Synovial Extracellular Vesicles: Structure and Role in Synovial Fluid Tribological Performances.

The quality of the lubricant between cartilaginous joint surfaces impacts the joint's mechanistic properties. In this study, we define the biochemical, ultrastructural, and tribological signatures of synovial fluids (SF) from patients with degenerative (osteoarthritis-OA) or inflammatory (rheumatoid arthritis-RA) joint pathologies in comparison with SF from healthy subjects. Phospholipid (PL) concentration in SF increased in pathological contexts, but the proportion PL relative to the overall lipids decreased. Subtle changes in PL chain composition were attributed to the inflammatory state. Transmission electron microscopy showed the occurrence of large multilamellar synovial extracellular vesicles (EV) filled with glycoprotein gel in healthy subjects. Synovial extracellular vesicle structure was altered in SF from OA and RA patients. RA samples systematically showed lower viscosity than healthy samples under a hydrodynamic lubricating regimen whereas OA samples showed higher viscosity. In turn, under a boundary regimen, cartilage surfaces in both pathological situations showed high wear and friction coefficients. Thus, we found a difference in the biochemical, tribological, and ultrastructural properties of synovial fluid in healthy people and patients with osteoarthritis and arthritis of the joints, and that large, multilamellar vesicles are essential for good boundary lubrication by ensuring a ball-bearing effect and limiting the destruction of lipid layers at the cartilage surface.

Nano/micro implant debris affect osteogenesis by chondrocytes: Comparison between ceramic and UHMWPE from hip walking simulator.

A new generation of ceramic on ceramic (BIOLOX ®delta) bearings has emerged more than 10 years ago proving a high resistance to wear and good clinical results. However, biological reactions to wear debris, particularly the nanoparticles, need to be evaluated. The first originality of this study is to start from real wear particles obtained by the hip walking simulator (CERsim). These particles were compared with particles obtained by usual methods to assess the biocompatibility of materials: press machine (CERpress). Two ranges of ceramic particles were thus observed: ceramic particles with micron (intergranular fractures) and nano sizes (intragranular fractures), and characterized compared to ultra-high molecular weight polyethylene (UHMWPE). The second originality of this work is to assess the cellular reaction using the primary joint chondrocyte cultures simulating the osteogenesis process and not the cell lines, which are used to simulate the biological reaction of osteolysis. The first results showed a significant difference in cell viability between the cells in contact with particles from the walking simulator and those obtained with the press machine. On the other hand, it was found that the way of extraction of the particles from the lubricant could significantly affect the biological reaction. More interestingly, nano-sized ceramic particles showed a significant impact on the secretion of functional inflammatory mediators, agreeing with recent results in vivo. These novel methods of characterizing the osteogenic impact of UHMWPE and ceramic wear debris can complement the conventional expertise method focusing previously on the osteolysis aspect.

Tribochemical Competition within a MoS2/Ti Dry Lubricated Macroscale Contact in Ultrahigh Vacuum: A Time-of-Flight Secondary Ion Mass Spectrometry Investigation.

Controlling and predicting the tribological behavior of dry lubricants is a necessity to ensure low friction, long life, and low particle generation. Understanding the tribochemistry of the materials as a function of the environment is of primary interest as synergistic effects exist between the mechanics, the physicochemistry, and the thermodynamics within a contact. However, in most studies the role of the coating internal contaminants in the process is often discarded to the benefit of a more common approach in which the performances of the materials are compared as a function of different atmospheric pressure environments. The study focuses on the understanding of the tribochemical processes occurring between the materials and their internal contaminants inside an AISI440C contact lubricated by a MoS2/Ti coating. Time-of-flight secondary ion mass spectrometry is used to study at the molecular level, the material before and after friction. Friction tests with different durations are performed in ultrahigh vacuum at the macroscale to stay relevant to the real application (space). The adsorption/desorption of gaseous species during friction is monitored by mass spectrometry to ensure reliable study of the tribochemical processes inside the contact. The study shows that a competition exists between the Ti- and MoS2-based materials to create the appropriate lubricating materials via (i) recrystallization of MoS2 materials with creation of a MoS xO y material via reactions with internal contaminants (presumably H2O), (ii) reaction of Ti-based materials with internal contaminants (mostly H2O and N2). The biphasic material created is highly similar to the one created in both humid air and dry N2 environments and providing low friction and low particle generation. However, the process is incomplete. The study thus brings insight into the possibility of controlling friction via a rational inclusion of reactants in a form of contaminants to control the tribochemical processes governing the low friction and long life.

Ultrastructural analysis of healthy synovial fluids in three mammalian species.

A better knowledge of synovial fluid (SF) ultrastructure is required to further understand normal joint lubrication and metabolism. The aim of the present study was to elucidate SF structural features in healthy joints from three mammalian species of different size compared with features in biomimetic SF. High-resolution structural analysis was performed using transmission electron microscopy (TEM) and scanning electron microscopy (SEM) and environmental SEM/wet scanning transmission electron microscopy mode complemented by TEM and SEM cryogenic methods. Laser-scanning confocal microscopy (LCM) was used to locate the main components of SF with respect to its ultrastructural organization. The present study showed that the ultrastructure of healthy SF is built from a network of vesicles with a size range from 100 to a few hundred nanometers. A multilayered organization of the vesicle membranes was observed with a thickness of about 5 nm. LCM study of biological SF compared with synthetic SF showed that the microvesicles consist of a lipid-based membrane enveloping a glycoprotein gel. Thus, healthy SF has a discontinuous ultrastructure based on a complex network of microvesicles. This finding offers novel perspectives for the diagnosis and treatment of synovial joint diseases.

Interleaflet sliding in lipidic bilayers under shear flow: comparison of the gel and fluid phases using reversed non-equilibrium molecular dynamics simulations.

The friction between two rubbing surfaces lubricated by water can be diminished if they are coated with phospholipidic bilayers or brushes of polyelectrolytes. In the case of a coating by lipid membranes, the friction is lower when the lipids are in the gel phase rather than in the liquid phase. We investigated the response of fluid or gel bilayers to a mechanical load or under shear using non-equilibrium molecular dynamics simulations (NEMD) to understand whether this difference could come from intermonolayer sliding. The system is composed of a single fully hydrated bilayer of coarse grained phospholipids under a parallel shear with vorticity parallel to the bilayer. In both the liquid and the gel phases, an intermonolayer slip was measured in the velocity profile. In the liquid phase this slip is proportional to the shear stress. In the tilted gel phase of our model the stress is not systematically linear and relaxes differently when the shear is in the direction of the tilt or perpendicular to it. The impact of surface tension (or load) on the friction is different for the liquid and gel phases, but grossly the slip remains of the same order of magnitude.

Alteration of cartilage mechanical properties in absence of ß1 integrins revealed by rheometry and FRAP analyses.

CONTEXT: Mechanical properties are essential for biological functions of the hyaline cartilage such as energy dissipation and diffusion of solutes. Mechanical properties are primarily dependent on the hierarchical organization of the two major extracellular matrix (ECM) macromolecular components of the cartilage: the fibrillar collagen network and the glycosaminoglycan (GAG)-substituted proteoglycan, mainly aggrecan, aggregates. Interaction of chondrocytes, the only cell type in the tissue, with the ECM through adhesion receptors is involved in establishing mechanical stability via bidirectional transduction of both mechanical forces and chemical signals. In this study, we aimed to determine the role of the transmembrane ß1 integrin adhesion receptors in cartilage biomechanical properties by the use of genetic modification in mice. METHODS: Costal cartilages of wild type and mutant mice lacking ß1 integrins in chondrocytes were investigated. Cartilage compressive properties and solute diffusion were characterized by rheometric analysis and Fluorescence Recovery After Photobleaching (FRAP), respectively. Cartilage tissue sections were analyzed by histology, immunohistochemistry and transmission electron microscopy (TEM). RESULTS: At the histological level, the mutant costal cartilage was characterized by chondrocyte rounding and loss of tissue polarity. Immunohistochemistry and safranin orange staining demonstrated apparently normal aggrecan and GAG levels, respectively. Antibody staining for collagen II and TEM showed comparable expression and organization of the collagen fibrils between mutant and control cartilages. Despite the lack of gross histological and ultrastructural abnormalities, rheological measurements revealed that the peak elastic modulus in compression of mutant cartilage was 1.6-fold higher than the peak elastic modulus of wild-type sample. Interestingly, the diffusion coefficient within the mutant cartilage tissue was found to be 1.2-fold lower in the extracellular space and 14-fold lower in the pericellular (PCM) space compared to control. CONCLUSION: The results demonstrate that the absence of ß1 integrins on the surface of chondrocytes increases the stiffness and modifies the diffusion properties of costal cartilage. Our data imply that ß1 integrins-mediated chondrocyte-matrix interactions directly affect cartilage biomechanics probably by modifying physical properties of individual cells. This study thus highlights the crucial role of ß1 integrins in the cartilage function.

Nanomechanical and tribological characterization of the MPC phospholipid polymer photografted onto rough polyethylene implants.

Grafting biomimetic polymers onto biomaterials such as implants is one of the promising approaches to increase their tribological performance and biocompatibility and to reduce wear. In this paper, poly(2-methacryloyloxyethyl phosphorylcholine) (p(MPC)) brushes were obtained by photografting MPC from the rough surface of ultra high molecular weight polyethylene (UHMWPE) joint implants. Such substrates have a high roughness (Ra∼650nm) which often has the same order of magnitude as the brush thickness, so it is very difficult to estimate the vertical density profile of the grafted content. The quality of the p(MPC) grafting was evaluated through a wide range of characterization techniques to reveal the effectiveness of the grafting: atomic force microcopy (AFM) imaging and force spectroscopy, contact angle, SEM/EDX, and confocal microscopy. After testing the methods on smooth glass substrate as reference, AFM nano-indentation proves to be a reliable non destructive method to characterize the thickness and the mechanical properties of the p(MPC) layer in liquid physiological medium. Tribological measurements using a homemade biotribometer confirm that, despite heterogeneity thickness (h=0.5-6µm), the p(MPC) layer covers the roughness of the UHMWPE substrate and acts as an efficient lubricant with low friction coefficient and no wear for 9h of friction.

Tenascin-X increases the stiffness of collagen gels without affecting fibrillogenesis.

Tenascin-X is an extracellular matrix protein whose absence leads to an Ehlers-Danlos Syndrome in humans, mainly characterised by connective tissue defects including the disorganisation of fibrillar networks, a reduced collagen deposition, and modifications in the mechanical properties of dense tissues. Here we tested the effect of tenascin-X on in vitro collagen fibril formation. We observed that the main parameters of fibrillogenesis were unchanged, and that the diameter of fibrils was not significantly different when they were formed in the presence of tenascin-X. Interestingly, mechanical analysis of collagen gels showed an increased compressive resistance of the gels containing tenascin-X, indicating that this protein might be directly involved in determining the mechanical properties of collagen-rich tissues in vivo.

Role of nanomechanical properties in the tribological performance of phospholipid biomimetic surfaces.

The role of phospholipid bilayers in controlling and reducing frictional forces between biological surfaces is investigated by three complementary experiments: friction forces are measured using a homemade tribometer, mechanical resistance to indentation is measured by AFM, and lipid bilayer degradation is controlled in situ during friction testing using fluorescence microscopy. DPPC lipid bilayers in the solid phase generate friction coefficients as low as 0.002 (comparable to that found for cartilage) that are stable through time. DOPC bilayers formed by the vesicle fusion method or the adsorption of mixed micelles generate higher friction coefficients. These coefficients increased through time, during which the bilayers degraded. The friction coefficient is correlated with the force needed to penetrate the bilayer with the AFM tip. With only one bilayer in the contact region, the friction increased to a similar value of about 0.08 for the DPPC and DOPC. Our study therefore shows that good mechanical stability of the bilayers is essential and suggests that the low friction coefficient is ensured by the hydration layers between adjacent lipid bilayers.