The efficacy of MRI-based statistical texture analysis (TA) in predicting chemotherapy response among patients with osteosarcoma was assessed. Forty patients (male: female = 31:9; age = 17.2 ± 5.7 years) with biopsy-proven osteosarcoma were analyzed in this prospective study. Patients were scheduled for three cycles of neoadjuvant chemotherapy (NACT) and diffusion-weighted MRI acquisition at three time points: at baseline (t0), after the first NACT (t1) and after the third NACT (t2) using a 1.5 T scanner. Eight patients (nonsurvivors) died during NACT while 34 patients (survivors) completed the NACT regimen followed by surgery. Histopathological evaluation was performed in the resected tumor to assess NACT response (responder [≤50% viable tumor] and nonresponder [>50% viable tumor]) and revealed nonresponder: responder = 20:12. Apparent diffusion coefficient (ADC) and intravoxel incoherent motion (IVIM) parameters, diffusion coefficient (D), perfusion coefficient (D*) and perfusion fraction (f) were evaluated. A total of 25 textural features were evaluated on ADC, D, D* and f parametric maps and structural T1-weighted (T1W) and T2-weighted (T2W) images in the entire tumor volume using 3D TA methods gray-level cooccurrence matrix (GLCM), neighborhood gray-tone-difference matrix (NGTDM) and run-length matrix (RLM). Receiver-operating-characteristic curve analysis was performed on the selected textural feature set to assess the role of TA features (a) as marker(s) of tumor aggressiveness leading to mortality at baseline and (b) in predicting the NACT response among survivors in the course of treatment. Findings showed that the NGTDM features coarseness, busyness and strength quantifying tumor heterogeneity in D, D* and f maps and T1W and T2W images were useful markers of tumor aggressiveness in identifying the nonsurvivor group (area-under-the-curve [AUC] = 0.82-0.88) at baseline. The GLCM features contrast and correlation, NGTDM features contrast and complexity and RLM feature short-run-low-gray-level-emphasis quantifying homogeneity/terogeneity in tumor were effective markers for predicting chemotherapeutic response using D (AUC = 0.80), D* (AUC = 0.80) and T2W (AUC = 0.70) at t0, and D* (AUC = 0.80) and f (AUC = 0.70) at t1. 3D statistical TA features might be useful as imaging-based markers for characterizing tumor aggressiveness and predicting chemotherapeutic response in patients with osteosarcoma.