Multimodal fluorescence lifetime imaging and optical coherence tomography for longitudinal monitoring of tissue-engineered cartilage maturation in a preclinical implantation model.
Multimodal fluorescence lifetime imaging and optical coherence tomography for longitudinal monitoring of tissue-engineered cartilage maturation in a preclinical implantation model.
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DOI：
10.1117/1.jbo.28.2.026003
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发表时间：
2023-02
影响因子：
3.5
通讯作者：
Marcu, Laura
中科院分区：
文献类型：
--
作者：
Zhou, Xiangnan;Haudenschild, Anne K.;Li, Cai;Marcu, Laura
关键词：
软骨组织工程是治疗骨关节炎的有效治疗方法。然而，组织工程师主要依靠耗时、昂贵和破坏性的技术来监控工程软骨的成熟。这种做法对于大规模生物制造是不切实际的，并且阻碍了对组织生长的空间和时间监测，而这对于制造临床相关大小的软骨结构是至关重要的。结合荧光寿命成像（FLIm）和光学相干断层扫描（OCT）的无损多模态成像技术在解决这一挑战方面具有巨大的潜力。研究了在临床前小鼠模型中使用多模态flm - oct对自组装软骨组织成熟进行无损、空间和时间监测的可行性。自组装软骨结构在体外培养4周，然后在裸鼠体内成熟4周。在组织发育的多个时间点（4周和8周）进行无菌无损原位多光谱胶片和OCT成像。重建薄膜和三维体积OCT图像，用于分析组织生化均匀性、形态学和结构完整性。计算生化均匀性指数来表征不同时间点的非均匀组织生长。根据组织学对OCT图像进行验证。薄膜检测异质细胞外基质（ECM）生长的组织工程软骨。组织结构的外缘在375 ~ 410和450 ~ 485 nm光谱通道中显示出更长的荧光寿命，表明胶原含量增加。结构同质性指数在周间显著降低。在体外培养（和4周）期间，胶片和OCT图像均显示组织结构中心存在缺陷（空洞）。在体内培养过程中，用OCT检测到囊肿形成，并用组织学证实。多模态flm - oct在原位无损监测工程组织异质生长的能力得到了证明。利用FLIm技术检测构造ECM成分的时空变化。OCT显示结构缺陷（空洞和囊肿）。这种多模式方法具有巨大的潜力，可以取代组织工程医疗产品制造中昂贵的破坏性测试，促进其临床转化。
Cartilage tissue engineering is a promising strategy for effective curative therapies for treatment of osteoarthritis. However, tissue engineers depend predominantly on time-consuming, expensive, and destructive techniques as quality control to monitor the maturation of engineered cartilage. This practice can be impractical for large-scale biomanufacturing and prevents spatial and temporal monitoring of tissue growth, which is critical for the fabrication of clinically relevant-sized cartilage constructs. Nondestructive multimodal imaging techniques combining fluorescence lifetime imaging (FLIm) and optical coherence tomography (OCT) hold great potential to address this challenge. The feasibility of using multimodal FLIm–OCT for nondestructive, spatial, and temporal monitoring of self-assembled cartilage tissue maturation in a preclinical mouse model is investigated. Self-assembled cartilage constructs were developed for 4 weeks in vitro followed by 4 weeks of in vivo maturation in nude mice. Sterile and nondestructive in situ multispectral FLIm and OCT imaging were carried out at multiple time points (, 4, and 8 weeks) during tissue development. FLIm and 3D volumetric OCT images were reconstructed and used for the analysis of tissue biochemical homogeneity, morphology, and structural integrity. A biochemical homogeneity index was computed to characterize nonhomogeneous tissue growth at different time points. OCT images were validated against histology. FLIm detects heterogenous extracellular matrix (ECM) growth of tissue-engineered cartilage. The outer edge of the tissue construct exhibited longer fluorescence lifetime in 375 to 410 and 450 to 485 nm spectral channels, indicating increase in collagen content. Significant () decrease of construct homogeneity index was observed between weeks and weeks. Both FLIm and OCT images revealed defects (voids) at the center of the tissue construct during in vitro culture ( and 4 weeks). Cyst formation during in vivo culture was detected by OCT and confirmed with histology. The ability of multimodal FLIm–OCT to nondestructively monitor the heterogenous growth of engineered tissue constructs in situ is demonstrated. Spatial and temporal variation of construct ECM component was detected by FLIm. OCT reveals structural defects (voids and cysts). This multimodal approach has great potential to replace costly destructive tests in the manufacturing of tissue-engineered medical products, facilitating their clinical translation.
影响因子：
2.8
作者：
Alfonso-Garcia A;Shklover J;Sherlock BE;Panitch A;Griffiths LG;Marcu L
通讯作者：
Marcu L