Digital holographic microscopy of spiropyran-based dynamic materials.

Spiropyran (SP)-based dynamic materials undergo structural changes in response to external stimuli. In this paper, we show that digital holographic microscopy (DHM) is an effective candidate for characterisation of SPs (embedded in polymer matrices) and for monitoring of their dynamical changes. The polymer matrices are polylactic acid (PLA) and poly(methyl methacrylate) (PMMA) films, which are decorated with SPs and immobilised on graphene quantum dots (GQDs). GQDs are modified by benzylamines prior to the loading of SP species because of the enhancement of hydrophobic characteristics. UV irradiation is used as the external stimulus and the dynamical changes of the samples before and after UV irradiation are measured. DHM is arranged on a novel self-referencing setup, which substantially reduces the sensitivity of DHM to environmental vibrations. Morphometric information for characterisation of the samples is obtained by analysis of the recorded digital holograms. The experimental results demonstrate the potential of the presented technique to serve as an alternative technique for surface measurement methodologies such as atomic force microscope and stylus profiler for surface characterisation of similar materials.

Digital holographic microscopy for real-time investigation of 3D microstructural dynamics of starch-kefiran-based nanocomposite.

Investigating real-time phenomena in bio-polymers has received much attention because of their increasing demands in polymer substitution. The 3D morphometry of polymer surfaces may be very impactful in such studies. Here, digital holographic microscopy (DHM) is applied for quantitative measurement of the rare morphological changes of UV-A and UV-C exposed nanocomposites during their incubation with excess water. By reconstructing the recorded successive digital holograms, the time evolution of the swelled regions of the samples is derived. Our results clearly show that the higher water swelling of UV-A irradiated starch/kefiran/ZnO may be attributed to its higher hydrophilicity.

Extracellular vesicles derived from dendritic cells loaded with VEGF-A siRNA and doxorubicin reduce glioma angiogenesis in vitro.

BACKGROUND: Numerous attempts have been devoted to designing anti-angiogenic agents as a strategy to slow tumor growth and progression. Clinical applications of conventional anti-angiogenic agents face some challenges, e.g., off-target effects for TKIs and also low solid tumor penetration for mAbs. Furthermore, although anti-angiogenic therapy provides a normalization window for better chemo-RT response, in long-term treatments, tumor hypoxia as a result of total removal of VEGF-A by mAbs from the TME or complete blockade of TK receptors induces over-activation of compensatory angiogenic pathways, causing escape. Herein, we investigate the efficacy of si-DOX-DC-EVs to reduce glioma angiogenesis and invasiveness. METHODS: Mature DCs were generated from PBMC and EVs were isolated from the DCs culture media. siRNA and Doxorubicin were loaded into EVs by EP and incubation. Afterward, the uptake of DC-EVs was assessed by flow cytometry, and the subcellular localization of EVs was tested by confocal imaging. Tube formation assay was performed to assess the efficacy of si-DOX-DC-EVs to reduce tumor angiogenesis which was analyzed by DHM. Morphometric analysis of apoptotic cells was performed by DHM and confocal imaging and further, ELISA was performed for hypoxia-related and angiogenic cytokines. The impact of our theranostic system "si-DOX-DC-MVs" on the formation of vascular mimics, colonies, and invasion of C6 cells was checked in vitro. Afterward, orthotropic rat models of glioma were generated and the optimal administration route was selected by in vivo fluorescent analysis. Then, the microvessel density, vimentin expression, and accumulation of immune cells in tumoral tissues were assessed by IHC. Finally, necropsy and autopsy analyses were performed to check the safety of our theranostic agent. RESULTS: DC-EVs loaded with si-DOX-DC-EVs were successfully uptaken by cells with different subcellular trafficking for MVs and exosomes, reduced tumor angiogenesis in DHM analysis, and induced apoptosis in tumoral cells. Moreover, using DHM, we performed a detailed label-free analysis of tip cells which suggested that the tip cells in si-DC-MV treatments lost their geometrical migration capacity to form tube-like structures. Furthermore, the ELISAs performed highlighted that there is a mild overactivation of compensatory Tie2/Ang2 pathway after VEGF-A blockade which confers with severe hypoxia and sustains normal angiogenesis which is the optimal goal of anti-angiogenesis therapy for cancer to avoid resistance.The results of our VM analyses indicated that si-DOX-DC-MVs completely inhibited VM process. Moreover, the invasion, migration, and colony formation of the C6 cells treated with si-DOX-MVs were the least among all treatments. IN was the optimal route of administration. The MVD analyses indicated that si-DOX-DC-MVs reduced the number of tumoral microvessels and normalized vessel morphology. Intense CD8+ T cells were observed near the tumoral vessels in the si-DOX-DC-MVs group and with minimal activation of MT (low Vimentin expression). Necropsy and toxicology results proved that the theranostic system proposed is safe. CONCLUSIONS: DC-EVs loaded with VEGF-A siRNA and Doxorubicin were more potent than BV alone as a multi-disciplinary strategy that combats glioma growth by cytotoxic impacts of DOX and inhibits angiogenesis by VEGF-A siRNAs with excess immunologic benefits from DC-EVs. This next-generation anti-angiogenic agent normalizes tumor vessel density rather than extensively eliminating tumor vessels causing hypoxia and mesenchymal transition.

Evaluation of pitting corrosion by dynamic speckle pattern analysis.

There is an increasing interest in non-destructive and real-time high-resolution approaches for corrosion studies in metals. In this paper, we propose the dynamic speckle pattern method as a low-cost, easy-to-implement, and quasi in-situ optical technique for the quantitative evaluation of pitting corrosion. This type of corrosion occurs in a specific area of a metallic structure and causes holes formation leading to structural failure. A Custom 450 stainless steel sample, placed in 3.5 wt% NaCl solution and applied to a [Formula: see text] potential to initiate the corrosion, is used as the sample. The speckle patterns formed by the scattering of a He-Ne laser light is changed over time due to any corrosion in the sample. The analysis of the time-integrate speckle pattern suggests that the growth rate of pitting decreases with time.

Detection of intralayer alignment in multicomponent lipids by dynamic speckle pattern analysis.

Multicomponent mixtures of bilayer lipids, thanks to the coexistence of liquid-crystalline phases in their structures, may be used in the development of functional membranes. In such membranes interlayer ordering distributes across membrane lamellae, resulting in long-range alignment of phase-separated domains. In this paper, we explore the dynamics of this phenomenon by laser speckle pattern analysis. We show that cholesterol content decreases the activity, and the rate of the domains size development is related to the change of physical roughness of the multicomponent lipid mixture. Our results are in agreement with the previous experimental reports. However, our experimental procedure is an easy-to-implement and effective methodology.

Clinical theranostics applications of photo-acoustic imaging as a future prospect for cancer.

Photoacoustic imaging (PAI) of biological tissue has been a fast developing biomedical multi-wave imaging modality, after its introduction in the mid90s. PAI couples laser excitation to acoustic detection. Especially, in recent years its significant advantages in onco-surgery has attracted much attention due to its ability to detect malignant tissues. Monitoring cancer angiogenesis, assessment of blood oxygen saturation, functional brain imaging, evaluation of cortical blood volume, detection of skin/conjunctival melanoma depth, assessment of met-hemoglobin, investigating tumor hypoxia andcancer lymph node metastases are some of its promising applications. Moreover, as a real-time monitoring strategy, PAI allows intraoperative imaging of micro-metastases and residual islands in onco-surgery. Herein, we provide a brief introduction to biophysics and fundamentals of PAI, potential novel endogenous and exogenous contrast agents, and novel techniques to develop engineered and targeted contrast agents with theranostic applications. We also summarize the clinical trial pipelines for PAI. Furthermore, we discuss the potential obstacles and limitation of PAI theranostic agents for further clinical applications and strategies to overcome these hurdles.

3D monitoring of the surface slippage effect on micro-particle sedimentation by digital holographic microscopy.

In several phenomena in biology and industry, it is required to understand the comprehensive behavior of sedimenting micro-particles in fluids. Here, we use the numerical refocusing feature of digital holographic microscopy (DHM) to investigate the slippage effect on micro-particle sedimentation near a flat wall. DHM provides quantitative phase contrast and three-dimensional (3D) imaging in arbitrary time scales, which suggests it as an elegant approach to investigate various phenomena, including dynamic behavior of colloids. 3D information is obtained by post-processing of the recorded digital holograms. Through analysis of 3D trajectories and velocities of multiple sedimenting micro-particles, we show that proximity to flat walls of higher slip lengths causes faster sedimentation. The effect depends on the ratio of the particle size to (1) the slip length and (2) its distance to the wall. We corroborate our experimental findings by a theoretical model which considers both the proximity and the particle interaction to a wall of different hydrophobicity in the hydrodynamic forces.