Photoacoustic (PA) techniques are potential alternatives to histopathology. The physicochemical spectrogram (PCS) generated by the PA measurement at multiple wavelengths can presents the morphology and chemical composition target at multi-biomarkers simultaneously. In this work, via multi-wavelength PA measurements performed on rabbit bone models, we investigated the feasibility of using PCSs for bone health assessment. A comprehensive analysis of the PCSs, termed PA physicochemical analysis (PAPCA), was conducted. The “ slope ” and “ relative content ” were used as the PAPCA-quantified parameters to characterize the changes in the physical and chemical properties of bone tissue, respectively. The findings are consistent well with the gold-standard imaging results. It demonstrated that the PAPCA can be used to characterize both the microstructure and content of multi-biomarkers which highly related with bone health. Considering the PA technique is noninvasive and radiation-free, it has great potential in the implementation and monitoring of bone diseases progression.

Collagen is an important biomarker of osteoporosis progression. Noninvasive, multispectral, photoacoustic (PA) techniques use pulsed laser light to induce PA signals to facilitate the visualization of chemical components that are strongly related to tissue health. In this study, the feasibility of multi-wavelength PA (MWPA) measurement of the collagen in bone, using the wavelength range of 1300–1800 nm, was investigated. First, the feasibility of this approach for detecting the collagen content of bone was demonstrated by means of numerical simulation. Then, ex vivo experiments were conducted on both animal and human bone specimens with different bone densities using the MWPA method. The relative collagen content was extracted and compared with the results of micro-computed tomography (micro-CT) and histology. The results showed that the “relative collagen content” parameter obtained using the MWPA approach correlated well with the bone volume ratio obtained from micro-CT images and histological analysis results. This study highlights the potential of the proposed PA technique for determining the collagen content of bones as a biomarker for bone health assessment.

In this study, the feasibility of assessing the chemical composition in bone using the multi-wavelength photoacoustic analysis (MWPA) method was investigated. By illuminating a bone specimen using laser light with a wavelength tunable over an optical spectrum from 680 nm to 950 nm, the optical absorption spectrum of the bone was acquired. Then, with the optical absorption spectra of all the optically absorbing chemical components in the bone known, a spectral unmixing procedure was performed to quantitatively assess the relative content of each chemical component. The experimental results from porcine rib bones demonstrated that the contents of the chemical components, including not only non-organic materials such as minerals and water but also organic materials including oxygenated hemoglobin, deoxygenated hemoglobin, lipid, and collagen, can all be assessed by MWPA. As the chemical composition in the bone is directly associated with functional and metabolic activities, the finding from this study suggests that the MWPA method could offer a new diagnostic tool for the non-invasive evaluation of bone health.

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Photoacoustic (PA) signal analysis based on ultrasonic wave detection can provide both high-sensitivity optical contrast information and micro-architectural information which is highly related with the chemical composition of tissue. In this study, the feasibility assessment of bone composition assessment was investigated using the multi-wavelength PA analysis (MWPA) method which could reflect the molecular information. By illuminating a bone specimen using a laser light with wavelength over an optical spectrum ranging from 680 to 950 nm, the optical absorption spectrum of the bone was acquired. Then, with the optical absorption spectra of all optical absorption chemical components in the known bone, a spectral unmixing procedure was performed to quantitatively assess the relative content of each chemical component. The experimental results from rabbit bones show that MWPA method can be used to assess chemical components related to bone metabolism. Our study confirmed that PA technique can be used as a novel bone diagnostic technique by providing new information about the quantity of bone and identifying biomarkers of bone that can improve the current diagnostic imaging techniques.