Noninvasive Optical Assessment of Implanted Engineered Tissues Correlates with Cytokine Secretion.
Noninvasive Optical Assessment of Implanted Engineered Tissues Correlates with Cytokine Secretion.
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植入工程组织的无创光学评估与细胞因子分泌相关。
DOI：
10.1089/ten.tec.2017.0516
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发表时间：
2018
期刊：
影响因子：
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通讯作者：
Mycek,Mary-Ann
中科院分区：
文献类型：
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作者：
Elahi,SakibF;Lee,SeungYup;Lloyd,WilliamR;Chen,Leng-Chun;Kuo,Shiuhyang;Zhou,Ying;Kim,HyungjinMyra;Kennedy,Robert;Marcelo,Cynthia;Feinberg,StephenE;Mycek,Mary-Ann
荧光寿命感测已被证明可以非侵入性地表征工程化组织构建体的植入前健康和活力。然而，目前监测植入后结构整合的实践是定性的（目视评估）或破坏性的（组织组织学）。我们采用无标记荧光寿命光谱法对植入小鼠模型的工程化组织结构进行定量、非侵入性光学评估。该便携式系统设计为适用于活体测量，包括手持式探头，可在每个结构的多个部位精确快速地采集数据。我们的模型组织构建体由原代人类细胞制造，以基于标准方案模拟患者变异性，并且一半制造的构建体受到应力以创建一系列健康状态。在体外测量与细胞活力相关的三种细胞因子的分泌量以评估植入前构建体的健康：白细胞介素-8（IL-8）、人β-防御素1（hBD-1）和血管内皮生长因子（VEGF）。植入前IL-8的细胞因子分泌范围为1.5 - 33.5 pg/mL，hBD-1为3.4 - 195.0 pg/mL，VEGF为0.1 - 154.3 pg/mL。我们发现，在植入后1周，体内光学参数与所有三种细胞因子的体外植入前分泌水平相关（p< 0.05）。在植入后3周的光学测量中未观察到这种相关性，此时组织学显示结构已上皮化，与植入前健康状态无关，支持缺乏相关性。这些结果表明，基于内源性组织荧光团的无标记荧光寿命感测的临床光学诊断工具可以无创地监测工程组织的植入后整合。
Fluorescence lifetime sensing has been shown to noninvasively characterize the preimplantation health and viability of engineered tissue constructs. However, current practices to monitor postimplantation construct integration are either qualitative (visual assessment) or destructive (tissue histology). We employed label-free fluorescence lifetime spectroscopy for quantitative, noninvasive optical assessment of engineered tissue constructs that were implanted into a murine model. The portable system was designed to be suitable for intravital measurements and included a handheld probe to precisely and rapidly acquire data at multiple sites per construct. Our model tissue constructs were manufactured from primary human cells to simulate patient variability based on a standard protocol, and half of the manufactured constructs were stressed to create a range of health states. Secreted amounts of three cytokines that relate to cellular viability were measuredin vitroto assess preimplantation construct health: interleukin-8 (IL-8), human β-defensin 1 (hBD-1), and vascular endothelial growth factor (VEGF). Preimplantation cytokine secretion ranged from 1.5 to 33.5 pg/mL for IL-8, from 3.4 to 195.0 pg/mL for hBD-1, and from 0.1 to 154.3 pg/mL for VEGF.In vivooptical sensing assessed constructs at 1 and 3 weeks postimplantation. We found that at 1 week postimplantation,in vivooptical parameters correlated within vitropreimplantation secretion levels of all three cytokines (p< 0.05). This correlation was not observed in optical measurements at 3 weeks postimplantation when histology showed that the constructs had re-epithelialized, independent of preimplantation 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 postimplantation integration of engineered tissues.