Proteoglycan 4 reduces friction more than other synovial fluid components for both cartilage-cartilage and cartilage-metal articulation.

OBJECTIVE: The clinical success of focal metallic resurfacing implants depends largely on the friction between implant and opposing cartilage. Therefore, the present study determines the lubricating ability of the synovial fluid components hyaluronic acid (HA), proteoglycan 4 (PRG4) and a surface-active phospholipid (1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine, POPC), on the articulation between cartilage and a Cobalt Chromium Molybdenum (CoCrMo) implant surface, compared with two cartilage surfaces. METHODS: A ring-on-disk geometry was used to perform repeated friction measurements at physiologically relevant velocities (6 and 60 mm/s) using lubricants with an increasing number of components present. Shear measurements were performed in order to evaluate the viscosity. To ensure that it is clinically relevant to explore the effect of these components, the presence of PRG4 in synovial fluid obtained from primary and revision knee and hip implant surgeries was examined. RESULTS: PRG4 in the presence of HA was found to significantly reduce the coefficient of friction for both cartilage-cartilage and cartilage-CoCrMo interface. This is relevant, as it was also demonstrated that PRG4 is still present at the time of revision surgeries. The addition of POPC had no effect for either configurations. HA increased the viscosity of the lubricating fluid by one order of magnitude, while PRG4 and POPC had no effect. CONCLUSION: The present study demonstrates the importance of selecting the appropriate lubrication solution to evaluate implant materials with biotribology tests. Because PRG4 is a key component for reducing friction between cartilage and an opposing surface, developing coatings which bind PRG4 is recommended for cartilage resurfacing implants.

Spectroscopic photoacoustic imaging of cartilage damage.

OBJECTIVE: Osteoarthritis (OA) is a chronic joint disease characterized by progressive degradation of cartilage. It affects more than 10% of the people aged over 60 years-old worldwide with a rising prevalence due to the increasingly aging population. OA is a major source of pain, disability, and socioeconomic cost. Currently, the lack of effective diagnosis and affordable imaging options for early detection and monitoring of OA presents the clinic with many challenges. Spectroscopic Photoacoustic (sPA) imaging has the potential to reveal changes in cartilage composition with different degrees of damage, based on optical absorption contrast. DESIGN: In this study, the capabilities of sPA imaging and its potential to characterize cartilage damage were explored. To this end, 15 pieces of cartilage samples from patients undergoing a total joint replacement were collected and were imaged ex vivo with sPA imaging at a wide optical spectral range (between 500 nm and 1,300 nm) to investigate the photoacoustic properties of cartilage tissue. All the PA spectra of the cartilage samples were analyzed and compared to the corresponding histological results. RESULTS: The collagen related PA spectral changes were clearly visible in our imaging data and were related to different degrees of cartilage damage. The results are in good agreement with histology and the current gold standard, i.e., the Mankin score. CONCLUSIONS: This study demonstrates the potential and possible clinical application of sPA imaging in OA.

Characterizing the invasion of different breast cancer cell lines with distinct E-cadherin status in 3D using a microfluidic system.

E-cadherin is a cell-cell adhesion protein that plays a prominent role in cancer invasion. Inactivation of E-cadherin in breast cancer can arise from gene promoter hypermethylation or genetic mutation. Depending on their E-cadherin status, breast cancer cells adopt different morphologies with distinct invasion modes. The tumor microenvironment (TME) can also affect the cell morphology and invasion mode. In this paper, we used a previously developed microfluidic system to quantify the three-dimensional invasion of breast cancer cells with different E-cadherin status, namely MCF-7, CAMA-1 and MDA-MB-231 with wild type, mutated and promoter hypermethylated E-cadherin, respectively. The cells migrated into a stable and reproducible microfibrous polycaprolactone mesh in the chip under a programmed stable chemotactic gradient. We observed that the MDA-MB-231 cells invaded the most, as single cells. MCF-7 cells collectively invaded into the matrix more than CAMA-1 cells, maintaining their E-cadherin expression. The CAMA-1 cells exhibited multicellular multifocal infiltration into the matrix. These results are consistent with what is seen in vivo in the cancer biology literature. In addition, comparison between complete serum and serum gradient conditions showed that the MDA-MB-231 cells invaded more under the serum gradient after one day, however this behavior was inverted after 3 days. The results showcase that the microfluidic system can be used to quantitatively assess the invasion behavior of cancer cells with different E-cadherin expression, for a longer period than conventional invasion models. In the future, it can be used to quantitatively investigate effects of matrix structure and cell treatments on cancer invasion.

A numerical model to study mechanically induced initiation and progression of damage in articular cartilage.

OBJECTIVE: Proteoglycan (PG) loss and surface roughening, early signs of osteoarthritis (OA), are likely preceded by softening of the ground substance and the collagen network. Insight in their relative importance to progression of OA may assist the development of treatment strategies for early OA. To support interpretation of experimental data, a numerical model is proposed that can predict damage progression in cartilage over time, as a consequence of excessive mechanical loading. The objective is to assess the interaction between ground substance softening and collagen fiber damage using this model. DESIGN: An established cartilage mechanics model is extended with the assumption that excessive strains may damage the ground substance or the collagen network, resulting in softening of the overstrained constituent. During subsequent loading cycles the strain may or may not cross a threshold, resulting in damage to stabilize or to progress. To evaluate how softening of the ground substance and collagen may interact, damage progression is computed when either one of them, or both together are allowed to occur during stepwise increased loading. RESULTS: Softening in the ground substance was predicted to localize in the superficial and transitional zone and resulted in cartilage softening. Collagen damage was most prominent in the superficial zone, with more diffuse damage penetrating deeper into the tissue, resulting in adverse strain gradients. Effects were more pronounced if both constituents developed damage in parallel. CONCLUSION: Ground substance softening and collagen damage have distinct effects on cartilage mechanopathology, and damage in either one of them may promote each other.

Is collagen fiber damage the cause of early softening in articular cartilage?

OBJECTIVE: Because collagen damage and cartilage softening have not yet been determined simultaneously in one study for the very early onset of osteoarthritis (OA), it remains questionable whether they are associated. The aim of the present study is therefore to evaluate whether indeed, initial collagen damage can be found when tissue softening occurs as a result of excessive mechanical loading. METHODS: To investigate this aim, a series of specific indentation loading protocols were designed to induce and monitor cartilage softening in osteochondral explants of bovine carpometacarpal joints. The experiment contained one control group (n = 6) in which no damage was induced and four experimental groups in which samples received either a constant load of 3 (n = 5), 6 (n = 5) or 15 N (n = 6), or an increasing load (n = 7) from 2 to 13 N in 11 steps. Moreover, to determine mechanically induced collagen damage, Col2-3/4M (cumulative collagen damage) and Col2-3/4C(short) (only enzymatic damage) staining were compared. RESULTS: The normalized peak and equilibrium reaction forces decreased in the groups that received increasing and 15 N peak loading. However, Col2-3/4M staining was negative in all samples, while enzymatic damage (Col2-3/4C(short)) appeared similar in experiments and in unloaded control groups. CONCLUSION: It was shown that a loading magnitude threshold exists above which softening occurs in cartilage. However, in samples that did show softening, we were unable to detect collagen damage. Thus, our results demonstrate that cartilage softening most likely precedes collagen damage.

Alterations to the subchondral bone architecture during osteoarthritis: bone adaptation vs endochondral bone formation.

OBJECTIVE: Osteoarthritis (OA) is characterized by loss of cartilage and alterations in subchondral bone architecture. Changes in cartilage and bone tissue occur simultaneously and are spatially correlated, indicating that they are probably related. We investigated two hypotheses regarding this relationship. According to the first hypothesis, both wear and tear changes in cartilage, and remodeling changes in bone are a result of abnormal loading conditions. According to the second hypothesis, loss of cartilage and changes in bone architecture result from endochondral ossification. DESIGN: With an established bone adaptation model, we simulated adaptation to high load and endochondral ossification, and investigated whether alterations in bone architecture between these conditions were different. In addition, we analyzed bone structure differences between human bone samples with increasing degrees of OA, and compared these data to the simulation results. RESULTS: The simulation of endochondral ossification led to a more refined structure, with a higher number of trabeculae in agreement with the finding of a higher trabecular number in osteochondral plugs with severe OA. Furthermore, endochondral ossification could explain the presence of a "double subchondral plate" which we found in some human bone samples. However, endochondral ossification could not explain the increase in bone volume fraction that we observed, whereas adaptation to high loading could. CONCLUSION: Based on the simulation and experimental data, we postulate that both endochondral ossification and adaptation to high load may contribute to OA bone structural changes, while both wear and tear and the replacement of mineralized cartilage with bone tissue may contribute cartilage thinning.

Friction reducing ability of a poly-l-lysine and dopamine modified hyaluronan coating for polycaprolactone cartilage resurfacing implants.

Frictional properties of cartilage resurfacing implants should be sufficiently low to limit damaging of the opposing cartilage during articulation. The present study determines if native lubricious molecule proteoglycan 4 (PRG4) can adsorb onto a layer-by-layer bioinspired coating composed of poly-l-lysine (PLL) and dopamine modified hyaluronic acid (HADN) and thereby can reduce the friction between implant and articular cartilage. An ELISA was developed to quantify the amount of immobilized human recombinant (rh)PRG4 after exposure to the PLL-HADN coating. The effect on lubrication was evaluated by comparing the coefficient of friction (CoF) of bare polycaprolactone (PCL) disks to that of PLL-HADN coated PCL disks while articulated against cartilage using a ring-on-disk geometry and a lubricant solution consisting of native synovial fluid components including rhPRG4. The PLL-HADN coating effectively immobilized rhPRG4. The surface roughness of PCL disks significantly increased while the water contact angle significantly decreased after application of the coating. The average CoF measured during the first minute of bare PCL against cartilage exceeded twice the CoF of the PLL-HADN coated PCL against cartilage. After 60 min, the CoF reached equilibrium values which were still significantly higher for bare PCL compared to coated PCL. The present study demonstrated that PCL can effectively be coated with PLL-HADN. Additionally, this coating reduces the friction between PCL and cartilage when a PRG4-rich lubricant is used, similar to the lubricating surface of native cartilage. This makes PLL-HADN coating a promising application to improve the clinical success of PCL-based cartilage resurfacing implants.

Local variations in mechanical properties of human hamstring tendon autografts for anterior cruciate ligament reconstruction do not translate to a mechanically inferior strand.

A ruptured anterior cruciate ligament (ACL) is often reconstructed with a multiple-strand autograft of a semitendinosus tendon alone or combined with a gracilis tendon. Up to 10% of patients experience graft rupture. This potentially results from excessive local tissue strains under physiological loading which could either result in direct mechanical failure of the graft or induce mechanobiological weakening. Since the original location in the hamstring tendon cannot be traced back from an autograft rupture site, this study explored whether clinical outcome could be further improved by avoiding specific locations or regions of human semitendinosus and/or gracilis tendons in ACL grafts due to potential mechanical or biochemical inferiority. Additionally, it examined numerically which clinically relevant graft configurations experience the lowest strains - and therefore the lowest rupture risk - when loaded with equal force. Remnant full-length gracilis tendons from human ACL reconstructions and full-length semitendinosus- and ipsilateral gracilis tendons of human cadaveric specimens were subjected to a stress-relaxation test. Locations at high risk of mechanical failure were identified using particle tracking to calculate local axial strains. As biochemical properties, the water-, collagen-, glycosaminoglycan- and DNA content per tissue region (representing graft strands) were determined. A viscoelastic lumped parameter model per tendon region was calculated. These models were applied in clinically relevant virtual graft configurations, which were exposed to physiological loading. Configurations that provided lower stiffness - i.e., experiencing higher strains under equal force - were assumed to be at higher risk of failure. Suitability of the gracilis tendon proper to replace semitendinosus muscle-tendon junction strands was examined. Deviations in local axial strains from the globally applied strain were of similar magnitude as the applied strain. Locations of maximum strains were uniformly distributed over tendon lengths. Biochemical compositions varied between tissue regions, but no trends were detected. Viscoelastic parameters were not significantly different between regions within a tendon, although semitendinosus tendons were stiffer than gracilis tendons. Virtual grafts with a full-length semitendinosus tendon alone or combined with a gracilis tendon displayed the lowest strains, whereas strains increased when gracilis tendon strands were tested for their suitability to replace semitendinosus muscle-tendon junction strands. Locations experiencing high local axial strains - which could increase risk of rupture - were present, but no specific region within any of the investigated graft configurations was found to be mechanically or biochemically deviant. Consequently, no specific tendon region could be indicated to provide a higher risk of rupture for mechanical or biochemical reasons. The semitendinosus tendon provided superior stiffness to a graft compared to the gracilis tendon. Therefore, based on our results it would be recommended to use the semitendinosus tendon, and use the gracilis tendon in cases where further reinforcement of the graft is needed to attain the desired length and cross-sectional area. All these data support current clinical standards.

Bone structural changes in osteoarthritis as a result of mechanoregulated bone adaptation: a modeling approach.

OBJECTIVE: There are strong indications that subchondral bone may play an important role in osteoarthritis (OA), making it an interesting target for medical therapies. The subchondral bone structure changes markedly during OA, and it has long been assumed that this occurs secondary to cartilage degeneration. However, for various conditions that are associated with OA, it is known that they may also induce bone structural changes in the absence of cartilage degeneration. We therefore aimed to investigate if OA bone structural changes can result from mechanoregulated bone adaptation, independent of cartilage degeneration. METHOD: With a bone adaptation model, we simulated various conditions associated with OA -without altering the articular cartilage- and we evaluated if mechanoregulated bone remodeling by itself could lead to OA-like bone structural changes. RESULTS: For each of the conditions, the predicted changes in bone structural parameters (bone fraction, trabecular thickness, trabecular number, and trabecular separation) were similar to those observed in OA. CONCLUSION: This indicates that bone adaptation in OA can be mechanoregulated with structural changes occurring independent of cartilage degeneration.

Tuning the differentiation of periosteum-derived cartilage using biochemical and mechanical stimulations.

OBJECTIVE: In this study, we aim at tuning the differentiation of periosteum in an organ culture model towards cartilage, rich in collagen type II, using combinations of biochemical and mechanical stimuli. We hypothesize that addition of TGF-ß will stimulate chondrogenesis, whereas sliding indentation will enhance collagen synthesis. DESIGN: Periosteum was dissected from the tibiotarsus of 15-day-old chick embryos. Explants were embedded in between two agarose layers, and cultured without stimulation (control), with biochemical stimulation (10 ng/ml TGF-ß1), with mechanical stimulation (sliding indentation), or both biochemical and mechanical stimulations. Sliding indentation was introduced as a method to induce tensile tissue strain. Analysis included quantification of DNA, collagen and GAG content, conventional histology, and immunohistochemistry for collagen type I and II at 1 or 2 weeks of culture. RESULTS: Embedding the periosteal explants in between agarose layers induced cartilage formation, confirmed by synthesis of sGAG and collagen type II. Addition of TGF-ß1 to the culture medium did not further enhance this chondrogenic response. Applying sliding indentation only to the periosteum in between agarose layers enhanced the production of collagen type I, leading to the formation of fibrous tissue without any evidence of cartilage formation. However, when stimulated by both TGF-ß1 and sliding indentation, collagen production was still enhanced, but now collagen type II, while sGAG was found to be similar to TGF-ß1 or unloaded samples. CONCLUSIONS: The type of tissue produced by periosteal explants can be tuned by combining mechanical stimulation and soluble factors. TGF-ß1 stimulated a chondrocyte phenotype and sliding indentation stimulated collagen synthesis. Such a combination may be valuable for improvement of the quality of tissue-engineered cartilage.

The performance of resurfacing implants for focal cartilage defects depends on the degenerative condition of the opposing cartilage.

BACKGROUND: Non-degradable resurfacing implants are being developed for treatment of focal cartilage defects. Performance of these implants has been investigated opposing intact cartilage. This study investigates whether implants would perform equally well when the opposing cartilage is fibrillated. METHODS: Human osteochondral strips (~2x1x1 cm) with a smooth (n = 9) or fibrillated (n = 17) cartilage surface were obtained from human tibial plateaus excised during total knee arthroscopy. A custom-made pin-on-plate sliding indenter was used to apply simultaneous compression (0.75-3 MPa) and movement (4 mm/s over 6 mm). Either metal implants, polycarbonate-urethane or healthy porcine osteochondral plugs with a diameter of 6 mm were used as indenter. FINDINGS: Cartilage roughness of the osteochondral strips was significantly higher for the fibrillated than the smooth group prior to sliding-indentation. Roughness of the indenters was not significantly altered by sliding indentation using either smooth or fibrillated cartilage. For all but one sample, sliding of smooth cartilage against any of the indenter surfaces did not cause damage. However, samples with fibrillated cartilage showed varied responses from seemingly unaffected to severe tissue wear as quantified by analysis of Indian ink staining and histology. INTERPRETATION: This study demonstrates that the opposing cartilage quality is relevant for the clinical success of implanting an artificial implant in a focal cartilage defect. Therefore it is essential to test the efficacy of newly developed implants against arthritic joint surfaces, and care should be taken when interpreting in vivo studies in which implants are inserted in healthy joints.

The PTHrP-Ihh feedback loop in the embryonic growth plate allows PTHrP to control hypertrophy and Ihh to regulate proliferation.

Growth plate and long bone development is governed by biochemical signaling pathways of which the PTHrP-Ihh system is the best known. Other factors, such as BMPs, FGFs and mechanical loading, may interact with this system. This study aims at elucidating the relative importance of PTHrP and Ihh for controlling proliferation, and hypertrophy in fetal growth plate cartilage. We assessed the question why reduced Ihh expression leads to more pronounced effects on the number of non-hypertrophic cells and total bone formation, compared to PTHrP down-regulation. Using few basic equations, constituted from literature data, this paper shows how the PTHrP-Ihh feedback system can control different aspects of tissue differentiation at distinct locations. In particular, it is shown that (mechanical or biochemical) perturbations will affect proliferation via Ihh-related parameters, whereas changes in PTHrP-related parameters selectively interact with hypertrophy. This is contra-intuitive, since PTHrP acts to keep cells proliferating. In this context, the critical PTHrP level for keeping cells proliferating has been reconsidered. In addition, an explanation is provided for the aforementioned difference in effect between reduced Ihh and PTHrP expression.

Depth-dependent compressive equilibrium properties of articular cartilage explained by its composition.

For this study, we hypothesized that the depth-dependent compressive equilibrium properties of articular cartilage are the inherent consequence of its depth-dependent composition, and not the result of depth-dependent material properties. To test this hypothesis, our recently developed fibril-reinforced poroviscoelastic swelling model was expanded to include the influence of intra- and extra-fibrillar water content, and the influence of the solid fraction on the compressive properties of the tissue. With this model, the depth-dependent compressive equilibrium properties of articular cartilage were determined, and compared with experimental data from the literature. The typical depth-dependent behavior of articular cartilage was predicted by this model. The effective aggregate modulus was highly strain-dependent. It decreased with increasing strain for low strains, and increases with increasing strain for high strains. This effect was more pronounced with increasing distance from the articular surface. The main insight from this study is that the depth-dependent material behavior of articular cartilage can be obtained from its depth-dependent composition only. This eliminates the need for the assumption that the material properties of the different constituents themselves vary with depth. Such insights are important for understanding cartilage mechanical behavior, cartilage damage mechanisms and tissue engineering studies.

The critical size of focal articular cartilage defects is associated with strains in the collagen fibers.

The size of full-thickness focal cartilage defect is accepted to be predictive of its fate, but at which size threshold treatment is required is unclear. Clarification of the mechanism behind this threshold effect will help determining when treatment is required. The objective was to investigate the effect of defect size on strains in the collagen fibers and the non-fibrillar matrix of surrounding cartilage. These strains may indicate matrix disruption. Tissue deformation into the defect was expected, stretching adjacent superficial collagen fibers, while an osteochondral implant was expected to prevent these deformations. Finite element simulations of cartilage/cartilage contact for intact, 0.5 to 8mm wide defects and 8mm implant cases were performed. Impact, a load increase to 2MPa in 1ms, and creep loading, a constant load of 0.5MPa for 900s, scenarios were simulated. A composition-based material model for articular cartilage was employed. Impact loading caused low strain levels for all models. Creep loading increased deviatoric strains and collagen strains in the surrounding cartilage. Deviatoric strains increased gradually with defect size, but the surface area at which collagen fiber strains exceeded failure thresholds, abruptly increased for small increases of defect size. This was caused by a narrow distribution of collagen fiber strains resulting from the non-linear stiffness of the fibers. We postulate this might be the mechanism behind the existence of a critical defect size. Filling of the defect with an implant reduced deviatoric and collagen fiber strains towards values for intact cartilage.

The effect of loading rate on the development of early damage in articular cartilage.

Experimental reports suggest that cartilage damage depends on strain magnitude. Additionally, because of its poro-viscoelastic nature, strain magnitude in cartilage can depend on strain rate. The present study explores whether cartilage damage may develop dependent on strain rate, even when the presented damage numerical model is strain-dependent but not strain-rate-dependent. So far no experiments have been distinguished whether rate-dependent cartilage damage occurs in the collagen or in the non-fibrillar network. Thus, this research presents a finite element analysis model where, among others, collagen and non-fibrillar matrix are incorporated as well as a strain-dependent damage mechanism for these components. Collagen and non-fibrillar matrix stiffness decrease when a given strain is reached until complete failure upon reaching a maximum strain. With such model, indentation experiments at increasing strain rates were simulated on cartilage plugs and damage development was monitored over time. Collagen damage increased with increasing strain rate from 21 to 42 %. In contrast, damage in the non-fibrillar matrix decreased with increasing strain rates from 72 to 34 %. Damage started to develop at a depth of approximately 20 % of the sample height, and this was more pronounced for the slow and modest loading rates. However, the most severe damage at the end of the compression step occurred at the surface for the plugs subjected to 120 mm/min strain rate. In conclusion, the present study confirms that the location and magnitude of damage in cartilage may be strongly dependent on strain rate, even when damage occurs solely through a strain-dependent damage mechanism.

Relative contribution of articular cartilage's constitutive components to load support depending on strain rate.

Cartilage is considered a biphasic material in which the solid is composed of proteoglycans and collagen. In biphasic tissue, the hydraulic pressure is believed to bear most of the load under higher strain rates and its dissipation due to fluid flow determines creep and relaxation behavior. In equilibrium, hydraulic pressure is zero and load bearing is transferred to the solid matrix. The viscoelasticity of the collagen network also contributes to its time-dependent behavior, and the osmotic pressure to load bearing in equilibrium. The aim of the present study was to determine the relative contributions of hydraulic pressure, viscoelastic collagen stress, solid matrix stiffness and osmotic pressure to load carriage in cartilage under transient and equilibrium conditions. Unconfined compression experiments were simulated using a fibril-reinforced poroviscoelastic model of articular cartilage, including water, fibrillar viscoelastic collagen and non-fibrillar charged glycosaminoglycans. The relative contributions of hydraulic and osmotic pressures and stresses in the fibrillar and non-fibrillar network were evaluated in the superficial, middle and deep zone of cartilage under five different strain rates and after relaxation. Initially upon loading, the hydraulic pressure carried most of the load in all three zones. The osmotic swelling pressure carried most of the equilibrium load. In the surface zone, where the fibers were loaded in tension, the collagen network carried 20 % of the load for all strain rates. The importance of these fibers was illustrated by artificially modifying the fiber architecture, which reduced the overall stiffness of cartilage in all conditions. In conclusion, although hydraulic pressure dominates the transient behavior during cartilage loading, due to its viscoelastic nature the superficial zone collagen fibers carry a substantial part of the load under transient conditions. This becomes increasingly important with higher strain rates. The interesting and striking new insight from this study suggests that under equilibrium conditions, the swelling pressure generated by the combination of proteoglycans and collagen reinforcement accounts cartilage stiffness for more than 90 % of the loads carried by articular cartilage. This finding is different from the common thought that load is transferred from fluid to solid and is carried by the aggregate modulus of the solid. Rather, it is transformed from hydraulic to osmotic swelling pressure. These results show the importance of considering both (viscoelastic) collagen fibers as well as swelling pressure in studies of the (transient) mechanical behavior of cartilage.

Can the growth factors PTHrP, Ihh and VEGF, together regulate the development of a long bone?

Endochondral ossification is the process of differentiation of cartilaginous into osseous tissue. Parathyroid hormone related protein (PTHrP), Indian hedgehog (Ihh) and vascular endothelial growth factor (VEGF), which are synthesized in different zones of the growth plate, were found to have crucial roles in regulating endochondral ossification. The aim of this study was to evaluate whether the three growth factors PTHrP, Ihh and VEGF, together, could regulate longitudinal growth in a normal human, fetal femur. For this purpose, a one-dimensional finite element (FE) model, incorporating growth factor signaling, was developed of the human, distal, femoral growth plate. It included growth factor synthesis in the relevant zones, their transport and degradation and their effects. Simulations ran from initial hypertrophy in the center of the bone until secondary ossification starts at approximately 3.5 months postnatal. For clarity, we emphasize that no mechanical stresses were considered. The FE model showed a stable growth plate in which the bone growth rate was constant and the number of cells per zone oscillated around an equilibrium. Simulations incorporating increased and decreased PTHrP and Ihh synthesis rates resulted, respectively, in more and less cells per zone and in increased and decreased bone growth rates. The FE model correctly reflected the development of a growth plate and the rate of bone growth in the femur. Simulations incorporating increased and decreased PTHrP and Ihh synthesis rates reflected growth plate pathologies and growth plates in PTHrP-/- and Ihh-/- mice. The three growth factors, PTHrP, Ihh and VEGF, could potentially together regulate tissue differentiation.

Computational study of culture conditions and nutrient supply in cartilage tissue engineering.

Different culture conditions for cartilage tissue engineering were evaluated with respect to the supply of oxygen and glucose and the accumulation of lactate. A computational approach was adopted in which the culture configurations were modeled as a batch process and transport was considered within constructs seeded at high cell concentrations and of clinically relevant dimensions. To assess the extent to which mass transfer can be influenced theoretically, extreme cases were distinguished in which the culture medium surrounding the construct was assumed either completely static or well mixed and fully oxygenated. It can be concluded that severe oxygen depletion and lactate accumulation can occur within constructs for cartilage tissue engineering. However, the results also indicate that transport restrictions are not insurmountable, providing that the medium is well homogenized and oxygenated and the construct's surfaces are sufficiently exposed to the medium. The large variation in uptake rates of chondrocytes indicates that for any specific application the quantification of cellular utilization rates, depending on the cell source and culture conditions, is an essential starting point for optimizing culture protocols.

A fibril-reinforced poroviscoelastic swelling model for articular cartilage.

From a mechanical point of view, the most relevant components of articular cartilage are the tight and highly organized collagen network together with the charged proteoglycans. Due to the fixed charges of the proteoglycans, the cation concentration inside the tissue is higher than in the surrounding synovial fluid. This excess of ion particles leads to an osmotic pressure difference, which causes swelling of the tissue. The fibrillar collagen network resists straining and swelling pressures. This combination makes cartilage a unique, highly hydrated and pressurized tissue, enforced with a strained collagen network. Many theories to explain articular cartilage behavior under loading, expressed in computational models that either include the swelling behavior or the properties of the anisotropic collagen structure, can be found in the literature. The most common tests used to determine the mechanical quality of articular cartilage are those of confined compression, unconfined compression, indentation and swelling. All theories currently available in the literature can explain the cartilage response occurring in some of the above tests, but none of them can explain these for all of the tests. We hypothesized that a model including simultaneous mathematical descriptions of (1) the swelling properties due to the fixed-change densities of the proteoglycans and (2) the anisotropic viscoelastic collagen structure, can explain all these test simultaneously. To study this hypothesis we extended our fibril-reinforced poroviscoelastic finite element model with our biphasic swelling model. We have shown that the newly developed fibril-reinforced poroviscoelastic swelling (FPVES) model for articular cartilage can simultaneously account for the reaction force during swelling, confined compression, indentation and unconfined compression as well as the lateral deformation during unconfined compression. Using this theory it is possible to analyze the link between the collagen network and the swelling properties of articular cartilage.

The role of computational models in the search for the mechanical behavior and damage mechanisms of articular cartilage.

Articular cartilage plays a vital role in the function of diarthrodial joints. Due to osteoarthritis degeneration of articular cartilage occurs. The initial event that triggers the pathological process of cartilage degeneration is still unknown. Cartilage damage due to osteoarthritis is believed to be mechanically induced. Hence, to investigate the initiation of osteoarthritis the stresses and strains in the cartilage must be determined. So far the most common method to accomplish that is finite element analysis. This paper provides an overview of computational descriptions developed for this purpose, and what they can be used for. Articular cartilage composition and structure are discussed in relation with degenerative changes, and how these affect mechanical properties.