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Low-intensity pulsed ultrasound partially reversed the deleterious effects of a severe spinal cord injury-induced bone loss and osteoporotic fracture healing in paraplegic rats



To evaluate the effects of low-intensity pulsed ultrasound (LIPUS) on the quality of femoral fracture callus formation in rats with severe osteoporosis secondary to spinal cord injury (SCI).


Forty-five male rats were equally divided into three groups: the Sham group underwent sham surgery for SCI followed by surgery for femoral fracture on day ten post-spine surgery; the SCI group sustained a complete transection of the spinal cord and a femoral fracture ten days post-SCI; and the SCI group treated with ultrasound (SCI + US), which also sustained a femoral fracture on day ten post-SCI, concomitant with daily application of LIPUS at the fracture site.


At the non-fractured tibias, LIPUS counteracted the SCI-induced bone loss by normalizing the osteoblastic-related gene expression, decreasing resorptive area, increasing trabecular area, and decreasing RANK and RANK-L-positive areas, which resulted in higher cortical volume and stronger tibias. Likewise, LIPUS was effective at restoring bone fracture healing in SCI rats; by promoting endochondral ossification, increasing collagen deposition and OPG-positive-area, decreasing resorptive area, which led to higher density and improved microarchitecture, ultimately resulting in stronger fracture callus.


At the tibias, LIPUS counteracted the SCI-induced bone loss effects by simultaneously increasing bone formation and decreasing bone resorption. We also evidenced the osteogenic effects of LIPUS at partially restoring the endochondral ossification during callus formation, leading to a newly formed tissue with improved microarchitecture and mechanical integrity. Therefore, LIPUS may be an efficient and non-invasive approach to prevent bone loss and osteoporotic fracture in SCI individuals.

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Fig. 1: Gene expression in non-fractured tibias and in femoral calluses.
Fig. 2: Bone histology and immunohistochemistry assessment in non-fractured tibias.
Fig. 3: Bone histology and immunohistochemistry assessment in femoral calluses.

Data availability

The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request. All data generated or analyzed during this study are included in this published article.


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We would like to thank all the investigators, staff, and trainees from the Laboratory of Bioengineering at the School of Medicine of Ribeirão Preto, University of São Paulo, Brazil, who assisted in the entire study. We also would like to thank the 3B’s Research Group (Portugal) as well as the School of Dentistry of Ribeirão Preto, University of São Paulo, Brazil, who assisted in the microstructural analysis. The São Paulo Research Foundation funded this study (FAPESP) - 2015/22126-1.

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Correspondence to Ariane Zamarioli.

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Zamarioli, A., Butezloff, M.M., Ximenez, J.P.B. et al. Low-intensity pulsed ultrasound partially reversed the deleterious effects of a severe spinal cord injury-induced bone loss and osteoporotic fracture healing in paraplegic rats. Spinal Cord 61, 145–153 (2023).

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