Counting Rate Characteristics and Image Distortion in Preclinical PET Imaging During Radiopharmaceutical Therapy.

PET may provide important information on the response during radiopharmaceutical therapy (RPT). Emission of radiation from the RPT radionuclide may disturb coincidence detection and impair image resolution. In this study, we tested the feasibility of performing intratherapeutic PET on 3 preclinical PET systems. METHODS: Using 22Na point sources and phantoms filled with 18F, as well as a phantom filled with either 99mTc or 177Lu, we evaluated the coincidence counting rate and spatial resolution when both a PET and a therapeutic radionuclide were in the PET system. Because 99mTc has a suitable half-life and is easy obtainable, we used it as a substitute for a generic therapeutic radionuclide. RESULTS: High activities of 99mTc deteriorated the coincidence counting rate from the 18F-filled phantom and the 22Na point source on all 3 systems. The counting rate could be corrected to a high degree on one of the systems by its dead-time correction. Spatial resolution was degraded at high 99mTc activities for all systems. On one of the systems, 177Lu increased the coincidence counting rate and slightly affected the spatial resolution. The results for high 177Lu activities were similar to those for 99mTc. CONCLUSION: Intratherapeutic imaging might be a feasible method of studying the response to RPT. However, some sensitive preclinical PET systems, unable to handle high counting rates, will have count losses and may also introduce image artifacts.

Online dosimetry for temoporfin-mediated interstitial photodynamic therapy using the canine prostate as model.

Online light dosimetry with real-time feedback was applied for temoporfin-mediated interstitial photodynamic therapy (PDT) of dog prostate. The aim was to investigate the performance of online dosimetry by studying the correlation between light dose plans and the tissue response, i.e., extent of induced tissue necrosis and damage to surrounding organs at risk. Light-dose planning software provided dose plans, including light source positions and light doses, based on ultrasound images. A laser instrument provided therapeutic light and dosimetric measurements. The procedure was designed to closely emulate the procedure for whole-prostate PDT in humans with prostate cancer. Nine healthy dogs were subjected to the procedure according to a light-dose escalation plan. About 0.15 mg/kg temoporfin was administered 72 h before the procedure. The results of the procedure were assessed by magnetic resonance imaging, and gross pathology and histopathology of excised tissue. Light dose planning and online dosimetry clearly resulted in more focused effect and less damage to surrounding tissue than interstitial PDT without dosimetry. A light energy dose-response relationship was established where the threshold dose to induce prostate gland necrosis was estimated from 20 to 30 J/cm2.

Cerenkov Luminescence Imaging for Accurate Placement of Radioactive Plaques in Episcleral Brachytherapy of Intraocular Tumors.

PURPOSE: The purpose of this study was to determine the feasibility of using Cerenkov luminescence imaging (CLI) to facilitate plaque placement during episcleral brachytherapy of intraocular tumors. METHODS: Ruthenium-106 (Ru-106) decays to rhodium-106, which in turn emits high-energy beta particles. When the electrons propagate through the eyewall, the so-called Cerenkov effect leads to emission of weak light, which can be captured by high-sensitivity charge-couple device (CCD) cameras. Enucleated porcine eyes were prepared with tumor phantoms made of melanin-containing gelatin. The anterior portion of the globe was removed, and different Ru-106 plaque types (designated CCA, CCB, COB, and CIA) with activities ranging from 6.8 to 16.7 MBq were sutured to the sclera overlying the tumor phantom. The globe was placed in a transparent container with saline. CLI was performed through the anterior opening of the eye using a cooled electron-multiplying CCD camera. RESULTS: Exposure times between 5 and 60 seconds produced good quality images of the Cerenkov light. There was a linear relationship between plaque activity and Cerenkov radiance. The perimeters of the CCA and CCB plaques could be seen clearly as circles of light symmetrically surrounding the tumor phantoms. Notched COB and CIA plaques led to images revealing their actual positions in relation to the optic disc and ciliary body, respectively. Simulated plaque tilting resulted in diffuse demarcation of the light. CONCLUSIONS: The study indicates that CLI is a feasible method to ensure accurate placement of Ru-106 plaques in brachytherapy of intraocular tumors. CLI may offer a new tool to improve and document plaque placement, both perioperatively and postoperatively.

Intratherapeutic biokinetic measurements, dosimetry parameter estimates, and monitoring of treatment efficacy using cerenkov luminescence imaging in preclinical radionuclide therapy.

UNLABELLED: In recent years, there has been increasing interest in noninvasive Cerenkov luminescence imaging (CLI) of in vivo radionuclide distribution in small animals, a method proven to be a high-throughput modality for confirmation of tracer uptake. 11B6 is an IgG1 monoclonal antibody that is specific to free human kallikrein-related peptidase 2, an antigen abundant in malignant prostatic tissue. Free human kallikrein-related peptidase 2 was targeted in prostate cancer xenografts using (177)Lu-labeled 11B6 in either murine or humanized forms for radionuclide therapy. In this setting, CLI was investigated as a tool for providing parameters of dosimetric importance during radionuclide therapy. First, longitudinal imaging of biokinetics using CLI and SPECT was compared. Second, the CLI signal was correlated to quantitative ex vivo tumor activity measurements. Finally, CLI was used to monitor the radionuclide treatment, and the integrated CLI radiance was found to correlate well with subject-specific tumor volume reduction. METHODS: 11B6 was radiolabeled with (177)Lu through the CHX-A″-DTPA chelator. In vivo CLI and SPECT imaging of (177)Lu-DTPA-11B6 uptake was performed on NMRI and BALB/c nude mice with subcutaneous LNCaP xenografts up to 14 d after injection. Tumor size was measured to assess response to radionuclide therapy. RESULTS: CLI correlated well with SPECT imaging and could be applied up to 14 d after injection of 20 MBq with the specific tracer used. Through integration of the CLI radiance as a function of time, a dose metric for the tumors could be formed that correlated exponentially with tumor volume reduction. CONCLUSION: CLI provided valuable intratherapeutic biokinetic measurements for treatment monitoring and could be used as a tool for subject-specific absorbed dose estimation.

Pharmacokinetic and biodistribution study following systemic administration of Fospeg®--a Pegylated liposomal mTHPC formulation in a murine model.

Fospeg® is a newly developed photosensitizer formulation based on meso-tetra(hydroxyphenyl)chlorin (mTHPC), with hydrophilic liposomes to carry the hydrophobic photosensitizer to the target tissue. In this study the pharmacokinetics and biodistribution of Fospeg® were investigated by high performance liquid chromatography at various times (0.5-18 hours) following systemic i.v. administration. As a model an experimental HT29 colon tumor in NMRI nu/nu mice was employed. Our study indicates a higher plasma peak concentration, a longer circulation time and a better tumor-to-skin ratio than those of Foslip®, another liposomal mTHPC formulation. Data from ex vivo tissue fluorescence and reflectance imaging exhibit good correlation with chemical extraction. Our results have shown that optical imaging provides the potential for fluorophore quantification in biological tissues.

Design and validation of a fiber optic point probe instrument for therapy guidance and monitoring.

Optical techniques for tissue diagnostics currently are experiencing tremendous growth in biomedical applications, mainly due to their noninvasive, inexpensive, and real-time functionality. Here, we demonstrate a hand-held fiber optic probe instrument based on fluorescence/reflectance spectroscopy for precise tumor delineation. It is mainly aimed for brain tumor resection guidance with clinical adaptation to minimize the disruption of the standard surgical workflow and is meant as a complement to the state-of-the-art fluorescence surgical microscopy technique. Multiple light sources with fast pulse modulation and detection enable precise quantification of protoporphyrin IX (PpIX), tissue optical properties, and ambient light suppression. Laboratory measurements show the system is insensitive to strong ambient light. Validation measurements of tissue phantoms using nonlinear least squares support vector machines (LS-SVM) regression analysis demonstrate an error of <5% for PpIX concentration ranging from 400 to 1000 nM, even in the presence of large variations in phantom optical properties. The mean error is 3% for reduced scattering coefficient and 5% for blood concentration. Diagnostic precision of 100% was obtained by LS-SVM classification for in vivo skin tumors with topically applied 5-aminolevulinic acid during photodynamic therapy. The probe could easily be generalized to other tissue types and fluorophores for therapy guidance and monitoring.

Nanoparticle uptake in tumors is mediated by the interplay of vascular and collagen density with interstitial pressure.

Nanoparticle delivery into solid tumors is affected by vessel density, interstitial fluid pressure (IFP) and collagen, as shown in this article by contrasting the in vivo macroscopic quantitative uptake of 40 nm fluorescent beads in three tumor types.The fluorescence uptake was quantified on individual animals by normalization with the transmitted light and then normalized to normal tissue uptake in each mouse. Mean data for uptake in individual tumor lines then showed expected trends with the largest uptake in the most vascularized tumor line. Tumor lines with increased collagen were also consistent with highest interstitial fluid pressure and correlated with lowest uptake of nanoparticles. The data is consistent with a delivery model indicating that while vascular permeability is maximized by neovascular growth, it is inhibited by collagen content and the resulting interstitial pressure. Imaging of these parameters in vivo can lead to better individual noninvasive methods to assess drug penetration in situ. FROM THE CLINICAL EDITOR: In this manuscript the dependence of nanoparticle delivery is addressed from the standpoint of vascular factors (the more vascularized, the better delivery) and as a function of collagen density and interstitial pressure (the higher these are, the worse the delivery).

(68)Ga-labeled superparamagnetic iron oxide nanoparticles (SPIONs) for multi-modality PET/MR/Cherenkov luminescence imaging of sentinel lymph nodes.

The aim of this study was to develop (68)Ga-SPIONs for use as a single contrast agent for dynamic, quantitative and high resolution PET/MR imaging of Sentinel Lymph Node (SLN). In addition (68)Ga enables Cherenkov light emission which can be used for optical guidance during resection of SLN. SPIONs were labeled with (68)Ga in ammonium acetate buffer, pH 5.5. The labeling yield and stability in human serum were determined using instant thin layer chromatography. An amount of 0.07-0.1 mL (~5-10 MBq, 0.13 mg Fe) of (68)Ga-SPIONs was subcutaneously injected in the hind paw of rats. The animals were imaged at 0-3 h and 25 h post injection with PET/CT, 9.4 T MR and CCDbased Cherenkov optical systems. A biodistribution study was performed by dissecting and measuring the radioactivity in lymph nodes, kidneys, spleen, liver and the injection site. The labeling yield was 97.3 ± 0.05% after 15 min and the (68)Ga-SPIONs were stable in human serum. PET, MR and Cherenkov luminescence imaging clearly visualized the SLN. Biodistribution confirmed a high uptake of the (68)Ga-SPIONs within the SLN. We conclude that generator produced (68)Ga can be labeled to SPIONs. Subcutaneously injected (68)Ga-SPIONs can enhance the identification of the SLNs by combining sensitive PET and high resolution MR imaging. Clinically, hybrid PET/MR cameras are already in use and (68)Ga-SPIONs have a great potential as a single-dose, tri-modality agent for diagnostic imaging and potential Cherenkov luminescent guided resection of SLN.

Quantitative Cherenkov emission spectroscopy for tissue oxygenation assessment.

Measurements of Cherenkov emission in tissue during radiation therapy are shown to enable estimation of hemoglobin oxygen saturation non-invasively, through spectral fitting of the spontaneous emissions from the treated tissue. Tissue oxygenation plays a critical role in the efficacy of radiation therapy to kill tumor tissue. Yet in-vivo measurement of this has remained elusive in routine use because of the complexity of oxygen measurement techniques. There is a spectrally broad emission of Cherenkov light that is induced during the time of irradiation, and as this travels through tissue from the point of the radiation deposition, the tissue absorption and scatter impart spectral changes. These changes can be quantified by diffuse spectral fitting of the signal. Thus Cherenkov emission spectroscopy is demonstrated for the first time quantitatively in vitro and qualitatively in vivo, and has potential for real-time online tracking of tissue oxygen during radiation therapy when fully characterized and developed.

Cerenkov emission induced by external beam radiation stimulates molecular fluorescence.

PURPOSE: Cerenkov emission is induced when a charged particle moves faster than the speed of light in a given medium. Both x-ray photons and electrons produce optical Cerenkov photons in everyday radiation therapy of tissue; yet, this phenomenon has never been fully documented. This study quantifies the emissions and also demonstrates that the Cerenkov emission can excite a fluorophore, protoporphyrin IX (PpIX), embedded in biological phantoms. METHODS: In this study, Cerenkov emission induced by radiation from a clinical linear accelerator is investigated. Biological mimicking phantoms were irradiated with x-ray photons, with energies of 6 or 18 MV, or electrons at energies 6, 9, 12, 15, or 18 MeV. The Cerenkov emission and the induced molecular fluorescence were detected by a camera or a spectrometer equipped with a fiber optic cable. RESULTS: It is shown that both x-ray photons and electrons, at MeV energies, produce optical Cerenkov photons in tissue mimicking media. Furthermore, we demonstrate that the Cerenkov emission can excite a fluorophore, protoporphyrin IX (PpIX), embedded in biological phantoms. CONCLUSIONS: The results here indicate that molecular fluorescence monitoring during external beam radiotherapy is possible.

Drug quantification in turbid media by fluorescence imaging combined with light-absorption correction using white Monte Carlo simulations.

Accurate quantification of photosensitizers is in many cases a critical issue in photodynamic therapy. As a noninvasive and sensitive tool, fluorescence imaging has attracted particular interest for quantification in pre-clinical research. However, due to the absorption of excitation and emission light by turbid media, such as biological tissue, the detected fluorescence signal does not have a simple and unique dependence on the fluorophore concentration for different tissues, but depends in a complex way on other parameters as well. For this reason, little has been done on drug quantification in vivo by the fluorescence imaging technique. In this paper we present a novel approach to compensate for the light absorption in homogeneous turbid media both for the excitation and emission light, utilizing time-resolved fluorescence white Monte Carlo simulations combined with the Beer-Lambert law. This method shows that the corrected fluorescence intensity is almost proportional to the absolute fluorophore concentration. The results on controllable tissue phantoms and murine tissues are presented and show good correlations between the evaluated fluorescence intensities after the light-absorption correction and absolute fluorophore concentrations. These results suggest that the technique potentially provides the means to quantify the fluorophore concentration from fluorescence images.

System for interstitial photodynamic therapy with online dosimetry: first clinical experiences of prostate cancer.

The first results from a clinical study for Temoporfin-mediated photodynamic therapy (PDT) of low-grade (T1c) primary prostate cancer using online dosimetry are presented. Dosimetric feedback in real time was applied, for the first time to our knowledge, in interstitial photodynamic therapy. The dosimetry software IDOSE provided dose plans, including optical fiber positions and light doses based on 3-D tissue models generated from ultrasound images. Tissue optical property measurements were obtained using the same fibers used for light delivery. Measurements were taken before, during, and after the treatment session. On the basis of these real-time measured optical properties, the light-dose plan was recalculated. The aim of the treatment was to ablate the entire prostate while minimizing exposure to surrounding organs. The results indicate that online dosimetry based on real-time tissue optical property measurements enabled the light dose to be adapted and optimized. However, histopathological analysis of tissue biopsies taken six months post-PDT treatment showed there were still residual viable cancer cells present in the prostate tissue sections. The authors propose that the incomplete treatment of the prostate tissue could be due to a too low light threshold dose, which was set to 5 J∕cm2.

Photodynamic therapy: superficial and interstitial illumination.

Photodynamic therapy (PDT) is reviewed using the treatment of skin tumors as an example of superficial lesions and prostate cancer as an example of deep-lying lesions requiring interstitial intervention. These two applications are among the most commonly studied in oncological PDT, and illustrate well the different challenges facing the two modalities of PDT-superficial and interstitial. They thus serve as good examples to illustrate the entire field of PDT in oncology. PDT is discussed based on the Lund University group's over 20 yr of experience in the field. In particular, the interplay between optical diagnostics and dosimetry and the delivery of the therapeutic light dose are highlighted. An interactive multiple-fiber interstitial procedure to deliver the required therapeutic dose based on the assessment of light fluence rate and sensitizer concentration and oxygen level throughout the tumor is presented.

A matrix-free algorithm for multiple wavelength fluorescence tomography.

In the recent years, there has been an increase in applications of non-contact diffusion optical tomography. Especially when the objective is the recovery of fluorescence targets. The non-contact acquisition systems with the use of a CCD-camera produce much denser sampled boundary data sets than fibre-based systems. When model-based reconstruction methods are used, that rely on the inversion of a derivative operator, the large number of measurements poses a challenge since the explicit formulation and storage of the Jacobian matrix could be in general not feasible. This problem is aggravated further in applications, where measurements at multiple wavelengths are used. We present a matrix-free model-based reconstruction method, that addresses the problems of large data sets and reduces the computational cost and memory requirements for the reconstruction. The idea behind the matrix-free method is that information about the Jacobian matrix could be available through matrix times vector products so that the creation and storage of big matrices can be avoided. We tested the method for multiple wavelength fluorescence tomography with simulated and experimental data from phantom experiments, and we found substantial benefits in computational times and memory requirements.

In vivo photosensitizer tomography inside the human prostate.

Interstitial photodynamic therapy (IPDT) provides a promising means to treat large cancerous tumors and solid organs inside the human body. The treatment outcome is dependent on the distributions of light, photosensitizer, and tissue oxygenation. We present a scheme for reconstructing the spatial distribution of a fluorescent photosensitizer. The reconstruction is based on measurements performed in the human prostate, acquired during an ongoing IPDT clinical trial, as well as in optical phantoms. We show that in an experimental setup we can quantitatively reconstruct a fluorescent inclusion in a fluorescent background. We also show reconstructions from a patient showing a heterogeneous distribution of the photosensitizer mTHPC in the human prostate.

Low ambulatory blood pressure is associated with lower cognitive function in healthy elderly men.

INTRODUCTION: Low blood pressure (BP) has been found to be associated with cerebrovascular damage in the elderly. Studies of the relation of ambulatory BP to cognitive function in elderly persons aged 80 years or above is lacking, however. METHODS: Ninety-seven 81-year-old men from the population study 'Men born in 1914' underwent ambulatory BP monitoring and were given a cognitive test battery, 79 subjects completing all six tests. Low ambulatory systolic blood pressure (SBP) was defined as <130 mmHg and low ambulatory diastolic blood pressure (DBP) as <80 mmHg (corresponding in terms of office BP to approximately <140 and <90 mmHg, respectively). Odds ratios (OR) for lower cognitive function were calculated using a forward stepwise logistic regression model, controlling for confounding factors. RESULTS: Subjects with ambulatory SBP <130 mmHg had higher OR values for daytime (OR 2.6; P=0.037), nighttime (OR 3.6; P=0.032) and 24h (OR 2.6; P=0.038) BP measurements. A lower cognitive function was associated with lower nighttime SBP and DBP levels and lower 24-h mean SBP compared to subjects with higher cognitive function. OR values connected to low nocturnal SBP, had a tendency to be particularly high among subjects on anti-hypertensive drugs (OR 9.1; P=0.067, n.s.). CONCLUSION: Ambulatory SBP levels <130 mmHg and lower nighttime SBP and DBP were associated with lower cognitive function in healthy elderly men. Further investigation is needed to ascertain the effects of the presently recommended treatment goal of <140 mmHg for office SBP also on elderly over 80 years of age.

Lower cognitive performance in 81-year-old men with greater nocturnal blood pressure dipping.

Abnormal day-to-night blood pressure (BP) pattern have been found to be associated with cerebrovascular damage, yet studies of the elderly 80 years of age and above, for whom the risk pattern may be different due to ageing and age-associated diseases, are lacking. Ninety-seven 81-year-old men underwent ambulatory BP monitoring and were given six cognitive tests, 79 of the men completing the cognitive test battery. The odds ratio (OR) for performing one standard deviation below the mean on any cognitive test was calculated using a forward stepwise logistic regression model, confounding factors being controlled for. Groups defined in terms of day-to-night changes in BP were compared in this respect. Cognitive performance was lower (OR 3.6; P = 0.017) in the group usually described as dippers (10%-20% nocturnal drop in systolic BP [SBP]) as compared with nondippers (<10% drop). The tertile with the greatest SBP fall (10.6%-19.8%, a range considered as normal among middle aged) showed lowest cognitive performance (OR 4.7; P = 0.008) as compared with the middle tertile (5.1%-10.5% drop). The mean nocturnal fall in SBP was 7.4%, significantly greater in those with lower rather than higher cognitive performance. A nocturnal drop in SBP of >/=10% was associated with lower cognitive performance in these elderly men. The limits to normal dipping appear to be shifted in the direction of a lesser drop in the very elderly.

Low nocturnal blood pressure is associated with reduced cerebral blood flow in the cohort "Men born in 1914".

BACKGROUND: "Men born in 1914" is a population-based cohort study of the epidemiology of cardiovascular and cerebral disease. Little is known about how diurnal variation in blood pressure (BP) levels influence cerebral perfusion in very elderly populations. OBJECTIVES: To study the association between systolic (SBP) and diastolic BP (DBP) levels, during the day and at night, expressed through 24 h ambulatory BP monitoring (ABPM) and regional cerebral blood flow (rCBF) disturbances. METHODS: A cross-sectional study from a population-based cohort of 108 men 81 years of age (born in 1914) was performed in an out-patient university clinic. Cerebral blood flow measurements using (99m)Tc-hexamethylpropyleneamine oxime single photon emission computed tomography and 24 h ABPM were performed. Eleven men were excluded due to incomplete ABPM data. RESULTS: Mean DBP at night for each tertile was correlated to rCBF for the medial temporal right (P=0.012) and left (P=0.039) regions. Also, DBP during the day was correlated to the medial temporal right region (P=0.025). When analyses were stratified for DBP during the day, subjects with high DBP during the day (greater than 70 mmHg) showed a stronger association between low medial temporal right rCBF and low mean DBP at night (r=0.32, P=0.009) compared with subjects who had a lower daytime DBP. A corresponding positive correlation was noted for the medial temporal left region and daytime SBP, whereas a negative correlation was noted for frontal left region blood flow and SBP at night. CONCLUSIONS: A significant association was seen between low BP levels, especially at night, and rCBF in subjects with otherwise normal daytime DBP that may indicate a risk for nocturnal cerebral ischemia.

Realtime light dosimetry software tools for interstitial photodynamic therapy of the human prostate.

Photodynamic therapy (PDT) for the treatment of prostate cancer has been demonstrated to be a safe treatment option capable of inducing tissue destruction and decreasing prostate specific antigen (PSA) levels. However, prostate-PDT results in large intra- and interpatient 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 has been developed that combines therapeutic light delivery and monitoring of light transmission via numerous bare-ended optical fibers. Here, we present algorithms that utilize data on the light distribution within the target tissue to provide realtime treatment feedback based on a light dose threshold model for PDT. This realtime dosimetry module is implemented to individualize the light dose and compensate for any treatment-induced variations in light attenuation. More specifically, based on the light transmission signals between treatment fibers, spatially resolved spectroscopy is utilized to assess the effective attenuation coefficient of the tissue. These data constitute input to a block-Cimmino optimization algorithm, employed to calculate individual fiber irradiation times provided the requirement to deliver a predetermined light dose to the target tissue while sparing surrounding sensitive organs. By repeatedly monitoring the light transmission signals during the entire treatment session, optical properties and individual fiber irradiation times are updated in realtime. The functionality of the algorithms is tested on diffuse light distribution data simulated by means of the finite element method (FEM). The feasibility of utilizing spatially resolved spectroscopy within heterogeneous media such as the prostate gland is discussed. Furthermore, we demonstrate the ability of the block-Cimmino algorithm to discriminate between target tissue and organs at risk (OAR). Finally, the realtime dosimetry module is evaluated for treatment scenarios displaying spatially and temporally varying light attenuation levels within the target tissue. We conclude that the realtime dosimetry module makes it possible to deliver a certain light dose to the target tissue despite spatial and temporal variations of the target tissue optical properties at the therapeutic wavelength.

Fluorescence monitoring of a topically applied liposomal Temoporfin formulation and photodynamic therapy of nonpigmented skin malignancies.

Meso-tetra(hydroxyphenyl)chlorin (mTHPC) (INN: Temoporfin) is a potent photodynamically active substance in clinical use today. Usually, the substance is given systemically and a known drawback with this administration route is a prolonged skin light sensitization. For the first time to our knowledge, a liposomal Temoporfin gel formulation for topical application was studied in connection with photodynamic therapy (PDT) of nonpigmented skin malignancies in humans. Intervals of 4 hr between drug administration and light irradiation were used. Sensitizer distribution within tumor and surrounding normal skin was investigated by means of point monitoring and imaging fluorescence spectroscopy before, during, and after PDT, showing high tumor selectivity. Furthermore, the bleaching of Temoporfin was studied during the PDT procedure by monitoring the fluorescence following excitation by using a therapeutic light. A 30-35% light-induced photometabolization was shown. No pain occurred during or after treatment. It was also observed that the treated area did not show any swollen tissue or reddening, as is often seen in PDT using topical delta-aminolevulinic acid. On controlling the patients one week after treatment, healing progress was observed in several patients and no complications were registered.