The Effect of Low-Intensity Pulsed Ultrasound on Temporomandibular Joint Arthritis in Juvenile Rats.

Juvenile idiopathic arthritis is an inflammatory disease that can affect the temporomandibular joint (TMJ) and lower jaw growth. Better treatment options are needed, so this study investigated the effect of low-intensity pulsed ultrasound (LIPUS) on TMJ arthritis. Seventy-two 3-week-old male Wistar rats were in vivo microcomputed tomography (micro-CT) scanned and divided into eight groups (n = 9). These groups were Group 1-TMJ arthritis and immediate LIPUS treatment (20 min/day, 4 weeks); Group 2-immediate LIPUS treatment and no TMJ arthritis; Group 3-TMJ arthritis and no LIPUS; Group 4-no TMJ arthritis and no LIPUS; Group 5-TMJ arthritis and LIPUS treatment with a delayed start by 4 weeks; Group 6-Delayed LIPUS and no TMJ arthritis; Group 7-TMJ arthritis and no (delayed) LIPUS; and Group 8-no TMJ arthritis and no (delayed) LIPUS. Ex vivo micro-CT scanning was completed, and samples were prepared for tissue analysis. Synovitis was observed in the TMJ arthritis (collagen-induced arthritis [CIA]) groups, but the severity appeared greater in the groups without LIPUS treatment. Fibrocartilage and hypertrophic cell layer thicknesses in the CIA group without LIPUS treatment were significantly greater (p < 0.05). Proteoglycan staining appeared greater in the LIPUS groups. Immediate LIPUS treatment increased the expression of type II collagen, type X collagen, and transforming growth factor-beta 1 (TGF-ß1) immunostaining, and CIA (no LIPUS) increased MMP-13, vascular endothelial growth factor, and interleukin-1 beta (IL-1ß) immunostaining. LIPUS treatment prevented growth disturbances observed in the CIA groups (no LIPUS) (p < 0.005). Our results have contributed to the understanding of the uses and limitations of the CIA juvenile rat model and have demonstrated the effects of LIPUS on the TMJ and mandibular growth. This information will help in designing future studies for investigating LIPUS and TMJ arthritis, leading to the development of new treatment options for children with juvenile arthritis in their TMJs.

Development and validation of a novel ankle joint musculoskeletal model.

An improved understanding of contact mechanics in the ankle joint is paramount for implant design and ankle disorder treatment. However, existing models generally simplify the ankle joint as a revolute joint that cannot predict contact characteristics. The current study aimed to develop a novel musculoskeletal ankle joint model that can predict contact in the ankle joint, together with muscle and joint reaction forces. We modelled the ankle joint as a multi-axial joint and simulated contact mechanics between the tibia, fibula and talus bones in OpenSim. The developed model was validated with results from experimental studies through passive stiffness and contact. Through this, we found a similar ankle moment-rotation relationship and contact pattern between our study and experimental studies. Next, the musculoskeletal ankle joint model was incorporated into a lower body model to simulate gait. The ankle joint contact characteristics, kinematics, and muscle forces were predicted and compared to the literature. Our results revealed a comparable peak contact force and the same muscle activation patterns in four major muscles. Good agreement was also found in ankle dorsi/plantar-flexion and inversion/eversion. Thus, the developed model was able to accurately model the ankle joint and can be used to predict contact characteristics in gait.

Comparison of wear on articular cartilage by titanium alloy, ultra-high-molecular-weight polyethylene, and carbon fibre reinforced polyether-ether-ketone: A pilot study.

Artificial implant materials may articulate against native articular cartilage in certain clinical scenarios and the selection of an implant material that results in the least wear on articular cartilage is preferred to maintain normal joint architecture and function. This project compared the wear on porcine femoral condyles induced by articulation against porcine patellae, titanium alloy (Ti6Al4V), ultra high molecular weight polyethylene (UHMWPE), and carbon fibre reinforced polyether-ether-ketone (CFR-PEEK) through an ex vivo experimental setup. A sinusoidal compressive load of 30-160 N, representing an approximate joint pressure of 0.19-1 MPa at a frequency of 3 Hz coupled with a rotational displacement of +/- 10° at 3 Hz was used to simulate physiological joint motion. Wear was characterized via gross examination and histologically using the OARSI scoring system after 43,200 cycles. CFR-PEEK resulted in the most significant wear on articular cartilage compared to titanium alloy and UHMWPE whereas titanium alloy and UHMWPE resulted in similar levels of wear. All materials caused more wear compared to cartilage-on-cartilage testing. The wear mechanism was characterized by progressive loss of proteoglycan content in cartilage in histology samples.

Design of a lightweight universal talus implant using topology optimization.

Total talus replacement is a promising alternative treatment for talus fractures complicated by avascular necrosis and collapse. This surgical option replaces the human talus bone with a customized talus implant and can maintain ankle joint functionality compared to traditional treatment (e.g., ankle fusion). However, the customized implant is costly and time-consuming due to its customized nature. To circumvent these drawbacks, universal talus implants were proposed. While they showed clinically satisfactory results, existing talus implants are heavier than biological talus bones as they are solid inside. This can lead to unequal weight between the implant and biological talus bone, and therefore leading to other complications. The reduction of the implants' weight without compromising its performance and congruency with surrounding bones is a potential solution. Therefore, this study aims to design a lightweight universal talus implant using topology optimization. This is done through establishing the loading and boundary conditions for three common foot postures: neutral, dorsi- and plantar-flexion. The optimized implant performance in terms of mass, contact characteristics with surrounding joint cartilage and stress distributions is studied using a 3D Finite Element (FE) model of the ankle joint. The mass of the optimized implant is reduced by approximately 66.6% and its maximum stresses do not exceed 70 MPa, resulting in a safety factor of 15.7. Moreover, the optimized and solid implants show similar contact characteristics. Both implants produced peak contact pressures that were approximately 19.0%-196% higher than those produced by the biological talus. While further mechanical testing under in-vivo loading conditions is required to determine clinical feasibility, preliminarily, the use of a lightweight universal implant is expected to provide the patient with a more natural feel, and a reduced waiting period until surgery.

Effect of storage media on chondrocyte viability during cold storage of osteochondral dowels.

Osteochondral allograft transplantation is a successfully proven method to repair articular cartilage defects and prevent the degenerative effects of osteoarthritis. The number of osteochondral transplantations that can be performed each year is limited by availability of donor cartilage tissue and storage time constraints. Osteochondral transplantation success has been linked to high chondrocyte viability of the donor cartilage tissue at the time of implantation. Determining optimal storage conditions for donor cartilage is essential for tissue banks to safely provide quality cartilage tissue. In this study, we compared three tissue/cell media (DMEM/F12, RPMI-1640 and X-VIVO 10) for their ability to maintain chondrocyte viability during hypothermic storage for 28 days. Porcine osteochondral dowels were stored in each media for 28 days and cell viability was assessed every 7 days. Over the 28 day storage period, the chondrocyte viability of dowels stored in DMEM/F12, RPMI-1640, and X-VIVO 10 media all declined in a similar fashion. Our results show that all three media were equivalent in their ability to maintain cell viability of the cartilage tissue and provides rationale for the use of lower cost cell media (DMEM/F12 and RPMI-1640) for hypothermic storage of articular cartilage tissue.

Chondrocyte Viability of Particulated Porcine Articular Cartilage Is Maintained in Tissue Storage After Cryoprotectant Exposure, Vitrification, and Tissue Warming.

OBJECTIVE: Vitrification of articular cartilage (AC) is a promising technique which may enable long-term tissue banking of AC allografts. We previously developed a 2-step, dual-temperature, multi-cryoprotectant agent (CPA) loading protocol to cryopreserve particulated AC (1 mm3 cubes). Furthermore, we also determined that the inclusion of ascorbic acid (AA) effectively mitigates CPA toxicity in cryopreserved AC. Prior to clinical translation, chondrocytes must remain viable after tissue re-warming and before transplantation. However, the effects of short-term hypothermic storage of particulated AC after vitrification and re-warming are not documented. This study evaluated the chondrocyte viability of post-vitrified particulated AC during a 7-day tissue storage period at 4 °C. We hypothesized that porcine particulated AC could be stored for up to 7 days after successful vitrification without significant loss of cell viability, and these results would be enhanced when cartilage is incubated in storage medium supplemented with clinical grade AA. DESIGN: Three experimental groups were examined at 5 time points: a fresh control (only incubated in medium), a vitrified - AA group, and a vitrified + AA group (N = 7). RESULTS: There was a mild decline in cell viability but both treatment groups maintained a viability of greater than 80% viable cells which is acceptable for clinical translation. CONCLUSION: We determined that particulated AC can be stored for up to 7 days after successful vitrification without a clinically significant decline in chondrocyte viability. This information can be used to guide tissue banks regarding the implementation of AC vitrification to increase cartilage allograft availability.

Mechanical Properties of Fresh, Frozen and Vitrified Articular Cartilage.

Osteochondral allograft transplantations are typically used to treat focal articular cartilage injuries where the damaged cartilage is replaced with fresh cadaveric donor grafts. Despite the notable success rate of this procedure, it is limited by fresh donor tissue availability which can only be stored for approximately 28 days after harvest. Vitrification, a form of cryopreservation, can extend the storage time of cartilage. Although it has shown to preserve chondrocyte viability, its effect on the mechanical properties of the tissue has not been thoroughly investigated. Therefore, in this study, the mechanical properties of fresh, frozen, and vitrified articular cartilage were evaluated through unconfined compression testing. Results showed that the peak modulus, equilibrium modulus, and relaxation time constants of the vitrified and control samples (tested one day after harvest) were similar and higher than the fresh (tested 21 days after harvest) and frozen samples. This demonstrated that vitrification does not adversely affect the mechanical properties of cartilage and can be used as an alternative to fresh allografts which are limited by storage time. The fresh samples also had inferior mechanical properties compared to the control samples suggesting that vitrified allografts could potentially improve clinical outcomes in addition to increasing donor tissue availability.

Compressive mechanical properties of vitrified porcine menisci are superior to frozen and similar to fresh porcine menisci.

The common practice of freezing meniscal allograft tissue is limited due to the formation of damaging ice crystals. Vitrification, which eliminates the formation of damaging ice crystals, may allow the mechanical properties of meniscal allograft tissue to be maintained during storage and long-term preservation. The primary objective of this study was to investigate the differences between fresh, frozen, and vitrified porcine lateral menisci examining compressive mechanical properties in the axial direction. Unconfined compressive stress-relaxation testing was conducted to quantify the mechanical properties of fresh, frozen and vitrified porcine lateral menisci. The compressive mechanical properties investigated were peak and equilibrium stress, secant, instantaneous and equilibrium modulus, percent stress-relaxation, and relaxation time constants from three-term Prony series. Frozen menisci exhibited inferior compressive mechanical properties in comparison with fresh menisci (significant differences in peak and equilibrium stress, and secant, instantaneous and equilibrium modulus) and vitrified menisci (significant differences in peak stress, and secant and instantaneous modulus). Interestingly, fresh and vitrified menisci exhibited comparable compressive mechanical properties (stress, modulus and relaxation parameters). These findings are significant because (1) vitrification was successful in maintaining mechanical properties at values similar to fresh menisci, (2) compressive mechanical properties of fresh menisci were characterized providing a baseline for future research, and (3) freezing affected mechanical properties confirming that freezing should be used with caution in future investigations of meniscal mechanical properties. Vitrification was superior to freezing for preserving compressive mechanical properties of menisci which is an important advance for vitrification as a preservation option for meniscal allograft transplantation.

Modeling the Simultaneous Transport of Multiple Cryoprotectants into Articular Cartilage Using a Triphasic Model.

Cryopreserving articular cartilage by vitrification can increase the availability of tissue for osteochondral allograft transplantation to treat cartilage defects. Developing well-optimized vitrification protocols can be supported by mathematical modeling to reduce the amount of trial-and-error experimentation needed. Fick's law has been used to model cryoprotectant diffusion, but it assumes ideal, dilute solution behavior, neglects water movement, and assumes diffusion of each cryoprotectant is independent of the presence of other cryoprotectants. The modified triphasic model addresses some of these shortcomings by accounting for water movement and the nonideal, nondilute nature of cryoprotectant vitrification solutions. However, it currently only exists for solutions containing a single cryoprotectant. As such, we extend the modified triphasic model to include two permeating cryoprotectants so that simultaneous diffusion occurring in vitrification protocols can be more accurately modeled. Using previously published experimental data, we determine suitable values for the fitting parameters of the new model. We then model a successful vitrification protocol for particulated cartilage cubes by calculating concentration, freezing point, vitrifiability, and strain profiles at the end of each loading step. We observe that Fick's law consistently underestimates cryoprotectant concentration throughout the cartilage compared to the modified triphasic model, leading to an underestimation of tissue vitrifiability. We additionally observe that simultaneous diffusion of cryoprotectants increases the permeation rate of each individual cryoprotectant, which Fick's law fails to consider. This suggests that using the two-cryoprotectant modified triphasic model to develop vitrification protocols could reduce excess exposure to cryoprotectants and improve preserved tissue outcomes.

Vitrified Particulated Articular Cartilage for Joint Resurfacing: A Swine Model.

BACKGROUND: The use of particulated articular cartilage for repairing cartilage defects has been well established, but its use is currently limited by the availability and short shelf life of donor cartilage. Vitrification is an ice-free cryopreservation technology at ultralow temperatures for tissue banking. An optimized vitrification protocol has been developed for particulated articular cartilage; however, the equivalency of the long-term clinical efficacy of vitrified particulated articular cartilage compared with fresh articular cartilage has not yet been determined. HYPOTHESIS: The repair effect of vitrified particulated cartilage from pigs would be equivalent to or better than that of fresh particulated cartilage stored at 4°C for 21 days. STUDY DESIGN: Controlled laboratory study. METHODS: A total of 19 pigs were randomly divided into 3 experimental groups: fresh particulated cartilage group (n = 8), vitrified particulated cartilage group (n = 8), and negative control group (no particulated cartilage in the defect; n = 3). An additional pig was used as the initial cartilage donor for the first set of surgical procedures. Pigs were euthanized after 6 months to obtain femoral condyles, and the contralateral condyle was used as the positive (no defect) control. Samples were evaluated for gross morphology using the Outerbridge and Osteoarthritis Research Society International (OARSI) scoring systems, histology (safranin O, collagen type I/II, DAPI), and chondrocyte viability using live-dead membrane integrity staining. RESULTS: There were no infections after surgery, and all 19 pigs were followed for the duration of the study. The OARSI grades for the fresh and vitrified particulated cartilage groups were 2.44 ± 1.35 and 2.00 ± 0.80, respectively, while the negative control group was graded significantly higher at 4.83 ± 0.29. Analysis of histological and fluorescent staining demonstrated that the fresh and vitrified particulated cartilage groups had equivalent regeneration within cartilage defects, with similar cell viability and densities and expression of proteoglycans and collagen type I/II. CONCLUSION: The implantation of fresh or vitrified particulated cartilage resulted in the equivalent repair of focal cartilage defects when evaluated at 6 months after surgery. CLINICAL RELEVANCE: The vitrification of particulated cartilage is a viable option for long-term storage for cartilage tissue banking and could greatly increase the availability of donor tissue for transplantation.

Tensile mechanical properties of vitrified porcine menisci are superior to frozen and similar to fresh porcine menisci.

Vitrification inhibits crystallization of ice and may allow the mechanical properties of menisci to be preserved for transplantation without the damaging consequences of ice crystals formed during freezing. The primary objective of this study was to investigate the differences between fresh, frozen, and vitrified porcine lateral menisci examining tensile mechanical properties along the circumferential-peripheral, circumferential-central, longitudinal, and radial orientations. The secondary objective was to investigate the variations in the tensile mechanical properties of menisci comparing the circumferential-peripheral orientation to the three other orientations: circumferential-central, longitudinal, and radial. Quasi-static tensile testing was conducted to quantify the tensile mechanical properties of fresh, frozen and vitrified menisci. Ultimate tensile strength of frozen menisci were significantly decreased compared with fresh and vitrified menisci along three orientations: circumferential-peripheral, longitudinal, and radial. Along the circumferential-central orientation, tensile modulus of frozen menisci was significantly decreased compared with fresh menisci. The mechanical properties of vitrified menisci were comparable to fresh menisci along all four orientations. For all menisci (fresh, frozen and vitrified), ultimate tensile strength and failure strain along the circumferential-peripheral orientation were significantly increased compared with the three other orientations. Freezing was detrimental to the mechanical properties of menisci but vitrification likely avoided the negative effects of freezing thereby preserving mechanical properties that were comparable to fresh menisci. The findings of this study revealed that vitrification was superior to freezing for preserving mechanical properties of meniscal tissue; hence, vitrification is likely to be a competitive alternative to freezing for meniscal transplantation in the future.

The effect of sucrose supplementation on chondrocyte viability in porcine articular cartilage following vitrification.

Vitrification can extend the banking life of articular cartilage (AC) and improve osteochondral transplantation success. Current vitrification protocols require optimization to enable them to be implemented in clinical practice. Sucrose as a non-permeating cryoprotective agent (CPA) and clinical grade chondroitin sulfate (CS) and ascorbic acid (AA) as antioxidants were investigated for their ability to improve a current vitrification protocol for AC. The aim of this study was to assess the impact of sucrose and CS/AA supplementation on post-warming chondrocyte viability in vitrified AC. Porcine osteochondral dowels were randomly vitrified and warmed with one established protocol (Protocol 1) and seven modified protocols (Protocols 2-8) followed by chondrocyte viability assessment. Sucrose supplementation in both vitrification and warming media (Protocol 4) resulted in significantly higher (p = 0.018) post-warming chondrocyte viability compared to the protocol without sucrose (Protocol 1). There was no significant difference (p = 0.298) in terms of post-warming chondrocyte viability between sucrose-supplemented DMEM + CS solution (Protocol 4) and Unisol-CV (UCV) + CS (Protocol 6) solution. Clinical grade CS and AA contributed to similar post-warming chondrocyte viability to previous studies using research grade CS and AA, indicating their suitability for clinical use. The addition of an initial step (step 0) to reduce the initial concentration of CPAs to minimize osmotic effects did not enhance chondrocyte viability in the superficial layer of AC. In conclusion, sucrose-supplemented DMEM + clinical grade CS (Protocol 4) could be an ideal protocol to be investigated for future use in clinical applications involving vitrified AC.

Effect of vitrification on mechanical properties of porcine articular cartilage.

Articular cartilage (AC) injuries do not heal primarily and large lesions progress to degenerative osteoarthritis. Osteochondral allograft transplantation is an effective surgical treatment but is limited by the lack of donor tissue availability. Fresh allografts can be stored hypothermically up to 28-45 days after which the tissue is no longer viable for transplantation. Vitrification is a method of cryopreservation with the potential to extend the storage time of AC. A specific protocol has been demonstrated to preserve high chondrocyte viability; however, its effect on various mechanical properties of the extracellular matrix (ECM) remains unknown and is the focus of this initial study. Porcine AC was subject to a defined vitrification protocol, using fresh and frozen samples as positive and negative controls, respectively; n = 20 for all three groups. Unconfined compression testing was used to assess mechanical properties of the tissue under rapid load, stress relaxation, and equilibrium conditions. The stress relaxation time constants (modeled with a 2-term Prony series) τ1 and τ2 were significantly lower for frozen (p = 0.014, p < 0.001) and vitrified (p = 0.009, p = 0.003) tissue compared to fresh, with no differences between frozen and vitrified samples (p = 0.848 and 0.105 for τ1 and τ2, respectively). These values indicate that frozen and vitrified samples relaxed more rapidly than fresh, which may suggest altered matrix composition and permeability post-treatment. These results represent the initial study in our experimental path to evaluate differences in mechanical properties of vitrified tissues.

Evaluation of the permeation kinetics of formamide in porcine articular cartilage.

Cryopreservation of articular cartilage will increase tissue availability for osteochondral allografting and improve clinical outcomes. However, successful cryopreservation of articular cartilage requires the precise determination of cryoprotectant permeation kinetics to develop effective vitrification protocols. To date, permeation kinetics of the cryoprotectant formamide in articular cartilage have not been sufficiently explored. The objective of this study was to determine the permeation kinetics of formamide into porcine articular cartilage for application in vitrification. The permeation of dimethyl sulfoxide was first measured to validate existing methods from our previously published literature. Osteochondral dowels from dissected porcine femoral condyles were incubated in 6.5 M dimethyl sulfoxide for a designated treatment time (1 s, 1 min, 2 min, 5 min, 10 min, 15 min, 30 min, 60 min, 120 min, 180 min, 24 h) at 22 °C (N = 3). Methods were then repeated with 6.5 M formamide at one of three temperatures: 4 °C, 22 °C, 37 °C (N = 3). Following incubation, cryoprotectant efflux into a wash solution occurred, and osmolality was measured from each equilibrated wash solution. Concentrations of effluxed cryoprotectant were calculated and diffusion coefficients were determined using an analytical solution to Fick's law for axial and radial diffusion in combination with a least squares approach. The activation energy of formamide was determined from the Arrhenius equation. The diffusion coefficient (2.7-3.3 × 10-10 m2/s depending on temperature) and activation energy (0.9±0.6 kcal/mol) for formamide permeation in porcine articular cartilage were established. The determined permeation kinetics of formamide will facilitate its precise use in future articular cartilage vitrification protocols.

The evaluation of artificial talus implant on ankle joint contact characteristics: a finite element study based on four subjects.

Customized talus implants have been regarded as a better treatment alternative to talus avascular necrosis than traditional surgical fusion because of its ability to maintain joint mobility while ameliorating pain. Despite the use of ankle hemiarthroplasty clinically, the cartilage contact characteristics of adjacent bones remain unclear. This study aims to use finite element modeling to evaluate the contact characteristics of three types of cobalt-chrome talus implants in three postures, in four subjects. This study also compared the contact area, contact pressure, and peak contact pressure of the implant models with a reference biological model. Among the various biological and implant models, our results showed that the biological models generally had the largest contact areas and smallest peak contact pressures, whereas the implant-type models had smaller contact areas and relatively larger peak contact pressure. Moreover, among the three implant types, customized-scale models showed a larger total contact area than that of the SSM-scale and universal-scale models, but their variation was relatively limited. The results from this study can have significance in future endeavors into ankle joint modeling, as well as being able to improve implant design to enhance recovery outcomes for patients who may benefit from talar replacement.

Effectiveness of Clinical-Grade Chondroitin Sulfate and Ascorbic Acid in Mitigating Cryoprotectant Toxicity in Porcine Articular Cartilage.

High concentrations of cryoprotective agents (CPAs) are required to achieve successful vitrification of articular cartilage; however, CPA cytotoxicity causes chondrocyte death. To reduce CPA toxicity, supplementation with research-grade additives, in particular chondroitin sulfate (CS) and ascorbic acid (AA), have previously been shown to improve chondrocyte recovery and metabolic function after exposure to CPAs at hypothermic conditions. However, it is necessary to evaluate the pharmaceutical equivalent clinical grade of these additives to facilitate the supplementation of additives into future vitrification protocols, which will be designed for vitrifying human articular cartilage in tissue banks. We sought to investigate the effectiveness of clinical-grade CS, AA, and N-acetylcysteine (NAC) in mitigating toxicity to chondrocytes during CPA exposure and removal, and determine whether a combination of two additives would further improve chondrocyte viability. We hypothesized that clinical-grade additives would exert chondroprotective effects comparable to those of research-grade additives, and that this protective effect would be enhanced if two additives were combined when compared with a single additive. The results indicated that both clinical-grade and research-grade additives significantly improved cell viability (p < 0.10) compared with the negative control (CPA with no additives). CS, AA, and NAC+AA increased cell viability significantly (p < 0.10) compared with the negative control. However, NAC, NAC+CS, and CS+AA did not improve cell viability when compared with the negative control (p > 0.10). We demonstrated that supplementation with clinical-grade CS or AA significantly improved chondrocyte viability in porcine cartilage subjected to high CPA concentrations, whereas supplementation with clinical-grade NAC did not benefit chondrocyte viability. Supplementation with clinical-grade additives in CPA solutions can mitigate CPA toxicity, which will be important in translating previously developed effective protocols for the vitrification of articular cartilage to human tissue banks.

A feature-based statistical shape model for geometric analysis of the human talus and development of universal talar prostheses.

Statistical data pertaining to anatomic variations of the human talus contain valuable information for advances in biological anthropology, diagnosis of the talar pathologies, and designing talar prostheses. A statistical shape model (SSM) can be a powerful data analysis tool for the anatomic variations of the talus. The main concern in constructing an SSM for the talus is establishing the true geometric correspondence between the talar geometries. The true correspondence complies with biological and/or mathematical homologies on the talar surfaces. In this study, we proposed a semi-automatic approach to establish a dense correspondence between talar surfaces discretized by triangular meshes. Through our approach, homologous salient surface features in the form of crest lines were detected on 49 talar surfaces. Then, the point-wise correspondence information of the crest lines was recruited to create posterior Gaussian process morphable models that non-rigidly registered the talar meshes and consequently established inter-mesh dense correspondence. The resultant correspondence perceptually represented the true correspondence as per our visual assessments. Having established the correspondence, we computed the mean shape using full generalized Procrustes analysis and constructed an SSM by means of principal component analysis. Anatomical variations and the mean shape of the talus were predicted by the SSM. As a clinically related application, we considered the mean shape and investigated the feasibility of designing universal talar prostheses. Our results suggest that the mean shape of (the shapes of) tali can be used as a scalable shape template for designing universal talar prostheses.

Time course of 3D fibrocartilage formation by expanded human meniscus fibrochondrocytes in hypoxia.

Adult human meniscus fibrocartilage is avascular and nonhealing after injury. Meniscus tissue engineering aims to replace injured meniscus with lab-grown fibrocartilage. Dynamic culture systems may be necessary to generate fibrocartilage of sufficient mechanical properties for implantation; however, the optimal static preculture conditions before initiation of dynamic culture are unknown. This study thus investigated the time course of fibrocartilage formation by human meniscus fibrochondrocytes on a three-dimensional biomaterial scaffold under various static conditions. Human meniscus fibrochondrocytes from partial meniscectomy were expanded to passage 1 (P1) or P2 (3.0 ± 0.4 and 6.5 ± 0.6 population doublings), seeded onto type I collagen scaffolds, and grown in hypoxia (HYP, 3% O2 ) or normoxia (NRX, 20% O2 ) for 3, 6, and 9 weeks. Mechanical properties were not different between P1 and P2 cell-based constructs. Mechanical properties were lower in HYP, increased continually in NRX only, and were positively correlated with glycosaminoglycan content and accumulation of hyaline cartilage-like matrix components. The most mechanically competent tissues (NRX/9 weeks) reached 1/5 of the native meniscus instantaneous compression modulus but had an increasingly hypertrophic matrix-forming phenotype. HYP consistently suppressed the hypertrophic phenotype. The results provide baselines of engineered meniscus fibrocartilage properties under static conditions, which can be used to select a preculture strategy for dynamic culture depending on the desired combination of mechanical properties, hyaline cartilage-like matrix abundance, and hypertrophic phenotype.

Polycarbonate-urethane coating can significantly improve talus implant contact characteristics.

Talus implants can be utilized in cases of talus avascular necrosis and has been regarded as a promising treatment method. However, existing implants are made of stiff materials that directly oppose natural cartilage. The risk of long-term cartilage wear and bone fracture from the interaction between the cartilage and stiff implant surfaces has been documented in post-hemiarthroplasty of the hip, knee and ankle joints. The aim is to explore the effects of adding a layer of compliant material (polycarbonate-urethane; PCU) over a stiff material (cobalt chromium) in talus implants. To do so, we obtained initial ankle geometry from four cadaveric subjects in neutral standing to create the finite element models. We simulated seven models for each subject: three different types of talus implants, each coated with and without PCU, and a biological model. In total, we constructed 28 finite element models. By comparing the contact characteristics of the implant models with their respective biological model counterparts, our results showed that PCU coated implants have comparable contact area and contact pressure to the biological models, whereas stiff material implants without the PCU coating all have relatively higher contact pressure and smaller contact areas. These results confirmed that adding a layer of compliant material coating reduces the contact pressure and increases the contact area which in turn reduces the risk of cartilage wear and bone fracture. The results also suggest that there can be clinical benefits of adding a layer of compliant material coating on existing stiff material implants, and can provide valuable information towards the design of more biofidelic implants in the future.

The effect of cryoprotectant vehicle solution on cartilage cell viability following vitrification.

Osteochondral allografts are often used to repair large articular cartilage defects to prevent or delay the onset of osteoarthritis. This approach is limited by the timely acquisition and use of allograft tissue since standard hypothermic protocols allow for a maximum storage of 4 weeks. Vitrification is a proven technique for the long-term preservation of cells and tissues, but requires careful determination of parameters to be successful, particularly for articular cartilage. One parameter that is infrequently considered is the choice of cryoprotectant vehicle solution. The aim of this study was to evaluate the impact of a subset of vehicle solutions on an established vitrification protocol for articular cartilage. These solutions were phosphate-buffered saline (PBS), Dulbecco's Modified Eagle's Medium: Nutrient Mixture F-12 (DMEM), X-VIVO, and Unisol-CV (UCV). Both the solution pH at various points throughout vitrification and the cell viability of porcine articular cartilage slices following vitrification were measured. Using randomized block ANOVA, it was found that the normalized cell viability of articular cartilage vitrified in UCV was significantly greater than that of PBS (p < 0.05) and may be greater than those of DMEM and X-VIVO (p < 0.1). There was no correlation between pH parameters and cell viability, although significant differences between calculated pH parameters were identified. These results provide information to guide the design of effective vitrification protocols for articular cartilage.