Lilian Tan Moriyama

Photodynamic therapy induced vascular damage: an overview of experimental PDT

Photodynamic therapy (PDT) has been developed as one of the most important therapeutic options in the treatment of cancer and other diseases. By resorting to the photosensitizer and light, which convert oxygen into cytotoxic reactive oxygen species (ROS), PDT will induce vascular damage and direct tumor cell killing. Another consequence of PDT is the microvascular stasis, which results in hypoxia and further produces tumor regression. To improve the treatment with PDT, three promising strategies are currently attracting much interest: (1) the combination of PDT and anti-angiogenesis agents, which more effectively prevent the proliferation of endothelial cells and the formation of new blood vessels; (2) the nanoparticle-assisted delivery of photosensitizer, which makes the photosensitizer more localized in tumor sites and thus renders minimal damage to the normal tissues; (3) the application of intravascular PDT, which can avoid the loss of energy during the transmission and expose the target area directly. Here we aim to review the important findings on vascular damage by PDT on mice. The combination of PDT with other approaches as well as its effect on cancer photomedicine are also reviewed.

Possibility for a full optical determination of photodynamic therapy outcome

The efficacy of photodynamic therapy (PDT) depends on a variety of parameters: concentration of the photosensitizer at the time of treatment, light wavelength, fluence, fluence rate, availability of oxygen within the illuminated volume, and light distribution in the tissue. Dosimetry in PDT requires the congregation of adequate amounts of light, drug, and tissue oxygen. The adequate dosimetry should be able to predict the extension of the tissue damage. Photosensitizer photobleaching rate depends on the availability of molecular oxygen in the tissue. Based on photosensitizers photobleaching models, high photobleaching has to be associated with high production of singlet oxygen and therefore with higher photodynamic action, resulting in a greater depth of necrosis. The purpose of this work is to show a possible correlation between depth of necrosis and the in vivo photosensitizer (in this case, Photogem®) photodegradation during PDT. Such correlation allows possibilities for the development of a real time ...

Comparative ablation rate from a Er: YAG laser on enamel and dentin of primary and permanent teeth

This study was conducted to analyze the ablation rate and micromorphological aspects of microcavities in enamel and dentin of primary and permanent teeth using a Er: YAG laser system. Micromorphological evaluation has been performed in terms of permanent teeth; however, little information about Er: YAG laser interaction with primary teeth can be found in the literature. Because children have been the most beneficiary patients with laser therapy in our offices, it is extremely necessary to compare the effects of this kind of laser system on the enamel and dentin of permanent and primary teeth. In this study, we used eleven intact primary anterior exfoliated teeth and six extracted permanent molar teeth. We used a commercial laser system: a Er: YAG Twin Light laser system (Fotona Medical Lasers, Slovenia) at 2940 nm, changing average energy levels per pulse (100, 200, 300, and 400 mJ) producing 48 microcavities in enamel and dentin of primary and permanent teeth. Primary teeth are more easily ablated than are permanent teeth, when related to enamel or dentin. However, while this laser system is capable of slowly revealing the enamel’s microstructure, in dentin only the lowest laser energies permit this kind of observation, more easily decomposing the original tissue aspect, when related to primary or permanent teeth. Statistically, the only different factor at the 5% level was an energy per pulse of 400 mJ, confirming the results found in SEM. Our results showed that dentin in both primary and permanent teeth is less resistant to Er: YAG laser ablation; this fact is easily observed under SEM observation and through the ablation rate evaluation.

Experience and BCC subtypes as determinants of MAL-PDT response: Preliminary results of a national Brazilian project

Non-melanoma skin cancer is the most prevalent cancer type in Brazil and worldwide. Photodynamic therapy (PDT) is a noninvasive technique with excellent cosmetic outcome and good curative results, when used for the initial stages of skin cancer. A Brazilian program was established to determine the efficacy of methyl aminolevulinate (MAL)-PDT, using Brazilian device and drug. The equipment is a dual device that combines the photodiagnosis, based on widefield fluorescence, and the treatment at 630nm. A protocol was defined for the treatment of basal cell carcinoma with 20% MAL cream application. The program also involves the training of the medical teams at different Brazilian regions, and with distinct facilities and previous PDT education. In this report we present the partial results of 27 centers with 366 treated BCC lesions in 294 patients. A complete response (CR) was observed in 76.5% (280/366). The better response was observed for superficial BCC, with CR 160 lesions (80.4%), when compared with nodular or pigmented BCC. Experienced centers presented CR of 85.8% and 90.6% for superficial and nodular BCC respectively. A high influence of the previous doctor experience on the CR values was observed, especially due to a better tumor selection.

Time-resolved fluorescence lifetime for cutaneous melanoma detection

Melanoma is the most aggressive skin cancer type. It is characterized by pigmented lesions with high tissue invasion and metastatic potential. The early detection of melanoma is extremely important to improve patient prognosis and survival rate, since it can progress to the deadly metastatic stage. Presently, the melanoma diagnosis is based on the clinical analysis of the macroscopic lesion characteristics such as shape, color, borders following the ABCD rules. The aim of this study is to evaluate the time-resolved fluorescence lifetime of NADH and FAD molecules to detect cutaneous melanoma in an experimental in vivo model. Forty-two lesions were analyzed and the data was classified using linear discriminant analysis, a sensitivity of 99.4%, specificity of 97.4% and accuracy of 98.4% were achieved. These results show the potential of this fluorescence spectroscopy for melanoma detection.

Evaluation of vascular effect of photodynamic therapy in chorioallantoic membrane using different photosensitizers

Photodynamic Therapy (PDT) is a local treatment that requires a photosensitizing agent, light and molecular oxygen. With appropriate illumination, the photosensitizer is excited and produces singlet oxygen that is highly reactive and cytotoxic. Tumor vascular network is essential for the tumor growth and the understanding of vascular response mechanisms enables an improvement in the PDT protocol for cancer treatment. Compounds of porphyrin (Photogem®) and chlorin (Photodithazine®) were the photosensitizers tested. The incubation times varied from 20 to 80 min and the concentration ranged between 0.1 and 100 μg/cm(2). Different light doses were used between 4.8 and 40 J/cm(2) with irradiance varying between 80 and 100 mW/cm(2). The light dose of 30 J/cm(2) was used in the intravenous photosensitizer application. The membrane images were made from 0 to 300 min after treatment. The vascular response was evaluated by the average vessel area. Different responses was observed depending on the photosensitizer concentration and administration form. Intravenous application has been more efficient to produce vessel constriction and the most pronounced effect was observed for the chlorin.

Photodynamic therapy: Progress toward a scientific and clinical network in Latin America.

Cancer is one of the major challenges for Latin America health services, since the skin cancer is the most frequent lesion. This manuscript addresses an initiative for the treatment of basal cell carcinomas (BCC) by photodynamic therapy (PDT) based on a government-funded national program in Brazil. The program provides clinical training and facilitates access to drugs/equipment and significantly reduces PDT costs. It also lays foundations for the establishment of a Latin American research network to improve prevention, early detection and treatment of diseases. Centers have been established by direct contact (conferences, visits to healthcare facilities and official departments). A local training was divided into complementary theoretical and practical parts. This is an ongoing project that has involved 10 countries: Brazil, Bolivia Chile, Ecuador, El Salvador, Colombia, Cuba, Mexico, Peru and Venezuela, The initial results are encouraging and have provided assessment of Latin America patients relating, for example, the most common skin phototypes with incidence of BCC in such countries. The network is expected to produce relevant scientific information for PDT introduction in many countries. The experience acquired by local teams shall enable them to innovate PDT protocols and increase the number of skilled contributors/researchers to broaden knowledge on the ever-crescent PDT field in Latin America. The establishment of a collaboration network and introduction of other projects and experience exchange shall become an easier process with time. This PDT clinical research network is a start for the strengthening of Science in South Hemisphere countries.

Fluorescence guided PDT for optimization of the outcome of skin cancer treatment

The photodynamic therapy (PDT) is an alternative technique that can be indicated for superficial basal cell carcinoma (sBCC), Bowen’s disease and actinic keratosis with high efficiency. The objective of this study is to present the importance of fluorescence imaging for PDT guidance and monitoring in real time. Confirming that the lesion is well prepared and the photosensitizer shows a homogenous distribution, the outcome after few PDT sessions will be positive and the recurrence should be lower. Our proposition in this study is use the widefield fluorescence imaging to evaluate the PDT protocol in situ and in real time for each lesion. This evaluation procedure is performed in two steps: first with the monitoring of the production of protoporphyrin IX (PpIX) induced by methyl aminolevulinate (MAL), an derivative of 5-aminolevulinic acid (ALA) and second with the detection of PpIX photobleaching after illumination. The fluorescence images provide information correlated with distinct clinical features and with the treatment outcome. Eight BCC lesions are presented and discussed in this study. Different fluorescence patterns of PpIX production and photobleaching could be correlated with the treatment response. The presented results show the potential of using widefield fluorescence imaging as a guidance tool to customized PDT.

Fluorescence spectroscopy in the visible range for the assessment of UVB radiation effects in hairless mice skin

Ultraviolet (UV) radiation may induce skin alterations as observed in photoaging. Some recognized modifications are epidermal hyperplasia, amorphous deposition of degraded elastic fibers and reduction in the number of collagen fibers. They alter the tissue biochemical properties that can be interrogated by steady state fluorescence spectroscopy (SSFS). In this study, we monitored the changes in endogenous fluorescence emission from hairless mice skin during a protocol of photoaging using UVB irradiation. To perform the fluorescence spectroscopy, it was used a violet laser (408nm) to induce the native fluorescence that is emitted in the visible range. Under 408nm excitation, the emission spectrum showed bands with peaks centered around 510, 633 and 668nm for irradiated and control groups. A relative increase of the fluorescence at 633nm emission on the flank was observed with time when compared to the ventral skin at the same animal and the non-irradiated control group. We correlated the emission at 633nm with protoporphyrin IX (PpIX), and our hypothesis is that the PpIX metabolism in the photoaged and aged skin are different. PpIX fluorescence intensity in the photoaged skin is higher and more heterogeneous than in the aged skin. Notwithstanding, more spectroscopic and biochemistry studies investigating the 510 and 633nm emission are needed to confirm this hypothesis.

Relationship between the chemical and morphological characteristics of human dentin after Er:YAG laser irradiation.

Abstract. The effects of laser etching on dentin are studied by microenergy-dispersive x-ray fluorescence spectrometry (μ-EDXRF) and scanning electron microscopy (SEM) to establish the correlation of data obtained. Fifteen human third molars are prepared, baseline μ-EDXRF mappings are performed, and ten specimens are selected. Each specimen received four treatments: acid etching (control-CG) or erbium:yttrium–aluminum–garnet (Er:YAG) laser irradiation (I—100 mJ, II—160 mJ, and III—220 mJ), and maps are done again. The Ca and P content are significantly reduced after acid etching (p<0.0001) and increased after laser irradiation with 220 mJ (Ca: p<0.0153 and P: p=0.0005). The Ca/P ratio increased and decreased after CG (p=0.0052) and GI (p=0.0003) treatments, respectively. CG treatment resulted in lower inorganic content (GI: p<0.05, GII: p<0.01, and GIII: p<0.01) and higher Ca/P ratios than laser etching (GI: p<0.001, GII: p<0.01, and GIII: p<0.01). The SEM photomicrographies revealed open (CG) and partially open dentin tubules (GI, GII, and GIII). μ-EDXRF mappings illustrated that acid etching created homogeneous distribution of inorganic content over dentin. Er:YAG laser etching (220 mJ) produced irregular elemental distribution and changed the stoichiometric proportions of hydroxyapatite, as showed by an increase of mineral content. Decreases and increases of mineral content in the μ-EDXRF images are correlated to holes and mounds, respectively, as found in SEM images.