Intra-articular hyaluronic acid (HA) and oral glucosamine (GA) are frequently used to decrease pain and increase function in osteoarthritis patients. Injectable chondroitin sulfate and HA improve effectiveness however a formulation that combines GA and HA is currently not available. Aims of this study were to synthesize and characterize N-acetyl-d-GA immobilized HA based nanoparticles [GA-PEG@nano(HA)] to increase extracellular matrix production and to decrease OA markers of healthy and OA chondrocytes. Mean size of synthesized GA-PEG@nano(HA) was 175 nm and 20% of GA released from these composites during 21 days. Released GA reduced the proliferation of chondrosarcoma cells. Chondrogenic marker expression of bone marrow mesenchymal stem cells decreased slightly but not significantly. osteoarthritis and extracellular matrix markers of healthy and osteoarthritic chondrocytes at seven days in culture remained unchanged. In conclusion, GA-PEG@ nano(HA) composites had limited effects on mesenchymal stem cells, healthy and osteoarthritic chondrocytes.