Low-field and variable-field NMR relaxation studies of H2O and D2O molecular dynamics in articular cartilage

Osteoarthritis (OA) as the main degenerative disease of articular cartilage in joints is accompanied by structural and compositional changes in the tissue. Degeneration is a consequence of a reduction of the amount of macromolecules, the so-called proteoglycans, and of a corresponding increase in water content, both leading to structural weakening of cartilage. NMR investigations of cartilage generally address only the relaxation properties of water. In this study, two-dimensional (T1-T2) measurements of bovine articular cartilage samples were carried out for different stages of hydration, complemented by molecular exchange with D2O and treatment by trypsin which simulates degeneration by OA. Two signal components were identified in all measurements, characterized by very different T2 which suggests liquid-like and solid-like dynamics. These measurements allow the quantification of separate hydrogen components and their assignment to defined physical pools which had been discussed repeatedly in the literature, i.e. bulk-like water and a combination of protein hydrogens and strongly bound water. The first determination of 2H relaxation dispersion in comparison to 1H dispersion suggests intramolecular interactions as the dominating source for the pronounced magnetic field dependence of the longitudinal relaxation time T1.

Intermittent Dynamic Compression Confers Anabolic Effects in Articular Cartilage

(1) Background: Mechanical loading is an essential part of the function and maintenance of the joint. Despite the importance of intermittent mechanical loading, this factor is rarely considered in preclinical models of cartilage, limiting their translatability. The aim of this study was to investigate the effects of intermittent dynamic compression on the extracellular matrix during long-term culture of bovine cartilage explants. (2) Methods: Bovine articular cartilage explants were cultured for 21 days and subjected to 20 min of 1 Hz cyclic compressive loading five consecutive days each week. Cartilage remodeling was investigated in the presence of IGF-1 or TGF-β1, as well as a TGF-β receptor 1 (ALK5) kinase inhibitor and assessed with biomarkers for type II collagen formation (PRO-C2) and fibronectin degradation (FBN-C). (3) Results: Compression of cartilage explants increased the release of PRO-C2 and FBN-C to the conditioned media and, furthermore, IGF-1 and compression synergistically increased PRO-C2 release. Inhibition of ALK5 blocked PRO-C2 and FBN-C release in dynamically compressed explants. (4) Conclusions: Dynamic compression of cartilage explants increases both type II collagen formation and fibronectin degradation, and IGF-1 interacts synergistically with compression, increasing the overall impact on cartilage formation. These data show that mechanical loading is important to consider in translational cartilage models.

The Lubricating Activity of Phospholipids on Charged and Neutral Surfaces

In this paper, the amphoteric character of cartilage surface is used to study friction between charged (+/+), (-/-) and uncharged (±/±) biological surfaces. The surface of the bovine articular cartilage at varying pH solutions is either charged positively or negatively. When examining the coefficient of friction of the (cartilage/cartilage) pair, depending on the pH, we observe surfaces charged positively, negatively and without charge. The uncharged surface refers to the isoelectric point (IEP) and shows greater friction than the charged surfaces of the cartilage. The measurement of surface energy versus pH of the spherical lipid bilayer will reveal the amphoteric nature of the membrane surface. The friction coefficient of the (cartilage/cartilage) pair and the surface energy of the spherical lipid bilayer versus pH are expressed by a bell-shaped curve. This is an interesting case that confirms the fact that the phospholipid bi-layer is embedded on the surface cartilage. This experimental confirmation of negative surface cartilage is insufficiently highlighted in the literature on natural lubrication.

The Time-dependent Effects of Bipolar Radiofrequency Energy on Bovine Articular Cartilage

Purpose: The purpose of this study was to compare the effect of bipolar radiofrequency energy (bRFE) on chondroplasty at the different time durations in an in vitro experiment that simulated an arthroscopic procedure. Methods: Six fresh bovine knees were used in our study. Six squares were marked on both the medical and lateral femoral condyles of each femur. Each square was respectively treated with bRFE for 0s, 10s, 20s, 30s, 40s and 50s. Full-thickness articular cartilage specimens were harvested from the treatment areas. Each specimen was divided into three distinct parts: one for hematoxylin/eosin staining histology, another for cartilage surface contouring assessment via scanning electron microscopy (SEM), and the last one for glycosaminoglycan (GAG) content measurement. Results: bRFE caused time-correlated damage to chondrocytes, and GAG content in the cartilage was negatively correlated to exposure time. bRFE caused time-correlated damage to chondrocytes. The GAG content in the cartilage negatively correlated with the exposure time. The sealing effect positively correlated with the exposure time. Additionally, it took at least 20 seconds of radiofrequency exposure to render a smooth cartilage surface and a score of 2 (normal) in the scoring system used. Conclusion: bRFE usage in chondroplasty could effectively trim and polish the cartilage lesion area; however, it induces a dose-dependent detrimental effect on chondrocytes and metabolic activity that negatively correlated with the treatment time. Therefore, cautions should be taken in the use of bRFE for treatment of articular cartilage injury.