Post-Operational Photodynamic Therapy of the Tumor Bed: Comparative Analysis for Cold Knife and Laser Scalpel Resection.

In this paper, we report on a study regarding the efficiency of the post-operational phototherapy of the tumor bed after resection with both a cold knife and a laser scalpel in laboratory mice with CT-26 tumors. Post-operational processing included photodynamic therapy (PDT) with a topically applied chlorin-based photosensitizer (PS), performed at wavelengths of 405 or 660 nm, with a total dose of 150 J/cm2. The selected design of the tumor model yielded zero recurrence in the laser scalpel group and 92% recurrence in the cold knife group without post-processing, confirming the efficiency of the laser scalpel in oncology against the cold knife. The application of PDT after the cold knife resection decreased the recurrence rate to 70% and 42% for the 405 nm and 660 nm procedures, respectively. On the other hand, the application of PDT after the laser scalpel resection induced recurrence rates of 18% and 30%, respectively, for the considered PDT performance wavelengths. The control of the penetration of PS into the tumor bed by fluorescence confocal microscopy indicated the deeper penetration of PS in the case of the cold knife, which presumably provided deeper PDT action, while the low-dose light exposure of deeper tissues without PS, presumably, stimulated tumor recurrence, which was also confirmed by the differences in the recurrence rate in the 405 and 660 nm groups. Irradiation-only light exposures, in all cases, demonstrated higher recurrence rates compared to the corresponding PDT cases. Thus, the PDT processing of the tumor bed after resection could only be recommended for the cold knife treatment and not for the laser scalpel resection, where it could induce tumor recurrence.

Monitoring of optical properties of tumors during laser plasmon photothermal therapy.

We studied grafted tumors obtained by subcutaneous implantation of kidney cancer cells into male white rats. Gold nanorods with a plasmon resonance of about 800 nm were injected intratumorally for photothermal heating. Experimental irradiation of tumors was carried out percutaneously using a near-infrared diode laser. Changes in the optical properties of the studied tissues in the spectral range 350-2200 nm under plasmonic photothermal therapy (PPT) were studied. Analysis of the observed changes in the absorption bands of water and hemoglobin made it possible to estimate the depth of thermal damage to the tumor. A significant decrease in absorption peaks was observed in the spectrum of the upper peripheral part and especially the tumor capsule. The obtained changes in the optical properties of tissues under laser irradiation can be used to optimize laboratory and clinical PPT procedures.

Digital diaphanoscopy of maxillary sinus pathologies supported by machine learning.

Maxillary sinus pathologies remain among the most common ENT diseases requiring timely diagnosis for successful treatment. Standard ENT inspection approaches indicate low sensitivity in detecting maxillary sinus pathologies. In this paper, we report on capabilities of digital diaphanoscopy combined with machine learning tools in the detection of such pathologies. We provide a comparative analysis of two machine learning approaches applied to digital diapahnoscopy data, namely, convolutional neural networks and linear discriminant analysis. The sensitivity and specificity values obtained for both employed approaches exceed the reported accuracy indicators for traditional screening diagnosis methods (such as nasal endoscopy or ultrasound), suggesting the prospects of their usage for screening maxillary sinuses alterations. The analysis of the obtained values showed that the linear discriminant analysis, being a simpler approach as compared to neural networks, allows one to detect the maxillary sinus pathologies with the sensitivity and specificity of 0.88 and 0.98, respectively.

VIS-NIR Diffuse Reflectance Spectroscopy System with Self-Calibrating Fiber-Optic Probe: Study of Perturbation Resistance.

We report on the comparative analysis of self-calibrating and single-slope diffuse reflectance spectroscopy in resistance to different measurement perturbations. We developed an experimental setup for diffuse reflectance spectroscopy (DRS) in a wide VIS-NIR range with a fiber-optic probe equipped with two source and two detection fibers capable of providing measurements employing both single- and dual-slope (self-calibrating) approaches. In order to fit the dynamic range of a spectrometer in the wavelength range of 460-1030 nm, different exposure times have been applied for short (2 mm) and long (4 mm) source-detector distances. The stability of the self-calibrating and traditional single-slope approaches to instrumental perturbations were compared in phantom and in vivo studies on human palm, including attenuations in individual channels, fiber curving, and introducing optical inhomogeneities in the probe-tissue interface. The self-calibrating approach demonstrated high resistance to instrumental perturbations introduced in the source and detection channels, while the single-slope approach showed resistance only to perturbations introduced into the source channels.

Multimodal Optical Monitoring of Auto- and Allografts of Skin on a Burn Wound.

The aim of the study was to investigate the dynamics of the state of allo- and autografts of skin on a wound using optical modalities: diffuse reflectance spectroscopy (DRS), optical coherence tomography (OCT), and laser Doppler flowmetry (LDF). A deep thermal burn was simulated in 24 rats covering 20% of the body surface. On day 3 after the injury, a fascial necrectomy of two 500 mm2 areas on the left and right sides of the midline of the animal body were excised. Allografts and autografts were placed in the centers of these areas. Optical measurements of grafts were performed on the 0, 3rd, 6th, 10th, and 13th days after transplantation. The allografts demonstrated a pronounced decrease in oxygenation, blood content, and perfusion compared to autografts on the 6th day; in the following days of observation, these values returned to the average values of autografts. Water content gradually decreased from the beginning to the end of observation. In conclusion, optical diagnostics revealed changes in the morphological microstructure, the rate of restoration of blood circulation, and oxygen exchange in the early stages, specific for the allo- and autograft.

Autoquenching of spherical photon density waves during propagation in a turbid medium.

The effect of the formation of deep minima in frequency characteristics of photon density waves (PDWs) during their propagation in scattering media with different optical characteristics has been studied by statistical Monte Carlo modeling. The simulation was performed for the Henyey-Greenstein scattering phase function with the anisotropy factor value varying in the range of 0-0.93. The dependence of the position and magnitude of the minimum in the frequency response of PDWs on the combination of the parameters of the scattering medium and the distance to the radiation source is demonstrated.

Angular distribution of photon density waves radiance in media with different scattering anisotropy.

Statistical modeling of pulsed frequency responses of the light field radiance by an isotropic point source was performed by Monte Carlo technique. Scattering properties of the medium were simulated by the Henyey-Greenstein phase function with different anisotropy factor values. Angular distributions of the pulsed field and amplitudes of the photon density waves in a certain range of parameters were shown to have a qualitatively different character for media with quasi-isotropic and strongly anisotropic scattering. A comparison of the impulse and frequency characteristics was performed for media with strongly anisotropic scattering with different scattering phase functions yet the same anisotropy factor. The main difference in the angular distributions of the fields is observed in the rear hemisphere.

Dual-Wavelength Fluorescence Monitoring of Photodynamic Therapy: From Analytical Models to Clinical Studies.

Fluorescence imaging modalities are currently a routine tool for the assessment of marker distribution within biological tissues, including monitoring of fluorescent photosensitizers (PSs) in photodynamic therapy (PDT). Conventional fluorescence imaging techniques provide en-face two-dimensional images, while depth-resolved techniques require complicated tomographic modalities. In this paper, we report on a cost-effective approach for the estimation of fluorophore localization depth based on dual-wavelength probing. Owing to significant difference in optical properties of superficial biotissues for red and blue ranges of optical spectra, simultaneous detection of fluorescence excited at different wavelengths provides complementary information from different measurement volumes. Here, we report analytical and numerical models of the dual-wavelength fluorescence imaging of PS-containing biotissues considering topical and intravenous PS administration, and demonstrate the feasibility of this approach for evaluation of the PS localization depth based on the fluorescence signal ratio. The results of analytical and numerical simulations, as well as phantom experiments, were translated to the in vivo imaging to interpret experimental observations in animal experiments, human volunteers, and clinical studies. The proposed approach allowed us to estimate typical accumulation depths of PS localization which are consistent with the morphologically expected values for both topical PS administration and intravenous injection.

Optoacoustic Sensing of Surfactant Crude Oil in Thermal Relaxation and Nonlinear Regimes.

We propose a laser optoacoustic method for the complex characterization of crude oil pollution of the water surface by the thickness of the layer, the speed of sound, the coefficient of optical absorption, and the temperature dependence of the Grüneisen parameter. Using a 532 nm pulsed laser and a 1-100 MHz ultra-wideband ultrasonic antenna, we have demonstrated the capability of accurate (>95%) optoacoustic thickness measurements in the 5 to 500-micron range, covering 88% of slicks observed during 2010 oil spill in the Gulf of Mexico. In the thermal relaxation regime of optoacoustic measurements, the value of optical absorption coefficient (30 mm-1) agreed with the data of independent spectrophotometric measurements, while the sound speed (1430 m/s) agreed with the tabular data. When operating in a nonlinear regime, the effect of local deformation of the surface of the oil film induced by heating laser radiation was revealed. The dose-time parameters of laser radiation ensuring the transition from the thermal relaxation regime of optoacoustic generation to nonlinear one were experimentally investigated. The developed OA method has potential for quantitative characterization of not only the volume, but also the degree and even the type of oil pollution of the water surface.

Red and blue light in antitumor photodynamic therapy with chlorin-based photosensitizers: a comparative animal study assisted by optical imaging modalities.

The goal of this study is a comparative analysis of the efficiency of the PDT protocols for CT26 tumor model treatment in Balb/c mice employing red and blue light with both topical and intravenous administration of chlorin-based photosensitizers (PSs). The considered protocols include the doses of 250 J/cm2 delivered at 660 nm, 200 J/cm2 delivered at 405 nm, and 250 J/cm2 delivered at both wavelengths with equal energy density contribution. Dual-wavelength fluorescence imaging was employed to estimate both photobleaching efficiency, typical photobleaching rates and the procedure impact depth, while optical coherence tomography with angiography modality (OCT-A) was employed to monitor the tumor vasculature response for up to 7 days after the procedure with subsequent histology inspection. Red light or dual-wavelength PDT regimes with intravenous PS injection were demonstrated to provide the most pronounced tumor response among all the considered cases. On the contrary, blue light regimes were demonstrated to be most efficient among topical application and irradiation only regimes. Tumor size dynamics for different groups is in good agreement with the tumor response predictions based on OCT-A taken in 24h after exposure and the results of histology analysis performed in 7 days after the exposure.

Combined Fluorescence and Optoacoustic Imaging for Monitoring Treatments against CT26 Tumors with Photoactivatable Liposomes.

The newly developed multimodal imaging system combining raster-scan optoacoustic (OA) microscopy and fluorescence (FL) wide-field imaging was used for characterizing the tumor vascular structure with 38/50 µm axial/transverse resolution and assessment of photosensitizer fluorescence kinetics during treatment with novel theranostic agents. A multifunctional photoactivatable multi-inhibitor liposomal (PMILs) nano platform was engineered here, containing a clinically approved photosensitizer, Benzoporphyrin derivative (BPD) in the bilayer, and topoisomerase I inhibitor, Irinotecan (IRI) in its inner core, for a synergetic therapeutic impact. The optimized PMIL was anionic, with the hydrodynamic diameter of 131.6 ± 2.1 nm and polydispersity index (PDI) of 0.05 ± 0.01, and the zeta potential between -14.9 ± 1.04 to -16.9 ± 0.92 mV. In the in vivo studies on BALB/c mice with CT26 tumors were performed to evaluate PMILs' therapeutic efficacy. PMILs demonstrated the best inhibitory effect of 97% on tumor growth compared to the treatment with BPD-PC containing liposomes (PALs), 81%, or IRI containing liposomes (L-[IRI]) alone, 50%. This confirms the release of IRI within the tumor cells upon PMILs triggering by NIR light, which is additionally illustrated by FL monitoring demonstrating enhancement of drug accumulation in tumor initiated by PDT in 24 h after the treatment. OA monitoring revealed the largest alterations of the tumor vascular structure in the PMILs treated mice as compared to BPD-PC or IRI treated mice. The results were further corroborated with histological data that also showed a 5-fold higher percentage of hemorrhages in PMIL treated mice compared to the control groups. Overall, these results suggest that multifunctional PMILs simultaneously delivering PDT and chemotherapy agents along with OA and FL multi-modal imaging offers an efficient and personalized image-guided platform to improve cancer treatment outcomes.

Effect of scattering anisotropy on the properties of photon density waves.

The frequency characteristics of spherical photon density waves excited in media with different degrees of scattering anisotropy are studied. Statistical modeling of the frequency and phase responses of the spatial irradiance of the light field emitted by a point-sized isotropic source were performed employing the Monte Carlo technique. The scattering anisotropy of the medium was determined by the Henyey-Greenstein phase function with different values of the mean scattering cosine. It is shown that the scattering anisotropy factor determines the frequency range, in which the effect of the photon path length distribution on the magnitude of the photon density wave dispersion is maximal. In media with quasi-isotropic scattering, dispersion effects are manifested at lower frequencies as compared to those for anisotropic media. The simulation results are compared with the analytical solution for the asymptotic regime of the light field in an isotropically scattering medium.

Nanoparticles Produced via Laser Ablation of Porous Silicon and Silicon Nanowires for Optical Bioimaging.

Modern trends in optical bioimaging require novel nanoproducts combining high image contrast with efficient treatment capabilities. Silicon nanoparticles are a wide class of nanoobjects with tunable optical properties, which has potential as contrasting agents for fluorescence imaging and optical coherence tomography. In this paper we report on developing a novel technique for fabricating silicon nanoparticles by means of picosecond laser ablation of porous silicon films and silicon nanowire arrays in water and ethanol. Structural and optical properties of these particles were studied using scanning electron and atomic force microscopy, Raman scattering, spectrophotometry, fluorescence, and optical coherence tomography measurements. The essential features of the fabricated silicon nanoparticles are sizes smaller than 100 nm and crystalline phase presence. Effective fluorescence and light scattering of the laser-ablated silicon nanoparticles in the visible and near infrared ranges opens new prospects of their employment as contrasting agents in biophotonics, which was confirmed by pilot experiments on optical imaging.

Nonstationary angular distribution of optical field radiance from an isotropic source in sea water.

The spatial-angular and temporal characteristics of the radiance of the light field emitted by a nonstationary point isotropic source in sea water are studied. Using the Monte Carlo method, we calculated the pulse transfer functions and frequency responses of the angular radiance distributions at various distances from the source. Particular integral characteristics of the angular radiance distributions are estimated. It is shown that with an increase in the delay time, measured from the time of arrival of ballistic photons, the angular radiance distribution asymptotically tends to be isotropic. The frequency and phase responses of the alternating radiance component from a source modulated by power at a high frequency are studied. It is shown that with an increase in the modulation frequency, the angular distribution of the alternating radiance component is concentrated close to the direction to the source.

Toward whole-brain in vivo optoacoustic angiography of rodents: modeling and experimental observations.

Cerebrovascular imaging of rodents is one of the trending applications of optoacoustics aimed at studying brain activity and pathology. Imaging of deep brain structures is often hindered by sub-optimal arrangement of the light delivery and acoustic detection systems. In our work we revisit the physics behind opto-acoustic signal generation for theoretical evaluation of optimal laser wavelengths to perform cerebrovascular optoacoustic angiography of rodents beyond the penetration barriers imposed by light diffusion in highly scattering and absorbing brain tissues. A comprehensive model based on diffusion approximation was developed to simulate optoacoustic signal generation using optical and acoustic parameters closely mimicking a typical murine brain. The model revealed three characteristic wavelength ranges in the visible and near-infrared spectra optimally suited for imaging cerebral vasculature of different size and depth. The theoretical conclusions are confirmed by numerical simulations while in vivo imaging experiments further validated the ability to accurately resolve brain vasculature at depths ranging between 0.7 and 7 mm.

Comparative analysis of single- and dual-wavelength photodynamic therapy regimes with chlorin-based photosensitizers: animal study.

Two pronounced absorption peaks in blue and red ranges of the chlorin-based photosensitizer (PS) absorption spectrum provide additional benefits in photodynamic therapy (PDT) performance. Differing optical properties of biological tissues in these ranges allow for both dual-wavelength diagnostics and PDT performance. We provide a comparative analysis of different PDT regimes performed with blue and red lights and their combination, with doses varying from 50 to 150 J / cm2. The study was performed on the intact skin of a rabbit ear inner surface, with the use of chlorin e6 as a PS. PDT procedure protocol included monitoring of the treated site with fluorescence imaging technique to evaluate PS accumulation and photobleaching, as well as with optical coherence tomography (OCT) to register morphological and functional responses of the tissue. Optical diagnostic observations were compared with the results of histopathology examination. We demonstrated that PDT procedures with the considered regimes induce weaker organism reaction manifested by edema in normal tissue as compared to irradiation-only exposures with the same light doses. The light doses delivered with red light induce weaker tissue reaction as compared to the same doses delivered with blue light only or with a combination of red and blue lights in equal parts. Results of in-vivo OCT monitoring of tissue reaction are in agreement with the results of histopathology study.

Time delay and width variation caused by temporal dispersion of a complex modulated signal in underwater lidar.

The parameters of an echo signal from the underwater lidar are studied for the case of modulation of a probing pulse by a high frequency signal with a frequency linearly varying with time. The analysis is based on the statistical Monte Carlo simulations of the frequency and phase responses of a signal propagating along the emitter-water-reflector-water-receiver path and an analytical representation of the signal as a pulse described by a Gaussian function with intrapulse modulation. Delays and pulse shape changes caused by temporal dispersion of the photon-density waves are estimated. It is shown that the temporal dispersion effect reduces the efficiency of the matched processing of a complex signal in the receiving system.

Hyperspectral imaging of human skin aided by artificial neural networks.

We developed a compact, hand-held hyperspectral imaging system for 2D neural network-based visualization of skin chromophores and blood oxygenation. State-of-the-art micro-optic multichannel matrix sensor combined with the tunable Fabry-Perot micro interferometer enables a portable diagnostic device sensitive to the changes of the oxygen saturation as well as the variations of blood volume fraction of human skin. Generalized object-oriented Monte Carlo model is used extensively for the training of an artificial neural network utilized for the hyperspectral image processing. In addition, the results are verified and validated via actual experiments with tissue phantoms and human skin in vivo. The proposed approach enables a tool combining both the speed of an artificial neural network processing and the accuracy and flexibility of advanced Monte Carlo modeling. Finally, the results of the feasibility studies and the experimental tests on biotissue phantoms and healthy volunteers are presented.

Special Section Guest Editorial: Topical Problems of Biophotonics: from Optical Bioimaging to Clinical Biophotonics.

This editorial provides an introductory overview for the Special Section on Topical Problems of Biophotonics.