Abstract: Objective To observe effect of open injury in the rabbit limb soft tissue on periosteum coupled with seawater immersion. Methods Forty-eight New Zealand rabbits were used for the experiment. The animal model was developed with the incision of the soft tissue of both hinder limbs. Then, the animals were randomly divided into 3 groups: group A, the pure injury group; group B, the 30-minute seawater immersion group; and group C, the 1-hour seawater immersion group. The rabbits were sacrificed on days 0, 1, 3, 7 respectively after surgery. Then, periosteous samples were collected for histopathology and measurement of the expression of bone morphogenetic proteins (BMPs)and vascular endothelial growth factor (VEGF) of periosteumResults (1) Periosteous inflammatory response for those animals with open injury in limb soft tissue coupled with seawater immersion was obviously much severer than those with pure injury. The longer the immersion time, the severer the inflammatory response. (2)Intramembranous ossification was not obviously delayed after 30-minute seawater immersion, while an obvious delay in intramembranous ossification could be noted following 1-hour seawater immersion. (3) The expression of BMPs and VEGF of periosteum for the animals in group C was delayed and seemed significantly weaker than that of group A and B. In spite of the fact that the expression of BMPs and VEGF for the animals in group B was not delayed, it was clearly weaker than that of the animals in group A. Conclusions Open injury in limb soft tissue coupled with 1-hour seawater immersion could obviously delay and even damage intramembranous ossification of periosteum.