Beata J. Osiecka

The application of Levulan-based photodynamic therapy with imiquimod in the treatment of recurrent basal cell carcinoma.

Summary Background Common skin tumors like basal- and squamous-cell carcinoma present a serious problem in modern medicine. Exposure to ultraviolet solar radiation is the main cause of these lesions. Since application of Aldara® and PDT separately is well documented, we decided to use both methods together. The aim of our study was to evaluate the effectiveness of local photodynamic therapy supplemented with topical application of Aldara® in basal-cell carcinoma. Material/Methods Thirty-four patients ages 50 to 68 years were enrolled to the trial and underwent PDT treatment. Each case of BCC was histopathologically confirmed. Ten patients were subjected to local Levulan®-PDT and placebo (Eucerin as vehicle cream), and 24 patients were subjected to Levulan®-PDT and imiquimod. Photodynamic diagnosis (PDD) was used to detect and visualize suspicious foci (including cancer lesions). Results In the group of patients who were treated using Levulan®-PDT and placebo, 6 patients (60%) were totally cured and 4 lesions (40%) significantly decreased in size. In the group of patients treated with Levulan®-PDT and imiquimod, 18 lesions totally disappeared (75%), 6 lesions significantly diminished, and in 1 patient small foci of previously excised BCC developed again in scar tissue 10 month after the first control examination. Conclusions Cure was achieved without any scarring and with very good cosmetic effects. Although this is the preliminary report, the presented modification of PDT seems to be reasonable and promising in treating basal-cell carcinoma.

In vitro and in vivo matrix metalloproteinase expression after photodynamic therapy with a liposomal formulation of aminolevulinic acid and its methyl ester

Photodynamic therapy (PDT) is a well-known method for the treatment of malignant tumors, and its principles have been well established over the past 30 years. This therapy involves the application of a chemical called a photosensitizer and its subsequent excitation with light at the appropriate wavelength and energy. Topical photodynamic therapy with aminolevulinic acid (5-ALA) is an alternative therapy for many malignant processes, including nonmelanoma skin cancers such as basal-cell carcinoma (BCC). Our novel approach for this study was to use a liposomal formulation of 5-ALA and its methyl ester (commercially available as metvix) both in vitro and in vivo, and to check whether the liposome-entrapped precursors of photosensitizers can induce the expression of metalloproteinases (MMPs) in animal tumor cells and in other tissues from tumor-bearing rats and in selected cell lines in vitro. We also checked whether the application of tissue inhibitors of matrix metalloproteinases (TIMPs) has any effect on MMPs in the above-mentioned experimental models, and if they can cause complete inhibition of MMP expression. Immunohistochemical studies revealed that after the PDT, the intensity of expression of MMPs in healthy animals was very low and seen in single cells only. After the PDT in tumor-bearing rats, MMP-3 was expressed in the tumor cells with the highest intensity of staining in the tissues directly adjacent to the tumors, while MMP-2 and -9 were not found. In the control groups, there was no observed expression of MMPs. In vitro studies showed that MMP-3 was expressed in MCF-7 cells after PDT, but MMP-9 was not observed and MMP-2 was only seen in single cases. Our studies confirmed that the application of an MMP-3 inhibitor may block an induction of MMP-3 expression which had previously been initiated by PDT. The preliminary data obtained from cancer patients revealed that new precursors are effective in terms of PDT, and that using MMP inhibitors should be considered as a potential enhancing factor in clinical PDT.

Photodynamic therapy of vulvar lichen sclerosus et atrophicus in a woman with hypothyreosis — Case report

Lichen sclerosus et atrophicus (LSA) is disease of skin and mucosa, its pathogenesis remains unknown. Itching, pain and burning sensations and atrophy of vulva impair quality of life. Treatment is symptomatic. We report case of 30-year old woman with lesions in vulva in which series of topical PDT were carried out. We applied Levulan®Kerastick® for 4h and after that lesions were illuminated with red light. Along with above treatment patient started receiving Euthyrox®, because of recently diagnosed hypothyreosis. Significant relief from subjective symptoms was achieved and lesions in vulvar region disappeared. Combination of topical PDT with hormonal therapy allowed controlling course of disease and minimizing symptoms, and thus improved quality of life.

Immunocytochemical studies on the nuclear ubiquitous casein and cyclin-dependent kinases substrate following 5-aminolevulinicacid-mediated photodynamic therapy on MCF-7 cells

BACKGROUND Recent data indicates that nuclear ubiquitous casein and cyclin-dependent kinases substrate (NUCKS) may play role in tumor growth. In present study authors examined whether photodynamic therapy with 5-aminolevulinic acid (5-ALA) induces NUCKS expression in breast cancer cell line, MCF-7. METHODS In the experiment concentration of 5-ALA was 6.5mM. Excitation wavelength was 630 ± 20 nm, total light dose of light 5 or 10 J/cm(2) and irradiance 60 mW/cm(2) was used. Cells were collected at established time points and Western blot and immunocytochemical studies were performed using antibody against NUCKS. RESULTS Studies proved strong cytotoxic effects in cells following PDT with 6.5mM of precursor and 10 J/cm(2). Western blot analysis revealed the strongest expression of NUCKS at 7h after PDT. At next time points, 18 and 24h, expression of NUCKS decreased and became similar to that of control group. Further immunocytochemical studies showed very strong expression of NUCKS following PDT with 5-ALA and light irradiation of 5 J/cm(2). Early, at 0 h, that expression was predominantly seen in nuclei, while at 7h expression of NUCKS was observed in disseminated manner within entire cells in both nuclei and cytoplasm, with prevalence of cytoplasmic staining. CONCLUSIONS Authors suggest that NUCKS is involved in cellular responses following PDT, and since parallel induction of NUCKS and proapoptotic marker Bax and inhibition of anti-apoptotic Bcl-2 was observed, this protein might also be involved in induction of apoptosis following PDT.

The Use of Fractal Dimension Analysis in Estimation of Blood Vessels Shape in Transplantable Mammary Adenocarcinoma in Wistar Rats after Photodynamic Therapy Combined with Cysteine Protease Inhibitors

Fractal dimension analysis (FDA) is modern mathematical method widely used to describing of complex and chaotic shapes when classic methods fail. The main aim of this study was evaluating the influence of photodynamic therapy (PDT) with cystein proteases inhibitors (CPI) on the number and morphology of blood vessels inside tumor and on increase of effectiveness of combined therapy in contrast to PDT and CPI used separately. Animals were divided into four groups: control, treated using only PDT, treated using only CPI and treated using combined therapy, PDT and CPI. Results showed that time of animal survival and depth of necrosis inside tumor were significantly higher in CPI+PDT group in contrast to other groups. The higher value of fractal dimension (FD) was observed in control group, while the lowest value was found in the group which was treated by cystein protease inhibitors. The differences between FD were observed in CPI group and PDT+CPI group in comparison to control group. Our results revealed that fractal dimension analysis is a very useful tool in estimating differences between irregular shapes like blood vessels in PDT treated tumors. Thus, the implementation of FDA algorithms could be useful method in evaluating the efficacy of PDT.

Immunohistochemical study of nuclear ubiquitous casein and cyclin-dependent kinase substrate 1 in invasive breast carcinoma of no special type

The aim of the present study was to investigate the immunohistochemical expression of nuclear ubiquitous casein and cyclin-dependent kinases substrate 1 (NUCKS1) in invasive breast carcinoma of no special type, in association with clinicopathological characteristics, including the tumor grade, frequency of lymph node involvement and distant metastasis. In addition, associations between NUCKS1 and other tumor subtype markers, including estrogen receptor (ER), progesterone receptor (PR), human epidermal growth factor receptor 2 (HER2), Ki-67 and cytokeratin 5/6 (CK 5/6), were investigated. NUCKS1 expression was shown to be associated with the formation of distant metastases and lymph node involvement. Furthermore, an association between the presence of NUCKS1 and histological grading was observed. The results confirmed that the expression of NUCKS1 in low grade invasive breast carcinoma of no special type was significantly less common compared with cases of high grade carcinoma. With regard to the additional tumor subtype markers, NUCKS1 expression was demonstrated to be significantly associated with Ki-67 and CK 5/6; however, no association was identified with ER, PR and HER2. Therefore, NUCKS1 may be a novel prognostic marker in the histopathological evaluation of invasive breast carcinoma of no special type.

Photodynamic therapy with green light for the treatment of vulvar lichen sclerosus – Preliminary results

INTRODUCTION The standard treatment for lichen sclerosus (LS) is symptomatic and is primarily based on the chronic use of corticosteroids, sometimes resulting in unsatisfactory effects. Therefore, other non-pharmacological methods are being sought, which are less aggravating for the patient. LS can be treated topically by using photodynamic therapy (PDT) based on 5-aminolevulinic acid (5-ALA). Unfortunately, therapy with the red light is often connected with severe local pain during the illumination. Green light can also be characterised by its ability to turn on photodynamic reactions in cells. MATERIALS AND METHODS The aim of this study was an evaluation into the efficacy and tolerance of 5-ALA-PDT with a green light (540nm±15nm) in 11 patients with chronic LS that were characterised by severe itching. The disease lasted from 1.5 to 4 years. All the patients were treated with three sessions of PDT. RESULTS Following treatment with PDT, a significant improvement of local status, as well as a reduction of the main symptom (pruritus), were observed. No patient complained of severe pain during the sessions that would have required an interruption of irradiation or local application of analgesics. CONCLUSIONS Our preliminary results of using green light in PDT for superficial skin non-oncological lesions are very promising but require further studies.

A brief history of photodynamic therapy in Wrocław

A brief history of photodynamic therapy in Wroclaw was presented in this paper.

Photodynamic detection in visualisation of cutaneous and oral mucosa premalignant and malignant lesions: two clinical cases

Photodynamic diagnosis (PDD) is promising method of visualisation of premalignant and malignant lesions. PDD is consisted of two main agents: special chemical compound which is called photosensitizer and light. Photosensitizer has affinity to fast proliferating cells such as pre- or malignant. During light irradiation (with proper wavelength - corresponding to absorption peak of photosensitizer) photosensitizer gains energy and passes into excited singlet state S1. Returning to basic singlet state Sn, leads to fluorescence. Due to difference between concentration of photosensitizer in lesion and normal tissue it is possible to obtain high contrast image of lesion. Case #1: 53 years old woman with basal cell carcinoma (BCC) in nasal region; 20% delta-aminolevulinic acid as a precursor of photosensitizer on eucerin base was used. Case #2: 57 years old woman with multifocal oral leukoplakia on cheek mucosa and tongue; 2% chlorophyll gel as photosesitizer was used. All photographs were taken in white light without any filter and in blue and UV light with orange filter: in both cases the total area of the lesions appeared to be larger than it has been clinically observed. Thus, the PDD might be helpful in evaluation of margins of surgical excision of such lesions.