Piotr Ziółkowski

New potent sensitizers for photodynamic therapy : 21-oxaporphyrin, 21-thiaporphyrin and 21,23-dithiaporphyrin induce extensive tumor necrosis

Abstract New sensitizers for photodynamic therapy (PDT) are reported. These compounds, namely 21-thiaporphyrin, 21,23-dithiaporphyrin and 21-oxaporphyrin, reveal some of the properties required for such therapy. Their physicochemical, chemical and pharmacological features meant that we could use them in the treatment of transplantable BFS1 fibrosarcoma in Balb/c mice. New sensitizers and the well-known chlorin e6 (Ce6) were used in doses of 2.5, 5.0, 7.5 and 10.0u2009mg/kg body weight, given intraperitoneally and followed by light irradiation, the total light doses being 50, 100 and 150u2009J/cm2 within 24u2009h after injection. The effectiveness of new sensitizers in PDT was evaluated with in terms of tumor necrosis intensity, the survival time of treated animals, the rate of tumor response (complete/partial/no response), and skin photosensitivity. These results were compared to results obtained in analogous conditions after Ce6-PDT. Distribution studies revealed that the highest concentration of new compounds occurred within 24u2009h after injection. The results of these experiments confirmed that 21-thiaporphyrin, 21,23-dithiaporphyrin and 21-oxaporphyrin can be considered as potent tumor photosensitizers that do not exert any unwanted effects, primarily skin photosensitization. We suggest that these porphyrins are possible sensitizers to be applied in clinical PDT.

Primary endometrioid carcinoma of Fallopian tube: Clinicomorphologic study

Twenty cases of primary Fallopian tube endometrioid carcinoma (PFTEC) are presented in the paper. This accounts for 42.5% of all histologic forms of primary Fallopian tube carcinoma (PFTC) found in our Department. The youngest patient was 38, and the oldest 68 years (mean: 56 years). Seven patients were nulliparas. Only two cases were bilateral. According to FIGO staging, 13 cases were evaluated as stage I, 4 as II, and 3 as stage III. Due to the histologic grading, 8 tumors were classified as well, 7 as moderately, and 5 as poorly differentiated. In the time of preparation of the manuscript, 12 women were still alive, 2 of them with recurrent disease. The follow-up of patients without recurrence ranged from 4 to 120 months (median: 63). Eight patients had died (survival time: from 4 to 65 months; median: 26). Metastases were found in 8 patients, especially to ovaries. In 14/20 cases of PFTEC various forms of tubal wall invasion were observed. Blood or lymphatic vessels involvement was found in 9 patients. Six of them had died and one is alive with the symptoms of disease. Immunohistochemical detection of the mutant form of p53 protein and oncogene product, c-erbB-2, was studied in 17 cases. Nine patients exhibited simultaneous p53 protein accumulation and c-erbB-2 expression. 2/9 of these patients are alive with recurrent tumors and 4/9 died. Endometrioid carcinoma of the Fallopian tube can be characterized by a tendency to superficial invasion of tubal wall and in a half of the cases by invasion of vessels. The majority of these tumors were diagnosed at an early stage tumors.

5,20-bis(4-sulphophenyl)-10,15-bis (2-methoxy-4-sulphophenyl)-21-thiaporphyrin as a new potent sensitizer in photodynamic therapy.

The main purpose of the study was to investigate the effectiveness of a new photosensitizer for photodynamic therapy. 5,20-bis(4-sulphophenyl)-10,15-bis(2-methoxy-4-sulphophenyl)-21-thiaporphyrin (21-thiaporphyrin) was compared to chlorin e6 and tetra(m-hydroxyphenyl)porphyrin (m-THPP) for its ability to sensitize tumors and skin to light. Chlorin e6 and m-THPP induced a strong tumor and skin photosensitization. In contrast, the same doses of 21-thiaporphyrin produced no skin sensitization and gave approximately 10 mm tumor necrosis after light exposure, in comparison to the 5-6 mm necrosis induced by chlorin e6 or m-THPP under identical conditions. 21-Thiaporphyrin, tested as a potential photosensitizer, induced no skin sensitization even at doses as high as 7.5 mg/kg body weight. 21-Thiaporphyrin presents a high potency in tumor sensitizing, i.e. a feature required for an efficient photosensitizer in photodynamic therapy applications.

In-vivo interleukin-1-alpha induction following chlorin-e6 photodynamic therapy in Balb/c mice

Photodynamic therapy may induce in in-vivo conditions the cytokine Interleukin-1-alpha in Balb/c mice. The sensitizer, i.e. chlorin e6, in the doses 2.5 and 5.0 mg/kg of body weight followed by light treatment with doses 50 and 100 J/sq.cm resulted in the increase in serum levels of the cytokine. The levels of Interleukin-1-alpha have been determined at different time points using enzyme-linked immunosorbent assay (ELISA). These levels in control animals did not exceed the mean value of 15 pg/ml, whereas in photodynamically treated mice the levels were almost 3 - 4 times higher. The entire experiment has been carried out on healthy animals.

In vitro human atherosclerotic plaque recognition by photosensitizer mono-L-aspartyl chlorin e6 assisted light induced fluorescence (PALIF)

The main aim of the presented experiments was to assess in vitro capabilities of Photosensitizer Assisted Light Induced Fluorescence (PALIF) to recognise different stages of atherosclerosis. Mono-L-asparyl-chlorin e-6 was used as a photosensitising agent and ultraviolet light (440 nm) as an excitation source to obtain spectra map of artery wall. The luminescence spectra were collected and analised. The specimans were histologically examined and classified into three groups: normal artery wall, atherosclerotic noncalcified plaque and calcified plaque. The ratio of green fluorescence (coming from collagen) to red fluorescence (coming from chlorin e6 bonded with lipid reach, noncalcified tissue) gies a chance to distinguish between normal artery or calcified plaque and noncalcified, lipid reach plaque. Further trials must be performed to evaluate in vitro athermoa detection algorithm as a feedback system for photoangioplasty.

The Results Of In Vivo 5-Aminolevulinic Acid Photodynamic Therapy Of Transplantable C51 Colon Adenocarcinoma

Results of in vivo studies on 5-aminolevulinic acid (5-ALA) - photodynamic therapy (PDT) are presented. The effectiveness of 5-ALA-PDT has been confirmed on BALB/c mice transplanted with mouse colon C51 adenocarcinoma. 5-ALA was administered intraperitoneally in the dose of 50 mg/kg followed by light exposure (6 hours later) at total light doses from 50 to 75 J/sq. cm and the fluence rate - 200 mW/sq.cm. PDT effects were evaluated with regard to tumor response and necrosis depth. The main finding in our tumor model was that the 5-ALA-PDT results in photodynamic dose dependent tumor response as well as in necrosis depth induction.

Immunohistochemical observations on tumor suppressor gene p53 status in mouse fibrosarcoma following in-vivo photodynamic therapy: the role of xanthine oxidase activity

Tumor suppressor gene p53 expression in a mouse fibrosarcoma following in-vivo photodynamic therapy has been studied using the immunohistochemical method. Photodynamic treatment involved injections of the well known sensitizer -- hematoporphyrin derivative at the doses 1.25 and 2.5 mg/kg of body weight and irradiations at the doses 25 and 50 J/sq cm. Glass slide preparations from PDT-treated tumors were obtained at different time points (15, 60 minutes, 2 and 24 hours) after therapy, subsequently stained for wild type/mutant p53, and assessed for positive reaction. High PDT doses (HpD -- 2.5 mg/kg; light dose -- 50 J/sq cm) correlated with decreased expression of p53 in tumor cells. The other part of the study was directed to measure the xanthine oxidase (XO) activity in the tumor cells. PDT included injections of HpD and light exposure at the same doses as for p53 study. We observed a complete inhibition of the enzyme activity. The slight increase in XO activity was found following treatment with either light or HpD alone.