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Treatment of early age-related macular degeneration in a cell culture model with femtosecond laser pulses Open Access


Other title
Femtosecond laser-tissue interaction
Age-related macular degeneration
Drusen ablation
Sub-RPE deposit
ARPE19 cell culture model
Ultrafast lasers
Type of item
Degree grantor
University of Alberta
Author or creator
Smith, Katherine N
Supervisor and department
Elezzabi, Dr. Abdulhakem (Department of Electrical and Computer Engineering)
Examining committee member and department
MacDonald, Dr. Ian (Department of Ophthalmology and Visual Sciences)
Shankar,Dr. Karthik (Department of Electrical and Computer Engineering)
Department of Electrical and Computer Engineering
Photonics and Plasmas
Date accepted
Graduation date
2017-11:Fall 2017
Master of Science
Degree level
This thesis work focuses on obtaining preliminary results demonstrating the isolated femtosecond laser ablation of sub-RPE drusen-like deposits. Age-related macular degeneration (AMD) is an age-onset degenerative condition of the retina that can lead to legal blindness. The early stages of the condition are characterized by the development of hemi-spherical cellular debris accumulations, known as drusen, under the retina. This project has two specific aims: demonstrating the laser ablation of AMD related sub-RPE deposits and assembling a prototype for a clinical therapeutic beam delivery system. An articulating arm is fitted with silver mirrors for 800 nm and coupled to a slit lamp ophthalmic device for femtosecond laser delivery to the retina. A novel cell culture model using the immortalized cell line ARPE-19 is developed. The cell culture model is evaluated by confocal and transmission electron microscopy for the presence of sub-RPE drusen-like deposits. The samples are stained for two known drusen components, apolipoprotein E and cholesterol, to verify the artificial drusen-like deposits share a similar biochemical composition to drusen in vivo. Sub-RPE drusen-like deposits in the ARPE-19 model were targeted with 10 femtosecond pulse trains, 1.9-4.2 nJ/pulse, centered at 800 nm wavelength, from a Ti:Sapphire laser. Fluorescence microscopy was used to qualitatively confirm successful deposit ablation. After laser ablation more fluorescent dye was added to the dish and monitored to see if the areas targeted by the laser regain fluorescence. Although areas of the sample did regain fluorescence, the targeted deposit accumulation never regained fluorescence. Thus, eliminating the possibility of laser photobleaching being responsible for deposit removal as opposed to plasma mediated breakdown.
This thesis is made available by the University of Alberta Libraries with permission of the copyright owner solely for the purpose of private, scholarly or scientific research. This thesis, or any portion thereof, may not otherwise be copied or reproduced without the written consent of the copyright owner, except to the extent permitted by Canadian copyright law.
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