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.

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.

Biodegradable and low-toxic silicon nanoparticles (SiNPs) have potential in different biomedical applications. Previous experimental studies revealed the efficiency of some types of SiNPs in tumor hyperthermia. To analyse the feasibility of employing SiNPs produced by the laser ablation of silicon nanowire arrays in water and ethanol as agents for laser tumor hyperthermia, we numerically simulated effects of heating a millimeter-size nodal basal-cell carcinoma with embedded nanoparticles by continuous-wave laser radiation at 633 nm. Based on scanning electron microscopy data for the synthesized SiNPs size distributions, we used Mie theory to calculate their optical properties and carried out Monte Carlo simulations of light absorption inside the tumor, with and without the embedded nanoparticles, followed by an evaluation of local temperature increase based on the bioheat transfer equation. Given the same mass concentration, SiNPs obtained by the laser ablation of silicon nanowires in ethanol (eSiNPs) are characterized by smaller absorption and scattering coefficients compared to those synthesized in water (wSiNPs). In contrast, wSiNPs embedded in the tumor provide a lower overall temperature increase than eSiNPs due to the effect of shielding the laser irradiation by the highly absorbing wSiNPs-containing region at the top of the tumor. Effective tumor hyperthermia (temperature increase above 42 °C) can be performed with eSiNPs at nanoparticle mass concentrations of 3 mg/mL and higher, provided that the neighboring healthy tissues remain underheated at the applied irradiation power. The use of a laser beam with the diameter fitting the size of the tumor allows to obtain a higher temperature contrast between the tumor and surrounding normal tissues compared to the case when the beam diameter exceeds the tumor size at the comparable power.

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.

Increase of the efficiency of photodynamic therapy (PDT) requires the development of advanced protocols employing both novel photosensitizer (PS) carriers and aids for online monitoring. Nanoconstructs may be comprised of a photosensitizer, chemotherapy drugs, or inhibitors of molecular pathways that support cancer growth. In this paper, we analyze the efficiency of a bimodal approach involving fluorescence and optoacoustic imaging in monitoring drug distribution and photobleaching. The study evaluates typical sensitivities of these techniques to the presence of the two key moieties of a nanoconstruct: benzoporphyrin derivatives (BPD) serving as a PS, and IRDye800 acting as a contrast agent. Both imaging modalities employ dual-wavelength probing at the wavelengths corresponding to absorption peaks of BPD and IRDye800, which enables their separate detection. In an experiment on a tissue-mimicking phantom with inclusions containing separate BPD and IRDye800 solutions, fluorescence imaging demonstrated higher contrast as compared to optoacoustic imaging for both components, though strong light scattering in the surrounding media restricted accurate localization of the markers. It was also sensitive to photobleaching, which is a measure of PDT efficiency. Optoacoustic imaging demonstrated sufficient sensitivity to both components, though less than that of fluorescence imaging, however, it enabled depth-resolved detection of an absorber and estimation of its relative content. Employment of the bimodal approach in monitoring of PS photobleaching adds to its potential in intraprocedural PDT monitoring.