Aside from the dependence on in vitro culture time, cytokines present in vivo can also influence integration. A study examined the effects of steroid hormones in bovine cartilage that is lacking a known inhibitor to integration, interleukin-1b. An increase of,50 kPa in mechanical integration was seen, as compared to the 700 kPa obtained in this study for the native-tonative controls. Also, it has also been shown that, without the assistance of exogenous agents, strength of half that which is seen in intact cartilage can be achieved in an equine model for chondrocyte transplantation. It is worth noting that, in the present study, by delivering cells to the interface in concert with LOX, integration strength can be increased to 1.6 MPa. Comparing this result to the stiffness of fibrin, which is clinically used as tissue glue and sealant, the stiffness of the LOXtreated interface is roughly fifty times higher. Averaged over various strain rates, the stiffness of fibrin alone is under 30 kPa. When fibrin is combined with chondrocytes to serve as a cartilage adhesive, the stiffness of the interface is increased over fibrin alone and also with time in vivo, to 0.645 MPa after 8 months. It is worth noting that LOX-treatment achieves twotimes the stiffness in a fraction of the time. It is expected that the stiffness of interfaces enhanced with LOX and chondrocytes will continue to improve in vivo as the cells remodel the matrix over time. Chondrocyte transplantation is a current therapy that, similar to the self-assembled constructs employed in this study, delivers metabolic cells to the wound edge using fibrin. It is conceivable for LOX to assist this clinical procedure, especially since the LOX treatment produces comparable results to fibrin at a shorter time. Of course, additional studies on 1) optimal dosing time, 2) cross-linker concentration, and 3) activity profile as related to not only the chondrocytes but also other cell types surrounding cartilage would need to be completed to ensure safety and efficacy, prior to deploying this technique clinically. A major component of articular cartilage ECM is the electronegative aggrecan. This electric charge is an obstacle to integration because the similar charges in two pieces of tissue would cause them to repel. Further studies need to be completed to fully understand the role which aggrecan��s electronegativity may play in blocking integration. Future studies might also include the combination of LOX with other bioactive agents that are known to influence cartilage behavior. Already, transforming growth factor b1 has shown efficacy when combined with a biomaterial, and it would be interesting to examine how LOX can assist this case. TFG-b1 may work in synergism with LOX, the cytokine and enzyme working in tandem to effect greater collagen production and cross-linking. It should be mentioned that, for this study, LOX concentration was based on a pilot study that examined LOX on native and engineered cartilages separately. Following this study’s exciting results, it may be prudent to conduct a systematic examination of various LOX concentrations to identify a minimum, yet effective, concentration between 0.015 and 0.15 mg/ml that enhances AbMole Halothane interfacial stiffness and strength to the levels of the engineered or native cartilages, or even for other tissues where cross-linking plays important functional roles. For instance, integrating engineered knee meniscus to native knee meniscus has shown dependence on maturation state, and therefore the extent of collagen crosslinks, and LOX may be used similarly for this tissue. Finding this minimum dose will be significant in not only reducing cost but also in mitigating any potential for this enzyme to interfere with other cellular processes, despite this being a naturally-occurring enzyme.