Genipin crosslinking decreases the mechanical wear and biochemical degradation of impacted cartilage in vitro.

High energy trauma to cartilage causes surface fissures and microstructural damage, but the degree to which this damage renders the tissue more susceptible to wear and contributes to the progression of post-traumatic osteoarthritis (PTOA) is unknown. Additionally, no treatments are currently available to strengthen cartilage after joint trauma and to protect the tissue from subsequent degradation and wear. The purposes of this study were to investigate the role of mechanical damage in the degradation and wear of cartilage, to evaluate the effects of impact and subsequent genipin crosslinking on the changes in the viscoelastic parameters of articular cartilage, and to test the hypothesis that genipin crosslinking is an effective treatment to enhance the resistance to biochemical degradation and mechanical wear. Results demonstrate that cartilage stiffness decreases after impact loading, likely due to the formation of fissures and microarchitectural damage, and is partially or fully restored by crosslinking. The wear resistance of impacted articular cartilage was diminished compared to undamaged cartilage, suggesting that mechanical damage that is directly induced by the impact may contribute to the progression of PTOA. However, the decrease in wear resistance was completely reversed by the crosslinking treatments. Additionally, the crosslinking treatments improved the resistance to collagenase digestion at the impact-damaged articular surface. These results highlight the potential therapeutic value of collagen crosslinking via genipin in the prevention of cartilage degeneration after traumatic injury. © 2016 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 35:558-565, 2017.

Genipin crosslinking of cartilage enhances resistance to biochemical degradation and mechanical wear.

Collagen crosslinking enhances many beneficial properties of articular cartilage, including resistance to chemical degradation and mechanical wear, but many crosslinking agents are cytotoxic. The purpose of this study was to evaluate the effectiveness of genipin, a crosslinking agent with favorable biocompatibility and cytotoxicity, as a potential treatment to prevent the degradation and wear of articular cartilage. First, the impact of genipin concentration and treatment duration on the viscoelastic properties of bovine articular cartilage was quantified. Next, two short-term (15 min) genipin crosslinking treatments were chosen, and the change in collagenase digestion, cartilage wear, and the friction coefficient of the tissue with these treatments was measured. Finally, chondrocyte viability after exposure to these genipin treatments was assessed. Genipin treatment increased the stiffness of healthy, intact cartilage in a dose-dependent manner. The 15-min crosslinking treatments improved cartilage's resistance to both chemical degradation, particularly at the articular surface, and to damage due to mechanical wear. These enhancements were achieved without sacrificing the low coefficient of friction of the tissue and at a genipin dose that preserved chondrocyte viability. The results of this study suggest that collagen crosslinking via genipin may be a promising preventative treatment to slow the degradation of cartilage.

The effect of collagen crosslinking on the biphasic poroviscoelastic cartilage properties determined from a semi-automated microindentation protocol for stress relaxation.

Given the important role of the collagenous structure in cartilage mechanics, there is considerable interest in the relationship between collagen crosslinking and the mechanical behavior of the cartilage matrix. While crosslink-induced alterations to the elastic modulus of cartilage have been described, changes to time-dependent behavior have not yet been determined. The objective of the study was to quantify changes to cartilage material properties, including viscoelastic coefficients, with crosslinking via indentation. To accomplish this, a semi-autonomous microindentation stress relaxation protocol was first developed, validated and then applied to cartilage specimens before and after crosslinking. The change in mechanical properties with crosslinking was analyzed both in the unloading portions of the test via the Oliver-Pharr method and in the holding portion with an inverse iterative finite element model that represented cartilage as a biphasic poroviscoelastic material. Although both techniques suggested a similar increase in equilibrium modulus in the crosslinked specimens as compared to the controls, distinct differences in the control specimens were apparent, suggesting that the two different techniques may be capturing different aspects of the material behavior. No differences in time-dependent properties were observed between the crosslinked and the control specimens. These results give further insight into the effects of crosslinking in cartilage mechanical behavior. Additionally, the microindentation stress relaxation protocol may enable increased automation for high-throughput testing.