Spatiotemporal Optoacoustic Mapping of Tumor Hemodynamics in a Clinically Relevant Orthotopic Rabbit Model of Head and Neck Cancer Laurie J. Rich, Sandra.

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Spatiotemporal Optoacoustic Mapping of Tumor Hemodynamics in a Clinically Relevant Orthotopic Rabbit Model of Head and Neck Cancer Laurie J. Rich, Sandra Sexton, Leslie Curtin, Mukund Seshadri Translational Oncology Volume 10, Issue 5, Pages 839-845 (October 2017) DOI: 10.1016/j.tranon.2017.08.004 Copyright © 2017 The Authors Terms and Conditions

Figure 1 Experimental setup for PAI with co-registered US of rabbits bearing orthotopic VX2 tumors. The system consists of a tunable NIR Nd:YAG laser connected to a fiber-optic 256-element linear array hybrid US transducer and processing workstation that creates co-registered PA and US images (Vevo 2100 LAZR; FujiFilm VisualSonics Corporation). Translational Oncology 2017 10, 839-845DOI: (10.1016/j.tranon.2017.08.004) Copyright © 2017 The Authors Terms and Conditions

Figure 2 Surgical procedure for establishing VX2 carcinomas in the neck of NZW rabbits. (A) Simplified view of rabbit anatomy. (B) Digital photograph of a naive rabbit showing the neck where an incision was made along the length of the dotted red line. (C, D) Subcutaneous tissue dissection and deposition of donor tumor tissue into the sternohyoid muscle. Clinical (E) and histologic (F) appearance of a rabbit bearing orthotopic VX2 carcinoma in the neck. Translational Oncology 2017 10, 839-845DOI: (10.1016/j.tranon.2017.08.004) Copyright © 2017 The Authors Terms and Conditions

Figure 3 Ultrasonography of VX2 tumor growth in vivo. (A) B-mode US images of a VX2 tumor in the neck of a rabbit at different times (days 7, 14, and 21 postimplantation). The extent of the tumor is outlined in white. (B) Bar graph shows US-based estimates of tumor volume in rabbits (n = 9) over a 3-week period. **P < .01; ***P < .001. Translational Oncology 2017 10, 839-845DOI: (10.1016/j.tranon.2017.08.004) Copyright © 2017 The Authors Terms and Conditions

Figure 4 Optoacoustic mapping of temporal changes in oxygenation of VX2 rabbit carcinomas. (A) Pseudocolorized PA map of tumor oxygenation saturation overlaid on a B-mode US image of an orthotopic VX2 tumor in a rabbit is shown on the left. Corresponding Doppler image of tumor vascularity is shown on the right. The tumor is outlined in white on both images. Bar graphs show temporal changes in tumor %sO2 (B) and HbT (C) detected by PAI (n = 9). Corresponding values of tumor vascularity (D) on Doppler sonographic examination (n = 8) of orthotopic rabbit VX2 carcinomas over a 3-week period. Plots illustrating the correlation between %sO2 and tumor volume (Spearman r = −0.80; P < .0001; E), HbT and tumor volume (Spearman r = −0.64, P = .0004; F), and PV and tumor volume (Spearman r = −0.72; P < .0001; G). *P < .05; **P < .01. Translational Oncology 2017 10, 839-845DOI: (10.1016/j.tranon.2017.08.004) Copyright © 2017 The Authors Terms and Conditions

Figure 5 PAI and Doppler sonography of spatial heterogeneity in oxygenation and vascularity of VX2 rabbit carcinoma. (A) Pseudocolorized PA %sO2 and Doppler vascularity maps of a VX2 tumor showing considerable heterogeneity within the tumor. Peripheral (rim) regions of the tumor showed consistently higher values of %sO2 (B) and HbT (C) than the tumor core. Consistent with the PAI data, Doppler sonography (D) also showed higher values of vascularity in the tumor rim compared to the core. *P < .05; **P < .01. Translational Oncology 2017 10, 839-845DOI: (10.1016/j.tranon.2017.08.004) Copyright © 2017 The Authors Terms and Conditions