Ryan M. Castile

Biaxial analysis of synthetic scaffolds for hernia repair demonstrates variability in mechanical anisotropy, non-linearity and hysteresis.

BACKGROUND Over the past 60 years, the soft tissue repair market has grown to include over 50 types of hernia repair materials. Surgeons typically implant these materials in the orientation that provides maximum overlap of the mesh over the defect, with little regard for mechanical properties of the mesh material. If the characteristics of the meshes were better understood, an appropriate material could be identified for each patient, and meshes could be placed to optimize integration with neighboring tissue and avoid the mechanical mis-match that can lead to impaired graft fixation. The purpose of this study was to fully characterize and compare the mechanical properties of thirteen types of hernia repair materials via planar biaxial tensile testing. METHODS Equibiaxial (i.e., equal simultaneous loading in both directions) and strip biaxial (i.e., loading in one direction with the other direction held fixed) tests were utilized as physiologically relevant loading regimes. After applying a 0.1N pre-load on each arm, samples were subjected to equibiaxial cyclic loading using a triangular waveform to 2.5mm displacement on each arm at 0.1Hz for 10 cycles. Samples were then subjected to two strip biaxial tests (using the same cyclic loading protocol), where extension was applied along a single axis with the other axis held fixed. RESULTS The thirteen evaluated mesh types exhibited a wide range of mechanical properties. Some were nearly isotropic (C-QUR™, DUALMESH(®), PHYSIOMESH™, and PROCEED(®)), while others were highly anisotropic (Ventralight™ ST, Bard™ Mesh, and Bard™ Soft Mesh). Some displayed nearly linear behavior (Bard™ Mesh), while others were non-linear with a long toe region followed by a sharp rise in tension (INFINIT(®)). These materials are currently utilized in clinical settings as if they are uniform and interchangeable, and clearly this is not the case. The mechanical properties most advantageous for successful hernia repairs are currently only vaguely described in the clinical literature. The characteristics of the human abdominal wall must be extensively characterized to provide a thorough understanding of the tissue being reinforced/replaced by these meshes. A better understanding of these mechanical differences would enable matching of patient characteristics to a specific mesh with the properties best suited to that particular repair.

Differences in the Microstructural Properties of the Anteromedial and Posterolateral Bundles of the Anterior Cruciate Ligament

Background: Tissue properties of the anteromedial (AM) and posterolateral (PL) bundles of the anterior cruciate ligament (ACL) have not been previously characterized with real-time dynamic testing. The current study used a novel polarized light technique to measure the material and microstructural properties of the ACL. Hypothesis: The AM and PL bundles of the ACL have similar material and microstructural properties. Study Design: Controlled laboratory study. Methods: The AM and PL bundles were isolated from 16 human cadaveric ACLs (11 male, 5 female; average age, 41 years [range, 24-53 years]). Three samples from each bundle were loaded in uniaxial tension, and a custom-built polarized light imaging camera was used to quantify collagen fiber alignment in real time. A bilinear curve fit was applied to the stress-strain data of a quasistatic ramp-to-failure to quantify the moduli in the toe and linear regions. Fiber alignment was quantified at zero strain, the transition point of the bilinear fit, and in the linear portion of the stress-strain curve by computing the degree of linear polarization (DoLP) and angle of polarization (AoP), which are measures of the strength and direction of collagen alignment, respectively. Data were compared using t tests. Results: The AM bundle exhibited significantly larger toe-region (AM 7.2 MPa vs PL 4.2 MPa; P < .001) and linear-region moduli (AM 27.0 MPa vs PL 16.1 MPa; P = .017) compared with the PL bundle. Average DoLP values were similar at low strain but were significantly larger (ie, more uniform alignment) for the AM bundle in the linear region of the stress-strain curve (AM 0.22 vs PL 0.19; P = .036) compared with the PL bundle. The standard deviation AoP values was larger for the PL bundle at both transition (P = .041) and linear-region strain (P = .014), indicating more disperse orientation. Conclusion: Material and microstructural properties of the AM and PL bundles of the ACL differ during loading. The AM bundle possessed higher tissue modulus and failure stress, as well as more uniform fiber alignment under load. Clinical Relevance: These insights into native ligament microstructure can be used to assess graft options for ACL reconstruction and optimize surgical reconstruction techniques.

The suture pullout characteristics of human and porcine linea alba

There is a substantial prevalence of post-operative incisional hernia for both laparoscopic and laparotomy procedures, but there have been few attempts at quantifying abdominal wound closure methodology in the literature. One method to ascertain a more robust method of wound closure is the identification of the influence of suture placement parameters on suture pullout force. Current surgical practice involves a recommended bite depth and bite separation of 10mm, but the evidence base for this is not clear. In this paper, the suture pullout characteristics of both porcine and human linea alba were investigated to ascertain a suture placement protocol for surgical wound closure. Uniaxial suture pullout force testing on fresh frozen porcine and human linea alba samples was performed using standard materials testing machines. The influence of the number of suture loops, the bite depth and the bite separation of the sutures and the orientation of the sutures with respect to the principal fibre direction in the linea alba were assessed. Results showed a clearly identifiable relationship between pullout force of the suture, bite separation and bite depth, with low suture separation and high suture depth as optimal parameters for increasing pullout force. Resistance to pullout could be improved by as much as 290% when optimizing test conditions. Both human and porcine tissue were observed to exhibit very similar pullout force characteristics, corroborating the use of a porcine model for investigations into wound closure methodology. Orientation of suture application was also found to significantly affect the magnitude of suture pullout, with suturing applied longitudinally across a transverse defect resulting in higher pullout forces for small suture bite separations. Although further assessment in an environment more representative of in vivo conditions is required, these findings indicate that increasing the bite depth and reducing the bite separation with respect to the current surgical recommendations may reduce the risk of post-operative incisional hernia.

Uniaxial and biaxial tensile stress–stretch response of human linea alba

There are few studies on the stress-stretch behaviour of human linea alba, yet understanding the mechanics of this tissue is important for developing better methods of abdominal wound closure. Published data focuses mainly on porcine linea alba and for human tissue there are conflicting results and no bi-axial data available. This variability is likely due to challenges with the physical dimensions of the tissue and differences in experimental methodology. This study focussed on the tensile mechanical characterisation of the human linea alba using uniaxial and equi-load biaxial testing performed using image-based strain measurement methods. Thirteen freshly frozen human cadaveric abdominal walls were obtained and used to prepare 7 samples in both the transverse and longitudinal directions for uniaxial testing, and 13 square samples for bi-axial testing. The results showed significant anisotropy and for the equi-load biaxial tests the deformation was heavily biased in the longitudinal direction. In comparison with similar tests on porcine tissue from a previous study, it was found that the response of porcine linea alba to uniaxial loading is similar to that of human linea alba, with no statistically significant differences observed. Under biaxial loading human and porcine linea showed no statistical significance in the difference between their means in the transverse direction. However, a significant difference was observed in the longitudinal direction, and further study of the respective tissue structures is needed to better understand this result. These results provide the first data on the biaxial tensile properties of human linea alba and can aid in an improved assessment of wound closure mechanics.

Regional Variation in the Mechanical and Microstructural Properties of the Human Anterior Cruciate Ligament

Background: The anteromedial (AM) bundle of the anterior cruciate ligament (ACL) has a higher modulus and failure stress than does the posterolateral (PL) bundle. However, it is unknown how these properties vary within each bundle. Purpose: To quantify mechanical and microstructural properties of samples within ACL bundles to elucidate any regional variation across the ligament. We hypothesized that there are no differences within each bundle in contrast to cross-bundle variation. Study Design: Descriptive laboratory study. Methods: Sixteen human ACLs were dissected into AM and PL bundles. Three samples were taken from each bundle in an ordered sequence from AM (region 1 AM bundle) to PL (region 6 PL bundle). Each sample was tested in uniaxial tension, using quantitative polarized light imaging (QPLI) to quantify collagen fiber alignment. After preconditioning, samples were subjected to a stress-relaxation (SR) test followed by quasistatic ramp-to-failure (RF). Peak and equilibrium stress values were computed from the SR test and modulus quantified in the toe- and linear-regions of the RF. QPLI values describing collagen orientation (angle of polarization [AoP]) and strength of alignment (degree of linear polarization [DoLP]) were computed for the SR test and at points corresponding to the zero, transition point, and linear region of the RF. Results: Toe- and linear-region modulus values decreased from region 1 to 6. Slopes of regression lines increased for the average DoLP during RF, with significance at higher strains. The standard deviation of AoP values decreased during RF, indicating tighter distribution of orientation angles, with significant correlations at all points of the RF. During SR, stress values uniformly decreased but did not show significant linear regression by region. DoLP and AoP values changed slightly during SR and demonstrated significant linear variation by region at both peak and equilibrium points. Conclusion: Most microstructural and material properties evaluated in this study appear to follow a linear gradient across the ACL, rather than varying by bundle. Clinical Relevance: This AM-to-PL variation provides a more accurate description of functional tissue anatomy and can be used to assess and guide techniques of ACL reconstruction.

Multi-directional mechanical analysis of synthetic scaffolds for hernia repair

Hernias remain one of the most common ailments to affect men and women worldwide. Surgical mesh materials were first used to reinforce hernia defects during surgery in the late 1950s (Laker, n.d.). Today, there are well over 50 prosthetic meshes available for hernia repair (Brown and Finch, 2010; Bryan et al., 2014; Hope and El-hayek, 2014). With the multitude of available options, surgeons are faced with the challenging task of optimizing mesh selection for each patient. If the mechanics of the mesh are not compatible with the surrounding tissue, mismatch can occur, which can lead to complications such as mesh failure and/or hernia recurrence. Unfortunately, many aspects of synthetic mesh mechanics remain poorly described. Therefore, the purpose of this study was to provide a more complete mechanical analysis of a variety of commercially available prosthetic meshes for hernia repair, including evaluation of meshes in a variety of orientations. Twenty different meshes were subjected to biaxial tensile tests at both 90° and 45° orientations, and results were analyzed for relative strength, strain behavior, and anisotropy. Peak tension and strain values varied dramatically across all mesh types for all directions, ranging between 4.08 and 25.74N/cm and -5% to 10% strain. Anisotropy ratios for the evaluated meshes ranged from 0.33 to 1.89, demonstrating a wide range in relative direction-dependence of mesh mechanics. While further study of prosthetic meshes and better characterization of properties of the human abdominal wall are needed, results of this study provide valuable data that may aid clinicians in optimizing mesh selection for specific patients and repair conditions.

Persistent motion loss after free joint mobilization in a rat model of post-traumatic elbow contracture

BACKGROUND Post-traumatic joint contracture (PTJC) in the elbow is a challenging clinical problem due to the anatomical and biomechanical complexity of the elbow joint. METHODS We previously established an animal model to study elbow PTJC, wherein surgically induced soft tissue damage, followed by 6 weeks of unilateral immobilization in Long-Evans rats, led to stiffened and contracted joints that exhibited features similar to the human condition. In this study, after 6 weeks of immobilization, we remobilized the animal (ie, external bandage removed and free cage activity) for an additional 6 weeks, after which the limbs were evaluated mechanically and histologically. The objective of this study was to evaluate whether this decreased joint motion would persist after 6 weeks of free mobilization (FM). RESULTS After FM, flexion-extension demonstrated decreased total range of motion (ROM) and neutral zone length, and increased ROM midpoint for injured limbs compared with control and contralateral limbs. Specifically, after FM total ROM demonstrated a significant decrease of approximately 22% and 26% compared with control and contralateral limbs for injury I (anterior capsulotomy) and injury II (anterior capsulotomy with lateral collateral ligament transection), respectively. Histologic evaluation showed increased adhesion, fibrosis, and thickness of the capsule tissue in the injured limbs after FM compared with control and contralateral limbs, which is consistent with patterns previously reported in human tissue. CONCLUSION Even with FM, injured limbs in this model demonstrate persistent joint motion loss and histologic results similar to the human condition. Future work will use this animal model to investigate the mechanisms responsible for PTJC and responses to therapeutic intervention.

Pronation–Supination Motion Is Altered in a Rat Model of Post-Traumatic Elbow Contracture

The elbow joint is highly susceptible to joint contracture, and treating elbow contracture is a challenging clinical problem. Previously, we established an animal model to study elbow contracture that exhibited features similar to the human condition including persistent decreased range of motion (ROM) in flexion-extension and increased capsule thickness/adhesions. The objective of this study was to mechanically quantify pronation-supination in different injury models to determine if significant differences compared to control or contralateral persist long-term in our animal elbow contracture model. After surgically inducing soft tissue damage in the elbow, Injury I (anterior capsulotomy) and Injury II (anterior capsulotomy with lateral collateral ligament transection), limbs were immobilized for 6 weeks (immobilization (IM)). Animals were evaluated after the IM period or following an additional 6 weeks of free mobilization (FM). Total ROM for pronation-supination was significantly decreased compared to the uninjured contralateral limb for both IM and FM, although not different from control limbs. Specifically, for both IM and FM, total ROM for Injury I and Injury II was significantly decreased by ∼20% compared to contralateral. Correlations of measurements from flexion-extension and pronation-supination divulged that FM did not affect these motions in the same way, demonstrating that joint motions need to be studied/treated separately. Overall, injured limbs exhibited persistent motion loss in pronation-supination when comparing side-to-side differences, similar to human post-traumatic joint contracture. Future work will use this animal model to study how elbow periarticular soft tissues contribute to contracture.

Microstructural and Mechanical Properties of the Posterior Cruciate Ligament: A Comparison of the Anterolateral and Posteromedial Bundles

BACKGROUND The microstructural organization (collagen fiber alignment) of the posterior cruciate ligament (PCL), which likely corresponds with its functional properties, has only been described qualitatively in the literature, to our knowledge. The goal of this study was to quantify the tensile mechanical and microstructural properties of the PCL and compare these qualities between the anterolateral and posteromedial bundles. METHODS Twenty-two knee specimens from 13 donors (8 male and 5 female; mean age [and standard deviation] at the time of death, 43.0 ± 4.1 years; mean body mass index, 30.0 ± 6.7 kg/m2) were dissected to isolate the PCL, and each bundle was split into 3 regions. Mechanical testing of each regional sample consisted of preconditioning followed by a ramp-and-hold stress-relaxation test and a quasi-static ramp-to-failure test. Microstructural analysis was performed with use of a high-resolution, division-of-focal-plane polarization camera to evaluate the average direction of collagen orientation and the degree to which the collagen fibers were aligned in that direction. Results were compared between the anterolateral and posteromedial bundles and across the regions of each bundle. RESULTS The anterolateral and posteromedial bundles demonstrated largely equivalent mechanical and microstructural properties. Elastic moduli in the toe and linear regions were not different; however, the posteromedial bundle did show significantly more stress relaxation (p = 0.004). There were also few differences in microstructural properties between bundles, which again were seen only in stress relaxation. Comparing regions within each bundle, several mechanical and microstructural parameters showed significant relationships across the posteromedial bundle, following a gradient of decreasing strength and alignment from anterior to posterior. CONCLUSIONS The PCL has relatively homogenous microstructural and mechanical properties, with few differences between the anterolateral and posteromedial bundles. This finding suggests that distinct functions of the PCL bundles result primarily from size and anatomical location rather than from differences in these properties. CLINICAL RELEVANCE These properties of the PCL can be used to assess the utility of graft choices and operative techniques for PCL reconstruction and may partly explain limited differences in the outcomes of single-bundle compared with double-bundle reconstruction techniques for the PCL.