Identification of chondrocyte cell division in intact rabbit cartilage grafts following photothermal heating using fluorescence-activated cell counting

Mai Thy Truong; Kenneth K. H. Chao; Nidhi S. Pandoh; Fang-Yao Hou; David M. Gardiner; Brian Jet-Fei Wong M.D.

doi:10.1117/12.441077

17 June 2002 Identification of chondrocyte cell division in intact rabbit cartilage grafts following photothermal heating using fluorescence-activated cell counting

Mai Thy Truong, Kenneth K. H. Chao, Nidhi S. Pandoh, Fang-Yao Hou, David M. Gardiner, Brian Jet-Fei Wong M.D.

Author Affiliations +

Proceedings Volume 4609, Lasers in Surgery: Advanced Characterization, Therapeutics, and Systems XII; (2002) https://doi.org/10.1117/12.441077
Event: International Symposium on Biomedical Optics, 2002, San Jose, CA, United States

Abstract

The lack of adequate cartilage for aesthetic and reconstructive head and neck surgery has prompted intense research in developing methods to stimulate chondrocyte proliferation in cartilage, a terminally differentiated tissue. In this study, we propose that laser heating can stimulate chondrocyte proliferation in rabbit nasal septal cartilage. Ex-vivo rabbit nasal septal cartilage grafts were irradiated using a CO2 laser (?=10.6 um, 5, 10, or 20 W, 0.1 or 0.5 seconds) and then placed in tissue culture for 7-21 days with 10 _M of 5-bromo-2’-deoxyuridine (BrdU), a thymidine analog that is incorporated into DNA during the S phase of the cell cycle. Chondrocytes were then isolated using enzymatic digestion, incubated with fluorescein conjugated anti-BrdU monoclonal antibodies (Roche Diagnostic, Basel, Switzerland) and counterstained with propidium iodide (PI), a DNA intercalater.? Fluorescent-activated cell counting was performed by flow cytometry (excitation wavelength ?=488 nm, Model BD LSR Flow Cytometer, Becton Dickson, Franklin Lakes, NJ) identifying cells with emissions measured at 515 nm (fluorescein labeled anti-BrdU) and 620 nm (PI).? Cellular debris and cell aggregates were gated out of the signal.?Cartilage grafts irradiated approximately 24 times with the CO2 laser were found to have a significant proliferating population by flow cytometry. Chondrocytes incorporating BrdU into their DNA were clearly identified and the signal intensity increased in integral multiples from baseline values indicating these cells had undergone cell division.? This provides further evidence that chondroctyes have a proliferative response to photothermal stimulation, which may be a response to injury to thermal heating by laser irradiation.

Citation Download Citation

Mai Thy Truong, Kenneth K. H. Chao, Nidhi S. Pandoh, Fang-Yao Hou, David M. Gardiner, and Brian Jet-Fei Wong M.D. "Identification of chondrocyte cell division in intact rabbit cartilage grafts following photothermal heating using fluorescence-activated cell counting", Proc. SPIE 4609, Lasers in Surgery: Advanced Characterization, Therapeutics, and Systems XII, (17 June 2002); https://doi.org/10.1117/12.441077

ACCESS THE FULL ARTICLE

INSTITUTIONAL
Select your institution to access the SPIE Digital Library.

SELECT YOUR INSTITUTION

PERSONAL
Sign in with your SPIE account to access your personal subscriptions or to use specific features such as save to my library, sign up for alerts, save searches, etc.

PERSONAL SIGN IN

No SPIE Account? Create one

PURCHASE THIS CONTENT

SUBSCRIBE TO DIGITAL LIBRARY

50 downloads per 1-year subscription

Members: $195

Non-members: $335 ADD TO CART

25 downloads per 1 - year subscription

Members: $145

Non-members: $250 ADD TO CART

PURCHASE SINGLE ARTICLE

Includes PDF, HTML & Video, when available

Members: $17.00

Non-members: $21.00 ADD TO CART

PROCEEDINGS
5 PAGES

DOWNLOAD PAPER SAVE TO MY LIBRARY

GET CITATION

RIGHTS & PERMISSIONS

Get copyright permission Get copyright permission on Copyright Marketplace

KEYWORDS

Cartilage

Carbon dioxide lasers

Nd:YAG lasers

Gas lasers

Head

Surgery

Laser irradiation

Show All Keywords

Keywords/Phrases

Search In:

Publication Years