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Inter-fragmentary rotational and axial instabilities are major challenges in nailing of complex or comminuted fractures. We aimed to compare the inter-fragmentary rotational and axial stability of novel anti-rotation interlocking nail and the conventional interlocking nail in complex or comminuted femur shaft fractures. Twenty composite femurs were divided into two groups, 30 mm was resected from the mid-portion of all composite femurs. The inter-fragmentary rotational and axial stabilities were assessed. Between 10-N m external and 6-N m internal rotation torques, mean maximum inter-fragmentary rotational arc motion in the novel nails was 1.63 mm and 291 % less than that of the conventional nails (6.38 mm, P = 0.000). Between 150 N distraction and 2300 N compression, mean axial motion in the novel nails was 0.8 mm and 257 % less than that of the conventional nails (2.86 mm, p = 0.000). An anti-rotational novel nail is superior to the conventional interlocking nail in terms of maximum inter-fragmentary rotational and axial stabilities in complex and comminuted femur shaft fractures.