The aim of the study was to develop a novel treatment method of knee osteochondral defects consisting in injecting platelet-rich plasma and crushed hyaline cartilage under a collagen membrane, and assess the technique in experiment. Materials and Methods. A prospective study was carried out on small cattle animals, 30 in number, aged 1.5-3 years weighing 20-30 kg. All subjects got a full-thickness defect to the subchondral bone, 4.5 mm in diameter. As a control, one of the joint defects was not replaced. Due to a replacement method, all animals were divided into three groups. One group animals underwent the replacement according to the developed technique: there were used an extracellular collagen matrix and the body resources (platelet-rich plasma and crushed autologous cartilage). Results. The results were assessed 1 month and 3 months after surgery analyzing the type and degree of defect filling. Best results were found in the group, where a defect was covered by an extracellular collagen matrix with platelet-rich plasma and crushed autologous cartilage. The results of the no replacement group were comparable with the findings of other researchers, according to which osteochondral defects almost have no self-regeneration. Conclusion. The suggested replacement technique for osteochondral defect using extracellular collagen matrix, autologous cartilage, and platelet-rich plasma is less aggressive compared to autochondroplasty, and the obtained results are more stable compared to microfracture or tunneling.