Monitoring bone changes due to calcium, magnesium, and phosphorus loss in rat femurs using Quantitative Ultrasound

Aldo Fontes-Pereira,Christiano Bittencourt Machado,Ricardo Erthal Santelli,Bernardo Ferreira Braz,Thiago Barboza,Paulo Rosa,Marco Antônio Von Krüger,Aline Soares Freire,Daniel Matusin,Wagner Coelho De Albuquerque Pereira,Sergio Augusto Lopes De Souza

doi:10.1038/s41598-018-30327-7

Abstract

Bone mineral density is an important parameter for the diagnosis of bone diseases, as well as for predicting fractures and treatment monitoring. Thus, the aim of the present study was to evaluate the potential of Quantitative Ultrasound (QUS) to monitor bone changes after calcium, phosphorus, and magnesium loss in rat femurs in vitro during a demineralization process. Four quantitative ultrasound parameters were estimated from bone surface echoes in eight femur diaphysis of rats. The echo signals were acquired during a decalcification process by Ethylenediaminetetraacetic Acid (EDTA). The results were compared to Quantitative Computed Tomography (QCT) and inductively coupled plasma optical emission spectrometry measurements for validation. Integrated Reflection Coefficient (IRC) reflection parameters and Frequency Slope of Reflection Transfer Function (FSRTF) during demineralization tended to decrease, while the backscattering parameter Apparent Integrated Backscatter (AIB) increased and Frequency Slope of Apparent Backscatter (FSAB) showed an oscillatory behavior with no defined trend. Results indicate a clear relation between demineralization and the corresponding decrease in the reflection parameters and increase in the scattering parameters. The trend analysis of the fall curve of the chemical elements showed a better relationship between IRC and QCT. It was possible to monitor bone changes after ions losses, through the QUS. Thus, it is an indication that the proposed protocol has potential to characterize bone tissue in animal models, providing consistent results towards standardization of bone characterization studies by QUS endorsing its use in humans.

Highlights

According to the World Health Organization, osteoporosis is responsible for more than 8.9 million fractures annually worldwide, of which approximately 4.5 million cases occur in America and Europe[1]
This work is the first to propose an in vitro mineral loss monitoring methodology using Quantitative Ultrasound (QUS) parameters and Quantitative Computed Tomography (QCT) together, and it constitutes an important contribution to ultrasound as an adjuvant tool for the diagnosis and monitoring of bone healing and bone disease
This is an important step toward preclinical trials and subsequent application of QUS in several diseases, such as osteoporosis[20], osteoarthritis[31], osteomyelitis[32], and Paget’s disease[31]

Summary

Introduction

According to the World Health Organization, osteoporosis is responsible for more than 8.9 million fractures annually worldwide, of which approximately 4.5 million cases occur in America and Europe[1] It is the most common osteometabolic disease among the elderly, posing great challenges for contemporary public health[2,3,4]. It is estimated that the lifetime risk for a wrist, hip, or vertebral fracture is up to 40% in developed countries, which is nearly equal to the lifetime risk for coronary heart disease[1] Another important fact is that the costs of osteoporotic fracture treatment are high[5,6]. Studies are ongoing to develop new measurement modes and model-based signal-processing techniques[19]

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Conclusion