FLIm for Tissue Engineering Applications

Tissue Engineering
Recellularization over time of both sides of antigen-removed bovine pericardium scaffolds with human aortic endothelial cells and human mesenchymal stem cells. Adapted from Alfonso-Garcia, A. et al. J Biophotonics 11, e201700391 (2018)

The Marcu Lab is developing FLIm as a non-destructive method to monitor engineered biomaterials. Growing and manipulating biomaterials to be used as tissue substitutes requires close monitoring of the tissue biochemical, structural and functional properties. Most tools and methods to measure the samples’ properties are destructive in nature, which render the tissue unusable after examination. As a result, a lot of samples are required to elaborate protocols and methods, which demands many resources and has an elevated cost. 

FLIm relies on the autofluorescence lifetime of cells and tissue components to provide information on biochemical and structural changes. Fluorescence lifetime is highly sensitive to the fluorophore microenvironment, which changes with different maturation stages of the engineered biomaterials. In addition, the label-free and non-destructive nature of the approach allows for in vivo imaging of the tissue in the host after implantation, providing a means to monitor the healing process and to predict possible failure events.

Our previous work has focused on vascular, cartilage, and bone tissue engineering. We have demonstrated the ability of FLIm to monitor scaffold structural properties such as the cross-linking and digestion of collagen networks, the recellularization of natural vascular scaffolds, and FLIm’s ability to detect the maturation stages of osteogenic grafts. 

The tissue engineering work is also a platform for technology development including multimodal imaging, see FLIm-OCT, FLIm-Raman, endo-exo platform.

 

Collaborators

Leigh G. Griffiths, Ph.D. (Mayo Clinic)

Prof. J. Kent Leach (UC Davis)

Kyriacos A. Athanasiou, Ph.D, Ph.M. (UC Davis/UC Irvine)

 

Funding

NIH (National Institutes of Health): R01HL121068

CIRM (California Institute for Regenerative Medicine): RT3-07879, RT3-07981

 

Publications
N. Vazquez-Portalatin, A. Alfonso-Garcia, J. C. Liu, L. Marcu, and A. Panitch, "Physical, Biomechanical, and Optical Characterization of Collagen and Elastin Blend Hydrogels," Ann Biomed Eng 48, 2924-2935 (2020).
J. Shklover, J. McMasters, A. Alfonso-Garcia, M. L. Higuita, A. Panitch, L. Marcu, and L. Griffiths, "Bovine pericardial extracellular matrix niche modulates human aortic endothelial cell phenotype and function," Sci Rep 9, 16688 (2019).
A. K. Haudenschild, B. E. Sherlock, X. Zhou, J. C. Hu, J. K. Leach, L. Marcu, and K. A. Athanasiou, "Non-destructive detection of matrix stabilization correlates with enhanced mechanical properties of self-assembled articular cartilage," J Tissue Eng Regen Med 13, 637-648 (2019).
J. N. Harvestine, C. S. Sheaff, C. Li, A. K. Haudenschild, M. A. Gionet-Gonzales, J. C. Hu, K. A. Athanasiou, L. Marcu, and J. K. Leach, "Multimodal Label-Free Imaging for Detecting Maturation of Engineered Osteogenic Grafts," Acs Biomater Sci Eng 5, 1956-1966 (2019).
X. Zhou, A. K. Haudenschild, B. E. Sherlock, J. C. Hu, J. K. Leach, K. A. Athanasiou, and L. Marcu, "Detection of glycosaminoglycan loss in articular cartilage by fluorescence lifetime imaging," J Biomed Opt 23, 1-8 (2018).
B. E. Sherlock, J. N. Harvestine, D. Mitra, A. Haudenschild, J. Hu, K. A. Athanasiou, J. K. Leach, and L. Marcu, "Nondestructive assessment of collagen hydrogel cross-linking using time-resolved autofluorescence imaging," J Biomed Opt 23, 1-9 (2018).
 C. Li, J. Shklover, M. Parvizi, B. E. Sherlock, A. Alfonso Garcia, A. K. Haudenschild, L. G. Griffiths, and L. Marcu, "Label-Free Assessment of Collagenase Digestion on Bovine Pericardium Properties by Fluorescence Lifetime Imaging," Ann Biomed Eng 46, 1870-1881 (2018).
A. K. Haudenschild, B. E. Sherlock, X. Zhou, J. C. Hu, J. K. Leach, L. Marcu, and K. A. Athanasiou, "Nondestructive fluorescence lifetime imaging and time-resolved fluorescence spectroscopy detect cartilage matrix depletion and correlate with mechanical properties," Eur Cell Mater 36, 30-43 (2018). 
A. Alfonso-Garcia, J. Shklover, B. E. Sherlock, A. Panitch, L. G. Griffiths, and L. Marcu, "Fiber-based fluorescence lifetime imaging of recellularization processes on vascular tissue constructs," J Biophotonics 11, e201700391 (2018).
A. Alfonso-Garcia, A. K. Haudenschild, and L. Marcu, "Label-free assessment of carotid artery biochemical composition using fiber-based fluorescence lifetime imaging," Biomed Opt Express 9, 4064-4076 (2018).