Search

Background and Objective: The goal of this paper is to perform a comparative study of the capabilities of optical coherence tomography (OCT) and fluorescence spectroscopy for detection of neoplastic processes and malignant tumor borders in the uterine cervix and vulva, and to preliminarily analyze the advisability of using these two methods complementarily.Study Design/Materials and Methods:Using the two methods-fluorescence spectroscopy, which is based on the detection of biochemical changes in tissue, and optical coherence tomography that detects morphologic or structural tissue information-borders of neoplasia in the uterine cervix and vulva were determined. A parallel analysis of optical properties of tissue and histologic material taken from the center of tumor and the OCT-detected boundary was performed; the visual and optically detected borders of the pathological zone were compared. A total of 12 women with neoplasia in the uterine cervix and vulva were enrolled in this study.Results: First results of this study show that an optically detected demarcation line of cervical neoplasia is farther from the tumor focus by an average of 2 mm than a colposcopically determined border. It was verified that the OCT-detected borders morphologically coincided with the real histologic borders. The OCT and fluorescence spectroscopy yielded the same results in only some experiments. In several clinical cases, it was difficult to interpret OCT images (combination with scar changes in the uterine cervix) and fluorescence spectroscopy data (combination with inflammatory changes).Conclusions: Experimental data presented here show the capability of OCT imaging and fluorescence spectroscopy to detect borders of neoplastic processes in the uterine cervix and vulva more accurately than conventional colposcopy. The results of this study provide a rationale for using these optical techniques in combination as complementary objective methods of cancer diagnostics. [ABSTRACT FROM AUTHOR]

Background: Therapeutic effects of PDT depend on many factors, including the amount of singlet oxygen, localization of photosensitizer and irradiation protocol. The present study was aimed to compare the cytotoxic mechanisms of PDT under continuous-wave (CW) and pulsed irradiation using a tumor spheroid model and a genetically encoded photosensitizer miniSOG., Methods: 1 O 2 detection in miniSOG and flavin mononucleotide (FMN) solutions was performed. Photobleaching of miniSOG in solution and in HeLa tumor spheroids was analyzed. Tumor spheroid morphology and growth and the cell death mechanisms after PDT in CW and pulsed modes were assessed., Results: We found a more rapid 1 O 2 generation and a higher photobleaching rate in miniSOG solution upon irradiation in pulsed mode compared to CW mode. Photobleaching of miniSOG in tumor spheroids was also higher after irradiation in the pulsed mode. PDT of spheroids in CW mode resulted in a moderate expansion of the necrotic core of tumor spheroids and a slight inhibition of spheroid growth. The pulsed mode was more effective in induction of cell death, including apoptosis, and suppression of spheroid growth., Conclusions: Comparison of CW and pulsed irradiation modes in PDT with miniSOG showed more pronounced cytotoxic effects of the pulsed mode. Our results suggest that the pulsed irradiation regimen enables enhanced 1 O 2 production by photosensitizer and stimulates apoptosis., General Significance: Our results provide more insights into the cellular mechanisms of anti-cancer PDT and open the way to improvement of light irradiation protocols., (Copyright © 2021 Elsevier B.V. All rights reserved.)

The introduction of technologically advanced methods of lithotripsy into medical practice changes the nature of postoperative complications. Among them, the main complications are inflammatory infections. This largely determines the search for new, improved methods of stone fragmentation avoiding small stone fragments and dissemination of the pelvicalyceal system of the kidney with stone-associated infection. The authors have developed a method for controlled stone fragmentation using a continuous-wave diode laser with a hot-spot effect at the optical fiber end. The aim of the study was to evaluate the efficacy of controlled urinary stone fragmentation using a continuous-wave diode laser with a highly heated distal end of the optical fiber light guide as a method of preventing inflammatory infections in clinical practice., Materials and Methods: We analyzed 1666 case histories of urolithiasis patients who underwent percutaneous nephrolithotripsy/ nephrolithoextraction and contact ureterolithotripsy/ureteroextraction, we also performed a prospective analysis of complications based on the Clavien-Dindo classification in 90 patients who underwent fine fragmentation of stones with various lithotripters: ultrasonic, pneumatic, and holmium laser. The method of controlled stone fragmentation by a diode laser with a hot-spot effect was tested on postoperative samples of 26 renal calculi. For the first time in clinical practice, this method was tested in the bladder cavity (n=10)., Results: In the percutaneous nephrolithotripsy group, postoperative infectious and inflammatory complications occurred in 34.1% of cases, in the percutaneous nephrolithoextraction group - in 24.6%, in the contact ureterolithotripsy group - in 7.8%, in the ureterolithoextraction group - in 2.5%. The analysis made it possible to identify factors promoting the development of infectious and inflammatory complications. For the first time in clinical practice, there were successfully performed ten operations of stone fragmentation using a continuous-wave diode laser with a hot-spot effect. Controlled coarse fragmentation of stones providing the possibility to reduce the number of infectious and inflammatory complications was performed in the bladder as a model for testing the method., Conclusion: The method of laser-induced controlled coarse fragmentation of stones with a hot-spot effect, developed and tested in clinical practice, is promising for the prevention of infectious and inflammatory complications in patients with potentially infected stones since their fine fragmentation and, consequently, spread of stone-associated toxins and microflora within the urinary system is avoided., Competing Interests: Conflicts of interest. The authors have no conflicts of interest to disclose.

Ensuring the complete removal of tumor tissue is the main challenge during resection operations. Recently, a technique of "indirect" contact laser surgery has been developed. In this study we assess the possibility of using such surgery for fluorescence image-guided tumor resection. Mouse colon adenocarcinoma CT-26 cells stably expressing the fluorescent protein mKate-2 was used as the tumor model. Resections of the tumor nodes were performed with either a scalpel blade, a laser scalpel with a bare tip, or a laser scalpel with a strongly absorbing coating on the fiber tip. Tumor-positive resection margins were detected using an IVIS Spectrum fluorescence imaging system. After tumor resection with the scalpel blade over half of the animals needed one additional resection to remove residual tumor cells. Animals in this group showed tumor recurrence within 7 days. Fluorescence imaging of the tumor bed, performed after resection to assess the presence of tumor cell clusters, was sufficiently effective only with a bloodless resection. The laser scalpels both with the bare tip and with the strongly absorbing coating on the tip provided such bloodless tumor resection in contact mode. Fewer animals required additional resections when the bare tipped scalpel was used and this also resulted in a reduction in tumor recurrence. After resections were carried out with the laser scalpel with the strongly absorbing coating on the tip, fluorescence was detected in the operative field and this led to undertaking additional resections, although subsequent investigation suggested that this was "false" fluorescence, resulting from the effects of the scalpel rather than the presence of residual tumor cells. The method of laser resection with a strongly absorbing coating on the tip therefore did not appear to demonstrate definite advantages over laser resection with a bare tip when removing tumors.

Background/purpose: The goal of the study is comparative analysis of the layers in OCT images and the morphological structure of skin with thick and thin epidermis., Methods: We analyzed the difference between skin with thin and thick epidermis in two ways. The first approach consisted in determination of the thicknesses of layers of skin with thin and thick epidermis of different localizations from experimental OCT images. The second approach was to develop numerical models fitting experimental OCT images based on Monte Carlo simulations revealing structure and optical parameters of layers of skin with thick and thin epidermis., Results: The correspondence between the OCT images of skin with thin and thick epidermis and the morphological structure was confirmed. OCT images of healthy skin comprise three layers in case of skin with thin epidermis and four layers in skin with thick epidermis. The OCT image of the zone of the transition from skin with thick to skin with thin epidermis features five layers., Conclusion: The revealed differences in the structure of horny and cellular layers of epidermis, as well as of papillary and reticular dermis in skin with thin and thick epidermis specify different optical properties of these layers in OCT images., (© 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

The purpose of this study was to evaluate photobleaching of the genetically encoded photosensitizer KillerRed in tumor spheroids upon pulsed and continuous wave (CW) laser irradiation and to analyze the mechanisms of cancer cell death after the treatment. We observed the light-dose dependent mechanism of KillerRed photobleaching over a wide range of fluence rates. Loss of fluorescence was limited to 80% at light doses of 150 J/cm(2) and more. Based on the bleaching curves, six PDT regimes were applied for irradiation using CW and pulsed regimes at a power density of 160 mW/cm(2) and light doses of 140 J/cm(2) , 170 J/cm(2) and 200 J/cm(2). Irradiation of KillerRed-expressing spheroids in the pulsed mode (pulse duration 15 ns, pulse repetition rate 10 Hz) induced predominantly apoptotic cell death, while in the case of CW mode the cancer cells underwent necrosis. In general, these results improve our understanding of photobleaching mechanisms in GFP-like proteins and show the importance of appropriate selection of treatment mode for PDT with KillerRed. Representative fluorescence image of two KillerRed-expressing spheroids before and immediately after CW irradiation., (© 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Background/purpose: The present study aimed to investigate the influence of topical corticosteroid therapy and tacrolimus on morphological indices of different skin phototypes and to optimize topical therapy using the OCT technique., Methods: Twenty healthy volunteers aging from 20 to 30 (14 men and 6 women) took part in the study: 10 persons with skin phototype I, II and 10 persons with skin phototype V, VI. Morphological state of the skin was assessed before and during application of topical steroids of different strength and calciumneurin inhibitors for 49 days. Morphological state was studied in vivo using the optical coherence tomograph., Results: Morphological manifestations of skin atrophy with the use of clobetasol propionate appear earlier than with the use of hydrocortisone 17-butyrate; this process was faster in representatives of groups V, VI. Epidermal thinning in the zone of tacrolimus application was not recorded in any phototype., Conclusion: Recording of early preclinical signs of epidermis thinning in the course of OCT follow-up may be an indication for changing the corticosteroid therapy by calciumneurin inhibitors, which will permit to individualize the therapy, to increase its efficacy, and to minimize the possibility of complications in each particular case., (© 2013 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

KillerRed is known to be a unique red fluorescent protein displaying strong phototoxic properties. Its effectiveness has been shown previously for killing bacterial and cancer cells in vitro. Here, we investigated the photototoxicity of the protein on tumor xenografts in mice. HeLa Kyoto cell line stably expressing KillerRed in mitochondria and in fusion with histone H2B was used. Irradiation of the tumors with 593 nm laser led to photobleaching of KillerRed indicating photosensitization reaction and caused significant destruction of the cells and activation of apoptosis. The portion of the dystrophically changed cells increased from 9.9% to 63.7%, and the cells with apoptosis hallmarks from 6.3% to 14%. The results of this study suggest KillerRed as a potential genetically encoded photosensitizer for photodynamic therapy of cancer., (Copyright © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Objective: The goal of the described study was complex assessment of age-specific morphofunctional features of skin of different anatomic localization using optical coherence tomography (OCT)., Methods: Forty-three healthy volunteers aging from 4 to 74 years were enrolled in the study. Optical and functional skin parameters were studied in 18 areas of the human body. All the studied areas were classified as open and closed, depending on external impact. Structural parameters of skin were determined by means of the OCT device equipped with a removable, flexible probe with microscanner (registration certificate no. 022a2035/2213-05) designed and fabricated at the IAP RAS (Nizhny Novgorod, Russia) with longitudinal and transverse resolution of about 20 μm., Results: Age-specific morphofunctional features of skin most evident in areas exposed to external action were found. Statistically significant difference in elasticity (-0.57, P = 0.00012) in areas closed to external action (further referred to as closed areas) and in useful signal depth, hydration and elasticity in areas exposed to external action (further referred to as open areas) was assessed for different age groups. Analysis of the dependence of parameters on age showed statistically significant correlations between age and thickness of cellular layers of the epidermis (-0.33, P = 0.035), pigmentation level (-0.22, P = 0.044) and elasticity in closed areas and highly significant correlations between age and hydration and elasticity in open areas., Conclusion: The current work is concerned with investigation of age-specific morpho-functional parameters of skin of different localizations., (© 2012 John Wiley & Sons A/S.)

The effects of non-ablative infrared (IR) laser treatment of collagenous tissue have been commonly interpreted in terms of collagen denaturation spread over the laser-heated tissue area. In this work, the existing model is refined to account for the recently reported laser-treated tissue heterogeneity and complex collagen degradation pattern using comprehensive optical imaging and calorimetry toolkits. Patella ligament (PL) provided a simple model of type I collagen tissue containing its full structural content from triple-helix molecules to gross architecture. PL ex vivo was subjected to IR laser treatments (laser spot, 1.6 mm) of equal dose, where the tissue temperature reached the collagen denaturation temperature of 60 ± 2°C at the laser spot epicenterin the first regime, and was limited to 67 ± 2°C in the second regime. The collagen network was analyzed versus distance from the epicenter. Experimental characterization of the collagenous tissue at all structural levels included cross-polarization optical coherence tomography, nonlinear optical microscopy, light microscopy/histology, and differential scanning calorimetry. Regressive rearrangement of the PL collagen network was found to spread well outside the laser spot epicenter (>2 mm) and was accompanied by multilevel hierarchical reorganization of collagen. Four zones of distinct optical and morphological properties were identified, all elliptical in shape, and elongated in the direction perpendicular to the PL long axis. Although the collagen transformation into a random-coil molecular structure was occasionally observed, it was mechanical integrity of the supramolecular structures that was primarily compromised. We found that the structural rearrangement of the collagen network related primarily to the heat-induced thermo-mechanical effects rather than molecular unfolding. The current body of evidence supports the notion that the supramolecular collagen structure suffered degradation of various degrees, which gave rise to the observed zonal character of the laser-treated lesion.

Modern optical diagnostic techniques often require deformations of the studied bio-tissues for image acquisition. This paper discusses the effect of mechanical compression on the formation of OCT images of human skin. The study was performed in vivo on human volunteers of different age. We show that application of compression to human skin induces changes in optical properties of the sample associated with elasticity of different skin layers. These changes induce an increase in the contrast of interlayer boundaries. Further application of compression causes the appearance of dark areas in the OCT images obtained, likely associated with interstitial or intracellular water inflow to the observed region. The effects studied are of importance for proper interpretation of obtained OCT images in diagnosis of skin pathologies., (Copyright © 2010 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

The capabilities of diffuse optical spectroscopy for noninvasive assessing of oxygen status in experimental tumors have been demonstrated. Specific features of the distribution of total hemoglobin, oxygenated hemoglobin, deoxygenated hemoglobin, and blood-oxygen saturation were shown on two tumor models having different histological structure and functional characteristics. The results obtained by the optical technique were verified by immunohistochemical study of tissue samples marked with exogenous marker of hypoxia--pimonidazole., (Copyright © 2010 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

The goal of this study is the development of a method of local laser hyperthermia with gold nanoparticles under noninvasive optical monitoring of nanoparticle accumulation in tumor tissue in vivo. Bifunctional plasmon resonant nanoparticles that are optimal for OCT diagnostics and laser heating at the wavelength of 810 nm were used in the study. The OCT examination showed that the accumulation of gold nanoparticles in the tumor invading into skin was maximal 4-5 h after intravenous injection. It was demonstrated that nanoparticle accumulation in tumor allowed more local heating and enhanced thermal sensitivity of tumor tissue. Laser hyperthermia that heated tumor up to 44-45 °C at maximum nanoparticle accumulation induced apoptotic death of tumor cells and inhibited tumor growth by 104% on the 5th day after treatment.

The possibility of using silica-gold nanoshells with 150 nm silica core size and 25 nm thick gold shell as contrasting agents for optical coherence tomography (OCT) is analyzed. Experiments on agar biotissue phantoms showed that the penetration of nanoshells into the phantoms increases the intensity of the optical coherence tomography (OCT) signal and the brightness of the corresponding areas of the OCT image. In vivo experiments on rabbit skin demonstrated that the application of nanoshells onto the skin provides significant contrasting of the borders between the areas containing nanoshells and those without. This effect of nanoshells on skin in vivo is manifested by the increase in intensity of the OCT signal in superficial parts of the skin, boundary contrast between superficial and deep dermis and contrast of hair follicles and glands. The presence of nanoshells in the skin was confirmed by electron microscopy. Monte Carlo simulations of OCT images confirmed the possibility of contrasting skin-layer borders and structures by the application of gold nanoshells. The Monte Carlo simulations were performed for two skin models and exhibit effects of nanoparticles similar to those obtained in the experimental part of the study, thus proving that the effects originate exactly from the presence of nanoparticles.

A fluorescence diffuse tomography (FDT) setup for monitoring tumor growth in small animals has been created. In this setup an animal is scanned in the transilluminative configuration by a single source and detector pair. To remove stray light in the detection system, we used a combination of interferometric and absorption filters. To reduce the scanning time, an experimental animal was scanned using the following algorithm: (1) large-step scanning to obtain a general view of the animal (source and detector move synchronously); (2) selection of the fluorescing region; and (3) small-step scanning of the selected region and different relative shifts between the source and detector to obtain sufficient information for 3D reconstruction. We created a reconstruction algorithm based on the Holder norm to estimate the fluorophore distribution. This algorithm converges to the solution with a minimum number of fluorescing zones. The use of tumor cell lines transfected with fluorescent proteins allowed us to conduct intravital monitoring studies. Cell lines of human melanomas Mel-P, Mel-Ibr, Mel-Kor, and human embryonic kidney HEK293 Phoenix were transfected with DsRed-Express and Turbo-RFP genes. The emission of red fluorescent proteins (RFPs) in the long-wave optical range permits detection of deep-seated tumors. In vivo experiments were conducted immediately after subcutaneous injection of fluorescing cells into small animals.

The purpose of this study was to characterize essential changes in the structure of annulus fibrosus (AF) after hydrothermal and infrared (IR) laser treatment and to correlate these results with alterations in tissue state. Polarization-sensitive optical coherence tomography imaging was used to measure collagen birefringence in AF. Differential scanning calorimetry was used as a complementary technique, providing detailed information on thermodynamic processes in the tissue. Birefringence, peak of the denaturation endotherm, and the enthalpy of denaturation (DeltaHm) were determined before and after hydrothermal heat treatment (85 degrees C for 15 min) and non-ablative Er:glass fiber laser exposures on AF in the whole disk (vertebrae-disk-vertebrae complex). Our data have demonstrated quantitative differences between results of laser and hydrothermal heating. Birefringence did not disappear and DeltaHm did not change after treatment in the water bath, but loss of birefringence and a decrease in the enthalpy did occur after laser exposure. These results could be explained by the photomechanical effect of laser irradiation. We suggest that thermo-mechanical stress played a dominant role in the disruption of the collagen network of AF under non-homogeneous laser heating.

A numerical algorithm based on a small-angle approximation of the radiative transfer equation (RTE) is developed to reconstruct scattering characteristics of biological tissues from optical coherence tomography (OCT) images. According to the algorithm, biological tissue is considered to be a layered random medium with a set of scattering parameters in each layer: total scattering coefficient, variance of a small-angle scattering phase function, and probability of backscattering, which fully describe the OCT signal behavior versus probing depth. The reconstruction of the scattering parameters is performed by their variation to fit the experimental OCT signal by the theoretical one using a time-saving genetic algorithm. The proposed reconstruction procedure is tested on model media with known scattering parameters. The possibility to estimate scattering parameters from OCT images is studied for various regimes of OCT signal decay. The developed algorithm is applied to reconstruct optical characteristics of epithelium and stroma for normal cervical tissue and its pathologies, and the potential to distinguish between the types of pathological changes in epithelial tissue by its OCT images is demonstrated.

The optical coherence tomography (OCT) capabilities of plants were evaluated using leaves of Tradescantia pallida (Rose) D. Hunt. The internal structure of the leaf tissues was visualized in vivo and the physiological and morphological states of the tissues under different water supply conditions were monitored using OCT. The OCT technique provides non-invasive two-dimensional images directly on intact plants. The acquisition time of a two-dimensional image with a size of 200x200 pixels and a spatial resolution of 15 microm is 1-3 s. It was shown that OCT is a useful tool for monitoring the physiological and morphological states of plant tissues supplied with varying amounts of water and under the influence of different chemical factors.

Background and Objectives: The goal of this study is to apply a new bioimaging modality, the Optical Coherence Tomography (OCT), for intraoperative control in laser surgery of laryngeal carcinoma., Study Design/materials and Methods: We studied 26 patients with laryngeal carcinoma in situ and in T(1), T(2) stage. We used an endoscopic OCT device for imaging at a wavelength of 0.83 microm with the acquisition rate of approximately 0.5 frames/s for a single (200 x 200 pixel) tomogram. All patients were operated with a surgical YAG:Nd laser at two switchable wavelengths of 1.44 microm and 1.32 microm by laryngofissure, direct microlaryngoscopy, and fibrolaryngoscopy., Results: Information on structural alterations in laryngeal mucosa to the depth of 2 mm, obtained by OCT, makes it possible to precisely locate tumor borders, thus giving an opportunity to control the surgical treatment of laryngeal carcinoma. The YAG:Nd laser scalpel with wavelengths of 1.32 microm and 1.44 microm is successful in surgical procedures both in open and closed larynx due to efficient coagulation and minimization of collateral tissue damage area. Combination of the two wavelengths in the single laser unit and intraoperative OCT monitoring result is a new modality for minimally invasive larynx surgery., Conclusions: OCT is promising to become a new diagnosing method of laryngeal carcinoma and a tool for laser treatment monitoring., (Copyright 2001 Wiley-Liss, Inc.)

A 3-mum laser-generation delivery instrument that uses chalcogenide fibers with a unique damage threshold and conical radiation input has been developed for medical applications. A new purification and synthesis scheme has been elaborated that yields fibers with a heterophase inclusion content of less than 10(4) cm(-3). In such fibers the damage threshold is 350 J/cm(2) at an average power density of 0.5 kW/cm(2) in a YAG:Er laser operating in the repetitive pulse free-running regime with a pulse duration of 350 ms. 1-3 x 10(4) laser pulses were transmitted at a repetition rate of 3 Hz and an average output power of 1 W under the condition of a 15% decrease in the output power.