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Proceedings Paper

Lifetime fluorescence spectroscopy for in situ investigation of osteogenic differentiation
Author(s): Laura Marcu; Amir Elbarbary; Patricia Zuk; Daniel A. De Ugarte; Prosper Benhaim; Hamza Kurt; Marc H. Hedrick; Peter Ashjian
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Paper Abstract

Time-Resolved Laser-Induced Fluorescence Spectroscopy (TR-LIFS) represents a potential tool for the in-situ characterization of bioengineered tissues. In this study, we evaluate the application of TR-LIFS to non-intrusive monitoring of matrix composition during osteogenetic differentiation. Human adipose-derived stem cells, harvested from 3 patients, were induced in osteogenic media for 3, 5, and 7 weeks. Samples were subsequently collected and probed for time-resolved fluorescence emission with a pulsed nitrogen laser. Fluorescence parameters, derived from both spectral- and time-domain, were used for sample characterization. The samples were further analyzed using Western blot analysis and computer-based densitometry. A significant change in the fluorescence parameters was detected for samples beyond 3 weeks of osteogenic differentiation. The spectroscopic observations: 1) show increase of collagen I when contrasted against the time-resolved fluorescence spectra of commercially available collagens; and 2) are in agreement with Western blot analysis that demonstrated significant increase in collagen I content between 3- vs. 5-weeks and 3- vs. 7-weeks and no changes for collagens III, IV, and V. Our results suggest that TR-LIFS can be used as a non-invasive means for the detection of specific collagens in maturing connective tissues.

Paper Details

Date Published: 27 August 2003
PDF: 8 pages
Proc. SPIE 4961, Laser-Tissue Interaction XIV, (27 August 2003); doi: 10.1117/12.477913
Show Author Affiliations
Laura Marcu, Cedars-Sinai Medical Ctr. (United States)
Univ. of Southern California (United States)
Amir Elbarbary, Univ. of California, Los Angeles School of Medicine (United States)
Patricia Zuk, Univ. of California, Los Angeles School of Medicine (United States)
Daniel A. De Ugarte, Univ. of California, Los Angeles School of Medicine (United States)
Prosper Benhaim, Univ. of California, Los Angeles School of Medicine (United States)
Hamza Kurt, Cedars-Sinai Medical Ctr. (United States)
Marc H. Hedrick, Univ. of California, Los Angeles School of Medicine (United States)
Peter Ashjian, Univ. of California, Los Angeles School of Medicine (United States)


Published in SPIE Proceedings Vol. 4961:
Laser-Tissue Interaction XIV
Donald Dean Duncan; Sean J. Kirkpatrick; Andres Kriete; Steven L. Jacques, Editor(s)

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