Because of its similar genetic makeup with humans, zebrafish are an available and well-established osteoporosis model in vivo for anti-osteoporosis drug development as well as the drug safety-evaluation process. However, few optical imaging methods could effectively visualize the bone of adult zebrafish due to their limited penetration depth. In this paper, in vivo high-resolution and long-term characterization of a prednisolone-induced osteoporotic zebrafish model was achieved with spectral-domain optical coherence tomography (SD-OCT). The capability of three-dimensional SD-OCT imaging was also demonstrated in this study. With SD-OCT images, we could non-destructively monitor the deforming process of adult zebrafish skull from several directions at any time. There is good correlation and agreement between SD-OCT and histology. Valuable phenomenon such as bone defects could be quantitatively evaluated using the SD-OCT images at different time points during a period of 21 days.

Background: As an innovative bone ablation tool, pulsed erbium: yttrium-aluminum-garnet (Er:YAG) laser is being used in clinical practice. It faces the same problems as traditional tools: the water content of bone usually changes with the position, while the amount of water spray in the process of laser irradiation is also uncertain. Real-time monitoring of the effects of laser bone ablation is necessary, but effective tools are still lacking. In this study, we examined the feasibility of rapidly and non-destructively evaluating the surface properties of bone after Er:YAG laser irradiation at different moisture contents by optical coherence tomography (OCT).Methods: Bone specimens (n=90) collected from pig cortical bone were used in this study. All bone specimens had similar volume and surface characteristics after machining. To display the baseline level before Er:YAG laser ablation, a control group (n=10) without dehydration or Er:YAG laser ablation was examined with OCT, scanning electron microscopy (SEM), and energy dispersive X-ray (EDX) spectroscopy. The remaining specimens were randomly divided into four groups (n=20/group) with different moisture content: dried (Group 2 and Group 4) or not dried (Group 1 and Group 3). Pulsed Er:YAG laser (120 mJ/pulse, 20 Hz, 100 μs) was used for perpendicular irradiation with (Group 1 and Group 2) or without (Group 3 and Group 4) water spray. The treated specimens were subjected to SEM, EDX spectroscopy, OCT, and then processed for histological evaluation.Results: After Er:YAG laser ablation, the melting effect was obvious in the SEM results of the dry groups and non-sprayed groups (Groups 2-4). EDX spectroscopy showed that the content of calcium and phosphorus and their ratios remained unchanged in the undried and with water spray group (Group 1) were the closest to the control group. Three-dimensional (3D) OCT could evaluate the depth and shape of ablation grooves. The roughness of the laser-ablated surface could be visualized by extracting the surface lines from cross-sectional OCT images. The results illustrated that the laser ablation with water spray could achieve a smoother surface. Furthermore, OCT results demonstrated that a layer with high image intensity was generated on the bone surface after laser irradiation. The thickness of these layers showed a correlation with whether or not the laser irradiation was sprayed with water. Histology showed that thin eosin-stained layers were created in all experimental groups, which matched well with OCT results of the layers with high image intensity. No denatured layer was observed in the non-irradiated areas of bone tissues.Conclusions: OCT could rapidly and non-destructively visualize the bone surface before and after Er:YAG laser ablation at four different moisture contents. The morphology of ablation grooves, as well as the roughness and thickness of the heterogeneous layer on the bone surface, could be characterized quantitatively with good correlation with SEM and histology. This study will promote the development ...

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