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Quantitative Analysis of Photodynamic Therapy Effects in Rat Mammary Tumor Vascular Density Using Image-Pro plus Software

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DOI: 10.4236/ojvm.2013.35041    3,102 Downloads   4,801 Views   Citations


Photodynamic therapy (PDT) is a treatment modality that has advanced rapidly in recent years. It causes tissue and vascular damage with the interaction of a photosensitizing agent (PS), light of a proper wavelength, and molecular oxygen. Evaluation of vessel damage usually relies on histopathology evaluation. Results are often qualitative or at best semi-quantitative based on a subjective system. The aim of this study was to evaluate, using CD31 immunohistochemistry and image analysis software, the vascular damage after PDT in a well-established rodent model of chemically induced mammary tumor. Fourteen Sprague-Dawley rats received a single dose of 7,12-dimethylbenz(a)anthraxcene (80 mg/kg by gavage), treatment efficacy was evaluated by comparing the vascular density of tumors after treatment with Photogem® as a PS, intraperitoneally, followed by interstitial fiber optic lighting, from a diode laser, at 200 mW/cm and light dose of 100 J/cm directed against his tumor (7 animals), with a control group (6 animals, no PDT). The animals were euthanized 30 hours after the lighting and mammary tumors were removed and samples from each lesion were formalin-fixed. Immunostained blood vessels were quantified by Image Pro-Plus version 7.0. The control group had an average of 3368.6 ± 4027.1 pixels per picture and the treated group had an average of 779 ± 1242.6 pixels per area (P < 0.01), indicating that PDT caused a significant decrease in vascular density of mammary tumors. The combined immunohistochemistry using CD31, with selection of representative areas by a trained pathology, followed by quantification of staining using Image Pro-Plus version 7.0 system was a practical and robust methodology for vessel damage evaluation, which probably could be used to assess other antiangiogenic treatments.

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The authors declare no conflicts of interest.

Cite this paper

I. Ferreira, C. Bulla, W. Baumgartner, V. Bagnato and N. Rocha, "Quantitative Analysis of Photodynamic Therapy Effects in Rat Mammary Tumor Vascular Density Using Image-Pro plus Software," Open Journal of Veterinary Medicine, Vol. 3 No. 5, 2013, pp. 259-262. doi: 10.4236/ojvm.2013.35041.


[1] V. H. Fingar, P. K. Kik, P. S. Haydon, P. B. Cerrito, M. Tseng, E. Abang and T. J. Wieman, “Analysis of Acute Vascular Damage after Photodynamic Therapy Using Benzoporphyrin Derivative (BPD),” British Journal of Cancer, Vol. 79, No. 11-12, 1999, pp. 1702-1708. doi:10.1038/sj.bjc.6690271
[2] V. H. Fingar, T. J. Wieman, S. A. Wiehle and P. B. Cerrito, “The Role of Microvascular Damage in Photodynamic Therapy: The Effect of Treatment on Vessel Constriction, Permeability, and Leukocyte Adhesion,” Cancer Research, Vol. 52, 1992, p. 4914.
[3] L. B. Li and R. C. Luo, “Effect of Drug-Light Interval on the Mode of Action of Photofrin Photodynamic Therapy in a Mouse Tumor Model,” Lasers in Medical Science, Vol. 24, No. 4, 2009, pp. 597-603. doi:10.1007/s10103-008-0620-9
[4] J. Folkman, “Angiogenesis in Cancer, Vascular, Rheumatoid and Other Disease,” Nature Medicine, Vol. 1, No. 1, 1995, pp. 27-31. doi:10.1038/nm0195-27
[5] D. L. Nielsen, M. Andersson, J. L. Andersen and C. Kamby, “Antiangiogenic Therapy for Breast Cancer,” Breast Cancer Research, Vol. 12, No. 5, 2010, p. 209. doi:10.1186/bcr2642
[6] J. Harper and M. A. Moses, “Molecular Regulation of Tumor Angiogenesis: Mechanisms and Therapeutic Implications,” EXS, Vol. 96, 2006, pp. 223-268.
[7] B. W. Henderson, S. O. Gollnick, J. W. Snyder, et al., “Choice of Oxygen-Conserving Treatment Regimen Determines the Inflammatory Response and Outcome of Photodynamic Therapy of Tumors,” Cancer Research, Vol. 64, 2004, pp. 2120-2126. doi:10.1158/0008-5472.CAN-03-3513
[8] R. Bhuvaneswari, Y. Y. Gan, S. S. Lucky, W. W. L. Chin, S. M. Ali, K. C. Soo and M. Olivo, “Molecular Profiling of Angiogenesis in Hypericin Mediated Photodynamic Therapy,” Molecular Cancer, Vol. 7, No. 1, 2008, p. 56. doi:10.1186/1476-4598-7-56
[9] A. Bottini, A. Berruti, A. Bersiga, M. P. Brizzi, G. Allevi, G. Bolsi, S. Aguggini, A. Brunelli, E. Betri, D. Generali, L. Scaratti, G. Bertoli, P. Alquati and L. Dogliotti, “Changes in Microvessel Density as Assessed by CD34 Antibodies after Primary Chemotherapy in Human Breast Cancer,” Clinical Cancer Research, Vol. 8, No. 6, 2002, pp. 1816-1821.
[10] D. C. Chhieng, S. O. Tabbara, E. F. Marley, L. I. Talley and A. R. Fros, “Microvessel Density and Vascular Endothelial Growth Factor Expression in Infiltrating Lobular Mammary Carcinoma,” Breast Journal, Vol. 9, No 3, 2003, pp. 200-207. doi:10.1046/j.1524-4741.2003.09311.x
[11] D. Wang, C. R. Stockard, L. Harkins, P. Lott, C. Salih, K. Yuan, D. Buchsbaum, A. Hashim, M. Zayzafoon, R. Hardy, O. Hameed, W. Grizzle and G. P. Siegal, “Immunohistochemistry for the Evaluation of Angiogenesis in Tumor Xenografts,” Biotechnic & Histochemistry, Vol. 83, No. 3, 2008, pp. 179-189. doi:10.1080/10520290802451085
[12] P. Mahzouni, F. Mohammadizadeh, K. Mougouei, N. A. Moghaddam, A. Chehrei and A. Mesbah, “Determining the Relationship between ‘Microvessel Density’ and Different Grades of Astrocytoma Based on Immunohistochemistry for ‘Factor VIII-Related Antigen’ (von Willebrand Factor) Expression in Tumor Microvessels,” Indian Journal of Pathology and Microbiology, Vol. 53, No. 4, 2010, pp. 605-610. doi:10.4103/0377-4929.71996
[13] M. Seshadri, J. A. Spernyak, R. Mazurchuk, S. H. Camacho, A. R. Oseroff, R. T. Cheney and D. A. Bellnie, “Tumor Vascular Response to Photodynamic Therapy and the Antivascular Agent 5,6-Dimethylxanthenone-4-Acetic Acid: Implications for Combination Therapy,” Clinical Cancer Research, Vol. 11, 2005, p. 4241. doi:10.1158/1078-0432.CCR-04-2703

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