Cartilage loss due to osteoarthritis (OA) in the patellofemoral joint provokes pain, stiffness, and restriction of joint motion, which strongly reduces quality of life. Early diagnosis is essential for prolonging painless joint function. Vibroarthrography (VAG) has been proposed in the literature as a safe, noninvasive, and reproducible tool for cartilage evaluation. Until now, however, there have been no strict protocols for VAG acquisition especially in regard to differences between the patellofemoral and tibiofemoral joints. The purpose of this study was to evaluate the proposed examination and acquisition protocol for the patellofemoral joint, as well as to determine the optimal examination protocol to obtain the best diagnostic results. Thirty-four patients scheduled for knee surgery due to cartilage lesions were enrolled in the study and compared with 33 healthy individuals in the control group. VAG acquisition was performed prior to surgery, and cartilage status was evaluated during the surgery as a reference point. Both closed (CKC) and open (OKC) kinetic chains were assessed during VAG. The selection of the optimal signal measures was performed using a neighborhood component analysis (NCA) algorithm. The classification was performed using multilayer perceptron (MLP) and radial basis function (RBF) neural networks. The classification using artificial neural networks was performed for three variants: I. open kinetic chain, II. closed kinetic chain, and III. open and closed kinetic chain. The highest diagnostic accuracy was obtained for variants I and II for the RBF 9-35-2 and MLP 10-16-2 networks, respectively, achieving a classification accuracy of 98.53, a sensitivity of 0.958, and a specificity of 1. For variant III, a diagnostic accuracy of 97.79 was obtained with a sensitivity and specificity of 0.978 for MLP 8-3-2. This indicates a possible simplification of the examination protocol to single kinetic chain analyses.