Although the benefits of topical sensitizer administration have been confirmed for photodynamic therapy (PDT), ALA-induced protoporphyrin IX is the only sensitizer clinically used with this administration route. Unfortunately, ALA-PDT results in poor treatment response for thicker lesions. Here, selectivity and depth distribution of the highly potent sensitizer meso-tetra(hydroxyphenyl)chlorin (mTHPC), supplied in a novel liposome formulation was investigated following topical administration for 4 and 6 h in a murine skin tumor model. Extraction data indicated an average [± standard deviation (SD)] mTHPC concentration within lesions of 6.0(±3.1) ng/mg tissue with no significant difference (p<0.05) between 4- and 6-h application times and undetectable levels of generalized photosensitivity. Absorption spectroscopy and chemical extraction both indicated a significant selectivity between lesion and normal surrounding skin at 4 and 6 h, whereas the more sensitive fluorescence imaging setup revealed significant selectivity only for the 4-h application time. Absorption data showed a significant correlation with extraction, whereas the results from the fluorescence imaging setup did not correlate with the other methods. Our results indicate that this sensitizer formulation and administration path could be interesting for topical mTHPC-PDT, decreasing the effects of extended skin photosensitivity associated with systemic mTHPC administration.
Meso-tetra(hydroxyphenyl)chlorin (mTHPC)(international generic name Temoporfin) is a potent photosensitizer used
for photodynamic therapy (PDT). In this study the pharmacokinetics of a systemically administered novel lipid
formulation of Temoporfin in a murine tumor model has been investigated. Fluorescence spectroscopy measurements
were performed at several time intervals following drug administration, yielding information on the Temoporfin
concentration within excised internal organs as a function of time after injection. Both point-monitoring and imaging
setups were used. The acquired fluorescence data were correlated to the concentration of Temoporfin obtained with High
Performance Liquid Chromatography (HPLC). There was a significant correlation between the fluorescence methods and
HPLC for most organs investigated. The pharmacokinetics of this new liposomal formulation of Temoporfin exhibited a
rather flat temporal profile in the time interval 2-8 hours in this study.
Photodynamic therapy (PDT) for the treatment of prostate cancer has been demonstrated to be a safe treatment
option capable of inducing tissue necrosis and decrease in prostate specific antigen (PSA). Research groups report
on large variations in treatment response, possibly due to biological variations in tissue composition and short-term
response to the therapeutic irradiation. Within our group, an instrument for interstitial PDT on prostate
tissue that incorporates realtime treatment feedback is being developed. The treatment protocol consists of
two parts. The first part incorporates the pre-treatment plan with ultrasound investigations, providing the
geometry for the prostate gland and surrounding risk organs, an iterative random-search algorithm to determine
near-optimal fiber positions within the reconstructed geometry and a Block-Cimmino optimization algorithm for
predicting individual fiber irradiation times. During the second part, the therapeutic light delivery is combined
with measurements of the light transmission signals between the optical fibers, thus monitoring the tissue effective
attenuation coefficient by means of spatially resolved spectroscopy. These data are then used as input for repeated
runs of the Block-Cimmino optimization algorithm. Thus, the irradiation times for individual fibers are updated
throughout the treatment in order to compensate for the influence of changes in tissue composition on the light
distribution at the therapeutic wavelength.
A system for interstitial photodynamic therapy is used in the treatment of thick skin tumors. The system allows simultaneous measurements of light fluence rate, sensitizer fluorescence, and tissue oxygen saturation by using the same fibers as for therapeutic light delivery. Results from ten tumor treatments using -aminolevulinic acid (ALA)-induced protoporphyrin IX show a significant, treatment-induced increase in tissue absorption at the therapeutic wavelength, and rapid sensitizer photobleaching. The changes in oxy- and deoxyhemoglobin content are monitored by means of near-infrared spectroscopy, revealing a varying tissue oxygenation and significant changes in blood volume during treatment. These changes are consistent with the temporal profiles of the light fluence rate at the therapeutic wavelength actually measured. We therefore propose the observed absorption increase to be due to treatment-induced deoxygenation in combination with changes in blood concentration within the treated volume. A higher rate of initial photobleaching is found to correlate with a less pronounced increase in tissue absorption. Based on the measured signals, we propose how real-time treatment supervision and feedback can be implemented. Simultaneous study of the fluence rate, sensitizer fluorescence, and local tissue oxygen saturation level may contribute to the understanding of the threshold dose for photodynamic therapy.
Measurements of concentration of sensitizers for photodynamic therapy can provide important information in the dosimetry planning and can also give input to the optimal time for treatment. There has been skepticism towards fluorescence techniques for this purpose, as the signal depends on the fluorescence yield and optical properties of the
tissue. Absorption based techniques, lack on the other hand, often the sensitivity required for many sensitizers with relative weak absorption in a wavelength region where hemoglobin absorption is dominant. A direct comparison between absorption and fluorescence techniques for measuring mTHPC concentration after topical application on hairless SKH-1 mice bearing skin carcinomas has been performed. 20 μl/cm2 of m-THPC thermogel (0.5 mg m-THPC/ml) were applied on normal and tumor area and the concentration of mTHPC was measured at 4 and 6 hours after drug application by two methods: 1. A fluorescence imaging system capturing images at two wavelengths (500 and 650 nm) following 405 nm excitation. Signals from different regions of interest were averaged and the intensity ratio at 650 to 500 was calculated. 2. A diffuse reflectance spectroscopy system with a fiber separation of 2 mm, providing the absorbance at 652 nm. Both
systems provided consistent results related to the photosensitizer concentration. The methods show a remarkable difference in the concentration of photosensitizer in normal skin and tumor. No significant difference in mTHPC concentration in tumor could be observed between the 4 and 6h groups after drug application.
A genetic algorithm for optimal placement of optical fibers in arbitrary geometries for interstitial photodynamic therapy has been developed. Based on calculated fiber positions, the diffuse light distribution is simulated by solving the diffusion equation by means of the finite element method and the treatment time is calculated. Utilizing an instrument for interstitial photodynamic therapy that is capable of both delivering the therapeutic irradiation and measuring
parameters of relevance to treatment monitoring, knowledge has been gained on temporal variations of tissue light absorption, sensitizer photobleaching and tissue oxygenation status. We speculate on how variations in these three parameters can be implemented in a crude pre-treatment dosimetry model for photodynamic therapy.
During δ-aminolevulinic acid (ALA) based Interstitial Photodynamic Therapy (IPDT) a high light fluence rate is present close to the source fibers. This might induce an unintentional tissue temperature increase of importance for the treatment outcome. In a previous study, we have observed, that the absorption in the tissue increases during the treatment. A system to measure the local tissue temperature at the source fibers during IPDT on tissue phantoms is presented. The temperature was measured by acquiring the fluorescence from small Cr3+-doped crystals attached to the tip of the illumination fiber used in an IPDT-system. The fluorescence of
the Alexandrite crystal used is temperature dependent. A ratio of the intensity of the fluorescence was formed between two different wavelength bands in the red region. The system was calibrated by immersing the fibers in an Intralipid solution placed in a temperature controlled oven. Measurements were then performed by placing the fibers interstitially in a pork chop as a tissue phantom. Measurements were also performed superficially on skin on a volunteer. A treatment was conducted for 10 minutes, and the fluorescence was measured each minute during the illumination. The fluorescence yielded the temperature at the fiber tip through the calibration curve. The measurements indicate a temperature increase of a few degrees during the simulated treatment.
Photodynamic therapy for the treatment of cancer relies on the presence of light, sensitizer and oxygen. By monitoring these three parameters during the treatment a better understanding and treatment control could possibly be achieved. Here we present data from in vivo treatments of solid skin tumors using an instrument for interstitial photodynamic therapy with integrated dosimetric monitoring. By using intra-tumoral ALA-administration and interstitial light delivery solid tumors are targeted. The same fibers are used for measuring the fluence rate at the treatment wavelength, the sensitizer fluorescence and the local blood oxygen saturation during the treatment. The data presented is based on 10 treatments in 8 patients with thick basal cell carcinomas. The fluence rate measurements at 635 nm indicate a major treatment induced absorption increase, leading to a limited light penetration at the treatment wavelength. This leads to a far from optimal treatment since the absorption increase prevents peripheral tumor regions from being fully treated. An interactive treatment has been implemented assisting the physician in delivering the correct light dose. The absorption increase can be compensated for by either prolonging the treatment time or increasing the output power of each individual treatment fiber. The other parameters of importance, i.e. the sensitizer fluorescence at 705 nm and the local blood oxygen saturation, are monitored in order to get an estimate of the amount of photobleaching and oxygen consumption. Based on the oxygen saturation signal, a fractionized irradiation can be introduced in order to allow for a re-oxygenation of the tissue.
A novel photodynamic therapy system based on interstitial illumination using multiple fibres is under development. The aim with this system is to enable treatment of large tumour volumes and also to utilise real-time measurements to allow on-line dosimetry. Important dosimetric parameters to measure are light fluence rate, sensitizer fluorescence intensity and local blood oxygenation. A construction which allows all functions to be readily performed with a single system is presented. We believe that interstitial PDT utilising this technique may be attractive in many clinical situations.
In this work a compact fluorosensor has been built for point-monitoring and imaging applications. The instrument has been applied in fluorescence studies on green vegetation and on malignant tissue. The instrument is based on a violet diode laser, an integrated spectrometer and optical fibers for light delivery and collection of the fluorescence signal. This combination makes the system very compact. The high laser output power allows for coupling of the laser light into a hyperspectral diagnostic imaging instrument, developed and built by Science and Technology International. In point-monitoring mode, the instrument has been tested on superficial skin tumors and when using δ-aminolevulinic acid induced protoporphyrin IX as a tumor sensitizer, good contrast between normal and malignant tissue was achieved, clearly demonstrating its feasibility in cancer diagnostics. In imaging mode, the instrument functioned solely as a light source, coupling the excitation light into the hyperspectral imaging instrument. The set-up was tested by studying chlorophyll fluorescence from vegetation. The fluorescence signal showed a low signal-to-noise ratio mainly because of
inefficient light coupling into the imaging instrument.
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