The temporal role of leptin within fracture healing and the effect of local application of recombinant leptin on fracture healing.

Abstract

We hypothesized that leptin is expressed in a specific time sequence during fracture healing, and its deficiency leads to impaired healing. Control (C57BL/6) mice and leptin -/- obese (ob/ob) mice were used. ARM 1:: Fracture callus was harvested at 1, 3, 5, 7, 10, 14, and 21 days (n = 8/time point) after closed middiaphyseal femur fractures were created in 56 C57BL/6 mice, and reverse transcriptase polymerase chain reaction analysis was then performed. Levels of leptin were tracked at each time point listed. ARM 2:: Forty-two C57BL/6 controls and 42 ob/ob mice underwent open stabilized middiaphyseal femur fractures, and tissues were harvested at 14, 21, and 42 days and radiographic, histologic, and quantitative computerized tomography analyses were performed. ARM 3:: Murine recombinant leptin was applied directly at the newly created fracture site in 2 separate groups (10 or 100 μg of leptin) of 42 ob/ob mice. Two-factor analysis of variance and the Student t-test were used for statistical analysis. The time course of Leptin mRNA expression within a fracture callus was detected. Delay in callus maturation was demonstrated radiographically and histologically in the ob/ob mice. ob/ob fractures had an increase in total callus volume by quantitative computerized tomography (P < 0.05). Application of local leptin at both doses reversed the delay in healing. Leptin is expressed in a unique time course during fracture healing and leptin deficiency leads to impaired fracture healing that reverses by local application of leptin.