Application of autofluorescence robotic histology for quantitative evaluation of the 3‐dimensional morphology of murine articular cartilage |
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| Authors: | Patricia Das Neves Borges Tonia L. Vincent Massimo Marenzana |
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| Affiliation: | 1. Department of Bioengineering, Imperial College London, London, United Kingdom;2. Kennedy Institute of Rheumatology, Nuffield Department of Orthopaedics, Rheumatology and Musculoskeletal Sciences, University of Oxford, Roosevelt Drive, Oxford, United Kingdom |
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| Abstract: | Murine models of osteoarthritis (OA) are increasingly important for understating pathogenesis and for testing new therapeutic approaches. Their translational potential is, however, limited by the reduced size of mouse limbs which requires a much higher resolution to evaluate their articular cartilage compared to clinical imaging tools. In experimental models, this tissue has been predominantly assessed by time‐consuming histopathology using standardized semi‐quantitative scoring systems. This study aimed to develop a novel imaging method for 3‐dimensional (3D) histology of mouse articular cartilage, using a robotic system—termed here “3D histocutter”—which automatically sections tissue samples and serially acquires fluorescence microscopy images of each section. Tibiae dissected from C57Bl/6 mice, either naïve or OA‐induced by surgical destabilization of the medial meniscus (DMM), were imaged using the 3D histocutter by exploiting tissue autofluorescence. Accuracy of 3D imaging was validated by ex vivo contrast‐enhanced micro‐CT and sensitivity to lesion detection compared with conventional histology. Reconstructions of tibiae obtained from 3D histocutter serial sections showed an excellent agreement with contrast‐enhanced micro‐CT reconstructions. Furthermore, osteoarthritic features, including articular cartilage loss and osteophytes, were also visualized. An in‐house developed software allowed to automatically evaluate articular cartilage morphology, eliminating the subjectivity associated to semi‐quantitative scoring and considerably increasing analysis throughput. The novelty of this methodology is, not only the increased throughput in imaging and evaluating mouse articular cartilage morphology starting from conventionally embedded samples, but also the ability to add the third dimension to conventional histomorphometry which might be useful to improve disease assessment in the model. |
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| Keywords: | automated image analysis destabilization of medial meniscus model fluorescence imaging high‐throughput 3D histomorphometry osteoarthritis |
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