In this study, carbon nanofibers (CNFs) having different amounts of defect were prepared by acid-treating for 0.5, 1 and 5 h and were combined with alumina. Then, the influence of the amount of defect on the CNFs on microstructure development of the CNFs/alumina composites and relationship between the fracture toughness and the average alumina grain size was investigated. The intensity ratio of D-band to G-band (D/G) in Raman spectra of the CNFs increased from 0.34 for pristine CNFs to 0.95 for the CNFs acid-treated for 5 h with the acid-treatment time, which indicates that the amount of defect on the CNFs increased with the acid-treatment time. The alumina grain growth in the dense composites sintered at 12001300°C was not influenced by the amount of defect on the CNFs, however, the composite containing CNFs having the moderate amount of defect (D/G = 0.56) showed the slowest alumina grain growth rate at 13501450°C. The fracture toughness of the composites containing the CNFs acid-treated for 0.5 h increased with a decrease in average alumina grain size and reached 5.6 MPa·m 0.5 at the average alumina grain size of 0.84¯m, which was 60% higher value compared to monolithic alumina (3.5 MPa·m 0.5 ). However, fracture toughness of the composites containing CNFs acid-treated for 1 and 5 h increased with a decrease in average alumina grain size, showed the maximum values of 5.0 and 4.5 MPa·m 0.5 at average alumina grain sizes of 1.3 and 1.6¯m, respectively, and decreased as the average alumina grain size decreased further. The maximum fracture toughness of the composite containing the CNFs acidtreated for 5 h was lower than that of the composite containing the CNFs acid-treated for 1 h.