INTRODUCTIONThe prevalence of bladder cancer is very high, due to its high recurrence rate in superficial bladder cancer (30 to 85%), which is the staging of approximately 80% of the patients at first diagnosis. Risk of recurrence and progression is associated with grade, stage, presence of concomitant carcinoma in situ, size and number of lesions, as well as time to first recurrence. Recurrences can be partly attributed to new occurrences but also to residual tumors after resection. Incomplete tumor removal has been observed in 30 to 50% of TUR's, especially when dealing with T 1 or poorly visible malignant or pre-malignant disease 1 . Fluorescence guided resection with 5 amino levulinic acid (ALA) or its hexyl ester derivative (Hexvix®), has now unequivocally been demonstrated to increase detection rate and a growing number of studies indicate this has a positive impact on recurrence and progression rates 2-3 . Implantation of viable tumor cells, dispersed during resection, is a third Photodynamic Therapy: Back to the Future, edited by David H. Kessel, Proc. of SPIE Vol. 7380, 73801V · © 2009 SPIE · CCC code: 1605-7422/09/$18 · Downloaded From: http://proceedings.spiedigitallibrary.org/ on 06/15/2016 Terms of Use: http://spiedigitallibrary.org/ss/TermsOfUse.aspx factor influencing bladder cancer recurrence 4-6 . The aim of early intravesical therapy is to interfere with cell viability and thus reduce implantation risks. Photodynamic therapy (PDT) is a treatment modality that combines the use of a photosensitising molecule and light in the presence of oxygen. The first approval for Photofrin® mediated PDT was obtained for the treatment of bladder cancer in 1982. Although encouraging oncological results, PDT was abandoned due to side effects such as skin sensitisation and bladder shrinking 7 . With the venue of ALA or derivatives thereof, which are administered intravesicaly and have better tumor specificity as opposed to muscle, interest for PDT in bladder cancer treatment has regained interest 8-12 .The aim of the current paper was to investigate whether PDT immediately after fluorescence guided TUR could reduce the amount of viable intraluminal tumor cells and whether this had an impact on tumor implantation rate.
MATERIAL AND METHODS
Hexvix ® solution and bladder cancer cell line preparationHexvix® (hALA) (Photocure ASA,Oslo, Norway) solution at 0.8 mM was prepared in RPMI-1640 (GIBCO, Invitrogen, UK) medium supplemented with 2 % foetal calf serum (FCS) (pH 7.4) before the experiment. The tumor cell line was AY27, initially derived from carcinomas of the urinary bladder induced in female Fischer (F344). Cells were cultivated in RPMI-1640 supplemented with 10 % FCS, 5 % L-glutamine (200 mM) and 5 % penicillin-streptomycin (Biotech, GmbH, Aidenbach, Germany). Subconfluent AY-27 cells in tissue culture flasks were washed twice with PBS (pH 7.4), trypsinized (trypsin EDTA, Gibco) and collected after centrifugation. Cells (3x10 6 ) were resuspended in 164 μl Nucleofector Solution V at room temperature followed by ...