Back
 OJVM  Vol.3 No.5 , September 2013
Quantitative Analysis of Photodynamic Therapy Effects in Rat Mammary Tumor Vascular Density Using Image-Pro plus Software
Abstract: 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.
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.
References

[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

 
 
Top