Abstract
Calcific aortic valve disease (CAVD) is the most common form of valve disease where the only available treatment strategy is surgical valve replacement. Technologies for the early detection of CAVD would benefit the development of prevention, mitigation and alternate therapeutic strategies. Two-photon excited fluorescence (TPEF) microscopy is a label-free, non-destructive imaging technique that has been shown to correlate with multiple markers for cellular differentiation and phenotypic changes in cancer and wound healing. Here we show how specific TPEF markers, namely, the optical redox ratio and mitochondrial fractal dimension, correlate with structural, functional and phenotypic changes occurring in the aortic valve interstitial cells (VICs) during osteogenic differentiation. The optical redox ratio, and fractal dimension of mitochondria were assessed and correlated with gene expression and nuclear morphology of VICs. The optical redox ratio decreased for VICs during early osteogenic differentiation and correlated with biological markers for CAVD progression. Fractal dimension correlated with structural and osteogenic markers as well as measures of nuclear morphology. Our study suggests that TPEF imaging markers, specifically the optical redox ratio and mitochondrial fractal dimension, can be potentially used as a tool for assessing early CAVD progression in vitro.
Highlights
Calcific aortic valve disease (CAVD) is the most common form of valve disease where the only available treatment strategy is surgical valve replacement
Our results showed that Two-photon excited fluorescence (TPEF) metrics correlated with the early markers of CAVD progression and suggest that TPEF microscopy can be utilized as a label-free non-destructive tool for assessing CAVD progression in vitro
Positive Alizarin Red S (ARS) staining on day 28 confirmed that the nodules present in the osteogenic cultures by day 28 were calcific in nature (Fig. 1b)
Summary
Calcific aortic valve disease (CAVD) is the most common form of valve disease where the only available treatment strategy is surgical valve replacement. We show how specific TPEF markers, namely, the optical redox ratio and mitochondrial fractal dimension, correlate with structural, functional and phenotypic changes occurring in the aortic valve interstitial cells (VICs) during osteogenic differentiation. Our study suggests that TPEF imaging markers, the optical redox ratio and mitochondrial fractal dimension, can be potentially used as a tool for assessing early CAVD progression in vitro. We seeded porcine aortic VICs as monolayers in quiescent versus osteogenic media on two dimensional soft or stiff substrates We examined these samples using TPEF microscopy to quantify ORR and mitochondrial FD and simultaneously characterized the CAVD progression in our model using traditional end-point biomarkers, such as calcific nodule quantification, gene expression, cell proliferation and apoptosis. Our results showed that TPEF metrics correlated with the early markers of CAVD progression and suggest that TPEF microscopy can be utilized as a label-free non-destructive tool for assessing CAVD progression in vitro
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Disclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.