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 | |
| 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 | United States |
| City | San Jose, CA |
| Period | 1/25/03 → 1/29/03 |
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Keywords
- Collagen
- Time-resolved fluorescence spectroscopy
- Tissue engineering osteogenesis
ASJC Scopus subject areas
- Electrical and Electronic Engineering
- Condensed Matter Physics
Cite this
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
}
TY - GEN
T1 - Lifetime fluorescence spectroscopy for in-situ investigation of osteogenic differentiation
AU - Marcu, Laura
AU - Elbarbary, Amir
AU - Zuk, Patricia
AU - De Ugarte, Daniel A.
AU - Benhaim, Prosper
AU - Kurt, Hamza
AU - Hedrick, Marc H.
AU - Ashjian, Peter
PY - 2003
Y1 - 2003
N2 - 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.
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.
KW - Collagen
KW - Time-resolved fluorescence spectroscopy
KW - Tissue engineering osteogenesis
UR - http://www.scopus.com/inward/record.url?scp=0345714665&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0345714665&partnerID=8YFLogxK
U2 - 10.1117/12.477913
DO - 10.1117/12.477913
M3 - Conference contribution
AN - SCOPUS:0345714665
VL - 4961
SP - 250
EP - 257
BT - Proceedings of SPIE - The International Society for Optical Engineering
A2 - Jacques, S.L.
A2 - Duncan, D.D.
A2 - Kirkpatrick, S.J.
A2 - Kriete, A.
ER -