12 February 2018 Assessment of post-implantation integration of engineered tissues using fluorescence lifetime spectroscopy
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Clinical translation of engineered tissue constructs requires noninvasive methods to assess construct health and viability after implantation in patients. However, current practices to monitor post-implantation construct integration are either qualitative (visual assessment) or destructive (tissue histology). As label-free fluorescence lifetime sensing can noninvasively characterize pre-implantation construct viability, we employed a handheld fluorescence lifetime spectroscopy probe to quantitatively and noninvasively assess tissue constructs that were implanted in a murine model. We designed the system to be suitable for intravital measurements: portability, localization with precise maneuverability, and rapid data acquisition. Our model tissue constructs were manufactured from primary human cells to simulate patient variability and were stressed to create a range of health states. Secreted amounts of three cytokines that relate to cellular viability were measured in vitro to assess pre-implantation construct health. In vivo optical sensing assessed tissue integration of constructs at one-week and three-weeks post-implantation. At one-week post-implantation, optical parameters correlated with in vitro pre-implantation secretion levels of all three cytokines (p < 0.05). This relationship was no longer seen at three-weeks post-implantation, suggesting comparable tissue integration independent of preimplantation health. Histology confirmed re-epithelialization of these constructs independent of pre-implantation health state, supporting the lack of a correlation. These results suggest that clinical optical diagnostic tools based on label-free fluorescence lifetime sensing of endogenous tissue fluorophores could noninvasively monitor post-implantation integration of engineered tissues.
Conference Presentation
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Sakib F. Elahi, Sakib F. Elahi, Seung Y. Lee, Seung Y. Lee, William R. Lloyd, William R. Lloyd, Leng-Chun Chen, Leng-Chun Chen, Shiuhyang Kuo, Shiuhyang Kuo, Ying Zhou, Ying Zhou, Hyungjin M. Kim, Hyungjin M. Kim, Robert Kennedy, Robert Kennedy, Cynthia Marcelo, Cynthia Marcelo, Stephen E. Feinberg, Stephen E. Feinberg, Mary-Ann Mycek, Mary-Ann Mycek, } "Assessment of post-implantation integration of engineered tissues using fluorescence lifetime spectroscopy", Proc. SPIE 10484, Advanced Biomedical and Clinical Diagnostic and Surgical Guidance Systems XVI, 104840G (12 February 2018); doi: 10.1117/12.2290280; https://doi.org/10.1117/12.2290280

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