Lifetime fluorescence spectroscopy for in-situ investigation of osteogenic differentiation

Laura Marcu; Amir Elbarbary; Patricia Zuk; Daniel A. De Ugarte; Prosper Benhaim; Hamza Kurt; Marc H. Hedrick; Peter Ashjian

doi:10.1117/12.477913

Lifetime fluorescence spectroscopy for in-situ investigation of osteogenic differentiation

Laura Marcu, Amir Elbarbary, Patricia Zuk, Daniel A. De Ugarte, Prosper Benhaim, Hamza Kurt, Marc H. Hedrick, Peter Ashjian

Biomedical Engineering

Research output: Chapter in Book/Report/Conference proceeding › Conference contribution

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.

Original language	English (US)
Title of host publication	Proceedings of SPIE - The International Society for Optical Engineering
Editors	S.L. Jacques, D.D. Duncan, S.J. Kirkpatrick, A. Kriete
Pages	250-257
Number of pages	8
Volume	4961
DOIs	https://doi.org/10.1117/12.477913
State	Published - 2003
Event	PROGRESS IN BIOMEDICAL OPTICS AND IMAGING: Laser-Tissue Interaction XIV - San Jose, CA, United States Duration: Jan 25 2003 → Jan 29 2003

Other

Other	PROGRESS IN BIOMEDICAL OPTICS AND IMAGING: Laser-Tissue Interaction XIV
Country/Territory	United States
City	San Jose, CA
Period	1/25/03 → 1/29/03

Keywords

Collagen
Time-resolved fluorescence spectroscopy
Tissue engineering osteogenesis

ASJC Scopus subject areas

Electrical and Electronic Engineering
Condensed Matter Physics

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10.1117/12.477913

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Marcu, L., Elbarbary, A., Zuk, P., De Ugarte, D. A., Benhaim, P., Kurt, H., Hedrick, M. H., & Ashjian, P. (2003). Lifetime fluorescence spectroscopy for in-situ investigation of osteogenic differentiation. In S. L. Jacques, D. D. Duncan, S. J. Kirkpatrick, & A. Kriete (Eds.), Proceedings of SPIE - The International Society for Optical Engineering (Vol. 4961, pp. 250-257) https://doi.org/10.1117/12.477913

Lifetime fluorescence spectroscopy for in-situ investigation of osteogenic differentiation. / Marcu, Laura; Elbarbary, Amir; Zuk, Patricia; De Ugarte, Daniel A.; Benhaim, Prosper; Kurt, Hamza; Hedrick, Marc H.; Ashjian, Peter.

Proceedings of SPIE - The International Society for Optical Engineering. ed. / S.L. Jacques; D.D. Duncan; S.J. Kirkpatrick; A. Kriete. Vol. 4961 2003. p. 250-257.

Research output: Chapter in Book/Report/Conference proceeding › Conference contribution

Marcu, L, Elbarbary, A, Zuk, P, De Ugarte, DA, Benhaim, P, Kurt, H, Hedrick, MH & Ashjian, P 2003, Lifetime fluorescence spectroscopy for in-situ investigation of osteogenic differentiation. in SL Jacques, DD Duncan, SJ Kirkpatrick & A Kriete (eds), Proceedings of SPIE - The International Society for Optical Engineering. vol. 4961, pp. 250-257, PROGRESS IN BIOMEDICAL OPTICS AND IMAGING: Laser-Tissue Interaction XIV, San Jose, CA, United States, 1/25/03. https://doi.org/10.1117/12.477913

Marcu, Laura ; Elbarbary, Amir ; Zuk, Patricia ; De Ugarte, Daniel A. ; Benhaim, Prosper ; Kurt, Hamza ; Hedrick, Marc H. ; Ashjian, Peter. / Lifetime fluorescence spectroscopy for in-situ investigation of osteogenic differentiation. Proceedings of SPIE - The International Society for Optical Engineering. editor / S.L. Jacques ; D.D. Duncan ; S.J. Kirkpatrick ; A. Kriete. Vol. 4961 2003. pp. 250-257

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AB - 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.

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Lifetime fluorescence spectroscopy for in-situ investigation of osteogenic differentiation

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