Histological Analysis of the Effect of a Vector Potential Generator on the Femur of a Hindlimb-Suspended Rat

Abstract

Disuse osteoporosis occurs due to rest and reduced mechanical stimulation. Under these conditions, bone resorption exceeds bone formation, leading to a decrease in bone density. Vector potential (VP) generators have been developed, and their ability to maintain cartilage thickness has been reported. However, their effects on bone tissue remain unstudied. In this study, experiments were conducted to test the effects of VP on bones that had undergone weight reduction due to hindlimb suspension as a model of disuse osteoporosis. Methods: In this study, 7-week-old male Wistar rats (N = 6 each) were classified into control (CO), hindlimb suspension (HS), and VP energization intervention groups. The tail was used to suspend the HS and VP to remove the load applied to the hindlimbs. The VP conditions were as follows: voltage, 67 mV; frequency, 20 kHz, 0.12 mA; experimental intervention, 30 min/day, 5 days/week, for 3 weeks. At the end of the experimental period, the rats were euthanized with carbon dioxide gas, and histological specimens were fixed in 4% paraformaldehyde (PFA) in the femur and analyzed by electron microscopy, bone morphometry, immunohistology, bone fracture testing, and gene expression analysis. Results: HS decreased trabecular bone density and strength. However, VP maintained a significantly higher bone mass than HS, and VP did not differ from CO in bone strength; more osteoclasts were observed on the bone surface in HS, but they were suppressed in VP, and gene expression of CTSK and MMP-9 was decreased. Conclusions: VP suppressed bone resorption by osteoclasts, suggesting that VP is useful in the treatment of disuse osteoporosis.