The invention provides a preparation method of a targeting chondrocyte exosome. The preparation method comprises the following steps of preparing an exosome carrier, preparing a targeting cartilage exosome, and preparing miRNA-140 wrapped by the targeting cartilage exosome. The preparation method has the beneficial effects that the miRNA medicine is delivered with treatment potential, high efficiency and tissue specificity for treating osteoarthritis, the application of the miRNA-140 wrapped by the targeting exosome in the medicine for treating osteoarthritis is mainly utilized, in an osteoarthritis animal model of a rat, the expression of MMP-13 can be inhibited by injecting the miRNA-140 into a knee joint cavity, the damage of cartilage matrix can be obviously improved, the cartilage damage process of an osteoarthritis animal model is delayed, and the cartilage protection effect is achieved, and therefore, the exosome is used for transporting miRNA-140 in a targeted manner, degradation of cartilage tissues can be slowed down, and the effect of inhibiting catabolism of cartilage matrix is achieved, so that the damage of articular cartilage is controlled, and cartilage repair is promoted.