Mirian D. Stringasci

Single LED-based device to perform widefield fluorescence imaging and photodynamic therapy

Photodynamic therapy (PDT) is a treatment modality that can be indicated for several cancer types and pre-cancer lesions. One of the main applications of PDT is the treatment of superficial skin lesions such as basal cell carcinoma, Bowen’s disease and actinic keratosis. Three elements are necessary in PDT, a photosensitizer (PS); light at specific wavelength to be absorbed by the PS, and molecular oxygen. A typical PS used for skin lesion is protoporphyrin IX (PpIX), which is an intrinsic PS; its production is stimulated by a pro-drug, such as 5-aminolevulinic acid (ALA). Before starting a treatment, it is very important to follow up the PpIX production (to ensure that enough PS was produced prior to a PDT application) and, during a PDT session, to monitor its photodegradation (as it is evidence of the photodynamic effect taking place). The aim of this paper is to present a unique device, LINCE (MMOptics - São Carlos, Brazil), that brings together two probes that can, respectively, allow for fluorescence imaging and work as a light source for PDT treatment. The fluorescence probe of the system is optically based on 400 nm LED (light emitting diodes) arrays that allow observing the fluorescence emission over 450 nm. The PDT illumination probe options are constituted of 630 nm LED arrays for small areas and, for large areas, of both 630 nm and 450 nm LED arrays. Joining both functions at the same device makes PDT treatment simpler, properly monitorable and, hence, more clinically feasible. LINCE has been used in almost 1000 PDT treatments of superficial skin lesions in Brazil, with 88.4% of clearance of superficial BCC.

Interstitial PDT using diffuser fiber—investigation in phantom and in vivo models

Photodynamic therapy (PDT) has been used for local treatment of several types of tumors. Light penetration of biological tissue is one limiting factor in PDT, decreasing the success rates of the treatment of invasive and solid tumors. In those cases, a possible solution is to use interstitial PDT, in which both diffuser optical fibers are inserted into the tumor. The uniformity of the diffuser emission plays a crucial role in planning the delivery of the appropriate light fluence and in ensuring treatment success. In this study, we characterized a diffuser optical fiber concerning its homogeneity. We showed that the diffuser emission can be inhomogeneous and that the necrosis generated by interstitial PDT using such a diffuser for illumination is asymmetrical in volume as a result. This observation has relevant consequences in achieving success in PDT and phototherapies in general, as the delivered light fluence depends on adequate previous knowledge of the irradiation profile.

Thermographic analysis of photodynamic therapy with intense pulsed light and needle-free injection photosensitizer delivery: an animal study

The photodynamic therapy (PDT) is a therapeutic modality that depends mostly on photosensitizer (PS), light and molecular oxygen species. However, there are still technical limitations in clinical PDT that are under constant development, particularly concerning PS and light delivery. Intense Pulsed Light (IPL) sources are systems able to generate pulses of high energy with polychromatic light. IPL is a technique mainly used in the cosmetic area to perform various skin treatments for therapeutic and aesthetic applications. The goals of this study were to determine temperature variance during the application of IPL in porcine skin model, and the PDT effects using this light source with PS delivery by a commercial high pressure, needle-free injection system. The PSs tested were Indocyanine Green (ICG) and Photodithazine (PDZ), and the results showed an increase bellow 10 °C in the skin surface using a thermographic camera to measure. In conclusion, our preliminary study demonstrated that IPL associated with needle-free injection PS delivery could be a promising alternative to PDT.

Discrimination of benign-versus-malignant skin lesions by thermographic images using support vector machine classifier

Skin cancer is the cancer type with the highest incidence in the world. Its diagnosis requires a specialist physician, with expertise in skin diagnostics. Thermography is a noninvasive technique based on the detection of infrared emission that is completely safe to humans. In this study, thermal images of clinically similar lesions were registered and analyzed aiming to provide a noninvasive diagnostic information for discrimination of: basal cell carcinoma versus intradermal nevus, squamous cell carcinoma versus actinic keratosis, and melanoma versus pigmented seborrheic keratosis. Thermal images were analyzed using a MATLAB® routine to evaluate statistical, histogram, and filtering metrics of each image, and a support vector machine classifier was used to discriminate the lesions based on those metrics values. Actinic keratoses and squamous cell carcinoma showed distinct average temperatures, whereas the other pairs of lesions presented similar temperatures. Nevertheless, the benign lesions showed higher definition of borders detection than malignant lesions, as a general rule. The results showed that support vector machine classifier has great ability for discrimination of clinically similar lesions based on their thermal images, suggesting that the thermography can be used as an auxiliary tool for the diagnosis of skin malignant lesions.Skin cancer is the cancer type with the highest incidence in the world. Its diagnosis requires a specialist physician, with expertise in skin diagnostics. Thermography is a noninvasive technique based on the detection of infrared emission that is completely safe to humans. In this study, thermal images of clinically similar lesions were registered and analyzed aiming to provide a noninvasive diagnostic information for discrimination of: basal cell carcinoma versus intradermal nevus, squamous cell carcinoma versus actinic keratosis, and melanoma versus pigmented seborrheic keratosis. Thermal images were analyzed using a MATLAB® routine to evaluate statistical, histogram, and filtering metrics of each image, and a support vector machine classifier was used to discriminate the lesions based on those metrics values. Actinic keratoses and squamous cell carcinoma showed distinct average temperatures, whereas the other pairs of lesions presented similar temperatures. Nevertheless, the benign lesions showed highe...

The relevance of light diffusion profiles for interstitial PDT using light-diffusing optical fibers

Photodynamic therapy (PDT) is a technique used for several tumor types treatment. Light penetration on biological tissue is one limiting factor for PDT applied to large tumors. An alternative is using interstitial PDT, in which optical fibers are inserted into tumors. Cylindrical diffusers have been used in interstitial PDT. Light emission of different diffusers depends on the manufacturing process, size and optical properties of fibers, which make difficult to establish an adequate light dosimetry, since usually light profile is not designed for direct tissue-fiber contact. This study discusses the relevance of light distribution by a cylindrical diffuser into a turbid lipid emulsion solution, and how parts of a single diffuser contribute to illumination. A 2 cm-long cylindrical diffuser optical fiber was connected to a diode laser (630 nm), and the light spatial distribution was measured by scanning the solution with a collection probe. From the light field profile generated by a 1 mm-long intermediary element of a 20 mm-long cylindrical diffuser, recovery of light distribution for the entire diffuser was obtained. PDT was performed in rat healthy liver for a real treatment outcome analysis. By using computational tools, a typical necrosis profile generated by the irradiation with such a diffuser fiber was reconstructed. The results showed that it was possible predicting theoretically the shape of a necrosis profile in a healthy, homogeneous tissue with reasonable accuracy. The ability to predict the necrosis profile obtained from an interstitial illumination by optical diffusers has the potential improve light dosimetry for interstitial PDT.

Monitoring of Ehrlich tumor growth using thermal image

In this study, we injected Ehrlich tumor cell in the dorsal region of nine Swiss mouse. The tumor in growth was assessed using thermography to monitor temperature changes and to observe the minimal tumor size that can be detected.

Onychomycosis diagnosis using fluorescence and infrared imaging systems

Onychomycosis is a common disease of the nail plate, constituting approximately half of all cases of nail infection. Onychomycosis diagnosis is challenging because it is hard to distinguish from other diseases of the nail lamina such as psoriasis, lichen ruber or eczematous nails. The existing methods of diagnostics so far consist of clinical and laboratory analysis, such as: Direct Mycological examination and culture, PCR and histopathology with PAS staining. However, they all share certain disadvantages in terms of sensitivity and specificity, time delay, or cost. This study aimed to evaluate the use of infrared and fluorescence imaging as new non-invasive diagnostic tools in patients with suspected onychomycosis, and compare them with established techniques. For fluorescence analysis, a Clinical Evince (MM Optics®) was used, which consists of an optical assembly with UV LED light source wavelength 400 nm ± 10 nm and the maximum light intensity: 40 mW/cm2 ± 20%. For infrared analysis, a Fluke® Camera FKL model Ti400 was used. Patients with onychomycosis and control group were analyzed for comparison. The fluorescence images were processed using MATLAB® routines, and infrared images were analyzed using the SmartView® 3.6 software analysis provided by the company Fluke®. The results demonstrated that both infrared and fluorescence could be complementary to diagnose different types of onychomycosis lesions. The simplicity of operation, quick response and non-invasive assessment of the nail patients in real time, are important factors to be consider for an implementation.

Thermographic diagnostics to discriminate skin lesions: a clinical study

Cancer is responsible for about 13% of all causes of death in the world. Over 7 million people die annually of this disease. In most cases, the survival rates are greater when diagnosed in early stages. It is known that tumor lesions present a different temperature compared with the normal tissues. Some studies have been performed in an attempt to establish new diagnosis methods, targeting this temperature difference. In this study, we aim to investigate the use of a handheld thermographic camera to discriminate skin lesions. The patients presenting Basal Cell Carcinoma, Squamous Cell Carcinoma, Actinic Keratosis, Pigmented Seborrheic Keratosis, Melanoma or Intradermal Nevus lesions have been investigated at the Skin Departament of Amaral Carvalho Hospital. Patients are selected by a dermatologist, and the lesion images are recorded using an infrared camera. The images are evaluated taken into account the temperature level, and differences into lesion areas, borders, and between altered and normal skin. The present results show that thermography may be an important tool for aiding in the clinical diagnostics of superficial skin lesions.