Sang-Kuy Han
University of Calgary
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Publication
Featured researches published by Sang-Kuy Han.
Journal of Orthopaedic Research | 2012
Sang-Kuy Han; Wim Wouters; A.L. Clark; Walter Herzog
Changes in intracellular calcium (Ca2+) concentration, also known as Ca2+ signaling, have been widely studied in articular cartilage chondrocytes to investigate pathways of mechanotransduction. Various physical stimuli can generate an influx of Ca2+ into the cell, which in turn is thought to trigger a range of metabolic and signaling processes. In contrast to most studies, the approach used in this study allows for continuous real time recording of calcium signals in chondrocytes in their native environment. Therefore, interactions of cells with the extracellular matrix (ECM) are fully accounted for. Calcium signaling was quantified for dynamic loading conditions and at different temperatures. Peak magnitudes of calcium signals were greater and of shorter duration at 37°C than at 21°C. Furthermore, Ca2+ signals were involved in a greater percentage of cells in the dynamic compared to the relaxation phases of loading. In contrast to the time‐delayed signaling observed in isolated chondrocytes seeded in agarose gel, Ca2+ signaling in situ is virtually instantaneous in response to dynamic loading. These differences between in situ and in vitro cell signaling responses might provide crucial insight into the role of the ECM in providing pathways of mechanotransduction in the intact cartilage that are absent in isolated cells seeded in gel constructs.
Medical Engineering & Physics | 2009
Sang-Kuy Han; Pina Colarusso; Walter Herzog
Chondrocytes synthesize extracellular matrix molecules, thus they are essential for the development, adaptation and maintenance of articular cartilage. Furthermore, it is well accepted that the biosynthetic activity of chondrocytes is influenced by the mechanical environment. Therefore, their response to mechanical stimuli has been studied extensively. Much of the knowledge in this area of research has been derived from testing of isolated cells, cartilage explants, and fixed cartilage specimens: systems that differ in important aspects from chondrocytes embedded in articular cartilage and observed during loading conditions. In this study, current model systems have been improved by working with the intact cartilage in real time. An indentation system was designed on a confocal microscope that allows for simultaneous loading and observation of chondrocytes in their native environment. Cell mechanics were then measured under precisely controlled loading conditions. The indentation system is based on a light transmissible cylindrical glass indentor of 0.17 mm thickness and 1.64 mm diameter that is aligned along the focal axis of the microscope and allows for real time observation of live cells in their native environment. The system can be used to study cell deformation and biological responses, such as calcium sparks, while applying prescribed loads on the cartilage surface. It can also provide novel information on the relationship between cell loading and cartilage adaptive/degenerative processes in the intact tissue.
Journal of the Royal Society Interface | 2010
Sang-Kuy Han; Ruth A. Seerattan; Walter Herzog
The aims of this study were (i) to quantify chondrocyte mechanics in fully intact articular cartilage attached to its native bone and (ii) to compare the chondrocyte mechanics for cells in healthy and early osteoarthritis (OA) tissue. We hypothesized that cells in the healthy tissue would deform less for given articular surface pressures than cells in the early OA tissue because of a loss of matrix integrity in early OA and the associated loss of structural integrity that is thought to protect chondrocytes. Chondrocyte dynamics were quantified by measuring the deformation response of the cells to controlled loading of fully intact cartilage using a custom-designed confocal indentation system. Early OA was achieved nine weeks following transection of the anterior cruciate ligament (ACL) in rabbit knees. Experiments were performed on the retropatellar cartilage of early OA rabbit knees (four joints and 48 cells), the corresponding intact contralateral control knees (four joints and 48 cells) and knees from normal control rabbits (four joints and 48 cells). Nine weeks following ACL transection, articular cartilage of the experimental joints showed substantial increases in thickness, and progression towards OA as assessed using histological grading. Local matrix strains in the superficial zone were greater for the experimental (38 ± 4%) compared with the contralateral (27 ± 5%) and normal (28 ± 4%) joints (p = 0.04). Chondrocyte deformations in the axial and depth directions were similar during indentation loading for all experimental groups. However, cell width increased more for the experimental cartilage chondrocytes (12 ± 1%) than the contralateral (6 ± 1%) and normal control chondrocytes (6 ± 1%; p < 0.001). On average, chondrocyte volume increased with indentation loading in the early OA cartilage (8 ± 3%, p = 0.001), while it decreased for the two control groups (−8 ± 2%, p = 0.002 for contralateral and −8 ± 1%, p = 0.004 for normal controls). We conclude from these results that our hypothesis of cell deformations in the early OA tissue was only partially supported: specifically, changes in chondrocyte mechanics in early OA were direction-specific with the primary axial deformations remaining unaffected despite vastly increased average axial matrix deformations. Surprisingly, chondrocyte deformations increased in early OA in specific transverse directions which have received little attention to date but might be crucial to chondrocyte signalling in early OA.
Biomechanics and Modeling in Mechanobiology | 2012
Eng Kuan Moo; Walter Herzog; Sang-Kuy Han; N. A. Abu Osman; Belinda Pingguan-Murphy; Salvatore Federico
Experimental findings indicate that in-situ chondrocytes die readily following impact loading, but remain essentially unaffected at low (non-impact) strain rates. This study was aimed at identifying possible causes for cell death in impact loading by quantifying chondrocyte mechanics when cartilage was subjected to a 5% nominal tissue strain at different strain rates. Multi-scale modelling techniques were used to simulate cartilage tissue and the corresponding chondrocytes residing in the tissue. Chondrocytes were modelled by accounting for the cell membrane, pericellular matrix and pericellular capsule. The results suggest that cell deformations, cell fluid pressures and fluid flow velocity through cells are highest at the highest (impact) strain rate, but they do not reach damaging levels. Tangential strain rates of the cell membrane were highest at the highest strain rate and were observed primarily in superficial tissue cells. Since cell death following impact loading occurs primarily in superficial zone cells, we speculate that cell death in impact loading is caused by the high tangential strain rates in the membrane of superficial zone cells causing membrane rupture and loss of cell content and integrity.
Journal of Biomechanics | 2013
Ryan Madden; Sang-Kuy Han; Walter Herzog
Articular cartilage and its native cells-chondrocytes-are exposed to a wide range of mechanical loading. Chondrocytes are responsible for maintaining the cartilage matrix, yet relatively little is known regarding their behavior under a complete range of mechanical loads or how cell mechanics are affected by region within the joint. The purpose of this study was to investigate chondrocyte deformations in situ under tissue loads ranging from physiological to extreme (0-80% nominal strain) in two regions of the rabbit knee joint (femoral condyles and patellae). Local matrix strains and cell compressive strains increased with increasing loads. At low loads the extracellular matrix (ECM) strains in the superficial zone were greater than the applied tissue strains, while at extreme loads, the local ECM strains were smaller than the applied strains. Cell compressive strains were always smaller than the applied tissue strains and, in our intact, in situ preparation, were substantially smaller than those previously found in hemi-cylindrical explants. This resulted in markedly different steady-state cell volume changes in the current study compared to those working with cartilage explants. Additionally, cells from different regions in the knee exhibited striking differences in deformation behavior under load. The current results suggest: (i) that the local extracellular and pericellular matrix environment is intimately linked to chondrocyte mechanobiology, protecting chondrocytes from potentially damaging strains at high tissue loads; and (ii) that cell mechanics are a function of applied load and local cartilage tissue structure.
Journal of Biomechanics | 2010
Rami K. Korhonen; Sang-Kuy Han; Walter Herzog
Osmotic loading is known to modulate chondrocyte (cell) height, width and volume in articular cartilage. It is not known how cartilage architecture, especially the collagen fibril orientation, affects cell shape changes as a result of an osmotic challenge. Intact patellae of New Zealand white rabbits (n=6) were prepared for fluorescence imaging. Patellae were exposed to a hypotonic osmotic shock and cells were imaged before loading and 5-60 min after the osmotic challenge. Cell volumes and aspect ratios (height/width) were analyzed. A fibril-reinforced poroelastic swelling model with realistic primary collagen fibril orientations, i.e. horizontal, random and vertical orientation in the superficial, middle and deep zones, respectively and cells in different zones was used to estimate cell aspect ratios theoretically. As the medium osmolarity was reduced, cell aspect ratios decreased and volumes increased in the superficial zone of cartilage both experimentally (p<0.05) and theoretically. Theoretically determined aspect ratios of middle zone cells remained virtually constant, while they increased for deep zone cells as osmolarity was reduced. Findings of this study suggest that osmotic loading modulates chondrocyte shapes in accordance with the primary collagen fibril directions in articular cartilage.
Computer Methods in Biomechanics and Biomedical Engineering | 2011
Sang-Kuy Han; Salvatore Federico; Walter Herzog
Experimental studies suggest that the magnitude of chondrocyte deformation is much smaller than expected based on the material properties of extracellular matrix (ECM) and cells, and that this result could be explained by a structural unit, the chondron, that is thought to protect chondrocytes from large deformations in situ. We extended an existing numerical model of chondrocyte, ECM and pericellular matrix (PCM) to include depth-dependent structural information. Our results suggest that superficial zone chondrocytes, which lack a pericellular capsule (PC), are relatively stiff, and therefore are protected from excessive deformations, whereas middle and deep zone chondrocytes are softer but are protected by the PC that limits cell deformations in these regions. We conclude that cell deformations sensitively depend on the immediate structural environment of the PCM in a depth-dependent manner, and that the functional stiffness of chondrocytes in situ is much larger than experiments on isolated cells would suggest.
Journal of Biomechanics | 2012
Sang-Kuy Han; Ryan Madden; Ziad Abusara; Walter Herzog
It has been proposed, based on theoretical considerations, that the strain rate-dependent viscoelastic response of cartilage reduces local tissue and cell deformations during cyclic compressions. However, experimental studies have not addressed the in situ viscoelastic response of chondrocytes under static and dynamic loading conditions. In particular, results obtained from experimental studies using isolated chondrocytes embedded in gel constructs cannot be used to predict the intrinsic viscoelastic responses of chondrocytes in situ or in vivo. Therefore, the purpose of this study was to investigate the viscoelastic response of chondrocytes in their native environment under static and cyclic mechanical compression using a novel in situ experimental approach. Cartilage matrix and chondrocyte recovery in situ following mechanical compressions was highly viscoelastic. The observed in situ behavior was consistent with a previous study on in vivo chondrocyte mechanics which showed that it took 5-7 min for chondrocytes to recover shape and volume following virtually instantaneous cell deformations during muscular loading of the knee in live mice. We conclude from these results that the viscoelastic properties of cartilage minimize chondrocyte deformations during cyclic dynamic loading as occurs, for example, in the lower limb joints during locomotion, thereby allowing the cells to reach mechanical and metabolic homeostasis even under highly dynamic loading conditions.
Journal of Orthopaedic Research | 2017
Sang-Kuy Han; Ari P. Ronkainen; Simo Saarakkala; Lassi Rieppo; Walter Herzog; Rami K. Korhonen
The structural integrity and mechanical environment of the articular cartilage matrix directly affect chondrocyte deformations. Rabbit models of early osteoarthritis at 9 weeks following anterior cruciate ligament transection (ACLT) have been shown to alter the deformation behavior of superficial zone chondrocytes in mechanically loaded articular cartilage. However, it is not fully understood whether these changes in cell mechanics are caused by changes in structural macromolecules in the extracellular matrix. Therefore, the purpose of this study was to characterize the proteoglycan content, collagen content, and collagen orientation at 9 weeks post ACLT using microscopic techniques, and relate these changes to the altered cell mechanics observed upon mechanical loading of cartilage. At 9 weeks following ACLT, collagen orientation was significantly (p < 0.05) altered and proteoglycan content was significantly (p < 0.05) reduced in the superficial zone cartilage matrix. These structural changes either in the extracellular or pericellular matrix (ECM and PCM) were also correlated significantly (p < 0.05) with chondrocyte width and height changes, thereby suggesting that chondrocyte deformation response to mechanical compression in early OA changes primarily because of alterations in matrix structure. However, compared to the normal group, proteoglycan content in the PCM from the ACLT group decreased less than that in the surrounding ECM. Therefore, PCM could play a key role to protect excessive chondrocyte deformations in the ACLT group.
Archive | 2011
Eng Kuan Moo; N. A. Abu Osman; Belinda Pingguan-Murphy; Sang-Kuy Han; Salvatore Federico; Walter Herzog
Previous findings indicated that in-situ chondrocytes die readily following impact loading, but remain essentially unaffected when loaded at the same magnitude but with a slow (non-impact) loading rate. The current study was aimed at identifying the causes for cell death in impact loading by quantifying chondrocyte mechanics when cartilage was compressed nominally by 5% at different loading rates.. Multi-scale modeling techniques were used. Cartilage was modeled accounting for collagen stiffening in tension. Chondrocytes were modelled to be including the cell membrane, pericellular matrix, and pericellular capsule. The results showed that cell deformations were lowest and cell fluid pressures were highest for the highest (impact) loading rate. Tangential strain rates on the cell membrane were highest at the highest loading rate and occurred primarily in superficial tissue cells. Since cell death following impact loading was primarily observed in superficial zone cells, we speculate that cell death in impact loading is caused by the high membrane strain rates observed in these cells for simulated impact loading conditions.