Polymer substrates obtained from poly(lactic acid) (PLA) nanofibres modified with carbon nanotubes (CNTs) and gelatin (GEL) for cartilage tissue engineering are studied. The work presents the results of physical, mechanical, and biological assessment. The hybrid structure of PLA and gelatine nanofibres, carbon nanotubes- (CNTs-) modified PLA nanofibres, and pure PLA-based nanofibres was manufactured in the form of fibrous membranes. The fibrous samples with different microstructures were obtained by electrospinning method. Microstructure, physical and mechanical properties of samples made from pure PLA nanofibres, CNTs-, and gelatin-modified PLA-nanofibres were studied. The scaffolds were also testedin vitroin cell culture of human chondrocytes collected from patients. To assess the influence of the nanofibrous scaffolds upon chondrocytes, tests for cytotoxicity and genotoxicity were performed. The work reveals that the nanofibrous structures studied were neither genotoxic nor cytotoxic, and their microstructure, physical and mechanical properties create promising scaffolds for potential use in cartilage repairing.