Due to the attractive properties of poly(L-lactic acid) (PLLA) and chitosan (CHT) for tissue engineering applications, this study is aimed at analyzing the chondrogenic potential of human bone marrow-derived mesenchymal stem cells (BM-MSCs) derived from osteoarthritic (OA) patients, in pure or CHT-coated PLLA, using different coating methodologies. Although PLLA scaffolds coated in one-step (PLLA-CHT1) yielded CHT smooth pellicles filling the PLLA macropores, a two-step strategy resulted in a CHT fiber-like thin coating covering PLLA pore walls (PLLA-CHT2). Both strategies led to the incorporation of similar content of CHT and a two-fold increase in the scaffold's water uptake capacity, providing elastic moduli values comparable to the ones found for human articular cartilage. After confirming OA-derived BM-MSCs, metabolic activity in the scaffolds, the chondrogenic potential was tested at 30 and 60 days, in a chondrogenic differentiation medium. PLLA scaffolds improved the chondrogenic differentiation of BM-MSCs, regarding cell pellet conventional culture and presented a typical cartilage zonal distribution, although was not able to revert a terminal differentiation. In PLLA-CHT1, on a short term, a rather heterogeneous tissue was formed, with confined areas of either slower cell infiltration or a faster maturation, with enhanced chondrogenic phenotype. In PLLA-CHT2, a similar tissue to PLLA was obtained, albeit on the long term, these scaffolds helped to maintain a hyaline-like phenotype and prevented the advance of the hypertrophic process. These results demonstrate the importance of the scaffolds microenvironment on the cellular events of chondrogenesis. The authors are thankful to the microscopy service of UPV for useful help and service; the Department of Orthopedic Surgery-CHUAC for providing human samples; T.H. Gomez and M.J. Sanchez for cell isolation and SCH of INIBIC for histological processing. The Biomedical Research Networking Center in Bioengineering, Biomaterials, and Nanomedicine (CIBER-BBN) is a national initiative of ISCIII. This study was supported by Program ACI-PRO-MOCIONA (ACI2010-1095), MAT2010-21611-CO3-01, Iniciativa Ingenio 2010, and Consolider Program. Magalhaes, J.; Lebourg, MM.; Deplaine, H.; Gómez Ribelles, JL.; Blanco, FJ. (2015). Effect of the physicochemical properties of pure chitosan-coated poly(lactic acid) scaffolds on the chondrogenic differentiation of mesenchymal stem cells from osteoarthritic patients. Tissue Engineering: Parts A, B, and C. 21(3-4):716-728. doi:10.1089/ten.tea.2014.0133