Cellular localization of Visudyne as a function of time after local injection in an in vivo model of squamous cell carcinoma: an investigation into tumor cell death

Vet Ophthalmol. 2010 May;13(3):158-65. doi: 10.1111/j.1463-5224.2010.00775.x.

Abstract

Objective: To evaluate the effects of time on cellular localization of Visudyne after local injection.

Animals: Twenty athymic nude mice.

Procedures: A squamous cell carcinoma (SCC) cell line (A-431) was injected into right and left dorsolumbar subcutaneous tissue of each mouse, representing treatment (T) and control (C) tumors. In experiment 1 (Exp 1; n = 10) and 2 (Exp 2; n = 10), the T tumors received a local injection of Visudyne (0.1 mg/cm(3)), and C tumors received an equal dose of 5% dextrose in water (D5W). Mice were randomly subdivided into two groups (A and B; n = 5 per group). Mice in Exp 1A and B were sacrificed 1 and 30 min after local injection, respectively. Experiment 1A and B tumors were evaluated by fluorescence microscopy to determine drug localization. Experiment 2A and B tumors were exposed to LED illumination 1 and 30 min after injection, respectively, and evaluated by transmission electron microscopy (TEM) to determine ultrastructural tumor cell damage.

Results: Fluorescence was detected within the cytoplasm of T tumors in both Exp 1A and B. Significance was detected in fluorescence intensity between T1 min vs. T30 min (P = 0.03) and between T1 min and C1 min tumors (P = 0.01), respectively. Tumors in Exp 2A and B demonstrated evidence of apoptotic cell death.

Conclusions: Fluorescence microscopy demonstrated higher Visudyne concentration within SCC cytoplasm of 1 min compared with 30-min tumors. Transmission electron microscopy results revealed that tumors treated by photodynamic therapy (PDT) within 30 min of local injection undergo cellular apoptosis.

MeSH terms

  • Animals
  • Biological Transport
  • Carcinoma, Squamous Cell / metabolism*
  • Cell Death / drug effects*
  • Cell Line, Tumor
  • Humans
  • Laser Therapy
  • Mice
  • Mice, Nude
  • Neoplasms, Experimental / drug therapy
  • Photosensitizing Agents / administration & dosage
  • Photosensitizing Agents / pharmacokinetics*
  • Porphyrins / administration & dosage
  • Porphyrins / pharmacokinetics*
  • Verteporfin

Substances

  • Photosensitizing Agents
  • Porphyrins
  • Verteporfin