Impact of Plasmonic Nanoparticles on Poikilocytosis and Microrheological Properties of Erythrocytes.

Plasmonic nanoparticles (NP) possess great potential in photothermal therapy and diagnostics. However, novel NP require a detailed examination for potential toxicity and peculiarities of interaction with cells. Red blood cells (RBC) are important for NP distribution and the development of hybrid RBC-NP delivery systems. This research explored RBC alterations induced by noble (Au and Ag) and nitride-based (TiN and ZrN) laser-synthesized plasmonic NP. Optical tweezers and conventional microscopy modalities indicated the effects arising at non-hemolytic levels, such as RBC poikilocytosis, and alterations in RBC microrheological parameters, elasticity and intercellular interactions. Aggregation and deformability significantly decreased for echinocytes independently of NP type, while for intact RBC, all NP except Ag NP increased the interaction forces but had no effect on RBC deformability. RBC poikilocytosis promoted by NP at concentration 50 µg mL-1 was more pronounced for Au and Ag NP, compared to TiN and ZrN NP. Nitride-based NP demonstrated better biocompatibility towards RBC and higher photothermal efficiency than their noble metal counterparts.

Photocatalytic activity of TiO2 nanoparticles: effect of thermal annealing under various gaseous atmospheres.

The structure, composition and photocatalytic activity of TiO(2) nanoparticles annealed in various gas atmospheres (N(2), NH(3) and H(2)) were studied in this work. The effect of treatment on crystal structure, particle size, chemical composition and optical absorbance were assessed by means of x-ray diffraction, transmission electron microscopy, x-ray photoelectron spectroscopy and diffuse optical reflectance/transmittance measurements, respectively. Photocatalytic properties of the materials were evaluated by three different methods: degradation of methyl orange in water, killing of Staphylococcus aureus bacteria and photogeneration of radicals in the presence of 3-carboxy-2,2,5,5-tetramethyl pyrrolidine-1-oxyl (PCA) marker molecules. The results indicate that the correlation between pretreatment and the photocatalytic performance depends on the photocatalytic processes and cannot be generalized.

Effect of size of TiO(2) nanoparticles applied onto glass slide and porcine skin on generation of free radicals under ultraviolet irradiation.

Titanium dioxide (TiO(2)) nanoparticles are extensively used today in sunscreens and coatings as protective compounds for human skin and material surfaces from UV radiation. In this paper, such particles are investigated by electron paramagnetic resonance spectroscopy as sources of free radicals under UV irradiation. The surface density of a placebo with embedded particles corresponds to the recommendations of dermatologists (2 mg cm(-2)). It is revealed that if applied onto glass, small particles 25 nm in diameter produce an increased amount of free radicals compared to the larger ones of 400 nm diam and the placebo itself. However, if applied onto porcine skin in vitro, there is no statistically distinct difference in the amount of radicals generated by the two kinds of particles on skin and by the skin itself. This proves that although particles as part of sunscreens produce free radicals, the effect is negligible in comparison to the production of radicals by skin in vitro.

Monitoring of temperature-mediated phase transitions of adipose tissue by combined optical coherence tomography and Abbe refractometry.

Observation of temperature-mediated phase transitions between lipid components of the adipose tissues has been performed by combined use of the Abbe refractometry and optical coherence tomography. The phase transitions of the lipid components were clearly observed in the range of temperatures from 24°C to 60°C, and assessed by quantitatively monitoring the changes of the refractive index of 1- to 2-mm-thick porcine fat tissue slices. The developed approach has a great potential as an alternative method for obtaining accurate information on the processes occurring during thermal lipolysis.

Delivery and reveal of localization of upconversion luminescent microparticles and quantum dots in the skin in vivo by fractional laser microablation, multimodal imaging, and optical clearing.

Delivery and spatial localization of upconversion luminescent microparticles [Y2O3:Yb, Er] (mean size ∼1.6 µm) and quantum dots (QDs) (CuInS2/ZnS nanoparticles coated with polyethylene glycol-based amphiphilic polymer, mean size ∼20 nm) inside rat skin was studied in vivo using a multimodal optical imaging approach. The particles were embedded into the skin dermis to the depth from 300 to 500 µm through microchannels performed by fractional laser microablation. Low-frequency ultrasound was applied to enhance penetration of the particles into the skin. Visualization of the particles was revealed using a combination of luminescent spectroscopy, optical coherence tomography, confocal microscopy, and histochemical analysis. Optical clearing was used to enhance the image contrast of the luminescent signal from the particles. It was demonstrated that the penetration depth of particles depends on their size, resulting in a different detection time interval (days) of the luminescent signal from microparticles and QDs inside the rat skin in vivo. We show that luminescent signal from the upconversion microparticles and QDs was detected after the particle delivery into the rat skin in vivo during eighth and fourth days, respectively. We hypothesize that the upconversion microparticles have created a long-time depot localized in the laser-created channels, as the QDs spread over the surrounding tissues.

Influence of probe pressure on diffuse reflectance spectra of human skin measured in vivo.

Mechanical pressure superficially applied on the human skin surface by a fiber-optic probe influences the spatial distribution of blood within the cutaneous tissues. Upon gradual load of weight on the probe, a stepwise increase in the skin reflectance spectra is observed. The decrease in the load follows the similar inverse staircase-like tendency. The observed stepwise reflectance spectra changes are due to, respectively, sequential extrusion of blood from the topical cutaneous vascular beds and their filling afterward. The obtained results are confirmed by Monte Carlo modeling. This implies that pressure-induced influence during the human skin diffuse reflectance spectra measurements in vivo should be taken into consideration, in particular, in the rapidly developing area of wearable gadgets for real-time monitoring of various human body parameters.

Nanoparticle-free tissue-mimicking phantoms with intrinsic scattering.

We present an alternative to the conventional approach, phantoms without scattering nanoparticles, where scattering is achieved by the material itself: spherical cavities trapped in a silicone matrix. We describe the properties and fabrication of novel optical phantoms based on a silicone elastomer polydimethylsiloxane (PDMS) and glycerol mixture. Optical properties (absorption coefficient µa , reduced scattering coefficient µs' , and anisotropy factor g) of the fabricated phantoms were retrieved from spectrophotometric measurements (in the 400-1100 nm wavelength range) using the inverse adding-doubling method. The internal structure of the phantoms was studied under a scanning electron microscope, and the chemical composition was assessed by Raman spectroscopy. Composition of the phantom material is reported along with the full characterization of the produced phantoms and ways to control their parameters.

Effect of size of TiO2 nanoparticles embedded into stratum corneum on ultraviolet-A and ultraviolet-B sun-blocking properties of the skin.

Recently there has been a strong demand to protect human skin against negative effects of the UV solar light. This problem is interesting due to the increased frequency of human diseases caused by such radiation. We aim to evaluate how the optical properties of the horny layer of skin can be effectively changed by imbedding TiO2 fine particles to achieve the maximal attenuation of the UV solar radiation. In-depth distribution of TiO2 particles embedded into the skin by multiple administration of sunscreens is determined experimentally using the tape-stripping technique. A computer code implementing the Monte Carlo method is developed to simulate photon migration within the 20-microm-thick horny layer filled with nanosized TiO2 spheres, 25 to 200 nm in diameter. Dependencies of the UV radiation of two wavelengths (310 and 400 nm) absorbed by and totally reflected from, as well as transmitted through the horny layer on the size of TiO2 particles are obtained and analyzed. The most attenuating particles are found to be 62 and 122 nm in diameter for 310- and 400-nm light, respectively. The former could be suggested as the main fraction to be used in sunscreens to prevent erythema.

Use of optical skin phantoms for preclinical evaluation of laser efficiency for skin lesion therapy.

Skin lesions are commonly treated using laser heating. However, the introduction of new devices into clinical practice requires evaluation of their performance. This study presents the application of optical phantoms for assessment of a newly developed 975-nm pulsed diode laser system for dermatological purposes. Such phantoms closely mimic the absorption and scattering of real human skin (although not precisely in relation to thermal conductivity and capacitance); thus, they can be used as substitutes for human skin for approximate evaluation of laser heating efficiency in an almost real environment. Thermographic imaging was applied to measure the spatial and temporal temperature distributions on the surface of laser-irradiated phantoms. The study yielded results of heating with regard to phantom thickness and absorption, as well as laser settings. The methodology developed can be used in practice for preclinical evaluations of laser treatment for dermatology.

Measurements of fundamental properties of homogeneous tissue phantoms.

We present the optical measurement techniques used in human skin phantom studies. Their accuracy and the sources of errors in microscopic parameters' estimation of the produced phantoms are described. We have produced optical phantoms for the purpose of simulating human skin tissue at the wavelength of 930 nm. Optical coherence tomography was used to measure the thickness and surface roughness and to detect the internal inhomogeneities. A more detailed study of phantom surface roughness was carried out with the optical profilometer. Reflectance, transmittance, and collimated transmittance of phantoms were measured using an integrating-sphere spectrometer setup. The scattering and absorption coefficients were calculated with the inverse adding-doubling method. The reduced scattering coefficient at 930 nm was found to be 1.57±0.14 mm(−1) and the absorption was 0.22±0.03 mm(−1) . The retrieved optical properties of phantoms are in agreement with the data found in the literature for real human tissues.

Titania nanofibers in gypsum composites: an antibacterial and cytotoxicology study.

Further developments of antibacterial coatings based on photocatalytic nanomaterials could be a promising route towards potential environmentally friendly applications in households, public buildings and health care facilities. Hereby we describe a simple chemical approach to synthesize photocatalytic nanomaterial-embedded coatings using gypsum as a binder. Various types of TiO2 nanofiber-based photocatalytic materials (nitrogen-doped and/or palladium nanoparticle decorated) and their composites with gypsum were characterized by means of scanning (SEM) and transmission (TEM) electron microscopy as well as electron and X-ray diffraction (XRD) and energy-dispersive X-ray spectroscopy (EDX) techniques. These gypsum-based composites can be directly applied as commercially available paints on indoor walls. Herein we report that surfaces coated with photocatalytic composites exhibit excellent antimicrobial properties by killing both methicillin-sensitive Staphylococcus aureus (MSSA) and methicillin-resistant Staphylococcus aureus (MRSA) under blue light. In the case of MSSA cells, the palladium nanoparticle-decorated and nitrogen-doped TiO2 composites demonstrated the highest antimicrobial activity. For the MRSA strain even pure gypsum samples were proven to be efficient in eradicating Gram-positive human pathogens. The cytotoxicity of freestanding TiO2 nanofibers was revealed by analyzing the viability of HeLa cells using MTT and fluorescent cell assays.

Feasibility study of the optical imaging of a breast cancer lesion labeled with upconversion nanoparticle biocomplexes.

Innovative luminescent nanomaterials, termed upconversion nanoparticles (UCNPs), have demonstrated considerable promise as molecular probes for high-contrast optical imaging in cells and small animals. The feasibility study of optical diagnostics in humans is reported here based on experimental and theoretical modeling of optical imaging of an UCNP-labeled breast cancer lesion. UCNPs synthesized in-house were surface-capped with an amphiphilic polymer to achieve good colloidal stability in aqueous buffer solutions. The scFv4D5 mini-antibodies were grafted onto the UCNPs via a high-affinity molecular linker barstar:barnase (Bs:Bn) to allow their specific binding to the human epidermal growth factor receptor HER2/neu, which is overexpressed in human breast adenocarcinoma cells SK-BR-3. UCNP-Bs:Bn-scFv4D5 biocomplexes exhibited high-specific immobilization on the SK-BR-3 cells with the optical contrast as high as 10:1 benchmarked against a negative control cell line. Breast cancer optical diagnostics was experimentally modeled by means of epi-luminescence imaging of a monolayer of the UCNP-labeled SK-BR-3 cells buried under a breast tissue mimicking optical phantom. The experimental results were analyzed theoretically and projected to in vivo detection of early-stage breast cancer. The model predicts that the UCNP-assisted cancer detection is feasible up to 4 mm in tissue depth, showing considerable potential for diagnostic and image-guided surgery applications.

Designing inorganic light-protective skin nanotechnology products.

In this review, we discuss the use of inorganic nanoparticles, mainly zinc oxide (ZnO) and titanium dioxide (TiO2), for sunscreen applications considering their intrinsic physical properties and the Mie theory. These properties cause, from one side, attenuation of the ultraviolet light by absorption and scattering (dependent on a particle size), which is the purpose sunscreens are designed for, and formation of free radicals (i.e., phototoxicity) during this process--from the other. Particle penetration into skin is also an important issue addressed in this review due to possible adverse effects associated with interaction between nanoparticles and skin living cells.