Objectives-Arthropod-borne arthritogenic alphaviruses such as chikungunya virus (CHIKV) and Ross River virus (RRV) cause large epidemics of severe musculoskeletal disease and have been progressively expanding their global distribution emerging in new regions of the world. It has been well-established that alphaviruses cause severe arthralgia/arthritis in the joints that can be both acute and chronic. The synovial space of joints is glycan rich, containing high levels of glycosaminoglycan (GAGs) frequently linked to protein backbones forming proteoglycan structures. Chondrocytes are the major cell type producing the proteoglycan rich articular cartilage matrix of joint. To date, there have been no studies to specifically elucidate the impact of alphaviruses on cartilage and cartilage producing chondrocytes. Therefore this study aims to characterise the complex interactions between alphaviruses and host cartilage components (chondrocytes and glycans) in disease pathogenesis. We examined the role of chondrocytes in alphavirus infection using in vitro, ex vivo and in vivo cells and tissues followed by molecular, imaging, bioplex, histology and glycan array assays. Results indicate that alphaviruses including CHIKV, RRV and Barmah Forest virus (BFV) can replicate in chondrocytes in cell lines and in primary human cells. This infection resulted in the production of critical soluble factors and degrading enzymes known to contribute to cartilage damage. Results correlated to in vivo where murine chondrocytes were found to be a site of alphavirus infection with RRV-RNA and antigen detected in chondrocytes of the cartilage. Furthermore this infection was found to produce effects, which caused in damage to the murine articular cartilage thereby contributing to disease pathogenesis. Overall these studies indicate that chondrocytes are a target for alphaviral infection and replication, contributing to the disease joint pathology. Detailed understanding of the role of chondrocytes in alphaviral pathogenesis and the specific role of glycans could ultimately lead to the development of safer and more targeted antiviral therapies against arthritis-causing viral pathogens.