Biomedical subjects

Lothar Lilge

Publications and source records attributed to Lothar Lilge.

At least 19 recordsLinked to original sources

PubMed2006

Towards conformal light delivery using tailored cylindrical diffusers: attainable light dose distributions.

Interstitial light delivery for therapeutic applications requires the use of fibre-based light diffusers. Such diffusers are presently manufactured to emit with a flat longitudinal power profile. Recently, diffusers with tailored longitudinal emission profiles have become available opening an avenue to improve conformal light delivery. This paper explores the ability of tailored diffusers to improve light dose confinement to the target volume. A formalism to calculate the light dose from an arbitrary source distribution is presented based on the convolution with an appropriate point source function. By choosing a source distribution corresponding to a cylindrical diffuser emitting with a sinusoidal profile, the set of attainable light dose distributions is characterized via a relationship between the diffuser's spatial frequency, the radial distance and the amplitude of the isodose contour.

Biophysical Phenomena↗

PubMed2006

Development and characterization of multi-sensory fluence rate probes.

Multi-sensory fluence rate probes (MSPs) yield several simultaneous measurements of photodynamic therapy (PDT) treatment light fluence from a single interstitial probe. Fluorescent sensors are embedded at desired positions along the axis of the optical fibre. A single fluorescence emission spectrum is obtained and decomposed using a partial least squares (PLS)-based analysis to yield the fluence at each sensor's location. The responsivity, linearity and possible photodegradation of each fluorophore chosen for the MSPs were evaluated using single-sensor probes. The performance of two- and three-sensor MSPs was evaluated experimentally. Individual fluorescence spectra collected from each sensor on the MSP were used to construct the training set necessary for the PLS-based analysis. The MSPs' responsivity, spatial resolution and accuracy were evaluated relative to a single scattering-tip detector. Three-fluorophore MSPs permitted three simultaneous measurements of the fluence rate gradient in a tissue-like phantom, with an average accuracy of 6.7%. No appreciable photodegradation or cross-talk was observed.

Equipment Design↗

PubMed2005

The distribution of the anticancer drug Doxorubicin in relation to blood vessels in solid tumors.

PURPOSE: Anticancer drugs gain access to solid tumors via the circulatory system and must penetrate the tissue to kill cancer cells. Here, we study the distribution of doxorubicin in relation to blood vessels and regions of hypoxia in solid tumors of mice. EXPERIMENTAL DESIGN: The distribution of doxorubicin was quantified by immunofluorescence in relation to blood vessels (recognized by CD31) of murine 16C and EMT6 tumors and human prostate cancer PC-3 xenografts. Hypoxic regions were identified by injection of EF5. RESULTS: The concentration of doxorubicin decreases exponentially with distance from tumor blood vessels, decreasing to half its perivascular concentration at a distance of about 40 to 50 mum, The mean distance from blood vessels to regions of hypoxia is 90 to 140 microm in these tumors. Many viable tumor cells are not exposed to detectable concentrations of drug following a single injection. CONCLUSIONS: Limited distribution of doxorubicin in solid tumors is an important and neglected cause of clinical resistance that is amenable to modification. The technique described here can be adapted to studying the distribution of other drugs within solid tumors and the effect of strategies to modify their distribution.

Animals↗

PubMed2005

Evidence for the direct binding of phosphorylated p53 to sites of DNA breaks in vivo.

Despite a clear link between ataxia-telangiectasia mutated (ATM)-dependent phosphorylation of p53 and cell cycle checkpoint control, the intracellular biology and subcellular localization of p53 phosphoforms during the initial sensing of DNA damage is poorly understood. Using G0-G1 confluent primary human diploid fibroblast cultures, we show that endogenous p53, phosphorylated at Ser15 (p53Ser15), accumulates as discrete, dose-dependent and chromatin-bound foci within 30 minutes following induction of DNA breaks or DNA base damage. This biologically distinct subpool of p53Ser15 is ATM dependent and resistant to 26S-proteasomal degradation. p53Ser15 colocalizes and coimmunoprecipitates with gamma-H2AX with kinetics similar to that of biochemical DNA double-strand break (DNA-dsb) rejoining. Subnuclear microbeam irradiation studies confirm p53Ser15 is recruited to sites of DNA damage containing gamma-H2AX, ATM(Ser1981), and DNA-PKcs(Thr2609) in vivo. Furthermore, studies using isogenic human and murine cells, which express Ser15 or Ser18 phosphomutant proteins, respectively, show defective nuclear foci formation, decreased induction of p21WAF, decreased gamma-H2AX association, and altered DNA-dsb kinetics following DNA damage. Our results suggest a unique biology for this p53 phosphoform in the initial steps of DNA damage signaling and implicates ATM-p53 chromatin-based interactions as mediators of cell cycle checkpoint control and DNA repair to prevent carcinogenesis.

Acid Anhydride Hydrolases↗

PubMed2005

Design and performance of thin cylindrical diffusers created in Ge-doped multimode optical fibers.

Cylindrical fiber diffusers have become common tools for various medical therapies. However, their large outer diameters and short lengths restrict their clinical application in some newly developed light therapies. Here, a 250-microm outer-diameter diffuser with an active length that exceeds 5 cm is presented. Diffusers are created in photosensitive optical fibers with outer cladding diameters of 140 microm by use of a structured beam from an excimer laser. Predetermined emission profiles can be achieved. Photometric characteristics, including longitudinal, polar, and azimuthal emission diagrams, were determined by use of a goniometer to assess the light-emission performance of the diffuser. Longitudinal isotropy of better than +/- 10% was achieved. Polar and azimuthal emissions were within +/- 15% of those of an ideal linear Lambertian emitter. Polar uniformity could be improved by an insignificant increase in the outer diameter by use of a diffusing recoating compound. The residual leakage of light at the distal end of the diffuser can be as little as 1%. Other physical parameters tested include minimal bending radius after recoating (< 5 mm) and maximum power handling (> 1.0W cm(-1)). All materials employed were biocompatible.

Journal Article↗

PubMed2005

Fluorescence image-guided brain tumour resection with adjuvant metronomic photodynamic therapy: pre-clinical model and technology development.

Fluorescence-guided resection (FGR) and photodynamic therapy (PDT) have previously been investigated separately with the objectives, respectively, of increasing the extent of brain tumour resection and of selectively destroying residual tumour post-resection. Both techniques have demonstrated trends towards improved survival, pre-clinically and clinically. We hypothesize that combining these techniques will further delay tumour re-growth. In order to demonstrate technical feasibility, we here evaluate fluorescence imaging and PDT treatment techniques in a specific intracranial tumour model. The model was the VX2 carcinoma grown by injection of tumour cells into the normal rabbit brain. An operating microscope was used for white light imaging and a custom-built fluorescence imaging system with co-axial excitation and detection was used for FGR. PDT treatment light was applied by intracranially-implanted light emitting diodes (LED). The fluorescent photosensitizer used for both FGR and PDT was ALA-induced PpIX. For PDT, ALA (100 mg kg(-1)) and low light doses (15 and 30 J) were administered over extended periods, which we refer to as metronomic PDT (mPDT). Eighteen tumour bearing rabbits were divided equally into three groups: controls (no resection); FGR; and FGR followed by mPDT. Histological whole brain sections (H&E stain) showed primary and recurrent tumours. No bacteriological infections were found by Gram staining. Selective tumour cell death through mPDT-induced apoptosis was demonstrated by TUNEL stain. These results demonstrate that the combined treatment is technically feasible and this model is a candidate to evaluate it. Further optimization of mPDT treatment parameters (drug/light dose rates) is required to improve survival.

Aminolevulinic Acid↗

PubMed2005

Thin cylindrical diffusers in multimode Ge-doped silica fibers.

BACKGROUND AND OBJECTIVES: Cylindrical fiber diffusers are common tools in photodynamic therapy (PDT). However, large outer diameter and short length limit the indications to which they can be applied. Here a diffuser with an outer diameter of 250 microm and an active length exceeding 5 cm was developed. STUDY DESIGN/MATERIALS AND METHODS: Diffusers based on photosensitive quartz optical fibers with a cladding diameter of 140 microm were developed using a structured beam from an excimer laser. A complete set of photometric characteristics, including longitudinal, polar, and azimuth radiance emission diagrams were collected. RESULTS AND CONCLUSIONS: The longitudinal radiance emission is homogenous to within +/-10%. The ability to create custom designed longitudinal emission profiles was demonstrated. The variations of the polar and azimuth radiance emission diagrams were within +/- 15% from an ideal Lambertian emitter. The polar uniformity can be improved with a slight increase of the outer diameter using a diffusing recoating compound. The residual light leakage at the distal end of the fiber diffuser was suppressed to 2%. The minimal bending radius after recoating is approximately 5 mm. Maximum power distribution is > 0.5 W cm(-1).

Equipment Design↗

PubMed2005

Pc 4 photodynamic therapy of U87-derived human glioma in the nude rat.

BACKGROUND AND OBJECTIVES: As a potential therapy for malignant glioma, we tested the phthalocyanine photosensitizer Pc 4 for: (1) rapid clearance from the vasculature, (2) specificity for glioma, and (3) tumoricidal photosensitizing capability. STUDY DESIGN/MATERIALS AND METHODS: Parenchymal injection of U87 cells into athymic rat brains (N = 100) was followed after 12 days by tail vein injection of 0.5 mg/kg Pc 4. After 1 day, the tumor was illuminated with either 5 (N = 11) or 30 (N = 16) J/cm(2) red light at 672 nm. Sacrifice was 1 day later. The brains from these 27 animals underwent H&E (necrosis) and TUNEL assay (apoptosis) histology. Pc 4 concentration of explanted brains and tumors (N = 16), and all blood samples (N = 52) were determined by HPLC-MS 1 day post Pc 4 administration. RESULTS: Tumor-specific apoptosis was almost uniformly seen; however, necrosis was found mostly in the high-light-dose group. Pc 4 concentration in bulk tumor averaged 3.8 times greater than in normal brain. CONCLUSIONS: These results warrant expanding this pre-clinical study to seek effective baseline Pc 4 drug- and light-doses and infusion-to-photoirradiation timing that would be necessary for a Pc 4-mediated PDT clinical trial for glioma patients.

Animals↗

PubMed2005

Diverse optical characteristic of the prostate and light delivery system: implications for computer modelling of prostatic photodynamic therapy.

OBJECTIVE: To explore the use of photodynamic therapy (PDT) as a minimally invasive form of treatment for organ-confined prostate cancer, for although there are several therapies, ablative treatments are associated with significant morbidity. MATERIALS AND METHODS: Using the photosensitizer tin etiopurpurin, dogs were treated with interstitially placed laser fibres in an effort to validate PDT for treating prostate cancer. Earlier models assumed a uniform distribution of light output from a cylindrical fibre and a uniform attenuation coefficient throughout the prostate. Subsequent observations show that this model was too simple and that light radiance is not linear. To overcome under-treatment, a computer program to complement real-time fibre placement was developed. RESULTS: As light radiance from interstitially placed laser fibres varies significantly from the commonly assumed ideal cylindrical emission, a predictive mathematical model of prostate PDT needs to consider the real emission. Also, the optical properties of the prostate, e.g. absorption and scattering of light, are anisotropic. Differences in the attenuation coefficient (combining absorption and scattering of light) also varied among different animals. Incorporating all these variables into a computer program produced a virtual model of the photo-ablated zone within +/- 2 mm of that observed in animals. CONCLUSION: PDT of the prostate is not trivial and should benefit from computer-aided methods as it is developed for clinical use.

Animals↗

PubMed2005

Partial least squares based decomposition of five spectrally overlapping factors.

The development of a multi-sensory fiber-optic based fluence rate probe (MSP) for light monitoring and dosimetry during photodynamic therapy (PDT) created the need for a robust multivariate signal analysis algorithm capable of quantifying the intensity of five component spectra, representing the sensors, which display a large degree of spectral overlap. Partial least squares (PLS) analysis, as an option for such an analysis algorithm, was evaluated through simulations in the presence of three types of noise, which experimentally may limit the accuracy of PLS quantification of component spectra contributions. Random, or white noise, background was varied over a range of 0-15% relative intensity. A non-random (Gaussian) background vector was simulated, as an experimentally relevant spectral interferent, over a range of 0-7% relative intensity. Spectral variation was modeled by a spectral shift of +/-1-5 nm. Procedures for selecting preferred combinations of fluorophores, with minimum possible spectral overlap, were developed. To illustrate the importance of this selection process, PLS performance results were compared for two possible combinations of five fluorophores, as well as for the combination of three fluorophores currently in experimental use with MSPs. Experimentally anticipated worst-case quantifications were identified for all three types of simulated noise (5% random background, 0.5% Gaussian background, and +/-2 nm spectral shift). The effects of single and combined types of noise were evaluated by independently varying each type of simulated noise over the experimentally relevant range. The mean percentage error in determining the fluorophore contributions and hence quantification of the fluence rate were compared with the required performance standard of better than 10% error for optical power meters in medical use. The PLS algorithm provided an accuracy of 7 +/- 2% for five fluorophores and 3.3 +/- 0.8% for three fluorophores, indicating that PLS is an appropriate choice for this application.

Algorithms↗

PubMed2004

Microfabricated system for parallel single-cell capillary electrophoresis.

Performing single-cell electrophoresis separations using multiple parallel microchannels offers the possibility of both increasing throughput and eliminating cross-contamination between different separations. The instrumentation for such a system requires spatial and temporal control of both single-cell selection and lysis. To address these problems, a compact platform is presented for single-cell capillary electrophoresis in parallel microchannels that combines optical tweezers for cell selection and electromechanical lysis. Calcein-labeled acute myloid leukemia (AML) cells were selected from an on-chip reservoir and transported by optical tweezers to one of four parallel microfluidic channels. Each channel entrance was manufactured by F2-laser ablation to form a 20- to 10-microm tapered lysis reservoir, creating an injector geometry effective in confining the cellular contents during mechanical shearing of the cell at the 10-microm capillary entrance. The contents of individual cells were simultaneously injected into parallel channels resulting in electrophoretic separation as recorded by laser-induced fluorescence of the labeled cellular contents.

Comet Assay↗

PubMed2004

Transperineal in vivo fluence-rate dosimetry in the canine prostate during SnET2-mediated PDT.

Advances in photodynamic therapy (PDT) treatment for prostate cancer can be achieved either by improving selectivity of the photosensitizer towards prostate gland tissue or improving the dosimetry by means of individualized treatment planning using currently available photosensitizers. The latter approach requires the ability to measure, among other parameters, the fluence rate at different positions within the prostate and the ability to derive the tissue optical properties. Here fibre optic probes are presented capable of measuring the fluence rate throughout large tissue volumes and a method to derive the tissue optical properties for different volumes of the prostate. The responsivity of the sensors is sufficient to detect a fluence rate of 0.1 mW cm(-2). The effective attenuation coefficient in the canine prostate at 660 nm is higher at the capsule (2.15+/-0.19 cm(-1)) than in proximity of the urethra (1.84+/-0.36 cm(-1)). Significant spatial and temporal intra- and inter-canine variability in the tissue optical properties was noted, highlighting the need for individualized monitoring of the fluence rate for improved dosimetry.

Algorithms↗

PubMed2004

Performance evaluation of cylindrical fiber optic light diffusers for biomedical applications.

BACKGROUND AND OBJECTIVES: Dosimetry for intracavity and interstitial light delivery requires next to the knowledge of tissue optical properties and models describing light propagation in tissue also exact knowledge of the spatial light source emission characteristics. However, the emission characteristics of cylindrical diffusers are often ill defined by the manufacturer, and not regularly determined by the end user, thus limiting the attainable dosimetry accuracy. STUDY DESIGN/MATERIALS AND METHODS: Commercial cylindrical diffusers, with active diffusive lengths of 1-2 cm and outer diameters of </=1 mm, were evaluated regarding their photometric emission characteristics. In addition to traditional tests of homogeneity such as light intensity emitted along the diffuser length and around its circumference (polar angle), azimuth emission was also measured. RESULTS: It was demonstrated that the light emission distribution is specific to the diffuser manufacturer, and while good polar isotropy is often attained, azimuth isotropy can vary by more than a factor of 5. In hollow organs the latter can result in over treatment of tissue distal to the actual placement of the cylindrical diffusers. CONCLUSIONS: To enable light dosimetry and treatment planning for applications such as photodynamic therapy or interstitial laser hyperthermia or photocoagulation, standardization of emission properties reporting for optical fiber diffusers is required.

Equipment Design↗

PubMed2004

Optical characteristics of the canine prostate at 665 nm sensitized with tin etiopurpurin dichloride: need for real-time monitoring of photodynamic therapy.

PURPOSE: Photodynamic therapy (PDT) is an emerging, minimally invasive therapy for prostate cancer that depends on the sequestration of a photosensitizing drug within targeted tissue. The photosensitizer is subsequently activated by light of a specific wavelength, resulting in destruction of the targeted tissue. Successful treatment requires knowledge of the optical properties of the target tissue, a critical element for therapy. MATERIALS AND METHODS: Adult canines were injected with tin etiopurpurin dichloride (1.0 mg/kg) as a liposome emulsion vehicle in saline 24 hours prior to light treatment. Laser light was delivered to the prostate via a 400 microm optical fiber fitted with a 2.0 cm cylindrical diffuser and optical properties of the prostate were measured. RESULTS: In this study we determined the attenuation coefficient and critical fluence in the canine prostate. Our studies shown that the attenuation coefficient is not uniform but higher at the base (average for all animals 2.59 to 2.79 cm-1) than in the mid section or apex of the prostate (1.71 to 1.90 cm-1). Significant differences among dogs (0.11 to 12.70 cm-1) were found. In some cases we observed a fluctuation of the attenuation coefficient during treatment. We also established experimentally the minimum energy (1449 mJ/cm2) needed (critical fluence) to produce necrosis. Experimentally establishing the values of effective attenuation and critical fluence is necessary to predict the area of ablation during PDT and protect surrounding organs from over treatment. CONCLUSIONS: Based on our results it is evident that for PDT of the prostate to be successful the optical parameters of the prostate must be measured and monitored during treatment. We suggest that the optimum way of doing this is real-time computerized monitoring combined with simulation PDT.

Animals↗

PubMed2004

Classification of breast tissue density by optical transillumination spectroscopy: optical and physiological effects governing predictive value.

Preventive oncology is in need of a risk assessment technique that can identify individuals at high risk for breast cancer and has the ability to monitor the efficacy of a risk reducing intervention. Optical transillumination spectroscopy (OTS) gives information about breast tissue composition and tissue density. OTS is noninvasive and in contrast to mammography, uses nonionizing radiation. It is safe and can be used frequently on younger women, potentially permitting early risk detection and thus increasing the time available for risk reduction interventions to assert their influence. Before OTS can be used as a risk assessment and/or monitoring technique, its predictive ability needs to be demonstrated and maximized through the construction of various mathematical models relating OTS and breast tissue density, and hence, risk. To establish a correlation between OTS and mammographic density principal components analysis (PCA), using risk classification, is calculated. The PCA scores are presented in three-dimensional cluster plots and a plane of differentiation that separates the high and low tissue densities is used to calculate the predictive value. Stratification of PCA for measurement position on the breast in cranial-caudal projection is introduced. Analysis of PCA scores as a function of the volunteer's age and body mass index (BMI) is examined. A small but significant correlation between the component scores and age or BMI is noted but the correlation is dependent on the tissue density category examined. Correction of the component scores for age and BMI is not recommended, since a priori knowledge of a woman's breast tissue density is required. Stratification for the center and distal measurement positions provide a predictive value for OTS above 96%.

Adult↗

PubMed2004

Photodynamic actinometry using microspheres: concept, development and responsivity.

Photodynamic therapy (PDT) relies on three main ingredients, oxygen, light and photoactivating compounds, although the PDT response is definitively contingent on the site and level of reactive oxygen species (ROS) generation. This study describes the development of a novel, fluorescent-based actinometer microsphere system as a means of discerning spatially resolved dosimetry of total fluence and ROS production. Providing a high resolution, localized, in situ measurement of fluence and ROS generation is critical for developing in vivo PDT protocols. Alginate-poly-L-lysine-alginate microspheres were produced using ionotropic gelation of sodium alginate droplets, ranging from 80 to 200 microm in diameter, incorporating two dyes, ADS680WS (ADS) and Rhodophyta-phycoerythrin (RPE), attached to the spheres' inside and outside layers, respectively. To test the responsivity and dynamic range of RPE for ROS detection, the production of ROS was initiated either chemically using increasing concentrations of potassium perchromate or photochemically using aluminum tetrasulphonated phthalocyanine. The generation of singlet oxygen was confirmed by phosphorescence at 1270 nm. The resulting photodegradation and decrease in fluorescence of RPE was found to correlate with increased perchromate or PDT treatment fluence, respectively. This effect was independent of pH (6.5-8) and could be inhibited using sodium azide. RPE was not susceptible to photobleaching with light alone (670 nm; 150 Jcm(-2)). ADS, which absorbs light between 600 and 750 nm, showed a direct correlation between radiant exposure (670 nm; 0-100 Jcm(-2)) and diminished fluorescence. Photobleaching was independent of irradiance (10-40 mW cm(-2)). We propose that actinometer microspheres may provide a means for obtaining high spatial resolution information regarding delivered PDT dose within model systems during investigational PDT development and dosimetric information for clinical extracorporeal PDT as in the case of ex vivo bone marrow purging.

Bone Marrow Purging↗

PubMed2003

Effects of low-level laser therapy (LLLT) of 810 nm upon in vitro growth of bacteria: relevance of irradiance and radiant exposure.

OBJECTIVE: The aim of this study was to investigate the irradiance-dependency of low-level laser therapy (LLLT) effects on bacterial growth. BACKGROUND: LLLT is applied to open wounds to improve healing; however, its effect on wound bacteria is not well understood. MATERIALS AND METHODS: Escherichia coli, Pseudomonas aeruginosa, and Staphylococcus aureus were irradiated using a wavelength of 810 nm at irradiances of 0.015 W/cm2 (0-50 J/cm2) and 0.03 W/cm2 (0-80 J/cm2). Bacteria were counted after 20 h of incubation. RESULTS: LLLT effects varied significantly with species. P.aeruginosa growth decreased overall dependent on an interaction of irradiance and radiant exposure; greatest inhibition was produced using high irradiance delivering radiant exposures in the range of 1-20 J/cm2 (p = 0.001-0.04). In contrast, E. coli growth increased overall (p = 0.01), regardless of irradiance; greatest effects were produced using low radiant exposures (1-20 J/cm2). There was a main effect for irradiance (p = 0.03) on S. aureus growth; however, growth was not different compared with controls. Additional analysis showed that there were differences in growth of P.aeruginosa when comparing samples that were matched by exposure times (66, 329, 658, 1316, 1974, and 2632 sec) rather than radiant exposure; this suggests that irradiance rather than exposure time was the significant factor in P. aeruginosa inhibition. CONCLUSION: These findings have immediate relevancy in the use of LLLT for infected wounds. Exposure to 810-nm irradiation (0.03 W/cm2) could potentially benefit wounds infected with P. aeruginosa. However, increased E. coli growth could further delay recovery.

Bacteria↗

PubMed2003

Spatial distribution of liposome encapsulated tin etiopurpurin dichloride (SnET2) in the canine prostate: implications for computer simulation of photodynamic therapy.

Photodynamic therapy (PDT) is a minimally invasive treatment that can be employed in many human diseases including prostate cancer. PDT for prostate cancer depends on the sequestration of a photosensitizing drug within the glandular tissue. The photosensitizer is subsequently activated by light (usually from a laser) and the active drug destroys tissue. Since prostate cancer is a multifocal disease, PDT must ablate the glandular prostate completely. This will depend on the precise placement of light sources in the prostate and delivery of a therapeutic light dose to the entire gland. Also, sources of light and their spatial distribution must be tailored to each individual patient. The uniform, therapeutic light distribution can be achieved by interstitial light irradiation. In this case, the light is delivered by diffusers placed within the substance of the prostate parallel to the urethra at a distance optimized to deliver adequate levels of light and to create the desired photodynamic effect. To help achieve the uniform light distribution throughout the prostate we have developed a computer program that can determine treatment effects. The program predicts the best set of parameters and the position of light diffusers in space, and displays them in graphical or in numerical form assuming a fixed attenuation coefficient. The two parameters of greatest importance in the computer simulation are attenuation coefficient and critical fluence. Both depend on the concentration of active drug within the prostate gland. It is necessary to know the nature of the spatial distribution of photosensitizer within the prostate to execute computer modeling of PDT with high precision. We found that the concentration of SnET2 is heterogeneous in nature, and is higher in the proximity of the glandular capsule. It is clear therefore that any future attempts of computerized modeling of this procedure must take into consideration the uneven sequestration of photosensitizer and the consequential asymmetrical necrosis of the prostate.

Algorithms↗