmarking parameters for characterizing cartilage health.

This article outlines the motivation and preliminary investigations into a novel method of characterizing cartilage health for potential in vivo application. Current in vivo indentation techniques, which primarily rely on stiffness measurements based on axial data, are unable to adequately distinguish between healthy and degraded tissue. The present in vitro study investigates the effects of controlled artificial degradation on the effective surface stretch, comparing the results with those obtained from the peripheral cartilage surrounding focal osteoarthritis. Results suggest that this technique is highly sensitive, showing a maximum range of 14% effective surface stretch in a normal joint compared with 42% for axial strain measurements. We further demonstrated that the technique can discriminate between degenerative changes and the intrinsic variations in cartilage properties across the normal joint. From these investigations we propose that the relationship between indentation and the in-plane strain field under the indenter can better distinguish degraded tissue than the currently used stiffness techniques.