aluminum-phthalocyanine-disulfonate and Mouth-Neoplasms

The use of photodynamic therapy (PDT) for the treatment of skin and oral cancer has been the subject of several clinical studies but there has been little scientific evaluation of its mechanism of action. Evidence to date suggests that whilst epithelial cell death may be secondary to vascular damage, direct cell killing may occur and may involve an apoptosis-like mechanism. To investigate the mechanism of epithelial cell death following PDT, two cell lines, human epidermal keratinocytes (UP) and oral squamous cell carcinoma-derived cells (H376) were subjected to PDT with aluminium disulphonated phthalocyanine (AlS2Pc) as the photosensitizer and red laser light at 675 nm. Control groups received red laser light, photosensitizer or neither. The effects of PDT were assessed using an MTS cell-proliferation assay, which showed a significant reduction in viability (p < 0.01) for PDT-treated cells compared to controls. For morphological analysis, cells were stained with haemotoxylin and eosin and the numbers showing typical apoptotic features counted. The treated cultures showed significantly increased numbers of apoptotic cells. Moreover, the H376 control cultures showed a baseline level of apoptosis of approx. 15%. Apoptosis was confirmed by ultrastructural analysis and by in situ end-labeling of DNA fragments. The results show that PDT using AlS2Pc as a photosensitizer promotes apoptotic cell death in UP and H376 cells in vitro and suggest that direct killing of epithelial cells may contribute to tumour necrosis in vivo.

Topics: Aluminum; Apoptosis; Carcinoma, Squamous Cell; Cell Count; Cell Division; Cell Line; Cell Survival; Coloring Agents; DNA Fragmentation; Eosine Yellowish-(YS); Epidermis; Epithelial Cells; Fluorescent Dyes; Hematoxylin; Humans; Indoles; Keratinocytes; Laser Therapy; Mouth Neoplasms; Necrosis; Organometallic Compounds; Photochemotherapy; Photosensitizing Agents; Tumor Cells, Cultured

Photosensitizer-induced fluorescence is studied as a technique for the detection of cancer. Therefore we investigated the ability of a photosensitizer, aluminum phthalocyanine disulfonate (AIPcS2), to localize in tumor tissue. In vivo endoscopic fluorescence imaging, fluorescence microscopy, conventional spectrofluorometry and high performance liquid chromatograpy combined with diode laser-induced fluorescence (HPLC-Dio-LIF) were used. Squamous cell carcinomas were induced with 4-nitro-quinoline-1-oxide (4NQO) in the mucosa of the palate of the rat. In vivo fluorescence images, taken after injection of 1.5 mumol/kg AIPcS2 intravenously, showed that 4NQO-treated palates had higher fluorescence signals than normal palates. Areas displaying locally high amounts of AIPcS2 fluorescence (hot spots) were present only in 4NQO-treated rats 2-8 h but had disappeared 24 h after injection. However, HPLC-Dio-LIF showed that the relative AIPcS2 content was highest at 24/48 h in biopsies taken in the areas of the hot spots. Fluorescence microscopy revealed that AIPcS2 was present only between 2 and 8 h in the epithelial layer, while in biopsies the connective tissue contained large quantities of AIPcS2 at 24/48 h. In vivo fluorescence imaging appears to show mainly fluorescence from the epithelial layer and the ex vivo analytical techniques mainly show the connective tissue fluorescence. Care should be taken when interpreting data using one technique only.

Topics: Animals; Carcinoma, Squamous Cell; Chromatography, High Pressure Liquid; Indoles; Male; Microscopy, Fluorescence; Mouth Neoplasms; Organometallic Compounds; Photosensitizing Agents; Rats; Rats, Wistar; Spectrometry, Fluorescence

In order to investigate the mechanism of action of photodynamic therapy (PDT) with sulfonated aluminum phthalocyanine (AlSPc) in squamous cell carcinoma, animal experiments were performed in induced carcinomas of the mucosa of the hamster's cheek pouch and skin of the laboratory mouse. Histological examinations revealed signs of massive interstitial bleeding, indicating a vascular response to PDT with AlSPc. It was also possible to induce similar change adjacent to newly formed vessels at the margin of an inflammatory reaction in the cheek pouch of five hamsters in the absence of tumor cells. Implanted human squamous cell carcinoma cells in athymic nude mice showed that carcinoma cells removed immediately following PDT remained viable, while tumors left in situ became necrotic. These results suggest that the primary effect of PDT with AlSPc in vivo is not the malignant cell itself, but the vascular stroma of the tumor or in the immediate vicinity of the latter.

Topics: Animals; Carcinoma, Squamous Cell; Cheek; Cricetinae; Indoles; Male; Mesocricetus; Mice; Mice, Nude; Mouth Mucosa; Mouth Neoplasms; Neoplasm Transplantation; Organometallic Compounds; Photochemotherapy; Radiation-Sensitizing Agents; Skin Neoplasms