Chen-Hua Yeow

Cartilage repair using hyaluronan hydrogel-encapsulated human embryonic stem cell-derived chondrogenic cells

Human embryonic stem cells (hESCs) have the potential to offer a virtually unlimited source of chondrogenic cells for use in cartilage repair and regeneration. We have recently shown that expandable chondrogenic cells can be derived from hESCs under selective growth factor-responsive conditions. In this study, we explore the potential of these hESC-derived chondrogenic cells to produce an extracellular matrix (ECM)-enriched cartilaginous tissue construct when cultured in hyaluronic acid (HA)-based hydrogel, and further investigated the long-term reparative ability of the resulting hESC-derived chondrogenic cell-engineered cartilage (HCCEC) in an osteochondral defect model. We hypothesized that HCCEC can provide a functional template capable of undergoing orderly remodeling during the repair of critical-sized osteochondral defects (1.5 mm in diameter, 1 mm depth into the subchondral bone) in a rat model. In the process of repair, we observed an orderly spatial-temporal remodeling of HCCEC over 12 weeks into osteochondral tissue, with characteristic architectural features including a hyaline-like neocartilage layer with good surface regularity and complete integration with the adjacent host cartilage and a regenerated subchondral bone. By 12 weeks, the HCCEC-regenerated osteochondral tissue resembled closely that of age-matched unoperated native control, while only fibrous tissue filled in the control defects which left empty or treated with hydrogel alone. Here we demonstrate that transplanted hESC-derived chondrogenic cells maintain long-term viability with no evidence of tumorigenicity, providing a safe, highly-efficient and practical strategy of applying hESCs for cartilage tissue engineering.

Sagittal knee joint kinematics and energetics in response to different landing heights and techniques.

Single-leg and double-leg landing techniques are common athletic maneuvers typically performed from various landing heights during intensive sports activities. However, it is still unclear how the knee joint responds in terms of kinematics and energetics to the combined effects of different landing heights and techniques. We hypothesized that the knee displays greater flexion angles and angular velocities, joint power and work in response to the larger peak ground reaction force from 0.6-m height, compared to 0.3-m height. We further hypothesized that the knee exhibits elevated flexion angles and angular velocities, joint power and work during double-leg landing, relative to single-leg landing. Ground reaction force, knee joint kinematics and energetics data were obtained from 10 subjects performing single-leg and double-leg landing from 0.3-m to 0.6-m heights, using motion-capture system and force-plates. Higher peak ground reaction force (p<0.05) was observed during single-leg landing and/or at greater landing height. We found greater knee flexion angles and angular velocities (p<0.05) during double-leg landing and/or at greater landing height. Elevated knee joint power and work were noted (p<0.05) during double-leg landing and/or at greater landing height. The knee joint is able to respond more effectively in terms of kinematics and energetics to a larger landing impact from an elevated height during double-leg landing, compared to single-leg landing. This allows better shock absorption and thus minimizes the risk of sustaining lower extremity injuries.

Effect of landing height on frontal plane kinematics, kinetics and energy dissipation at lower extremity joints

Lack of the necessary magnitude of energy dissipation by lower extremity joint muscles may be implicated in elevated impact stresses present during landing from greater heights. These increased stresses are experienced by supporting tissues like cartilage, ligaments and bones, thus aggravating injury risk. This study sought to investigate frontal plane kinematics, kinetics and energetics of lower extremity joints during landing from different heights. Eighteen male recreational athletes were instructed to perform drop-landing tasks from 0.3- to 0.6-m heights. Force plates and motion-capture system were used to capture ground reaction force and kinematics data, respectively. Joint moment was calculated using inverse dynamics. Joint power was computed as a product of joint moment and angular velocity. Work was defined as joint power integrated over time. Hip and knee joints delivered significantly greater joint power and eccentric work (p<0.05) than the ankle joint at both landing heights. Substantial increase (p<0.05) in eccentric work was noted at the hip joint in response to increasing landing height. Knee and hip joints acted as key contributors to total energy dissipation in the frontal plane with increase in peak ground reaction force (GRF). The hip joint was the top contributor to energy absorption, which indicated a hip-dominant strategy in the frontal plane in response to peak GRF during landing. Future studies should investigate joint motions that can maximize energy dissipation or reduce the need for energy dissipation in the frontal plane at the various joints, and to evaluate their effects on the attenuation of lower extremity injury risk during landing.

An investigation of lower extremity energy dissipation strategies during single-leg and double-leg landing based on sagittal and frontal plane biomechanics.

There is limited understanding of the differences in lower extremity energy dissipation strategies between single-leg and double-leg landing maneuvers. This study sought to investigate these differences in sagittal and frontal planes, and explain the differences using kinematics and kinetics. We hypothesized that single-leg and double-leg landing maneuvers involve different lower extremity energy dissipation strategies in both planes. Ten recreational athletes were recruited and instructed to perform double-leg and single-leg landing from 0.60-m height. Force-plates and motion-capture system were used to obtain kinetics and kinematics data respectively. Joint power was taken as product of joint moment and angular velocity. Joint work was computed as integral of joint power over time, whereby negative work represented energy dissipation. In the sagittal plane, the hip and knee showed major contributions to energy dissipation during double-leg landing; the hip and ankle were the dominant energy dissipaters during single-leg landing. In the frontal plane, the hip acted as the key energy dissipater during double-leg landing; the knee contributed the most energy dissipation during single-leg landing. The knee also exhibited greater frontal plane joint ROM, moment and energy dissipation during single-leg landing than double-leg landing. Our findings indicated that different energy dissipation strategies were adopted for double-leg and single-leg landing in sagittal and frontal planes. Considering the prominent frontal plane biomechanics exhibited by the knee during single-leg landing, we expect that this maneuver may have greater likelihood of leading to traumatic knee injuries, particularly non-contact ACL injuries, compared to the double-leg landing maneuver.

Anterior cruciate ligament failure and cartilage damage during knee joint compression: a preliminary study based on the porcine model.

Background Anterior cruciate ligament (ACL) injury incurred from high-impact activities leads to an increased risk of osteoarthritis. Hypothesis Impact forces that cause ACL failure can also inflict cartilage damage, whereby its extent and distribution may be influenced by the ligament failure mechanism. Study Design Descriptive laboratory study. Methods Six porcine knee specimens were mounted to a material testing system at 70° of flexion. During compression, rotational and translational data of the specimens were recorded with a motion-capture system. Compression was successively repeated with increasing actuator displacement until a significant drop in compressive force response was observed; ligament failure was assessed by dissection. Osteocartilage expiants were extracted from the meniscus-covered sites (anterior, exterior, and posterior) and exposed (interior) sites on both tibial compartments. The expiants were sectioned, stained, and histologically scored using the modified Mankin grading system. Results Five of the 6 specimens incurred ACL failure. On failure, a significant compressive force drop (1812.5–2659.3 N) was observed together with considerable posterior femoral translation; 2 specimens underwent external rotation, while 2 had internal rotation and 1 had no substantial rotation. Generally, the meniscus-covered sites displayed significant surface fraying and occasional deep clefts; the exposed site did not present substantial surface irregularities but indicated more tidemark disruption. Higher Mankin scores observed at certain sites illustrated a localized presence of contact and shear forces, which may be caused by pivoting and sliding of the femoral condyles during rotation. Conclusion The porcine model can be a tenable preliminary option for assessing the role of the human ACL during joint compression. Impact loads that result in ligament failure can potentially inflict considerable cartilage damage; the damage profile may be affected by the type of failure mechanism. Clinical Relevance Cartilage injury arising at the time of ACL injury may lead to an accelerated risk of joint degeneration.

Regression relationships of landing height with ground reaction forces, knee flexion angles, angular velocities and joint powers during double-leg landing

Ground reaction forces (GRF), knee flexion angles, angular velocities and joint powers are unknown at large landing heights, which are infeasible for laboratory testing. However, this information is important for understanding lower extremity injury mechanisms. We sought to determine regression relationships of landing height with these parameters during landing so as to facilitate estimation of these parameters at large landing heights. Five healthy male subjects performed landing tasks from heights of 0.15-1.05 m onto a force-plate. Motion capture system was used to obtain knee flexion angles during landing via passive markers placed on the lower body. An iterative regression model, involving simple linear/exponential/natural logarithmic functions, was used to fit regression equations to experimental data. Peak GRF followed an exponential regression relationship (R(2)=0.90-0.99, p<0.001; power=0.987-0.998). Peak GRF slope and impulse also had an exponential relationship (R(2)=0.90-0.96, p<0.001; power=0.980-0.997 and R(2)=0.90-0.99, p<0.001; power=0.990-1.000 respectively) with landing height. Knee flexion angle at initial contact and at peak GRF had an inverse-exponential regression relationship (R(2)=0.81-0.99, p<0.001-p=0.006; power=0.834-0.978 and R(2)=0.84-0.97, p<0.001-p=0.004; power=0.873-0.999 respectively). There was also an inverse-exponential relationship between peak knee flexion angular velocity and landing height (R(2)=0.86-0.96, p<0.001; power=0.935-0.994). Peak knee joint power demonstrated a substantial linear relationship (R(2)=0.98-1.00, p<0.001; power=0.990-1.000). The parameters analyzed in this study are highly dependent on landing height. The exponential increase in peak GRF parameters and the relatively slower increase in knee flexion angles, angular velocities and joint power may synergistically lead to an exacerbated lower extremity injury risk at large landing heights.

Early detection of biomolecular changes in disrupted porcine cartilage using polarized Raman spectroscopy

We evaluate the feasibility of applying polarized Raman spectroscopy in probing the early biochemical compositions and orientation changes in impacted porcine cartilage explants. We divide 100 fresh tibial cartilage explants into four groups: control (unimpacted) and 3 groups of single impact at 15, 20, and 25 MPa. Each group is examined for biochemical changes using Raman microscopy, cell viability changes using confocal fluorescence microscopy, and histological changes using the modified Mankin score. For the 15-MPa impact group, the modified Mankin score (p>0.05, n=15) and cell viability test (p>0.05, n=5) reveal no significant changes when compared to the control, but polarized Raman spectroscopy detects significant biochemical changes. A significant decrease in the parallel polarized intensity of the pyranose ring band at 1126 cm(-1) suggests a possible decrease in the glycoaminoglycan content in early cartilage damage (one-way analysis of variance with a post hoc Bonferonni test, p<0.05, n=10). For impacts greater than 15 MPa, cell viability and modified Mankin score are consistent with the changes in the observed polarized Raman signals. This suggests that the polarized Raman spectroscopy technique has potential for diagnosis and detection of early cartilage damage at the molecular level.

Temporal Activation of β-Catenin Signaling in the Chondrogenic Process of Mesenchymal Stem Cells Affects the Phenotype of the Cartilage Generated

Adult mesenchymal stem cells (MSCs) are an attractive cell source for cartilage tissue engineering. In vitro predifferentiation of MSCs has been explored as a means to enhance MSC-based articular cartilage repair. However, there remain challenges to control and prevent the premature progression of MSC-derived chondrocytes to the hypertrophy. This study investigated the temporal effect of transforming growth factor (TGF)-β and β-catenin signaling co-activation during MSC chondrogenic differentiation and evaluated the influence of these predifferentiation conditions to subsequent phenotypic development of the cartilage. MSCs were differentiated in chondrogenic medium that contained either TGFβ alone, TGFβ with transient β-catenin coactivation, or TGFβ with continuous β-catenin coactivation. After in vitro differentiation, the pellets were transplanted into SCID mice. Both coactivation protocols resulted in the enhancement of chondrogenic differentiation of MSCs. Compared with TGFβ activation, transient coactivation of TGFβ-induction with β-catenin activation resulted in heightened hypertrophy and formed highly ossified tissues with marrow-like hematopoietic tissue in vivo. The continuous coactivation of the 2 signaling pathways, however, resulted in inhibition of progression to hypertrophy, marked by the suppression of type X collagen, Runx2, and alkaline phosphatase expression, and did not result in ossified tissue in vivo. Chondrocytes of the continuous co-activation samples secreted significantly more parathyroid hormone-related protein (PTHrP) and expressed cyclin D1. Our results suggest that temporal co-activation of the TGFβ signaling pathway with β-catenin can yield cartilage of different phenotype, represents a potential MSC predifferentiation protocol before clinical implantation, and has potential applications for the engineering of cartilage tissue.

A fabric-regulated soft robotic glove with user intent detection using EMG and RFID for hand assistive application

This paper presents a soft robotic glove designed to assist individuals with functional grasp pathologies in performing activities of daily living. The glove utilizes soft fabric-regulated pneumatic actuators that are low-profile and require lower pressure than previously developed actuators. They are able to support fingers and thumb motions during hand closure. Upon pressurization, the actuators are able to generate sufficient force to assist in hand closing and grasping during different manipulation tasks. In this work, experiments were conducted to evaluate the performances of the actuators as well as the glove in terms of its kinetic and kinematic assistance on a healthy participant. Additionally, surface electromyography and radio-frequency identification techniques were adopted to detect user intent to activate or deactivate the glove. Lastly, we present preliminary results of a healthy participant performing different manipulation tasks with the soft robotic glove controlled by surface electromyography and radio-frequency identification techniques.

Shod landing provides enhanced energy dissipation at the knee joint relative to barefoot landing from different heights

Athletic shoes can directly provide shock absorption at the foot due to its cushioning properties, however it remains unclear how these shoes may affect the level of energy dissipation contributed by the knee joint. This study sought to investigate biomechanical differences, in terms of knee kinematics, kinetics and energetics, between barefoot and shod landing from different heights. Twelve healthy male recreational athletes were recruited and instructed to perform double-leg landing from 0.3-m and 0.6-m heights in barefoot and shod conditions. The shoe model tested was Brooks Maximus II. Markers were placed on the subjects based on the Plug-in Gait Marker Set. Force-plates and motion-capture system were used to capture ground reaction force (GRF) and kinematics data respectively. 2×2-ANOVA (barefoot/shod condition×landing height) was performed to examine differences in knee kinematics, kinetics and energetics between barefoot and shod conditions from different landing heights. Peak GRF was not significantly different (p=0.732-0.824) between barefoot and shod conditions for both landing heights. Knee range-of-motion, flexion angular velocity, external knee flexion moment, and joint power and work were higher during shod landing (p<0.001 to p=0.007), compared to barefoot landing for both landing heights. No significant interactions (p=0.073-0.933) were found between landing height and barefoot/shod condition for the tested parameters. While the increase in landing height can elevate knee energetics independent of barefoot/shod conditions, we have also shown that the shod condition was able to augment the level of energy dissipation contributed by the knee joint, via the knee extensors, regardless of the tested landing heights.