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The objective of the current study is to evaluate using of platelet-rich fibrin (PRF) or adipose-derived stem cells (ADSCs) for the therapy of rabbit ear cartilage defects. Nine adult New Zealand white rabbits were divided into three groups. The rabbit ears developed two defects in the cartilage. Phosphate-buffered saline (PBS) was injected subperichondrially into Group I, adipose-derived MSCs (ADMSCs) were injected into Group II, and platelet-rich fibrin (PRF) was injected into Group III in the defective ear. After 4 weeks, the auricular defect was examined morphologically, histologically, and immunohistochemically. In addition, the gene expression of the collagen type II (Col II) and aggrecan. Anatomically, the auricles of all treatments seemed to be fully healed with smooth surfaces. Histopathologically, a small region of immature cartilage was seen in the control positive group's defects. While the group treated with PRF and ADSCs demonstrated the normal characteristics of new cartilage development, including mature chondrocytes within their lacunae and rich extracellular matrix (ECM). The PRF-treated group responded to Masson's trichrome and orcein stains, while produced a faint staining in the case of the control positive and ADMSC groups. Immunohistochemically, there was a strong positive S100 expression in ADSCs and PRF but mild reaction in the control group. The expression of relative col II and aggrecan was substantially highest in PRF (± 0.72 and ± 0.57, respectively). While, Control positive and ADMSC groups recorded (± 0.41 : ± 0.21, ± 0.6 : ± 0.44) respectively. The PRF had the highest rate of proliferation and chondrogenic potential when compared to the ADMSCs and control groups. [ABSTRACT FROM AUTHOR]