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Proceedings Paper

Localization of hypericin-induced fluorescence after Hypericum perforatum polar fraction instillation in normal rat urinary bladder
Author(s): Nikos E. Stavropoulos M.D.; Dimitris Skalkos; Ioannis Tsimaris; D. Kalogeras; Unyime O. Nseyo M.D.; A. Batistatou; N. J. Agnantis
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Paper Abstract

The photodynamic action of the Hypericum perforatum L. extract, mainly its polar methanolic fraction (PMF) has recently been substantiated by our group. The herb contains a number of naphthodianthrones - photosensitizers mainly hypericin and pseudohypericin. The concentration of hypericins in PMF was found to be 1.37 %. The distribution of hypericins fluorescence in sections of normal rat bladder tissues after the intravesical instillation of the polar methanolic fraction of hypericum (PMF) was studied by the use of fluorescence microscopy. PMF was dissolved in normal saline containing 0.5 μg/ml concentration of hypericins, and was then instilled in rat bladder for 15, 30, 60 and 120 minutes respectively. PMF solutions were withdrawn, bladders were rinsed through the catheter with normal saline and rats were sacrificed. Bladders were then removed, cut open and immediately mounted in medium, and immersed in liquid nitrogen. Two consecutive 3-μm frozen sections were cut with a cryostat. The first section was examined by fluorescence microscopy and the second section was stained with hematoxylin and eosin. For fluorescence imaging the filter set used included a 535/50 nm bandpass excitation filter and a 610/75 nm emission filter. Fluorescence images were acquired and documented using photography. Fluorescene could be detected in bladder samples after only 15 minutes of instillation with the above described solution. The urothelium / muscle fluorescence ratio ranged from 5/1 to 11/1 in various sites of the samples examined. No fluorescence originating from the muscle could be detected. PMF should be further studied towards the direction of its use in photodynamic therapy.

Paper Details

Date Published: 8 April 2005
PDF: 7 pages
Proc. SPIE 5689, Optical Methods for Tumor Treatment and Detection: Mechanisms and Techniques in Photodynamic Therapy XIV, (8 April 2005); doi: 10.1117/12.588496
Show Author Affiliations
Nikos E. Stavropoulos M.D., Hatzikosta General Hospital (Greece)
Dimitris Skalkos, Univ. of Ioannina (Greece)
Ioannis Tsimaris, Hatzikosta General Hospital (Greece)
D. Kalogeras, Hatzikosta General Hospital (Greece)
Unyime O. Nseyo M.D., Virginia Commonwealth Univ. (United States)
A. Batistatou, Univ. of Ioannina (Greece)
N. J. Agnantis, Univ. of Ioannina (Greece)

Published in SPIE Proceedings Vol. 5689:
Optical Methods for Tumor Treatment and Detection: Mechanisms and Techniques in Photodynamic Therapy XIV
David Kessel, Editor(s)

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