Department of Biomedical Engineering, University of Michigan College of Engineering, Ann Arbor, Michigan, USA.
Department of Oral and Maxillofacial Surgery, University of Michigan School of Dentistry, Ann Arbor, Michigan, USA.
Tissue Eng Part C Methods. 2021 May;27(5):287-295. doi: 10.1089/ten.TEC.2021.0018.
Quantitative diffuse reflectance spectroscopy (DRS) was developed for label-free, noninvasive, and real-time assessment of implanted tissue-engineered devices manufactured from primary human oral keratinocytes (six batches in two 5-patient cohorts). Constructs were implanted in a murine model for 1 and 3 weeks. DRS evaluated construct success using optical absorption (hemoglobin concentration and oxygenation, attributed to revascularization) and optical scattering (attributed to cellular density and layer thickness). Destructive pre- and postimplantation histology distinguished experimental control from stressed constructs, whereas noninvasive preimplantation measures of keratinocyte glucose consumption and residual glucose in spent culture media did not. In constructs implanted for 1 week, DRS distinguished control due to stressed and compromised from healthy constructs. In constructs implanted for 3 weeks, DRS identified constructs with higher postimplantation success. These results suggest that quantitative DRS is a promising, clinically compatible technology for rapid, noninvasive, and localized tissue assessment to characterize tissue-engineered construct success . Impact statement Despite the recent advance in tissue engineering and regenerative medicine, there is still a lack of nondestructive tools to longitudinally monitor the implanted tissue-engineered devices. In this study, we demonstrate the potential of quantitative diffuse reflectance spectroscopy as a clinically viable technique for noninvasive, label-free, and rapid characterization of graft success .
定量漫反射光谱(DRS)技术用于非侵入式、实时评估源自原代人口腔角质细胞的组织工程植入物(两个五名患者队列中的六批),无需标记。构建体在小鼠模型中植入 1 周和 3 周。DRS 使用光学吸收(归因于新生血管化的血红蛋白浓度和氧合)和光学散射(归因于细胞密度和层厚度)来评估构建体的成功。破坏性的植入前和植入后组织学将实验对照与应激构建体区分开来,而非侵入性的植入前角质细胞葡萄糖消耗和废弃培养介质中的残留葡萄糖测量则无法区分。在植入 1 周的构建体中,DRS 区分了由于应激和受损导致的对照和健康构建体。在植入 3 周的构建体中,DRS 识别出具有更高植入后成功率的构建体。这些结果表明,定量 DRS 是一种很有前途的、临床兼容的技术,可用于快速、非侵入性和局部组织评估,以表征组织工程构建体的成功。 尽管组织工程和再生医学最近取得了进展,但仍缺乏非破坏性工具来纵向监测植入的组织工程设备。在这项研究中,我们证明了定量漫反射光谱作为一种可行的临床技术,可用于非侵入性、无标记和快速表征移植物的成功。