Introduction: Cryoablation with liquid nitrogen has shown promise in treating bone sarcomas, but its effect on fibroblast morphology and viability in ligament tissues remains unclear. This study aims to evaluate the impact of liquid nitrogen on fibroblasts in the patellar tendon ligament of male Wistar rats. Methods: This experimental study utilized 28 male Wistar rats (6 months old, 300-400 grams) that were randomly divided into four groups. The control group was not exposed to liquid nitrogen, while the experimental groups were exposed to liquid nitrogen for 1, 5, and 10 minutes. After a 7-day acclimatization, the rats were exposed to liquid nitrogen and euthanized. The patellar tendon ligaments were collected for histological analysis, stained with Hematoxylin-Eosin, and examined under a microscope. Statistical analysis was performed using One-Way ANOVA, followed by Tukey post hoc tests to determine the significant differences between the exposure times. Results: Significant changes were observed in fibroblast morphology and count with increasing exposure times. One-way ANOVA analysis revealed significant differences (p < 0.001) in fibroblast count and morphological alterations across exposure groups. Tukey post hoc tests revealed that 10-minute exposure resulted in substantial reductions in fibroblast count and viability, accompanied by progressive morphological changes as exposure time increased. Discussion: Liquid nitrogen exposure significantly impacts fibroblast cells, with longer exposure times causing more severe cellular damage. The exposure duration should be carefully controlled in cryotherapy to optimize therapeutic benefits. Conclusion: Exposure to liquid nitrogen adversely affects fibroblast cells in the patellar tendon ligament, with prolonged exposure resulting in decreased cesll viability and morphological changes.