Identification of chondrocyte proliferation with the flourescent-activated cell sorter following laser irradiation and thermal and mechanical treatments

Nidhi S. Pandoh; Mai Thy Truong; Sergio H. Diaz; Kenneth Chao; Stephen Hou; David M. Gardiner; Brian J. F. Wong M.D.

doi:10.1117/12.485127

12 September 2003 Identification of chondrocyte proliferation with the fluorescent-activated cell sorter following laser irradiation and thermal and mechanical treatments

Nidhi S. Pandoh, Mai Thy Truong, Sergio H. Diaz, Kenneth Chao, Stephen Hou, David M. Gardiner, Brian J. F. Wong M.D.

Author Affiliations +

Proceedings Volume 4949, Lasers in Surgery: Advanced Characterization, Therapeutics, and Systems XIII; (2003) https://doi.org/10.1117/12.485127
Event: Biomedical Optics, 2003, San Jose, CA, United States

Abstract

Laser irradiation may cause a proliferative response in cartilage leading to new, less invasive treatment modalities for diseases such as osteoarthritis. Our previous studies have shown that laser irradiation causes chondrocytes to proliferate on the periphery of the laser-irradiated region and this effect is dose dependent. In this study, flow cytometry was used for cell counting because the colorimetric assay used previously was relatively insensitive to small proliferative responses. Ex-vivo rabbit nasal septal cartilages were harvested and subjected to one of three treatments: laser irradiation, heating, or mechanical modification. Specimens were irradiated with an Nd:YAG laser (λ=1.32μm, 4-16 sec, 6 W/cm²), heated by immersion in saline or contact heating, and mechanically modified by scoring with a scalpel or crushing with a metal stamp. Specimens were incubated for 7-21 days in growth media containing 10μM Bromodeoxyuridine (BrdU) then chondrocytes were isolated with enzymatic digestion. Cells were incubated with fluorescein conjugated anti-BrdU monoclonal antibody (Roche Diagnostic, Basel, Switzerland) and counterstained with propidium iodide (PI). Fluorescent-activated cell counting was performed by flow cytometry (λ=488 nm, Model BD LSR Flow Cytometer, Becton Dickson) identifying cells with emissions measured at 515 nm (anti-BrdU) and 620 nm (PI). Analysis showed a population of cells in S-phase of the cell cycle indicating these cells had undergone cell division. This provides further evidence that laser irradiation causes a proliferative response in chondrocytes and may lead to new treatments for degenerative articular diseases and disorders.

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Nidhi S. Pandoh, Mai Thy Truong, Sergio H. Diaz, Kenneth Chao, Stephen Hou, David M. Gardiner, and Brian J. F. Wong M.D. "Identification of chondrocyte proliferation with the fluorescent-activated cell sorter following laser irradiation and thermal and mechanical treatments", Proc. SPIE 4949, Lasers in Surgery: Advanced Characterization, Therapeutics, and Systems XIII, (12 September 2003); https://doi.org/10.1117/12.485127

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KEYWORDS

Laser irradiation

Cartilage

Metals

Flow cytometry

Cell death

Iron

Tissues

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