Quantitative Analysis of Contrast-Enhanced Ultrasound That Can Be Used to Evaluate Angiogenesis during Patellar Tendon Healing in Rats

Abstract

Objective. To investigate the efficacy of contrast-enhanced ultrasound (CEUS) in quantitatively evaluating angiogenesis during patellar tendon healing in rats. Methods. A total of 40 Sprague-Dawley rats were used in this study. The patellar tendons of 30 rats (60 limbs) that underwent incision and suture were treated as the operation group and monitored after 7, 14, and 28 days. The normal patellar tendons of 10 rats (20 limbs) were treated as the control group and monitored on day 0. The ultrasound examination was used to evaluate the structure and blood perfusion of the patellar tendon. Immunohistochemistry was used to assess angiogenesis, and the biomechanical test was used to verify functional recovery of the patellar tendon. Results. The tendons in the operation group were significantly thickened compared with those in the control group (p < 0.01). The peak intensity (PI) in CEUS of the tendons showed a clear difference at each time point after the surgery (p < 0.01). PI increased in the operation group with a maximum on day 7, and then gradually decreased until day 28 when PI was close to the basic intensity (BI) in the control group (p > 0.05). It was consistent with the change of the CD31-positive staining areas representing angiogenesis of the injured patellar tendons. The PI was positively correlated with the CD31-positive staining area fraction (R = 0.849, p < 0.001). The failure load and tensile strength of the repaired patellar tendons in the operation group increased over time. The PI showed negative correlations with the failure load (R = -0.787, p < 0.001) and tensile strength (R = -0.714, p < 0.001). Conclusion. The PI in CEUS could quantitatively reflect the time-dependent change in the blood supply of the healing site, and the PI correlated with histologic and biomechanical properties of the healing tendon. Quantitative analysis of contrast-enhanced ultrasound could be a useful method to evaluate angiogenesis in healing tendons.