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Background: Several tissues contribute to the onset and advancement of knee osteoarthritis (OA). One tissue type that is worthy of closer evaluation, particularly in the context of sex, is the infrapatellar fat pad (IFP). We previously demonstrated that removal of the IFP had short-term beneficial effects for a cohort of male Dunkin-Hartley guinea pigs. The present project was designed to elucidate the influence of IFP removal in females of this OA-prone strain. It was hypothesized that resection of the IFP would reduce the development of OA in knees of a rodent model predisposed to the disease., Methods: Female guinea pigs (n=16) were acquired at an age of 2.5 months. Surgical removal of the IFP and associated synovium complex (IFP/SC) was executed at 3 months of age. One knee had the IFP/SC resected; a comparable sham surgery was performed on the contralateral knee. All animals were subjected to voluntary enclosure monitoring and dynamic weight-bearing, as well as compulsory treadmill-based gait analysis monthly; baseline data was collected prior to surgery. Guinea pigs were euthanized at 7 months. Knees from eight animals were evaluated via histology, mRNA expression, and immunohistochemistry (IHC); knees from the remaining eight animals were allocated to microcomputed tomography (microCT), biomechanical analyses (whole joint testing and indentation relaxation testing), and atomic absorption spectroscopy (AAS)., Results: Fibrous connective tissue (FCT) replaced the IFP/SC. Mobility/gait data indicated that unilateral IFP/SC removal did not affect bilateral hindlimb movement. MicroCT demonstrated that osteophytes were not a significant feature of OA in this sex; however, trabecular thickness (TbTh) in medial femorae decreased in knees containing the FCT. Histopathology scores were predominantly influenced by changes in the lateral tibia, which demonstrated that histologic signs of OA were increased in knees containing the native IFP/SC versus those with the FCT. Similarly, indentation testing demonstrated higher instantaneous and equilibrium moduli in the lateral tibial articular cartilage of control knees with native IFPs. AAS of multiple tissue types associated with the knee revealed that zinc was the major trace element influenced by removal of the IFP/SC., Conclusions: Our data suggest that the IFP/SC is a significant component driving knee OA in female guinea pigs and that resection of this tissue prior to disease has short-term benefits. Specifically, the formation of the FCT in place of the native tissue resulted in decreased cartilage-related OA changes, as demonstrated by reduced Osteoarthritis Research Society International (OARSI) histology scores, as well as changes in transcript, protein, and cartilage indentation analyses. Importantly, this model provides evidence that sex needs to be considered when investigating responses and associated mechanisms seen with this intervention., Competing Interests: Conflicts of Interest: All authors have completed the ICMJE uniform disclosure form (available at https://atm.amegroups.com/article/view/10.21037/atm-23-1886/coif). The authors have no conflicts of interest to declare., (2024 Annals of Translational Medicine. All rights reserved.)

Background: The infrapatellar fat pad (IFP) is the largest adipose deposit in the knee; however, its contributions to the homeostasis of this organ remain undefined. To determine the influence of the IFP and its associated synovium (IFP/synovium complex or IFP/SC) on joint health, this study evaluated the progression of osteoarthritis (OA) following excision of this unit in a rodent model of naturally-occurring disease., Methods: Male Dunkin-Hartley guinea pigs (n=18) received surgical removal of the IFP in one knee at 3 months of age; contralateral knees received sham surgery as matched internal controls. Mobility and gait assessments were performed prior to IFP/SC removal and monthly thereafter. Animals were harvested at 7 months of age. Ten set of these knees were processed for microcomputed tomography (microCT), histopathology, transcript expression analyses, and immunohistochemistry (IHC); 8 sets of knees were dedicated to microCT and biomechanical testing (material properties of knee joints tissues and anterior drawer laxity)., Results: Fibrous connective tissue (FCT) developed in place of the native adipose depot. Gait demonstrated no significant differences between IFP/SC removal and contralateral hindlimbs. MicroCT OA scores were improved in knees containing the FCT. Quantitatively, IFP/SC-containing knees had more osteophyte development and increased trabecular volume bone mineral density (vBMD) in femora and tibiae. Histopathology confirmed maintenance of articular cartilage structure, proteoglycan content, and chondrocyte cellularity in FCT-containing knees. Transcript analyses revealed decreased expression of adipose-related molecules and select inflammatory mediators in FCTs compared to IFP/SCs. This was verified via IHC for two key inflammatory agents. The medial articular cartilage in knees with native IFP/SCs showed an increase in equilibrium modulus, which correlated with increased amounts of magnesium and phosphorus., Discussion/conclusion: Formation of the FCT resulted in reduced OA-associated changes in both bone and cartilage. This benefit may be associated with: a decrease in inflammatory mediators at transcript and protein levels; and/or improved biomechanical properties. Thus, the IFP/SC may play a role in the pathogenesis of knee OA in this strain, with removal prior to disease onset appearing to have short-term benefits., (© 2022. The Author(s).)

Meniscal root repairs are susceptible to unrecoverable loosening that may displace the meniscus from the initial position reduced during surgery. Despite this, the effects of a loosened meniscal root repair on knee mechanics are unknown. We hypothesized that anatomic root repairs without loosening would restore knee mechanics to the intact condition better than loosened anatomic root repairs, but that loosened repairs would restore mechanics better than untreated meniscal root tears. Finite element knee models were used to evaluate changes in cartilage and meniscus mechanics due to repair loosening. The mechanical response from loosened anatomic root repairs was compared to anatomic repairs without loosening and untreated root tears. All conditions were evaluated at three flexion angles, 0 deg, 30 deg, and 60 deg, and a compressive force of 1000 N to simulate return-to-activity loading. The two-simple suture method was represented within the models to simulate posteromedial meniscal root repairs and the loosening of repairs was derived from previous biomechanical experimental data. Loosening decreased hoop stresses throughout the meniscus, increased posterior extrusion, and shifted loading through the meniscus-cartilage region to the cartilage-cartilage region compared to the anatomic root repair without loosening. Despite differences between repairs and loosened repairs, the changes from loosened repairs more closely resembled the anatomic repair without loosening than the untreated root repair condition. Therefore, meniscal root repairs are susceptible to loosening that will prevent a successful initial repair from remaining in the intended position and will alter cartilage and meniscus mechanics, although repairs that loosen appear better than leaving tears untreated., (Copyright © 2022 by ASME.)

Previous studies have shown that reconstructive surgery alone following injury to the anterior cruciate ligament (ACL) does not prevent the development of post-traumatic osteoarthritis (PTOA). Poloxamer 188 (P188) has been shown to prevent cell death following trauma in both articular cartilage and meniscal tissue. This study aims to test the efficacy of single or multiple administrations of P188 in conjunction with reconstructive surgery to help prevent or delay the onset of the disease. Thirty skeletally mature rabbits underwent closed-joint trauma that resulted in ACL rupture and meniscal damage and were randomly assigned to one of four treatment groups with varying doses of P188. ACL reconstruction was then performed using an autograft from the semitendinosus tendon. Animals were euthanized 1-month following trauma, meniscal tissue was assessed for changes in morphology, mechanical properties, and proteoglycan content. Femurs and tibias were scanned using microcomputed tomography to determine changes in bone quality, architecture, and osteophyte formation. The medial meniscus experienced more damage and a decrease in the instantaneous modulus regardless of treatment group, while P188 treatment tended to limit degenerative changes in the lateral meniscus. Both lateral and medial menisci had documented decreases in the equilibrium modulus and inconsistent changes in proteoglycan content. Minimal changes were documented in the tibias and femurs, with the only significant change being the formation of osteophytes in both bones regardless of treatment group. The data suggest that P188 was able to limit some degenerative changes in the meniscus associated with PTOA and may warrant future studies., (Copyright © 2022 by ASME.)

Post-traumatic osteoarthritis (PTOA) is a debilitating disease that is a result of a breakdown of knee joint tissues following traumatic impact. The interplay of how these tissues influence each other has received little attention because of complex interactions. This study was designed to correlate the degeneration of the menisci, cartilage and subchondral bone following an acute traumatic event that resulted in anterior cruciate ligament (ACL) and medial meniscus tears. We used a well-defined impact injury animal model that ruptures the ACL and tears the menisci. Subsequently, the knee joints underwent ACL reconstruction and morphological analyses were performed on the menisci, cartilage and subchondral bone at 1-, 3- and 6-months following injury. The results showed that the morphological scores of the medial and lateral menisci worsened with time, as did the tibial plateau and femoral condyle articular cartilage scores. The medial meniscus was significantly correlated to the medial tibial subchondral bone at 1 month ( p = 0.01), and to the medial tibial cartilage at 3 months ( p = 0.04). There was only one significant correlation in the lateral hemijoint, i.e., the lateral tibial cartilage to the lateral tibial subchondral bone at 6 months ( p = 0.05). These data may suggest that, following trauma, the observed medial meniscal damage should be treated acutely by means other than a full or partial meniscectomy, since that procedure may have been the primary cause of degenerative changes in the underlying cartilage and subchondral bone. In addition to potentially treating meniscal damage differently, improvements could be made in optimizing treatment of acute knee trauma., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Haut Donahue, Narez, Powers, Dejardin, Wei and Haut.)

Purpose/aim: The knee menisci are vital for maintaining the stability of the joint, allowing for force distribution, and protecting the underlying articular cartilage during loading. Each meniscus is attached to the underlying bone via two ligamentous entheses composed of collagen fibers that are continuous throughout all four zones of the attachment: ligament, uncalcified fibrocartilage, calcified fibrocartilage, and subchondral bone. The collagen fibers of the meniscal entheses are important for proper functionality of the entheses, particularly in preventing meniscal extrusion which is a common hallmark of osteoarthritis. The goal of this work was to assess changes in collagen fiber orientation present in osteoarthritic knee joints., Materials and Methods: Entheses were harvested from patients undergoing total knee arthroplasties and prepared histological sections were stained with picrosirius red to identify collagen fiber angle and fiber deviation., Results: In the calcified fibrocartilage the collagen fibers of the lateral anterior enthesis inserted at significantly (p < 0.1) shallower angles, and the fiber deviation was significantly (p < 0.1) less compared to the lateral posterior enthesis. These differences in the calcified fibrocartilage may occur as an adaptation to loading regimes of the osteoarthritic joint. When compared to the collagen fiber orientation of healthy entheses, collagen fibers in osteoarthritic tissue inserted at shallower insertion angles and demonstrated higher levels of deviation., Conclusions: Changes to meniscal enthesis collagen fiber orientation with end stage osteoarthritis could offer an explanation for the change in functionality of diseased tissue and may contribute to meniscal extrusion and ultimately the degeneration of articular cartilage.

Surgical reconstruction of the torn ACL is performed to restore native contact mechanics. Drawbacks to traditional ACL repair techniques motivate the development of a tissue engineered ACL scaffold. Our group has developed a hierarchical electrospun polycaprolactone (PCL) scaffold that consists of rolled nanofiber bundles attached at each end with solvent-case blocks of PCL. The goal of this study was to compare ovine cadaver tibiofemoral contact mechanics after ACL reconstruction with the electrospun scaffold to a clinically applicable ACL reconstruction with a soft tissue graft and the ACL transected condition (ACLX). In the ACLX group and after ACL reconstruction with either the electrospun scaffold or soft tissue graft, pressure sensors were inserted under the menisci. Loads up to 890 N were applied at various flexion angles. The scaffold performed the best at restoring contact mechanics in the medial hemijoint to that of the native ACL. The scaffold was good at maintaining a medial-lateral balance of pressures as in the native joint whereas the ACLX shifted pressure off the lateral and on to the medial hemijoint. While the ACL scaffold didn't restore mechanics to that of the native condition, it improved contact mechanics compared to the standard graft replacement and ACLX condition., (© 2021. Biomedical Engineering Society.)

Despite reconstruction surgery to repair a torn anterior cruciate ligament (ACL), patients often still show signs of post-traumatic osteoarthritis (PTOA) years following the procedure. The goal of this study was to document changes in the meniscus and subchondral bone due to closed-joint impact and surgical reconstruction in a lapine model. Animals received insult to the joint followed by surgical reconstruction of the ACL and partial meniscectomy. Following euthanasia of the animals at 1, 3, and 6-months post-impact, meniscal tissue was assessed for changes in morphology, mechanical properties and proteoglycan content. Femurs and tibias were scanned via micro-computed tomography to determine changes in bone quality, morphometry, and formation of osteophytes. Both the lateral and medial menisci showed severe degradation and tearing at all-time points, with higher degree of degeneration being observed at 6-months. Decreases in both the instantaneous and equilibrium modulus were documented in both menisci. Minimal changes were found in bone quality and morphometry, with most change documented in the tibia. Bones from the reconstructed limbs showed large volumes of osteophyte formations, with an increase in volume over time. The initial changes that were representative of PTOA may have been limited to the meniscus, but at later time points consistent changes due to the disease were seen in both tissues. This study, which builds on a previous study by this laboratory, suggests that the addition of surgical reconstruction of the ACL to our model was not sufficient to prevent the development of PTOA., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Objective: To determine the efficacy of single and multiple administrations of Poloxamer 188 (P188) in saving meniscal cells following an injurious impact., Methods: Meniscal explants were harvested from both the lateral and medial menisci of Flemish Giant rabbits. After a 24-h incubation period, explants were subjected to 50% impact strain to simulate traumatic joint injury, and the explants were then placed in media with or without supplemented P188. Temporal administrations of P188 over a 14-day period were given based on one of 6 different treatments regimes. Over the 14-day period, explants were cyclically loaded to 10% strain at 1 Hz for 1 h per day, five days a week. Cell viability was assessed on day 14, with the remainder of the tissue being fixed to determine cell apoptosis levels and proteoglycan changes via histology., Results: The injurious impact proved to produce significant levels of cell death in meniscal explants. The ability of P188 to prevent cell death was not affected by the number of P188 doses (single versus multiple). P188 treatment proved to maintain cell viability levels comparable to those from unimpacted explants. There were no significant changes in cell apoptosis or proteoglycan coverage in the tissues over a 14-day period for any group, all treatment groups were statistically similar to the unimpacted explants., Conclusion: A single dose of P188 following impact is all that is necessary to inhibit cell death in the meniscus following a traumatic impact. Thus, orthopaedic surgeons may choose to administer P188 in addition to treating any other acute damage due to a traumatic load to the knee, such as anterior cruciate ligament rupture, although more in depth in vivo studies are necessary., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Nonanatomic placement of posteromedial meniscal root repairs alters knee mechanics; however, little is known about how the position and magnitude of misplacement affect knee mechanics. Finite element knee models were developed to assess changes in cartilage and meniscus mechanics for anatomic and various nonanatomic repairs with respect to intact. In total, 25 different repair locations were assessed at loads of 500 N and 1000 N. The two-simple-suture method was represented within the models to simulate posteromedial meniscal root repairs. Anatomic repairs nearly restored total contact area; however, meniscal hoop stress decreased, meniscal extrusion increased, and cartilage-cartilage contact area increased. Repairs positioned further posterior altered knee mechanics the most and repairs positioned further anterior restored knee mechanics for posteromedial root repairs. Despite this, repair tension increased with further anterior placement. Anterior placement of repairs results in more restorative contact mechanics than posterior placement; however, anterior placement also increased the risk of suture cut-out or failure following repairs. Anatomic placement of repairs remains the best option because of the risks involved with anterior placement; however, suture methods need to be improved to better restore the strength of repairs to that of the native insertion. Proper placement of repairs is important to consider with meniscal root repairs because misplacement may negatively affect cartilage and meniscus mechanics in patients., (Copyright © 2020 by ASME.)

Objective: Despite immense popularity of Jaipur foot as low cost prosthetic, not much work has been reported on its design for manufacturing standardization. Without manufacturing standardization, it cannot be mass produced using contemporary manufacturing technologies. The objective of this work is to carry out its computer aided design (CAD) followed by computer aided engineering (CAE) based on the material properties obtained from the previous work [1] of the authors. This may lead to the possible use of modern manufacturing processes for the Jaipur foot design. Design: After modelling using CAD tool including its organic surfaces, the designed foot was analysed using a CAE tool for balanced standing load conditions to determine maximum stresses and deformation in its various parts. The bending analysis was done to check the dorsiflexion movement so that the strained sections could be identified for more reliable and durable prosthetic foot. For the static load analysis, base of the foot was constrained and 300-500 N load was applied through the bolt whereas for bending, the part near the bolt was fixed and pressure was applied at junction of front foot and toes. Results: The results show that the maximum stress and deformation occur at the bolt, while the skin undergoes maximum strain. CAE analysis also proves the robustness of the Jaipur foot design and a well manufactured Jaipur foot as per standardized design should be able to withstand the real life conditions without failure. The CAD model is also used for FDM based printing for a nonfunctional prototype of Jaipur foot.Implications for rehabilitationThe results of this study will serve as an important guideline for further research regarding equivalent material replacement, material optimization and obtaining an optimized design after studying the foot for dynamic analysis.

Purpose: To determine whether meniscal root repairs recover from displacement due to rehabilitative loading., Methods: Transtibial pullout repairs of the posteromedial meniscal root were performed in 16 cadaveric ovine knees. Single- and double-tunnel repairs using the 2-simple suture technique were cyclically loaded in tension to 10,000 cycles, allowed to rest, and loaded in tension again. Paired differences in displacement with rest were recorded to evaluate recoverability. Displacement of repairs at cycles of interest was recorded, and the response of repairs to 10,000 cycles was assessed., Results: All outcomes were not significantly different between the single- and double-tunnel techniques; therefore, the results were pooled. The difference in displacement between the first cycle and the first cycle after rest was 1.59 ± 0.69 mm. Repair displacement did not reach an equilibrium within 10,000 cycles and instead resulted in a steady increase in displacement of 0.05 ± 0.02 mm per additional 1,000 cycles. Sutures macroscopically began to cut out of the meniscus in both single- and double-tunnel repairs., Conclusions: This study showed that significant, unrecoverable loosening from rehabilitative loading occurred in single- and double-tunnel meniscal root repairs. Root repairs also gradually displaced with continued loading instead of reaching an equilibrium displacement after 10,000 cycles. This progressive, unrecoverable loosening needs to be studied further to better understand the resultant impact on knee mechanics. In addition, the quality and quantity of meniscal root repair healing at the time of rehabilitation should be studied to determine how susceptible patients are to repair loosening., Clinical Relevance: Rehabilitative loading caused unrecoverable and progressive loosening of root repairs, showing the importance of healing before loading. Investigations on the effects of loosening on mechanics and the quality of repair healing at weight bearing are necessary to better understand the clinical implications., (Copyright © 2019 Arthroscopy Association of North America. Published by Elsevier Inc. All rights reserved.)

The anterior cruciate ligament (ACL) acts to stabilize the knee and prevent excessive motion of the tibia relative to the femur. Tears of the ACL are common and can result in pain and damage to surrounding tissues. Thus a torn ACL is often surgically replaced with an autograft or allograft material. Drawbacks to clinically available ACL grafts motivate the development of a tissue engineered ACL replacement. Our group has previously developed a polycaprolactone electrospun scaffold that mimics the hierarchical structure of the ACL. The goal of this study was to investigate the mechanical properties of the electrospun scaffold as an ACL replacement. Scaffold mechanical properties were assessed prior to implantation via stress relaxation and pull to failure testing. Following in vitro characterization, electrospun scaffolds and soft tissue grafts were implanted into ovine cadaver stifle joints as ACL replacements. Stifle joints with ACL replacements were tested via a simulated anterior drawer test as well as in situ stress relaxation and pull to failure tests and compared to stifle joints with the native ACL intact. Prior to implantation the scaffold matched the native ovine ACL well in the range of functional strains as evidenced by stress relaxation measures and the toe region stiffness. After implantation the scaffold was more similar to the native ACL than the soft tissue graft, particularly when it came to reducing joint laxity and matching stress relaxation measures. These results demonstrate that the electrospun scaffold has the potential to be a suitable material for ACL replacement. © 2018 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 37:421-430, 2019., (© 2018 Orthopaedic Research Society. Published by Wiley Periodicals, Inc.)

Synthesis of hydrogel networks capable of accurately replicating the biomechanical demands of musculoskeletal soft tissues continues to present a formidable materials science challenge. Current systems are hampered by combinations of limited moduli at biomechanically relevant strains, inefficiencies driven by undesirable hysteresis and permanent fatigue, and recovery dynamics too slow to accommodate rapid cycling prominent in most biomechanical loading profiles. Here, we report on a novel paradigm in hydrogel design based on prefabrication of an efficient nanoscale network architecture using the melt-state self-assembly of amphiphilic block copolymers. Rigorous characterization and mechanical testing reveal that swelling of these preformed networks produces hydrogels with physiologically relevant moduli and water compositions, negligible hysteresis, subsecond elastic recovery rates, and unprecedented resistance to fatigue over hundreds of thousands of compression cycles. Furthermore, by relying only on simple thermoplastic processing to form these nanostructured networks, the synthetic complexities common to most solution-based hydrogel fabrication strategies are completely avoided.

Objective: The purpose of this study was to examine effects of usage and demographics on damage to the Jaipur Foot prosthesis as well as the epidemiology and etiology of amputations performed at Santokba Durlabjhi Memorial Hospital (SDMH) in Jaipur, India., Design: Total time spent standing, total time spent wearing and total distance walked were compared against severity and location of damage to the prosthesis. Time between initial fitting and follow-up visit for damaged prosthetic was also considered in this analysis. A novel damage severity scale based on prosthesis functionality is presented along with a damage location legend., Results: Patients from 10 different states and two territories throughout India were included in the study. No main effects were found to be statistically significant in predicting severity or location of damage. Only the interaction between a patient's total time spent standing and their total time spent wearing the prosthesis as well as the interaction between a patient's total time spent standing and total distance walked was significant in predicting location of damage to the Jaipur Foot (p = .0327, p = .0278, respectively)., Conclusions: The lack of significant usage factor effect on damage severity or location could support previous findings that lack standardization in materials and manufacturing processes, which is the major drawback of the Jaipur Foot. Implications for Rehabilitation The Jaipur Foot is a safe, reliable and stable product as no abrupt breakage or sudden falls causing injury to the patient were noted. Hence, it is a safe rehabilitation device for lost limbs. The population can squat and sit cross-legged while wearing the prosthetic foot and it does not affect damage severity or location of damage, allowing for these activities to be performed while rehabilitating. The manufacturing of the foot needs to be standardized to improve life of foot. Total time spent standing, total time spent wearing and total distance walked were not predictive of severity or location of damage to the prosthesis, hence providing patient guidelines for activity during rehabilitation.

Pressure distribution of the native ovine knee meniscus was compared to a medial meniscectomy and three treatment conditions including a suture reattachment of the native tissue, an allograft, and a novel thermoplastic elastomer hydrogel (TPE) construct. The objective of this study was to assess the efficacy of a novel TPE hydrogel construct at restoring joint pressure and distribution. Limbs were loaded in uniaxial compression at 45°, 60°, and 75° flexion and from 0 to 181 kg. The medial meniscectomy decreased contact area by approximately 50% and doubled the mean and maximum pressure reading for the medial hemijoint. No treatment condition tested within this study was able to fully restore medial joint contact area and pressures to the native condition. A decrease in lateral contact area and increase in pressures with the meniscectomy was also seen; and to some degree, all reattachment and replacement conditions including the novel TPE hydrogel replacement helped to restore lateral pressures. Although the TPE construct did not perform as well as hoped in the medial compartment, it performed as well as, if not better, than the other reattachment and replacement options in the lateral. Further work is necessary to determine the best anchoring and attachment methods.

Clinical treatments of skeletal muscle weakness are hindered by a lack of an approach to evaluate individual muscle force. Intramuscular pressure (IMP) has shown a correlation to muscle force in vivo, but patient to patient and muscle to muscle variability results in difficulty of utilizing IMP to estimate muscle force. The goal of this work was to develop a finite element model of whole skeletal muscle that can predict IMP under passive and active conditions to further investigate the mechanisms of IMP variability. A previously validated hypervisco-poroelastic constitutive approach was modified to incorporate muscle activation through an inhomogeneous geometry. Model parameters were optimized to fit model stress to experimental data, and the resulting model fluid pressurization data were utilized for validation. Model fitting was excellent (root-mean-square error or RMSE <1.5 kPa for passive and active conditions), and IMP predictive capability was strong for both passive (RMSE 3.5 mmHg) and active (RMSE 10 mmHg at in vivo lengths) conditions. Additionally, model fluid pressure was affected by length under isometric conditions, as increases in stretch yielded decreases in fluid pressurization following a contraction, resulting from counteracting Poisson effects. Model pressure also varied spatially, with the highest gradients located near aponeuroses. These findings may explain variability of in vivo IMP measurements in the clinic, and thus help reduce this variability in future studies. Further development of this model to include isotonic contractions and muscle weakness would greatly benefit this work.

Hydrogels are a class of synthetic biomaterials composed of a polymer network that swells with water and as such they have both an elastic and viscous component making them ideal for soft tissue applications. This study characterizes the compressive, tensile, and shear properties of a thermoplastic elastomer (TPE) hydrogel and compares the results to published literature values for soft tissues such as articular cartilage, the knee meniscus, and intervertebral disc components. The results show the TPE hydrogel material is viscoelastic, strain rate dependent, has similar surface and bulk properties, displays minimal damping under dynamic load, and has tension-compression asymmetry. When compared to other soft tissues it has a comparable equilibrium compressive modulus of approximately 0.5MPa and shear modulus of 0.2MPa. With a tensile modulus of only 0.2MPa though, the TPE hydrogel is inferior in tension to most collagen based soft tissues. Additional steps may be necessary to reinforce the hydrogel system and increase tensile modulus depending on the desired soft tissue application. It can be concluded that this material could be a viable option for soft tissue replacements., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Understanding how human meniscal tissue responds to loading regimes mimetic of daily life as well as how it compares to larger animal models is critical in the development of a functionally accurate synthetic surrogate. Seven human and eight ovine cadaveric meniscal specimens were regionally sectioned into cylinders 5 mm in diameter and 3 mm thick along with 10 polystyrene-b-polyethylene oxide block copolymer-based thermoplastic elastomer (TPE) hydrogels. Samples were compressed to 12% strain at 1 Hz for 5000 cycles, unloaded for 24 h, and then retested. No differences were found within each group between test one and test two. Human and ovine tissue exhibited no regional dependency (p < 0.05). Human samples relaxed quicker than ovine tissue or the TPE hydrogel with modulus values at cycle 50 not significantly different from cycle 5000. Ovine menisci were found to be similar to human menisci in relaxation profile but had significantly higher modulus values (3.44 MPa instantaneous and 0.61 MPa after 5000 cycles compared with 1.97 and 0.11 MPa found for human tissue) and significantly different power law fit coefficients. The TPE hydrogel had an initial modulus of 0.58 MPa and experienced less than a 20% total relaxation over the 5000. Significant differences in the magnitude of compressive modulus between human and ovine menisci were observed, however the relaxation profiles were similar. Although statistically different than the native tissues, modulus values of the TPE hydrogel material were similar to those of the human and ovine menisci, making it a material worth further investigation for use as a synthetic replacement. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 2722-2728, 2017., (© 2017 Wiley Periodicals, Inc.)

The anterior cruciate ligament (ACL) of the knee is vital for proper joint function and is commonly ruptured during sports injuries or car accidents. Due to a lack of intrinsic healing capacity and drawbacks with allografts and autografts, there is a need for a tissue-engineered ACL replacement. Our group has previously used aligned sheets of electrospun polycaprolactone nanofibers to develop solid cylindrical bundles of longitudinally aligned nanofibers. We have shown that these nanofiber bundles support cell proliferation and elongation and the hierarchical structure and material properties are similar to the native human ACL. It is possible to combine multiple nanofiber bundles to create a scaffold that attempts to mimic the macroscale structure of the ACL. The goal of this work was to develop a hierarchical bioactive scaffold for ligament tissue engineering using connective tissue growth factor (CTGF)-conjugated nanofiber bundles and evaluate the behavior of mesenchymal stem cells (MSCs) on these scaffolds in vitro and in vivo. CTGF was immobilized onto the surface of individual nanofiber bundles or scaffolds consisting of multiple nanofiber bundles. The conjugation efficiency and the release of conjugated CTGF were assessed using X-ray photoelectron spectroscopy, assays, and immunofluorescence staining. Scaffolds were seeded with MSCs and maintained in vitro for 7 days (individual nanofiber bundles), in vitro for 21 days (scaled-up scaffolds of 20 nanofiber bundles), or in vivo for 6 weeks (small scaffolds of 4 nanofiber bundles), and ligament-specific tissue formation was assessed in comparison to non-CTGF-conjugated control scaffolds. Results showed that CTGF conjugation encouraged cell proliferation and ligament-specific tissue formation in vitro and in vivo. The results suggest that hierarchical electrospun nanofiber bundles conjugated with CTGF are a scalable and bioactive scaffold for ACL tissue engineering.

Proteoglycans have vital biochemical and biomechanical functions. Their proteolytic degradation results in loss of these functions. We have previously reported nonprotein proteoglycan-mimetic graft copolymers that stabilize and deliver growth factors and are not subject to proteases. Here we expand our investigation of these proteoglycan mimics by also investigating their effects on hydrogel mechanical properties. Four polysaccharide side chains, chondroitin sulfate, heparin, dextran, and dextran sulfate, are each grafted to a hyaluronan backbone. The polysaccharides and graft copolymers are added to agarose hydrogels. Cyclic compression and stress relaxation tests reveal how the addition of the polysaccharides and graft copolymers influence hydrogel modulus. Cells encapsulated in agarose hydrogels containing chondroitin sulfate and the chondroitin sulfate graft copolymer have decreased cell viability and metabolic activity compared to cells in unmodified agarose hydrogels. These multifunctional additives can be used to improve both the biochemistry and biomechanics of materials, warranting further optimization to overcome the potentially negative effects these may have on cell viability and activity.

The passive properties of skeletal muscle are often overlooked in muscle studies, yet they play a key role in tissue function in vivo. Studies analyzing and modeling muscle passive properties, while not uncommon, have never investigated the role of fluid content within the tissue. Additionally, intramuscular pressure (IMP) has been shown to correlate with muscle force in vivo and could be used to predict muscle force in the clinic. In this study, a novel model of skeletal muscle was developed and validated to predict both muscle stress and IMP under passive conditions for the New Zealand White Rabbit tibialis anterior. This model is the first to include fluid content within the tissue and uses whole muscle geometry. A nonlinear optimization scheme was highly effective at fitting model stress output to experimental stress data (normalized mean square error or NMSE fit value of 0.993) and validation showed very good agreement to experimental data (NMSE fit values of 0.955 and 0.860 for IMP and stress, respectively). While future work to include muscle activation would broaden the physiological application of this model, the passive implementation could be used to guide surgeries where passive muscle is stretched.

Finite element models of skeletal muscle typically ignore the biphasic nature of the tissue, associating any time dependence with a viscoelastic formulation. In this study, direct experimental measurement of permeability was conducted as a function of specimen orientation and strain. A finite element model was developed to identify how various permeability formulations affect compressive response of the tissue. Experimental and modeling results suggest the assumption of a constant, isotropic permeability is appropriate. A viscoelastic only model differed considerably from a visco-poroelastic model, suggesting the latter is more appropriate for compressive studies.

Background: Meniscal root tears cause menisci and their insertions to inadequately distribute loads and potentially leave underlying articular cartilage unprotected. Untreated meniscal root tears are becoming increasingly recognized to induce joint degradation; however, little information is known about anterior meniscal root tears and how they affect joint tissue., Purpose: To observe the early degenerative changes within the synovial fluid, menisci, tibial articular cartilage, and subchondral bone after arthroscopic creation of untreated anterior meniscal root tears., Study Design: Controlled laboratory study., Methods: Anterolateral meniscal root tears were created in 1 knee joint of 5 adult Flemish Giant rabbits, and anteromedial meniscal root tears were created in 4 additional rabbits. The contralateral limbs were used as nonoperated controls. The animals were euthanized at 8 weeks postoperatively; synovial fluid was aspirated, and tissue samples of menisci and tibial articular cartilage were collected and processed for multiple analyses to detect signs of early degeneration., Results: Significant changes were found within the synovial fluid, meniscal tissue, and tibial subchondral bone of the knees with anterior meniscal root tears when compared with controls. There were no significant changes identified in the tibial articular cartilage when comparing the tear groups with controls., Conclusion: This study demonstrated early degenerative changes within the synovial fluid, menisci, and tibial subchondral bone when leaving anterior meniscal root tears untreated for 8 weeks. The results suggest that meniscal tissue presents measurable, degenerative changes prior to changes within the articular cartilage after anterior meniscal root tears. Anterior destabilization of the meniscus arthroscopically may lead to measurable degenerative changes and be useful for future in vivo natural history and animal repair studies., Clinical Relevance: The present study is the first to investigate various tissue changes after anterior meniscal root tears of both the medial and lateral menisci. The results from this study suggest that degenerative changes occur within the synovial fluid, meniscus, and tibial subchondral bone prior to any measurable changes to the tibial articular cartilage. Further studies should expand on this study to evaluate how these components continue to progress when left untreated for long periods., Competing Interests: One or more of the authors has declared the following potential conflict of interest or source of funding: R.F.L. receives royalties from Arthrex, Ossur, and Smith & Nephew; is a paid consultant for Arthrex, Ossur, and Smith & Nephew; and receives research support from Arthrex, Smith & Nephew, Ossur, and Linvatec. Funding for this project was provided by the Steadman Philippon Research Institute in Vail, Colorado.

Objective: The objective of this study was to monitor the progression of joint damage in two animal models of knee joint trauma using two non-invasive, clinically available imaging modalities., Methods: A 3-T clinical magnet and micro-computed tomography (μCT) was used to document changes immediately following injury (acute) and post-injury (chronic) at time points of 4, 8, or 12 weeks. Joint damage was recorded at dissection and compared to the chronic magnetic resonance imaging (MRI) record. Fifteen Flemish Giant rabbits were subjected to a single tibiofemoral compressive impact (ACLF), and 18 underwent a combination of anterior cruciate ligament (ACL) and meniscal transection (mACLT)., Results: All ACLF animals experienced ACL rupture, and 13 also experienced acute meniscal damage. All ACLF and mACLT animals showed meniscal and articular cartilage damages at dissection. Meniscal damage was documented as early as 4 weeks and worsened in 87% of the ACLF animals and 71% of the mACLT animals. Acute cartilage damage also developed further and increased in occurrence with time in both models. A progressive decrease in bone quantity and quality was documented in both models. The MRI data closely aligned with dissection notes suggesting this clinical tool may be a non-invasive method for documenting joint damage in lapine models of knee joint trauma., Conclusions: The study investigates the acute to chronic progression of meniscal and cartilage damage at various time points, and chronic changes to the underlying bone in two models of posttraumatic osteoarthritis (PTOA), and highlights the dependency of the model on the location, type, and progression of damage over time., (Copyright © 2016 Osteoarthritis Research Society International. Published by Elsevier Ltd. All rights reserved.)

A range of bone regeneration strategies, from growth factor delivery and/or mesenchymal stem cell (MSC) transplantation to endochondral tissue engineering, have been developed in recent years. Despite their tremendous promise, the clinical translation and future use of many of these strategies is being hampered by concerns such as off target effects associated with growth factor delivery. Therefore the overall objective of this study was to investigate the influence of alpha-tricalcium phosphate (α-TCP) nanoparticle delivery into MSCs using an amphipathic cell penetrating peptide RALA, on osteogenesis in vitro and both intramembranous and endochondral bone formation in vivo. RALA complexed α-TCP nanoparticle delivery to MSCs resulted in an increased expression of bone morphogenetic protein-2 (BMP-2) and an upregulation in a number of key osteogenic genes. When α-TCP stimulated MSCs were encapsulated into alginate hydrogels, enhanced mineralization of the engineered construct was observed over a 28 day culture period. Furthermore, the in vivo bone forming potential of RALA complexed α-TCP nanoparticle delivery to MSCs was found to be comparable to growth factor delivery. Recognizing the potential and limitations associated with endochondral bone tissue engineering strategies, we then sought to explore how α-TCP nanoparticle delivery to MSCs influences early mineralization of engineered cartilage templates in vitro and their subsequent ossification in vivo. Despite accelerating mineralization of engineered cartilage templates in vitro, RALA complexed α-TCP nanoparticle delivery did not enhance endochondral bone formation in vivo. Therefore the potential of RALA complexed α-TCP nanoparticle delivery appears to be as an alternative to growth factor delivery as a single stage strategy for promoting bone generation.

Background: The anterolateral meniscal root (ALMR) has been reported to intricately insert underneath the tibial insertion of the anterior cruciate ligament (ACL). Previous studies have begun to evaluate the relationship between the insertion areas and the risk of iatrogenic injuries; however, the overlap of the insertions has yet to be quantified in the sagittal and coronal planes., Purpose: To investigate the insertions of the human tibial ACL and ALMR using scanning electron microscopy (SEM) and to quantify the overlap of the ALMR insertion in the coronal and sagittal planes., Study Design: Descriptive laboratory study., Methods: Ten cadaveric knees were dissected to isolate the tibial ACL and ALMR insertions. Specimens were prepared and imaged in the coronal and sagittal planes. After imaging, fiber directions were examined to identify the insertions and used to calculate the percentage of the ACL that overlaps with the ALMR instead of inserting into bone., Results: Four-phase insertion fibers of the tibial ACL were identified directly medial to the ALMR insertion as they attached onto the tibial plateau. The mean percentage of ACL fibers overlapping the ALMR insertion instead of inserting into subchondral bone in the coronal and sagittal planes was 41.0% ± 8.9% and 53.9% ± 4.3%, respectively. The percentage of insertion overlap in the sagittal plane was significantly higher than in the coronal plane ( P = .02)., Conclusion: This study is the first to quantify the ACL insertion overlap of the ALMR insertion in the coronal and sagittal planes, which supplements previous literature on the insertion area overlap and iatrogenic injuries of the ALMR insertion. Future studies should determine how much damage to the ALMR insertion is acceptable to properly restore ACL function without increasing the risk for tears of the ALMR., Clinical Relevance: Overlap of the insertion areas on the tibial plateau has been previously reported; however, the results of this study demonstrate significant overlap of the insertions superior to the insertion sites on the tibial plateau as well. These findings need to be considered when positioning for tibial tunnel creation in ACL reconstruction to avoid damage to the ALMR insertion.

Purpose: To further elucidate the direct and indirect fibre insertion morphology within the human ACL femoral attachment using scanning electron microscopy and determine where in the footprint each fibre type predominates. The hypothesis was that direct fibre attachment would be found centrally in the insertion site, while indirect fibre attachment would be found posteriorly adjacent to the posterior articular cartilage., Methods: Ten cadaveric knees were dissected to preserve and isolate the entirety of the femoral insertion of the ACL. Specimens were then prepared and evaluated with scanning electron microscopy to determine insertional fibre morphology and location., Results: The entirety of the fan-like projection of the ACL attachment site lay posterior to the lateral intercondylar ridge. In all specimens, a four-phase architecture, consistent with previous descriptions of direct fibres, was found in the centre of the femoral attachment site. The posterior margin of the ACL attachment attached directly adjacent to the posterior articular cartilage with some fibres coursing into it. The posterior portion of the ACL insertion had a two-phase insertion, consistent with previous descriptions of indirect fibres. The transition from the ligament fibres to bone had less interdigitations, and the interdigitations were significantly smaller (p < 0.001) compared to the transition in the direct fibre area. The interdigitations of the direct fibres were 387 ± 81 μm (range 282-515 μm) wide, while the interdigitations of indirect fibres measured 228 ± 75 μm (range 89-331 μm)., Conclusions: The centre of the ACL femoral attachment consisted of a direct fibre structure, while the posterior portion had an indirect fibre structure. These results support previous animal studies reporting that the centre of the ACL femoral insertion was comprised of the strongest reported fibre type. Clinically, the femoral ACL reconstruction tunnel should be oriented to cover the entirety of the central direct ACL fibres and may need to be customized based on graft type and the fixation device used during surgery.

Unlabelled: Bighorn sheep (Ovis canadensis) routinely experience violent impacts to the head as part of intraspecific fighting. Dynamic 3D finite element models of the skull and horns of a male bighorn sheep were developed to gain an understanding of the roles that the horn and bone materials and structure play in absorbing the impact that occurs during ramming. The geometry and volume mesh of the model were derived from CT scan images. The models included the horn, bony horn core, and bone of the skull. The horn core fills a portion of the hollow horn and consists of a thin cortical bone shell filled with foam-like trabecular bone. Two modified models were also created: one with the distal half of the horn length removed to assess the effects of the tapered spiral geometry of the horn, and one with the internal trabecular bone material of the horn core removed. The trabecular bone material stored three times more strain energy during impact than the horn material in the intact model. Removing half of the horn length had the effect of increasing translational accelerations in the brain cavity by 49%. Removing the trabecular bone in the horn core resulted in a 442% increase in rotational accelerations within the brain cavity. These findings support the investigation of novel bioinspired materials and designs that could be used in mitigating brain injuries and in other applications involving high-impact collisions., Statement of Significance: Bighorn sheep routinely experience violent impacts to the head and horns without apparent negative consequences to the brain or horns. A portion of the horn is filled with a thin cortical bone shell containing foam-like trabecular bone. We developed novel dynamic finite element models of the skull and horns of bighorn sheep to gain an understanding of the roles that the horn and bone materials play in absorbing the impact that occurs during ramming. The study revealed that both horn and bone materials and the structures made from these materials (i.e., tapered spiral horns and foam-like trabecular bone struts) are important for absorbing impact energy and reducing brain cavity accelerations., (Copyright © 2016 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.)

Electrospun nanofibers are a promising material for ligamentous tissue engineering, however weak mechanical properties of fibers to date have limited their clinical usage. The goal of this work was to modify electrospun nanofibers to create a robust structure that mimics the complex hierarchy of native tendons and ligaments. The scaffolds that were fabricated in this study consisted of either random or aligned nanofibers in flat sheets or rolled nanofiber bundles that mimic the size scale of fascicle units in primarily tensile load bearing soft musculoskeletal tissues. Altering nanofiber orientation and geometry significantly affected mechanical properties; most notably aligned nanofiber sheets had the greatest modulus; 125% higher than that of random nanofiber sheets; and 45% higher than aligned nanofiber bundles. Modifying aligned nanofiber sheets to form aligned nanofiber bundles also resulted in approximately 107% higher yield stresses and 140% higher yield strains. The mechanical properties of aligned nanofiber bundles were in the range of the mechanical properties of the native ACL: modulus=158±32MPa, yield stress=57±23MPa and yield strain=0.38±0.08. Adipose derived stem cells cultured on all surfaces remained viable and proliferated extensively over a 7 day culture period and cells elongated on nanofiber bundles. The results of the study suggest that aligned nanofiber bundles may be useful for ligament and tendon tissue engineering based on their mechanical properties and ability to support cell adhesion, proliferation, and elongation., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Understanding the behavior of skeletal muscle is critical to implementing computational methods to study how the body responds to compressive loading. This work presents a novel approach to studying the fully nonlinear response of skeletal muscle in compression. Porcine muscle was compressed in both the longitudinal and transverse directions under five stress relaxation steps. Each step consisted of 5% engineering strain over 1 s followed by a relaxation period until equilibrium was reached at an observed change of 1 g/min. The resulting data were analyzed to identify the peak and equilibrium stresses as well as relaxation time for all samples. Additionally, a fully nonlinear strain energy density-based Prony series constitutive model was implemented and validated with independent constant rate compressive data. A nonlinear least squares optimization approach utilizing the Levenberg-Marquardt algorithm was implemented to fit model behavior to experimental data. The results suggested the time-dependent material response plays a key role in the anisotropy of skeletal muscle as increasing strain showed differences in peak stress and relaxation time (p < 0.05), but changes in equilibrium stress disappeared (p > 0.05). The optimizing procedure produced a single set of hyper-viscoelastic parameters which characterized compressive muscle behavior under stress relaxation conditions. The utilized constitutive model was the first orthotropic, fully nonlinear hyper-viscoelastic model of skeletal muscle in compression while maintaining agreement with constitutive physical boundaries. The model provided an excellent fit to experimental data and agreed well with the independent validation in the transverse direction.

Introduction: Computational modeling of skeletal muscle requires characterization at the tissue level. While most skeletal muscle studies focus on hyperelasticity, the goal of this study was to examine and model the nonlinear behavior of both time-independent and time-dependent properties of skeletal muscle as a function of strain., Materials and Methods: Nine tibialis anterior muscles from New Zealand White rabbits were subject to five consecutive stress relaxation cycles of roughly 3% strain. Individual relaxation steps were fit with a three-term linear Prony series. Prony series coefficients and relaxation ratio were assessed for strain dependence using a general linear statistical model. A fully nonlinear constitutive model was employed to capture the strain dependence of both the viscoelastic and instantaneous components., Results: Instantaneous modulus (p<0.0005) and mid-range relaxation (p<0.0005) increased significantly with strain level, while relaxation at longer time periods decreased with strain (p<0.0005). Time constants and overall relaxation ratio did not change with strain level (p>0.1). Additionally, the fully nonlinear hyperviscoelastic constitutive model provided an excellent fit to experimental data, while other models which included linear components failed to capture muscle function as accurately., Conclusions: Material properties of skeletal muscle are strain-dependent at the tissue level. This strain dependence can be included in computational models of skeletal muscle performance with a fully nonlinear hyperviscoelastic model., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Subchondral bone is thought to play a significant role in the initiation and progression of the post-traumatic osteoarthritis. The goal of this study was to document changes in tibial and femoral subchondral bone that occur as a result of two lapine models of anterior cruciate ligament injury, a modified ACL transection model and a closed-joint traumatic compressive impact model. Twelve weeks post-injury bones were scanned via micro-computed tomography. The subchondral bone of injured limbs from both models showed decreases in bone volume and bone mineral density. Surgical transection animals showed significant bone changes primarily in the medial hemijoint of femurs and tibias, while significant changes were noted in both the medial and lateral hemijoints of both bones for traumatic impact animals. It is believed that subchondral bone changes in the medial hemijoint were likely caused by compromised soft tissue structures seen in both models. Subchondral bone changes in the lateral hemijoint of traumatic impact animals are thought to be due to transmission of the compressive impact force through the joint. The joint-wide bone changes shown in the traumatic impact model were similar to clinical findings from studies investigating the progression of osteoarthritis in humans., (© 2015 Orthopaedic Research Society. Published by Wiley Periodicals, Inc.)

Traumatic injury to the knee leads to the development of post-traumatic osteoarthritis. The objective of this study was to characterize the effects of a single intra-articular injection of a non-ionic surfactant, Poloxamer 188 (P188), in preservation of meniscal tissue following trauma through maintenance of meniscal glycosaminoglycan (GAG) content and mechanical properties. Flemish Giant rabbits were subjected to a closed knee joint, traumatic compressive impact with the joint constrained to prevent anterior tibial translation. The contralateral limb served as an un-impacted control. Six animals (treated) received an injection of P188 in phosphate buffered saline (PBS) post trauma, and another six animals (sham) received a single injection of PBS to the impacted limb. Histological analyses for GAG was determined 6 weeks post trauma, and functional outcomes were assessed using stress relaxation micro-indentation. The impacted limbs of the sham group demonstrated a significant decrease in meniscal GAG coverage compared to non-impacted limbs (p<0.05). GAG coverage of the impacted P188 treated limbs was not significantly different than contralateral non-impacted limbs in all regions except the medial anterior (p<0.05). No significant changes were documented in mechanics for either the sham or treated groups compared to their respective control limbs. This suggests that a single intra-articular injection of P188 shows promise in prevention of trauma induced GAG loss., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Inverse finite element (FE) analysis is an effective method to predict material behavior, evaluate mechanical properties, and study differences in biological tissue function. The meniscus plays a key role in load distribution within the knee joint and meniscal degradation is commonly associated with the onset of osteoarthritis. In the current study, a novel transversely isotropic hyper-poro-viscoelastic constitutive formulation was incorporated in a FE model to evaluate changes in meniscal material properties following tibiofemoral joint impact. A non-linear optimization scheme was used to fit the model output to indentation relaxation experimental data. This study is the first to investigate rate of relaxation in healthy versus impacted menisci. Stiffness was found to be decreased (p=0.003), while the rate of tissue relaxation increased (p=0.010) at twelve weeks post impact. Total amount of relaxation, however, did not change in the impacted tissue (p=0.513)., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Healthy menisci function within the joint to prevent the underlying articular cartilage from excessive loads. Understanding how mechanical properties of menisci change with degeneration can drive future therapeutic studies to prevent this degeneration. Thus, the goal of this study was to characterize both compressive and tensile moduli of human menisci with varying degrees of gross damage due to osteoarthritis (OA). Twenty four paired menisci were collected from total knee joint replacement patients and the menisci were graded on a scale from 0-4 according to level of gross meniscal degeneration with 0=normal and 4=full tissue maceration. Each meniscus was then sectioned into anterior and posterior regions and subjected to indentation relaxation tests. Samples were sliced into 1mm thick strips, made into dumbbells using a custom punch, and pulled to failure. Significant decreases in instantaneous compressive modulus were seen in the lateral posterior region between grades 0 and 1 (36% decrease) and in the medial anterior regions between grades 1 and 2 (67% decrease) and 1 and 3 (72% decrease). Changes in equilibrium modulus where seen in the lateral anterior region between grades 1 and 2 (35% decrease), lateral posterior region between grades 0-2 (41% decrease), and medial anterior regions between grades 1 and 2 (59% decrease), 1 and 3 (67% decrease), 2 and 4 (54% decrease), and 3 and 4 (42% decrease). No significant changes were observed in tensile modulus across all regions and degenerative grades. The results of this study demonstrate the compressive moduli are affected even in early stages of gross degeneration, and continue to decrease with increased deterioration. However, osteoarthritic menisci retain a tensile modulus similar to that of previously reported healthy menisci. This study highlights progressive changes in meniscal mechanical compressive integrity as level of gross tissue degradation increases, and thus, early interventions should focus on restoring or preserving compressive integrity., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

The objective of this study was to induce anterior cruciate ligament (ACL) and meniscal damage, via a single tibiofemoral compressive impact, in order to document articular cartilage and meniscal changes post-impact. Tibiofemoral joints of Flemish Giant rabbits were subjected to a single blunt impact that ruptured the ACL and produced acute meniscal damage. Animals were allowed unrestricted cage activity for 12 weeks before euthanasia. India ink analysis of the articular cartilage revealed higher degrees of surface damage on the impacted tibias (p=0.018) and femurs (p<0.0001) compared to controls. Chronic meniscal damage was most prevalent in the medial central and medial posterior regions. Mechanical tests revealed an overall 19.4% increase in tibial plateau cartilage thickness (p=0.026), 34.8% increase in tibial plateau permeability (p=0.054), 40.8% increase in femoral condyle permeability (p=0.029), and 20.1% decrease in femoral condyle matrix modulus (p=0.012) in impacted joints compared to controls. Both instantaneous and equilibrium moduli of the lateral and medial menisci were decreased compared to control (p<0.02). Histological analyses revealed significantly increased presence of fissures in the medial femur (p=0.036). In both meniscus and cartilage there was a significant decrease in GAG coverage for the impacted limbs. Based on these results it is clear that an unattended combined meniscal and ACL injury results in significant changes to the soft tissues in this experimental joint 12 weeks post-injury. Such changes are consistent with a clinical description of mid to late stage PTOA of the knee., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

The body has approximately 434 muscles, which makes up 40-50% of the body by weight. Muscle is hierarchical in nature and organized in progressively larger units encased in connective tissue. Like many soft tissues, muscle has nonlinear visco-elastic behavior, but muscle also has unique characteristics of excitability and contractibility. Mechanical testing of muscle has been done for crash models, pressure sore models, back pain, and other disease models. The majority of previous biomechanical studies on muscle have been associated with tensile properties in the longitudinal direction as this is muscle's primary mode of operation under normal physiological conditions. Injury conditions, particularly high rate injuries, can expose muscle to multiple stress states. Compressive stresses can lead to tissue damage, which may not be reversible. In this study, we evaluate the structure-property relationships of porcine muscle tissue under compression, in both the transverse and longitudinal orientations at 0.1 s-1, 0.01 s-1, or 0.001 s-1. Our results show an initial toe region followed by an increase in stress for muscle in both the longitudinal and transverse directions tested to 50% strain. Strain rate dependency was also observed with the higher strain rates showing significantly more stress at 50% strain. Muscle in the transverse orientation was significantly stiffer than in the longitudinal orientation indicating anisotropy. The mean area of fibers in the longitudinal orientation shows an increasing mean fiber area and a decreasing mean fiber area in the transverse orientation. Data obtained in this study can help provide insight on how muscle injuries are caused, ranging from low energy strains to high rate blast events, and can also be used in developing computational injury models.

Background: Traumatic impaction is known to cause acute cell death and macroscopic damage to cartilage and menisci in vitro. The purpose of this study was to investigate cell viability and macroscopic damage of the medial and lateral menisci using an in situ model of traumatic loading. Furthermore, the release of nitric oxide from meniscus, synovium, cartilage, and subchondral bone was also documented., Methods: The left limbs of five rabbits were subjected to tibiofemoral impaction resulting in anterior cruciate ligament (ACL) rupture and meniscal damage. Meniscal tear morphology was assessed immediately after trauma and cell viability of the lateral and medial menisci was assessed 24 hrs post-injury. Nitric oxide (NO) released from joint tissues to the media was assayed at 12 and 24 hrs post injury., Results: ACL and meniscal tearing resulted from the traumatic closed joint impact. A significant decrease in cell viability was observed in the lateral menisci following traumatic impaction compared to the medial menisci and control limbs. While NO release was greater in the impacted joints, this difference was not statistically significant., Conclusion: This is the first study to investigate acute meniscal viability following an in situ traumatic loading event that results in rupture of the ACL. The change in cell viability of the lateral menisci may play a role in the advancement of joint degeneration following traumatic knee joint injury.

Post-traumatic osteoarthritis (PTOA) develops as a result of traumatic loading that causes tears of the soft tissues in the knee. A modified transection model, where the anterior cruciate ligament (ACL) and both menisci were transected, was used on skeletally mature Flemish Giant rabbits. Gross morphological assessments, elastic moduli, and glycosaminoglycan (GAG) coverage of the menisci were determined to quantify the amount of tissue damage 12 weeks post injury. This study is one of the first to monitor meniscal changes after inducing combined meniscal and ACL transections. A decrease in elastic moduli as well as a decrease in GAG coverage was seen.

This study investigated the internal fluid pressure of human cadaver meniscal root attachments. A pressure micro-sensor was implanted inside each attachment site. Tibiofemoral joints were compressed to 2× body weight at various flexion angles and pressure recorded for 20 min. The anterior cruciate ligament (ACL) was then transected and joints retested. Lastly, a longitudinal incision of the lateral posterior (LP) horn was made and the joint retested. Ramp pressure was defined as the pressure when 2× body weight was reached, and equilibrium pressure was recorded at the end of the hold period. The medial posterior (MP) attachment was subjected to greater ramp pressure than the medial anterior (p = 0.002) and greater equilibrium pressure than all other root attachment sites (p < 0.001). Flexion angle had a significant effect on pressure as full extension was greatest at ramp (p = 0.040). Transection of the ACL decreased ramp pressure in the LP attachment (p = 0.025) and increased equilibrium pressure (p = 0.031) in the MP attachment. The results suggest that repair strategies should be developed which reconstruct the MP attachments to be sufficient to withstand large pressures. Furthermore, since meniscal pressure is highest at full extension, this fact should be considered when prescribing rehabilitation following repair of an attachment., (Copyright © 2013 Orthopaedic Research Society.)

Menisci are crescent shaped fibrocartilaginous structures which support load distribution of the knee. The menisci are specifically designed to fit the contour of the femoral condyles, aiding to disperse the stresses on the tibial plateau and in turn safeguarding the underlying articular cartilage. The importance of the meniscal superficial layer has not been fully revealed and it is suspected that this layer plays a pivotal role for meniscal function. In this study, both femoral (proximal) and tibial (distal) contacting meniscal surfaces were mechanically examined on the nano-level among three distinct regions (anterior, central and posterior) of the lateral and medial menisci. Nanoindentation testing showed no significant differences among regions, surfaces or anatomical locations, possibly elucidating on the homogeneity of the meniscal superficial zone structure (E(instantaneous): 3.17-4.12MPa, E(steady-state): 1.47-1.69MPa). Nanomechanical moduli values were approximately an order of magnitude greater than micro-scale testing derived moduli values. These findings validate the structural homogeneity of the meniscal superficial zone, showing that material properties are statistically similar regardless of meniscal surface and region. Understanding the mechanical behavior of meniscal surfaces is imperative to properly design an effective meniscal replacement., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Background: Damage to the knee meniscus may result in tears that are difficult or unable to heal, and are often treated by partial removal of the damaged tissue. In vitro, 20% dynamic compressive strains on meniscal tissue explants have resulted in an increase in the release of sulfated glycosaminoglycans (GAG) and nitric oxide (NO) from the tissue explants and increased expression of matrix metalloproteinases (MMP) and interleukin-1α (IL-1α). The objective of this study was to explore the efficacy of IL-1 blockade on the expression of a wide range of genes, as well as NO and GAG release, following dynamic compression of porcine meniscal explants., Methods: Explants were dynamically compressed for 2 h at 1 Hz to 0, 10, or 20% strain with and without a pre-treatment of 500 ng/ml interleukin-1 receptor antagonist (IL-1RA). Relative changes in gene expression of IL-1α, MMP-1, -3, -13, A Disintegrin and Metalloproteinase with ThromboSpondin 4 (ADAMTS-4), ADAMTS-5, iNOS, aggrecan, and COX-2, as well as changes in NO and GAG release, were measured with standard biochemical assays., Results: Expression of IL-1α, MMP-3, MMP-13, and ADAMTS-4 in superficial explants was significantly downregulated at 20% dynamic strain compared to 10% strain following treatment with IL-1RA. GAG and NO release were not significantly influenced by IL-1RA treatment., Conclusions: Treatment of meniscal explants with IL-1RA inhibited the expression of many catabolic genes following a single bout of high dynamic strain. IL-1RA may therefore be a potential therapy option during the acute phase of meniscal tear or meniscectomy treatment.

Meniscal attachments are ligamentous tissues anchoring the menisci to the underlying subchondral bone. Currently little is known about the behavior of meniscal attachments, with only a few studies quantitatively documenting their properties. The objective of this study was to quantify and compare the tensile mechanical properties of human meniscal attachments in the transverse direction, curve fit experimental Cauchy stress-stretch data to evaluate the hyperelastic behavior, and couple these results with previously obtained longitudinal data to generate a more complete constitutive model. Meniscal attachment specimens were tested using a uniaxial tension test with the collagen fibers oriented perpendicular to the loading axis. Tests were run until failure and load-optical displacement data was recorded for each test. The medial posterior attachment was shown to have a significantly greater elastic modulus (6.42±0.78 MPa) and ultimate stress (1.73±0.32 MPa) when compared to the other three attachments. The Mooney-Rivlin material model was selected as the best fit for the transverse data and used in conjunction with the longitudinal data. A novel computational approach to determining the transition point between the toe and linear regions is presented for the hyperelastic stress-stretch curves. Results from piece-wise non-linear longitudinal curve fitting correlate well with previous linear elastic and SEM findings. These data can be used to advance the design of meniscal replacements and improve knee joint finite element models., (Copyright © 2010 Elsevier Ltd. All rights reserved.)

The menisci have crucial weight-bearing roles in the knee. Regional variations in structure and cellularity of the meniscus have only been minimally investigated. Therefore, the goal of this study was to illustrate the regional cell density, tissue area, and structure of healthy lapine menisci. Skeletally mature Flemish Giant rabbits were used for this study. Upon sacrifice, menisci were removed, fixed in formalin, and cryosectioned. Histological analysis was performed for the detection of sulfated glycosaminoglycans (GAG), collagen Types I and II, cellular density, and tissue area. ANOVA and paired t tests were used for testing of statistical significance. Glycosaminoglycan coverage of the medial meniscus significantly varied between regions, with the anterior region demonstrating significantly more GAG coverage than the posterior region. Inter- and intra-meniscal comparisons revealed variations between zones, with trends that outer zones of the medial menisci had less GAG coverage. Collagen Types I and II had marked characteristics and varying degrees of coverage across regions. Tissue area varied between regions for both medial and lateral menisci. Cellular density was dependent on region in the lateral meniscus. This is the first study to illustrate regional and zonal variation in glycosaminoglycan coverage, size, and cellular density for healthy lapine meniscal tissue. This data provides baseline information for future investigations in meniscal injury models in rabbits.

Meniscectomies have been shown to lead to osteoarthritis and the success of meniscal replacements remains questionable. It has been suggested that the success of a meniscal replacement is dependent on several factors, one of which is the secure fixation and firm attachment of the replacement to the tibial plateau at the horn locations. To aid in the development of meniscal replacements, the objectives of the current study were to determine the time-dependent and failure properties of human meniscal attachments. In contrast to the time-dependent tests, during uniaxial failure testing a charge-coupled video camera was used to document the local strain and linear modulus distribution across the surface of the attachments. The lateral attachments were statistically smaller in cross-sectional area and longer than the medial attachments. The anterior attachments were statistically longer and had a smaller cross-sectional area than the posterior attachments. From the stress relaxation tests, the load and stress relaxation rates of the medial anterior attachment were statistically greater than the medial posterior attachment. There were no significant differences in the creep, structural properties or the ultimate stress between the different attachments. Ultimate strain varied between attachments, as well as along the length of the attachment. Ultimate strain in the meniscus region (10.4+/-6.9%) and mid-substance region (12.7+/-16.4%) was smaller than the bony insertion region (32.2+/-21.5%). The lateral and anterior attachments were also found to have statistically greater strain than the medial and posterior attachments, respectively. The linear modulus was statistically weaker in the bony insertion region (69.7+/-33.7MPa) compared to the meniscus region (153+/-123MPa) and mid-substance region (195+/-121MPa). Overall the anterior attachments (169+/-130MPa) were also found to be statistically stronger than the posterior attachments (90.8+/-64.9MPa). These results can be used to help design tissue-engineered replacement menisci and their insertions and show the differences in material properties between attachments, as well as within an attachment., (Copyright 2009 Elsevier Ltd. All rights reserved.)

Of the plethora of work performed analyzing skeletal muscle tissue, relatively little has been done in the examination of its passive material properties. Previous studies of the passive properties of skeletal muscle have been primarily performed along the longitudinal material direction. In order to ensure the accuracy of the predictions of computational models of skeletal muscles, a better understanding of the tensile three-dimensional material properties of muscle tissue is necessary. To that end, the purpose of this study was to collect a comprehensive set of tensile stress-strain data from skeletal muscle tissue. Load-deformation data was collected from eighteen extensor digitorum longus muscles, dissected free of aponeuroses, from nine New Zealand White rabbits tested under longitudinal extension (LE), transverse extension (TE), or longitudinal shear (LS). The linear modulus, ultimate stress, and failure strain were calculated from stress-strain results. Results indicate that the linear modulus under LE is significantly higher than the modulus of either TE or LS. Additionally, the ultimate stress of muscle was seen to be significantly higher under LE than TE. Conversely, the failure strain was significantly higher under TE than under LE.