Spatial and temporal changes of subchondral bone proceed to articular cartilage degeneration in rats subjected to knee immobilization

This study was aimed to investigate the spatial and temporal changes of subchondral bone and its overlying articular cartilage in rats following knee immobilization. A total of 36 male Wistar rats (11–13 months old) were assigned randomly and evenly into 3 groups. For each group, knee joints in 6 rats were immobilized unilaterally for 1, 4, or 8 weeks, respectively, while the remaining rats were allowed free activity and served as external control groups. For each animal, femurs at both sides were dissected after sacrificed. The distal part of femur was examined by micro‐CT. Subsequently, femoral condyles were collected for further histological observation and analysis. For articular cartilage, significant changes were observed only at 4 and 8 weeks of immobilization. The thickness of articular cartilage and chondrocytes numbers decreased with time. However, significant changes in subchondral bone were defined by micro‐CT following immobilization in a time‐dependent manner. Immobilization led to a thinner and more porous subchondral bone plate, as well as a reduction in trabecular thickness and separation with a more rod‐like architecture. Changes in subchondral bone occurred earlier than in articular cartilage. More importantly, immobilization‐induced changes in subchondral bone may contribute, at least partially, to changes in its overlying articular cartilage. Microsc. Res. Tech. 79:209–218, 2016. © 2016 Wiley Periodicals, Inc.     

A viscoelastic analysis of the tensile weakening of deep femoral head articular cartilage

Articular cartilage from below the surface of the femoral head of the hip joint shows a profound age-dependent weakening in its tensile mechanical properties. This ageing is also associated with a reduced viscoelastic response in the older tissue. A constitutive model of the viscoelastic behaviour of deep articular cartilage (as discussed by Egan in 1988) is used to generate a graphical pattern which represents the mechanical behaviour. This constitutive approach suggests that the tensile weakening of the older cartilage is due to an age-related reduction in the recruitment of load-carrying structures as the tissue is deformed. The viscoelastic constitutive model also predicts a reduction in the tensile strength of deep articular cartilage with rate of deformation. This prediction is supported by experimental fracture stress data. A weakening of the tensile integrity of the microstructure of articular cartilage could make the tissue less able to sustain normal compressive physiological loading without damage and thus make the tissue more susceptible to osteoarthritic degeneration. The constitutive approach indicates that the weakening of the older tissue may be related to changes within the microstructure which determine how applied mechanical energy is stored and dissipated.     

Development of artificial articular cartilage

Attempts have been made to develop an artificial articular cartilage on the basis of a new viewpoint of joint biomechanics in which the lubrication and load-bearing mechanisms of natural and artificial joints are compared. Polyvinyl alcohol hydrogel (PVA-H), 'a rubber-like gel', was investigated as an artificial articular cartilage and the mechanical properties of this gel were improved through a new synthetic process. In this article the biocompatibility and various mechanical properties of the new improved PVA-H is reported from the perspective of its usefulness as an artificial articular cartilage. As regards lubrication, the changes in thickness and fluid pressure of the gap formed between a glass plate and the specimen under loading were measured and it was found that PVA-H had a thicker fluid film under higher pressures than polyethylene (PE) did. The momentary stress transmitted through the specimen revealed that PVA-H had a lower peak stress and a longer duration of sustained stress than PE, suggesting a better damping effect. The wear factor of PVA-H was approximately five times that of PE. Histological studies of the articular cartilage and synovial membranes around PVA-H implanted for 8-52 weeks showed neither inflammation nor degenerative changes. The artificial articular cartilage made from PVA-H could be attached to the underlying bone using a composite osteochondral device made from titanium fibre mesh. In the second phase of this work, the damage to the tibial articular surface after replacement of the femoral surface in dogs was studied. Pairs of implants made of alumina, titanium or PVA-H on titanium fibre mesh were inserted into the femoral condyles. The two hard materials caused marked pathological changes in the articular cartilage and menisci, but the hydrogel composite replacement caused minimal damage. The composite osteochondral device became rapidly attached to host bone by ingrowth into the supporting mesh. The clinical implications of the possible use of this material in articular resurfacing and joint replacement are discussed.     

Comparison of human and animal femoral head chondral properties and geometries

Investigations into tissue-preserving orthopaedic treatments should consider the tribology of articular cartilage; where simulations using animal joints are a predominant choice. However, very few studies have investigated the differences between human and animal cartilage. The aim of the present study was to characterise the differences in geometry and mechanical properties of human, porcine, bovine and ovine articular cartilage. Creep indentation was performed on osteochondral plugs taken from the superior region of femoral heads of all these species. Cartilage thickness was measured via the resistive force change of a needle descending through cartilage and bone. A biphasic finite element model was used to derive equilibrium elastic modulus and permeability. Results showed that human cartilage was significantly thicker than all other species tested. A positive correlation was found between femoral head diameter and cartilage thickness when comparing between species of quadrupeds. Human cartilage had the largest equilibrium elastic modulus, which was significant when comparing against porcine and bovine. However, porcine cartilage had significantly lower permeability. Significant differences in geometry and mechanical properties of articular cartilage were found between all species tested. It is necessary to consider these variations when choosing animal tissue to represent human.     

Application of design‐based stereology for estimation of absolute volume and surface area of the articular and calcified cartilage compartments of undecalcified human femoral heads

Design‐based stereological methods using systematic uniform random sampling, the Cavalieri estimator and vertical sections are used to investigate undecalcified human femoral heads. Ten entire human femoral heads, obtained from normal women and normal men, were systematically sampled and thin undecalcified vertical sections were obtained. Absolute volumes and surface areas of the entire femoral head, the articular cartilage and the calcified cartilage compartments were estimated. In addition, the average thickness of the articular cartilage and the calcified cartilage were calculated. The stereological procedures applied to the human femoral heads resulted in average coefficient of errors, which were 0.03–0.06 for the volume estimates and 0.03–0.04 for the surface area estimates. We conclude that design‐based stereology using the Cavalieri estimator and vertical sections can successfully be used in large undecalcified tissue specimens, like the human femoral head, to estimate the absolute volume and surface area of macroscopic as well as of microscopic tissue compartments. The application of well‐known design‐based stereological methods carries potential advantage for investigating the pathology in inflammatory and degenerative joint diseases.     

关节软骨手术切除机理研究现状

在关节软骨切除机理和临床医学手术效果评价等方面,采用的研究方法不同,对同一问题所得出的结论也不完全一致,各评价指标尚未有明确的统一标准。因此,充分了解手术器械对关节软骨的切除机理,有助于对关节软骨损伤进行合理治疗,并减少手术过程中手术刀具对其造成的二次损伤。本文在总结关节软骨生物力学特性的基础上,综述了软骨切除机理及损伤评估以及手术中软骨与不锈钢刀具间的摩擦磨损问题的研究进展,指出手术切除病变关节软骨相关研究存在的主要问题。     

Design, validation, and utilization of an articular cartilage impact instrument

This paper describes the development and use of an instrument mechanically to impact bovine articular cartilage and record the event using a piezoelectric accelerometer, as well as to carry out post-impact characterization of the tissue. Two levels of impact (low: 6 cm drop height, 18.4 N tup; high: 10 cm drop height, 27.8 N tup) were chosen such that the former did not show gross damage upon inspection, while the latter showed substantial gross damage. Peak stress, time to peak stress, and impact duration were taken from data recorded by the instrument. Three cartilage biomechanical properties (aggregate modulus, Poisson's ratio, and permeability) were acquired by creep indentation, and tissue morphology rated on a standardized scale was also determined. When subjected to the high level of impact, articular cartilage showed statistically significant (p < 0.05) differences in all three impact metrics and morphology. This high level of impact also resulted in a 37 per cent decrease in the aggregate modulus of the tissue. Lower drop heights resulted in more consistent impact curves, demonstrated less standard deviation, and did not change the biomechanical properties of the tissues. With the instrument and techniques described in this study, articular cartilage can be subjected to specific levels of impact in order to study injury biomechanics of the tissue at specific levels of mechanical damage.     

A finite element model of an idealized diarthrodial joint to investigate the effects of variation in the mechanical properties of the tissues

The stiffness of articular cartilage increases dramatically with increasing rate of loading, and it has been hypothesized that increasing the stiffness of the subchondral bone may result in damaging stresses being generated in the articular cartilage. Despite the interdependence of these tissues in a joint, little is understood of the effect of such changes in one tissue on stresses generated in another. To investigate this, a parametric finite element model of an idealized joint was developed. The model incorporated layers representing articular cartilage, calcified cartilage, the subchondral bone plate and cancellous bone. Taguchi factorial design techniques, employing a two-level full-factorial and a four-level fractional factorial design, were used to vary the material properties and thicknesses of the layers over the wide range of values found in the literature. The effects on the maximum values of von Mises stress in each of the tissues are reported here. The stiffness of the cartilage was the main factor that determined the stress in the articular cartilage. This, and the thickness of the cartilage, also had the largest effect on the stresses in all the other tissues with the exception of the subchondral bone plate, in which stresses were dominated by its own stiffness. The stiffness of the underlying subchondral bone had no effect on the stresses generated in the cartilage. This study shows how stresses in the various tissues are affected by changes in their mechanical properties and thicknesses. It also demonstrates the benefits of a structured, systematic approach to investigating parameter variation in finite element models.     

Mechanical anisotropy of the human knee articular cartilage in compression

Articular cartilage exhibits anisotropic mechanical properties when subjected to tension. However, mechanical anisotropy of mature cartilage in compression is poorly known. In this study, both confined and unconfined compression tests of cylindrical cartilage discs, taken from the adult human patello-femoral groove and cut either perpendicular (normal disc) or parallel (tangential disc) to the articular surface, were utilized to determine possible anisotropy in Young's modulus, E, aggregate modulus, Ha, Poisson's ratio, v and hydraulic permeability, k, of articular cartilage. The results indicated that Ha was significantly higher in the direction parallel to the articular surface as compared with the direction perpendicular to the surface (Ha = 1.237 +/- 0.486 MPa versus Ha = 0.845 +/- 0.383 MPa, p = 0.017, n = 10). The values of Poisson's ratio were similar, 0.158 +/- 0.148 for normal discs compared with 0.180 +/- 0.046 for tangential discs. Analysis using the linear biphasic model revealed that the decrease of permeability during the offset compression of 0-20 per cent was higher (p = 0.015, n = 10) in normal (from 25.5 x 10(-15) to 1.8 x 10(-15) m4/N s) than in tangential (from 12.3 x 10(-15) to 1.3 x 10(-15) m4/N s) discs. Based on the results, it is concluded that the mechanical characteristics of adult femoral groove articular cartilage are anisotropic also during compression. Anisotropy during compression may be essential for normal cartilage function. This property has to be considered when developing advanced theoretical models for cartilage biomechanics.     

Influence of proteoglycan contents and of tissue hydration on the frictional characteristics of articular cartilage

In this work, the hypothesis that water content and substances present on the articular surface play an important role in lubrication through the formation of a layer with a high content of water on the articular surface is analysed. The hydrophilic properties of proteoglycans exposed at the articular surface and hydration of tissue are the main responsible factors for the formation of this layer. The role of the articular surface in the frictional characteristics of articular cartilage was examined using specimens (femoral condyles of pigs) with intact and wiped surfaces tested in intermittent friction tests. Results indicated that the intact condition presented low friction in comparison with the wiped condition. The measured water loss of the articular cartilage after sliding and loading indicated a gradual decrease in the water content as the time evolved, and rehydration was observed after the submersion of unloaded specimens in the saline bath solution. Micrographic analyses indicated the presence of a layer covering the articular surface, and histological analyses indicated the presence of proteoglycans in this superficial layer. The hydration of the cartilage surface layer and proteoglycan in this layer influence lubrication.     

Alteration of cartilage matrix morphology with histological processing

An interlacunar network in the extracellular matrix of femoral head articular cartilage of neontal rats was seen by light microscopy to: (1) consist of elements, 0·5 μm thick, which occurred as individual elements, as bundles of elements, and as fused elements, (2) stain intensely with toluidine blue, methylene blue, and safranin O, and (3) connect chondrocytes by inserting on the chondrocyte capsules which were composed of morphologically and cytochemically similar material. By electron microscopy, the single elements were seen to be composed of thicker, denser staining areas of the honeycomb appearing matrix and the fused elements appeared as non-membrane bound channels containing granular material. Articular cartilage was processed using combinations of fixatives, dehydrating agents, and embedding media. Regardless of fixation, demineralization, or embedding, the network was not seen after dehydration of the cartilage with methanol, ethanol, acetone or tert-butanol but was seen after dehydration with aqueous solutions of glycol methacrylate, propylene oxide, 2-propanol or 2,2-dimethoxypropane. Network visualization following a variety of methods demonstrated that no single fixative, dehydrating agent, or embedding medium caused its formation. The presence of the network in different cartilage zones, its consistent morphology by light and electron microscopy, the uniformity of the elements in their connection with the chondrocytes, and presence in fresh-frozen sections suggest the network may be real, but rigorous evidence for its existence in vivo is still required. Since cartilage morphology was altered by histological methods, especially dehydration, common methods used in studying connective tissue matrix should be evaluated to determine their effect on matrix morphology.     

The development of a model for in vitro testing of femoral stems in impaction bone grafting

Comprehension of the biomechanical behaviour of orthopaedic implants is essential. This paper describes the development of an in vitro model to investigate the behaviour of femoral implants in the revision setting. The development of a femoral model and a bone graft substitute is described. The properties of human, bovine, ovine morselized bone graft, and a graft substitute were compared. On measuring hoop strain after impaction bone grafting there was no significant difference between the ovine bone graft and graft substitute with the size 1 Exeter stem. The results suggest that this bone graft substitute is a viable alternative for in vitro testing. The authors recommend the use of the graft substitute and the femoral model to predict femoral stem biomechanics.     

An investigation of the performance of Biostop G and Hardinge bone plugs.

Although distal plugging is a common procedure to prevent distal flow of polymethyl-methacrylate (PMMA) cement during cementing of femoral prostheses, there is little biomechanical testing to confirm that (a) the plugs do not displace under cementing pressure, and (b) they do in fact occlude cement flow. Two designs of femoral intramedullary plugs, the Biostop G (Bioland, France) and Hardinge (De Puy, Leeds, UK) were examined to determine their performance under cement pressurization in a biomechanical test. A testing rig was fabricated in which distal migration could be measured as a function of cement pressurization. Sectioning of the samples after polymerization of the cement revealed the extent of cement flow. The results show that, even in this well controlled test, there is significant variability in plug performance. It is shown that the Biostop G displaces less than the Hardinge for similar cement pressures. Sectioning reveals that cement can escape around the Hardinge plug at high pressures. Furthermore, a pore forming effect of the Biostop G plug was occasionally observed indicating that design improvements may be possible for this plug.     

Behavior of graft and host cells in underlying subchondral bone after transplantation of osteochondral autograft

Transplantation of osteochondral autograft is widely used as a therapeutic strategy for the defect of articular cartilage. In the repair process, although underlying subchondral bone becomes necrotic and then is followed by bone reconstruction, the fate of graft and host cells during remodeling of underlying subchondral bone has not been elucidated. The objectives of this study were to establish a method to follow graft and host cells after transplantation of osteochondral autograft, and to elucidate the fate of both graft and host cells during remodeling of underlying subchondral bone. For these purposes, autologous transplantation models employing transgenic rats and wild-type rats, which were genetically identical to each other except for transgenes, were used. Two transplantation models were designed so that either the graft or the host cells had transgenes. Model I: transgenic rats were the donor, and wild-type rats were the recipient; model II: conversely, wild-type rats were the donor, and transgenic rats were the recipient. The grafted bone marrow cells and osteocytes in the trabeculae survived in the graft at 3 weeks after transplantation. Invasion of the host bone marrow cells into the graft was also found. Thus, bone marrow cells in the host as well as both bone marrow cells and osteocytes in the graft could potentially participate in the remodeling of underlying subchondral bone. Furthermore, the interface between graft and host was consisted with both graft and host derived cells. Since new bone formation was found in this space, both graft and host cells could have the potential to contribute to remodeling of underlying subchondral bone. The two models of the transplantations using the transgenic rats were found to be beneficial in following graft cells as well as host cells and in understanding their function on healing after autologous transplantation.     

Mathematical representation of articular surfaces using influence surface theory

Many mathematical techniques have been developed to determine the geometry of articular joint surfaces, because of its so importance to the study of human joint biomechanics. However, a three-dimensional geometric model of the articular joint, which is essential to solid modelling, contact area measurement, and load bearing analyses, has not been well developed. This study proposes to define the articular geometry of the distal femoral joint of the human knee. A mathematical method based on the influence surface theory of plates is established to generate representations of three-dimensional articular surfaces. A mathematical cone and the surface of the human distal femur are accurately recreated, allowing their geometric properties to be determined. Results suggest that this method can be an effective tool for representing articular surfaces.     

Response of the donor and recipient cells in mesenchymal cell transplantation to cartilage defect

To facilitate the repair of articular cartilage defects, autologous mesenchymal cells from bone marrow or periosteum were transplanted in a rabbit model. Two weeks after the transplantation of the mesenchymal cells, the whole area of the original defect was occupied by cartilage. From the deep area of the reparative cartilage, which contacted with host bone, chondrocytes became hypertrophic and the invasion of bone with vasculature started, until the replacement reached the natural junction of the host cartilage and the subchondral bone about 4 weeks after transplantation. Twelve weeks after the transplantation, the repair cartilage in the defect became a little thinner than the adjacent normal cartilage, which became a little thinner 24 weeks after the transplantation (the longest observation period in the study). Large, full-thickness defects of the weight-bearing region of the articular cartilage were repaired with hyaline-like cartilage after implantation of autologous mesenchymal cells. The repair process by mesenchymal cell transplantation was explained as follows: The donor transplanted cell differentiated into cartilage and the defects were completely filled with cartilage. Then, mesenchymal cells that entered the chondrogenic lineage rapidly progressed through this lineage to the hypertrophic state, which was then the target for erosion and vascular invasion. Although this vasculature and the newly formed bone were considered to be host-derived, there was no evidence to that effect. To prove this, suitable experimental marking of these donor cells is needed. In the case of chondrocyte transplantation, the repair cartilage maintained its thickness to the full depth of the original defect; the tissue derived from the implanted chondrocytes was not invaded by vessels or replaced by subchondral bone.     

Bioengineering activities at the New York Orthopaedic Hospital Research Laboratory.

The studies described above are representative of the Laboratory's long-standing dedication to the philosophy of multi-disciplinary research. Results from these studies will continue to provide new insights as to the function of diarthrodial joints, articular cartilage and other hydrated soft tissues. It appears reasonable to expect that in the years ahead we will have a better understanding of the mechanisms behind histomorphogenesis, growth, functional adaptation, regeneration, ageing and degeneration of connective tissues and osteoarthritis resulting from these investigations being conducted at the New York Orthopaedic Hospital Research Laboratory.     

Investigation of the friction and surface degradation of innovative chondroplasty materials against articular cartilage

Understanding the wear of the biomaterial-cartilage interface is vital for the development of innovative chondroplasty. The aim of this study was to investigate a number of biphasic materials as potential chondroplasty biomaterials. Simple geometry friction and wear studies were conducted using bovine articular cartilage pins loaded against a range of single-phase and biphasic materials. The frictions of each biomaterial was compared within simple and protein-containing lubricants. Longer-term continuous sliding tests within a protein containing lubricant were also conducted at various loading conditions to evaluate the friction and degradation for each surface. All single-phase materials showed a steady rise in friction, which was dependent on the loss of interstitial fluid load support from the opposing cartilage pin. All biphasic materials demonstrated a marked reduction in friction when compared with the single-phase materials. It is postulated that the biphasic nature of each material allowed an element of fluid load support to be maintained by fluid rehydration and expulsion. In the longer-term study, significant differences in the articular cartilage pin (surface damage) between the positive control (stainless steel) and the negative control (articular cartilage) was found. The potential biphasic chondroplasty materials produced a reduction in articular cartilage pin damage when compared with the single-phase materials. The changes in surface topography of the cartilage pin were associated with increased levels of friction achieved during the continuous wear test. The study illustrated the importance of the biphasic properties of potential chondroplasty materials, and future work will focus on the optimization of biphasic properties as well as long-term durability, such that materials will more closely mimic the biotribology of natural articular cartilage.     

The effect of glycosaminoglycan depletion on the friction and deformation of articular cartilage

Glycosaminoglycans (GAGs) have been shown to be responsible for the interstitial fluid pressurization of articular cartilage and hence its compressive stiffness and load-bearing properties. Contradictory evidence has been presented in the literature on the effect of depleting GAGs on the friction properties of articular cartilage. The aim of this study was to investigate the effect of depleting GAGs on the friction and deformation characteristics of articular cartilage under different tribological conditions. A pin-on-plate machine was utilized to measure the coefficient of friction of native and chondroitinase ABC (CaseABC)-treated articular cartilage under two different models: static (4 mm/s start-up velocity) and dynamic (4 mm/s sliding velocity; 4 mm stroke length) under a load of 25 N (0.4 MPa contact stress) and with phosphate-buffered saline as the lubricant. Indentation tests were carried out at 1 N and 2 N loads (0.14 MPa and 0.28 MPa contact stress levels) to study the deformation characteristics of both native and GAG-depleted cartilage samples. CaseABC treatment rendered the cartilage tissue soft owing to the loss of compressive stiffness and a sulphated-sugar assay confirmed the loss of GAGs from the cartilage samples. CaseABC treatment significantly increased (by more than 50 per cent) the friction levels in the dynamic model (p < 0.05) at higher loading times owing to the loss of biphasic lubrication. CaseABC treatment had no effect on friction in the static model in which the cartilage surfaces did not have an opportunity to recover fluid because of static loading unlike the cartilage tissue in the dynamic model, in which translation of the cartilage surfaces was involved, ensuring effective biphasic lubrication. Therefore the depletion of GAGs had a smaller effect on the coefficient of friction for the static model. Indentation tests showed that GAG-depleted cartilage samples had a lower elastic modulus and higher permeability than native tissue. These results corroborate the role of GAGs in the compressive and friction properties of articular cartilage and emphasize the need for developing strategies to control GAG loss from diseased articular cartilage tissue.     

Mechanical and tribological properties of poly(hydroxyethyl methacrylate) hydrogels as articular cartilage substitutes

Poly(hydroxyethyl methacrylate) (p(HEMA)) hydrogels have been proposed as promising biomaterials to replace damaged articular cartilage. A major obstacle to their use as replacement bearing tissue is their poor mechanical properties in comparison with healthy articular cartilage. The purpose of this study was to obtain p(HEMA) hydrogels with physicochemical and mechanical properties close to healthy articular cartilage, by introducing a hydrophilic monomer, namely acrylic acid (AA). Formulations of hydrogels with different amounts of hydrophilic monomer (acrylic acid, AA) were synthesized and tested: p(HEMA), p(HEMA-co-5%AA), p(HEMA-co-25%AA). The macro-mechanical tests were reproduced at nanoscale in order to verify if the superficial properties of the hydrogels are similar to the bulk ones.