Claes af Klinteberg

Photodynamic therapy vs. cryosurgery of basal cell carcinomas: results of a phase III clinical trial

Background A previously reported randomized clinical trial showed treatment of Bowens disease using photodynamic therapy (PDT) with topically applied δ‐aminolaevulinic acid (ALA) to be at least as effective as cryosurgery and to be associated with fewer adverse effects.

In vivo fluorescence imaging for tissue diagnostics

Non-invasive fluorescence imaging has the potential to provide in vivo diagnostic information for many clinical specialties. Techniques have been developed over the years for simple ocular observations following UV excitation to sophisticated spectroscopic imaging using advanced equipment. Much of the impetus for research on fluorescence imaging for tissue diagnostics has come from parallel developments in photodynamic therapy of malignant lesions with fluorescent photosensitizers. However, the fluorescence of endogenous molecules (tissue autofluorescence) also plays an important role in most applications. In this paper, the possibilities of imaging tissues using fluorescence spectroscopy as a mean of tissue characterization are discussed. The various imaging techniques for extracting diagnostic information suggested in the literature are reviewed. The development of exogenous fluorophores for this purpose is also presented. Finally, the present status of clinical evaluation and future directions are discussed.

Preliminary evaluation of two fluorescence imaging methods for the detection and the delineation of basal cell carcinomas of the skin

Fluorescence techniques can provide powerful noninvasive means for medical diagnosis, based on the detection of either endogenous or exogenous fluorophores. The fluorescence of δ‐aminolevulinic acid (ALA)‐induced protoporphyrin IX (PpIX) has already shown promise for the diagnosis of tumors. The aim of the study was to investigate the localization of skin tumors after the topical application of ALA, by detecting the PpIX fluorescence either in the spectral or in the time domain.

Feasibility study of a system for combined light dosimetry and interstitial photodynamic treatment of massive tumors

A system for the photodynamic laser treatment of massive tumors that employs multiple optical fibers to be inserted into the tumor mass is described. The light flux through the tumor can be assessed by use of the individual fibers both as transmitters and as receivers. With a computer model that describes the diffusive light propagation, optical dosimetry is under development. The system has been tested in an experimental animal tumor model in preparation for clinical work. Currently, delta-aminolevulinic acid is used as a sensitizer, activated by 635-nm radiation from a 2.0-W compact diode laser system. With the availability of future, highly selective drugs absorbing approximately 750 nm, larger tumor volumes should be treatable, and surrounding, sensitive normal tissue should be spared.

Concentration measurement of gas embedded in scattering media by employing absorption and time-resolved laser spectroscopy

Diode-laser-based absorption spectroscopy for the evaluation of embedded gas concentrations in porous materials is demonstrated in measurements of molecular oxygen dispersed throughout scattering polystyrene foam, used here as a generic test material. The mean path length of light scattered in the material is determined with the temporal characteristics of the radiation transmitted through the sample. This combined with sensitive gas-absorption measurements employing wavelength-modulation spectroscopy yields an oxygen concentration in polystyrene foam of 20.4% corresponding to a foam porosity of 98%, which is consistent with manufacturing specifications. This feasibility study opens many possibilities for quantitative measurements by using the method of gas-in-scattering-media absorption spectroscopy.

Compact medical fluorosensor for minimally invasive tissue characterization

A compact fiber-optic point-measuring fluorosensor fully adapted to clinical studies is described. The system can use two excitation wavelengths, 337 and 405nm, obtained from a nitrogen laser directly, or after dye laser conversion, respectively. The image intensifier used in the spectrometer can be gated with a variable time delay, allowing also time-resolved spectra to be extracted, with a time resolution of about 4ns. Moreover, diffusely scattered white light can be spectrally recorded. The system is fully computer controlled enabling short recording times in clinical application, which are illustrated.

In vivo absorption spectroscopy of tumor sensitizers with femtosecond white light

A system based on a femtosecond white-light continuum and a streak camera was used for recordings of the in vivo absorption spectra of the tumor-seeking agent disulphonated aluminum phthalocyanine. Measurements for different drug doses were performed on tumor tissue (muscle-implanted adenocarcinoma) and normal muscle tissue in rats. It was found that the shape of the spectrum is tissue dependent. The peak of the absorption spectrum is blueshifted in tumor tissue as compared with the muscle. Thus the contrast in the drug-related absorption can be altered by up to a factor of 2 from the primary drug molecular-concentration contrast between normal muscle and tumor by the proper selection of the illumination wavelength.

Diffusely scattered femtosecond white-light examination of breast tissue in vitro and in vivo

Multispectral studies of light propagation in female breast tissue have been performed. Short pulses of white light were generated by using self-phase modulation of a high-power laser pulse focused into a cuvette filled with water. The white light pulses illuminated the tissue and the scattered light was recorded with time- and wavelength dispersion by a streak camera. Measurements were performed on breast mastectomies in vitro and measurements on healthy breast tissue in vivo. The reduced scattering coefficient and the absorption coefficient of breast tissue were obtained in different wavelength regions by fitting solutions of the diffusion equation to the experimental data. Significant variations in the magnitude of the optical properties could be seen between the different individuals. No characteristic spectral discrepancy for tumor tissue was found.

Laser-induced fluorescence studies of premalignant and benign lesions in the female genital tract

Laser-induced fluorescence (LIF) was studied in vivo from premalignant and benign lesions in the female genital tract, in particular the cervix. The aim of the study was to investigate the possibilities to differentiate cervical intraepithelial neoplasia (CIN) from normal tissue by means of two different fluorescence modalities. Most of the patients were given a low dose (5 mg/kg bw) of (delta) -amino levulinic acid (ALA). The ALA was orally administered 2 - 4 hours prior to the investigation. During this time, the ALA is transformed to the strongly fluorescent protoporphyrin IX (PpIX) via the haem cycle. Excitation light with a wavelength of 405 nm was used to excite the PpIX fluorescence. Excess amounts of PpIX were accumulated preferentially in diseased tissue. However, the variability in the PpIX accumulation from patient to patient was large. By using excitation light at 337 nm, the endogenous fluorophores are more efficiently excited. Therefore, this excitation modality was exploited for studying spectral characteristics of the autofluorescence in different tissue types. The spectra obtained were evaluated by forming fluorescence intensity ratios. The tissue types were grouped according to the histopathological examination. A correlation with the fluorescence ratios was performed. Some problems with the classification remain, mostly due to the difficulties in obtaining histopathologic evaluation of the biopsies at the exact location of the LIF measurements.

Interstitial photodynamic therapy - diagnostic measurements and treatment in rat malignant experimental tumours

A recently developed multiple fiber system for treating malignant tumors with interstitial photodynamic therapy was used in studies on rats with colon adenocarcinoma inoculated into the muscles of the hind legs. The animals were intraperitonially administrated (delta) -aminolevulinic acid (ALA), which is metabolized to protoporphyrin IX (PpIX) in the tissue. The treatment system consists of a laser light source, a beam-splitting system dividing the light into three or six output fibers and a dosimetry program calculating the optimal fiber position within the tumor as well as the treatment time needed to obtain a given threshold value of the light dose. One aim of the study was to compare the treatment outcome with the modelled dosimetry predictions. Tumor reduction was examined three days post treatment. A volume decrease was found in 85% of the treated tumors. The mean volume reduction was 44%, with one tumor completely disappearing. Histopathological examination three days post treatment showed substantial necrotic parts which, however, to a smaller extent were present also for non-treated tumors. These results indicated that the tumors have been under treated and the light dose has to be increased. Measurements of the build-up and photo-induced bleaching of PpIX using laser-induced fluorescence were also performed during the experiments.