Photoacoustic study of percutaneous absorption of Carbopol and transdermic gels for topic use in skin

Topical medicine application has been used to treat a good number of pathological processes. Its efficacy is associated to an efficient penetration of the drug in the internal skin layers, promoting systemic effects and excluding the possibility of drug degradation by the digestive tract and hepatic elimination. This work analyzes the penetration kinetics of two soluble bases employed as vehicles for topic application: superficial gel (Carbopol 940) and transdermic (transdermal) gel. Analysis was performed with the photoacoustic technique, based upon the absorption of modulated light by a sample with subsequent conversion of the absorbed energy in heat, generating acoustic waves in the air layer adjacent to the sample. Each of the two vehicles was evaluated through in vivo (human skin) and in vitro application. Measurements in vitro employed samples of VitroSkin (synthetic material with properties similar to those of real skin, employed in the pharmaceutical industry research). Results show that the permeation was faster for the transdermal gel, both for in vivo and in vitro measurements, indicating that in vitro measurements may be utilized in qualitative, comparative permeation studies.     

Time-Resolved Measurements of in vivo Optical Properties of Piglet Brain

The in vivo optical properties of a piglet brain were measured using a time-resolved system at three different wavelengths (759, 794, 824 nm). To separate the contribution of different head layers to the detected signals, the measurements were acquired from the surfaces of skin, skull, dura mater and brain. The source detector distance was chosen to assure a mean penetration depth within each layer. Measurements were analyzed and compared with the results of other in vivo measurements reported in literature.     

Characterization of reconstructed human skin using Photoacoustic Spectroscopy

Recent progress in skin culture techniques has led to the development of systems in which the reconstructed human skin obtained exhibits morphologic characteristics similar to those observed in vivo. Reconstructed human skin may be the best substrate for pharmacological tests of topically applied drugs; besides, it can be employed in the treatment of burns wounds and chronic skin ulcers. However, this newly developed material must be validated by comparison with human skin, in order to show that reconstructed skin presents characteristics similar to those of human skin. This was accomplished in the present work, through photoacoustic spectroscopy (PAS) measurements. Results show similarity between reconstructed skin and ex-vivo human skin, validating possible therapeutic and cosmetic treatments to be developed using the reconstructed human skin analyzed in this work.     

Simultaneous grayscale and subharmonic ultrasound imaging on a modified commercial scanner

Objective

To demonstrate the feasibility of simultaneous dual fundamental grayscale and subharmonic imaging on a modified commercial scanner.

Motivation

The ability to generate signals at half the insonation frequency is exclusive to ultrasound contrast agents (UCA). Thus, subharmonic imaging (SHI; transmitting at f₀ and receiving at f₀/2) provides improved visualization of UCA within the vasculature via suppression of the surrounding tissue echoes. While this capability has proven useful in a variety of clinical applications, the SHI suppression of surrounding tissue landmarks (which are needed for sonographic navigation) also limits it use as a primary imaging modality. In this paper we present results using a commercial ultrasound scanner modified to allow imaging in both grayscale (f₀ = 4.0 MHz) and SHI (f₀ = 2.5 MHz, f₀/2 = 1.25 MHz) modes in real time.

Methods

A Logiq 9 ultrasound scanner (GE Healthcare, Milwaukee, WI) with a 4C curvilinear probe was modified to provide this capability. Four commercially available UCA (Definity, Lantheus Medical Imaging, North Billerica, MA; Optison, GE Healthcare, Princeton, NJ; SonoVue, Bracco Imaging, Milan, Italy; and Sonazoid, GE Healthcare, Oslo, Norway) were all investigated in vitro over an acoustic output range of 3.34 MPa. In vivo the subharmonic response of Sonazoid was investigated in the portal veins of four canines (open abdominal cavity) and four patients with suspected portal hypertension.

Results

In vitro, the four UCA showed an average maximum subharmonic amplitude of 44.1 ± 5.4 dB above the noise floor with a maximum subharmonic amplitude of 48.6 ± 1.6 dB provided by Sonazoid. The average in vivo maximum signal above the noise floor from Sonazoid was 20.8 ± 2.3 dB in canines and 33.9 ± 5.2 dB in humans. Subharmonic amplitude as a function of acoustic output in both groups matched the S-curve behavior of the agent observed in vitro. The dual grayscale imaging provided easier sonographic navigation, while the degree of tissue suppression in SHI mode varied greatly on a case by case basis.

Conclusions

These results demonstrate the feasibility of dual grayscale and SHI on a modified commercial scanner. The ability to simultaneously visualize both imaging modes in real time should improve the applicability of SHI as a future primary clinical imaging modality.     

Diagnostic assessment of human iron stores by measurement of hepatic magnetic susceptibility

Summary A magnetic method for direct noninvasive measurements of human hepatic storage iron with a SQUID susceptometer has been developed.In vivo magnetic andin vitro chemical determinations of liver nonheme iron are closely correlated. Magnetic measurements of iron stores are clinically useful in the diagnosis of disorders of iron metabolism. Supported in part by research grants AM 25105, AM 14370 and HL 24198 from the National Institutes of Health.     

Change of Color Appearance in Photopic,Mesopic and Scotopic Vision

Mesopic vision describes a range of light levels where vision is mediated by both cones and rods. The appearance of color in mesopic vision differs drastically from that in photopic vision, where only cones mediate visual information. We used a haploscopic color matching technique to investigate the color appearance under various illuminance levels, ranging from photopic to scotopic via mesopic levels. The observers did color matching between a test color chip under various illuminance levels and a matching color stimulus presented on the Cathode-Ray Tube (CRT) display under the photopic illuminance condition. The results showed that not only chroma and lightness but hue of most color chips changed with illuminance. The manner of the hue changed depended on the color of the test chip, while matching points approached a neutral gray with decrease in illuminance level for all test chips. Chroma reduced continuously with decrease of the illuminance level until 0.1 lx for reddish and yellowish color chips or until 1 lx for greenish and bluish ones. Beyond those illuminance levels, chroma was approximately constant. Lightness decreased with decreasing illuminance level for all test chips except bluish color chips, for which lightness did not decrease much in general and even increased in some cases as predicted by the Purkinje shift. The experimental results obtained in the present study provide critical features that should be considered in predicting the appearance of color at low light levels.     

Effect of storage conditions of skin samples on their optical characteristics

The optical characteristics of skin samples are experimentally studied ex vivo and in vitro at different storage conditions. The experiments are performed on a Cary-2415 spectrophotometer in the spectral range 400–700 nm. Based on the measured diffuse reflectance and total transmittance spectra, the spectra of the absorption and reduced scattering coefficients are calculated in terms of the inverse adding-doubling method. It is shown that the method of storage of samples mainly affects the reduced scattering coefficient of biotissue. Thus, upon storage of skin in an isotonic solution and in its absence, the reduced scattering coefficient increases ∼1.5 and ∼2 times, respectively, compared to the value of this parameter for the intact sample. The differences in the absorption spectra of ex vivo samples and samples stored under different conditions are the most noticeable in the absorption range of blood and are significant above 600 nm.     

Tunable structural color of anodic tantalum oxide films

Tantalum(Ta) oxide films with tunable structural color were fabricated easily using anodic oxidation.The structure,components,and surface valence states of the oxide films were investigated by using gazing incidence X-ray diffractometry,X-ray photoelectron microscopy,and surface analytical techniques.Their thickness and optical properties were studied by using spectroscopic ellipsometry and total reflectance spectrum.Color was accurately defined using L*a*b* scale.The thickness of compact Ta2O5 films was linearly dependent on anodizing voltage.The film color was tunable by adjusting the anodic voltage.The difference in color appearance resulted from the interference behavior between the interfaces of air-oxide and oxide-metal.     

Identification of epidermal L‐tryptophan and its oxidation products by in vivo FT‐Raman Spectroscopy further supports oxidative stress in patients with vitiligo

In the past, non‐invasive in vivo FT‐Raman spectroscopy has been used to detect H₂O₂‐mediated oxidation of methionine to methionine sulfoxide and methionine sulfone, as well as cysteine to cysteic acid, in the sequence of proteins in the epidermis of patients with vitiligo. L ‐tryptophan (Trp) is another potential target for this oxidation. Owing to the presence of 10⁻³M epidermal albumin which contains one Trp residue, it was tempting to follow the oxidation of this amino acid. Using in vivo and in vitro FT‐Raman spectroscopy, we show for the first time that epidermal Trp is oxidised in patients with vitiligo, yielding 5‐OH‐Trp at 930 cm⁻¹ and other oxidation products (i.e. N‐formyl kynurenine and kynurenine) from indole ring oxidation peaking at 1050 cm⁻¹. On the basis of detailed in vitro results, we could conclude that 5‐OH‐Trp as well as formyl kynurenine and kynurenine are formed via H₂O₂‐mediated Fenton chemistry. These results once again bring out the strength of non‐invasive in vivo FT‐Raman Spectroscopy in dermatology to follow the effect of oxidative stress in the skin of patients with vitiligo. Copyright © 2008 John Wiley & Sons, Ltd.     

The effects of contact area and applied load on the morphology of in vitro worn ultra‐high molecular weight knee prostheses: a micro‐Raman and gravimetric study

Ultra‐high molecular weight polyethylene knee prostheses of two different sizes (i.e. characterised by different contact areas) were run on a knee wear simulator in two tests differing for the applied load. Gravimetric and micro‐Raman spectroscopic analyses were performed to investigate at macroscopic and molecular levels the effects of contact area and applied load on the morphology of the knee components. The larger prostheses showed a higher mass loss in both tests, while Raman spectroscopy showed a more significant polymer degradation in the smaller prostheses, suggesting that a lower contact area is responsible for a higher wear at the molecular wear, but for a lower gravimetric wear. Raman measurements showed that in the second test (using a higher applied load), the wear mechanism changed with respect to the first test: the less severe conditions used in the latter resulted in an increase in the orthorhombic content, while the more severe conditions used in the former determined an increase and rearrangement of the amorphous phase. The results here reported allow to gain more insights into the effects of contact area, applied load and contact stress on wear. Preliminary micro‐Raman analyses on in vivo worn knee prostheses confirmed the results obtained in vitro: the retrievals of smaller size were found more degraded at molecular level than those of larger size. These findings showed that the conditions used in the in vitro tests well simulate the knee kinematics of the knee joint and thus the in vivo wear. Copyright © 2014 John Wiley & Sons, Ltd.     

Raman and fluorescence investigations on retrieved Biolox® delta femoral heads

Since the introduction of ceramic‐on‐ceramic couplings in total hip arthroplasty, continuous efforts have been performed to improve the performance of the prosthetic components. To expand the applications of ceramics, in 2000 an innovative alumina–zirconia composite was introduced in the market. The present study represents the first investigation aimed at characterising at the molecular level Biolox® delta retrievals implanted between 1999 and 2009. Fluorescence and Raman results showed that a progressive improvement of the material properties has occurred in the period between 1999 and 2009. Raman spectroscopy showed that wear was the main cause of the in vivo tetragonal‐to‐monoclinic zirconia transformation. Our findings validated the in vitro accelerated ageing protocols proposed in the literature to simulate the effects of the in vivo wear, because the mechanism operating in vivo was found to be the same active in vitro. The in vitro fracture of a new femoral head appeared to be an extreme wear condition that determined the most significant changes in the residual stress state and monoclinic content both in the section of the fragments and on their surface. The micro‐Raman mapping of the fractured articulating surface showed that the tetragonal‐to‐monoclinic transformation involved a region much more extended than as reported in the literature. Copyright © 2012 John Wiley & Sons, Ltd.     

Kinetics of oxygenation of skin tissue exposed to low-intensity laser radiation

We present the results of modeling the action spectrum and an experimental investigation of the effect of laser-induced photodissociation of oxyhemoglobin in vivo on the increase in the degree of oxygenation of skin tissue in the exposed area. We have shown that controlling the local concentration of free molecular oxygen in biological tissues together with the possibility of eliminating tissue hypoxia using laser radiation makes it possible to stimulate aerobic cell metabolism and to achieve the needed therapeutic effect. __________ Translated from Zhurnal Prikladnoi Spektroskopii, Vol. 74, No. 1, pp. 120–125, January–February, 2007.     

Evidence for neuroprotective properties of human umbilical cord blood cells after neuronal hypoxia <Emphasis Type="Italic">in vitro</Emphasis>

Background  

One of the most promising options for treatment of stroke using adult stem cells are human umbilical cord blood (HUCB) cells that were already approved for therapeutic efficacy in vivo. However, complexity of animal models has thus far limited the understanding of beneficial cellular mechanisms. To address the influence of HUCB cells on neuronal tissue after stroke we established and employed a human in vitro model of neuronal hypoxia using fully differentiated vulnerable SH-SY5Y cells. These cells were incubated under an oxygen-reduced atmosphere (O₂< 1%) for 48 hours. Subsequently, HUCB mononuclear cells (MNC) were added to post-hypoxic neuronal cultures. These cultures were characterized regarding to the development of apoptosis and necrosis over three days. Based on this we investigated the therapeutic influence of HUCB MNC on the progression of apoptotic cell death. The impact of HUCB cells and hypoxia on secretion of neuroprotective and inflammatory cytokines, chemokines and expression of adhesion molecules was proved.     

Mapping phosphorylation rate of fluoro-deoxy-glucose in rat brain by F chemical shift imaging

¹⁹F magnetic resonance spectroscopy (MRS) studies of 2-fluoro-2-deoxy-d-glucose (FDG) and 2-fluoro-2-deoxy-d-glucose-6-phosphate (FDG-6P) can be used for directly assessing total glucose metabolism in vivo. To date, ¹⁹F MRS measurements of FDG phosphorylation in the brain have either been achieved ex vivo from extracted tissue or in vivo by unusually long acquisition times. Electrophysiological and functional magnetic resonance imaging (fMRI) measurements indicate that FDG doses up to 500 mg/kg can be tolerated with minimal side effects on cerebral physiology and evoked fMRI-BOLD responses to forepaw stimulation. In halothane-anesthetized rats, we report localized in vivo detection and separation of FDG and FDG-6P MRS signals with ¹⁹F 2D chemical shift imaging (CSI) at 11.7 T. A metabolic model based on reversible transport between plasma and brain tissue, which included a non-saturable plasma to tissue component, was used to calculate spatial distribution of FDG and FDG-6P concentrations in rat brain. In addition, spatial distribution of rate constants and metabolic fluxes of FDG to FDG-6P conversion were estimated. Mapping the rate of FDG to FDG-6P conversion by ¹⁹F CSI provides an MR methodology that could impact other in vivo applications such as characterization of tumor pathophysiology.     

In vitro determination of the sun protection factor of sunscreens through photoacoustic spectroscopy: A new approach

The investigation of the sun protection factor (SPF) and the efficacy of ultraviolet absorber ingredients are extremely important items for the cosmetic industry. The development of new efficient in vitro methodologies for these investigations is an essential condition for the evolution of sunscreens. The most accepted test for assessing the efficacy of sunscreens is the in vivo (in humans) SPF determination, following validated and worldwide adopted methodologies like those regulated by FDA (USA), COLIPA (Europe) and JCIA (Japan). However, due to the high costs and time consumption of in vivo SPF determination, the constant development of in vitro methods able to predict the in vivo SPF is very important. The present study evaluates different experimental methods for in vitro SPF determination – photoacoustic spectroscopy (PAS) and transmission spectroscopy (TS), employing different substrates (PMMA and VitroSkin^?) and concentrations (0.8 mg/cm² and 2 mg/cm²). Results are compared to the labeled SPF. The high correlation between the labeled SPF and the SPF determined by PAS expands the application of this technique in the photobiology field.     

Effect of electron beam irradiation on the structural,thermal and optical properties of poly(vinyl alcohol) thin film

Poly(vinyl alcohol) (PVA) polymer was prepared using the casting technique. The obtained PVA thin films have been irradiated with electron beam doses ranging from 20 to 300 kGy. The resultant effect of electron beam irradiation on the structural properties of PVA has been investigated using X-ray diffraction (XRD) and Fourier transform infrared spectroscopy (FTIR), while the thermal properties have been investigated using thermo-gravimetric analysis and differential thermal analysis (DTA). The onset temperature of decomposition T ₀ and activation energy of thermal decomposition E _a were calculated, results indicate that the PVA thin film decomposes in one main weight loss stage. Also, the electron beam irradiation in dose range 95–210 kGy led to a more compact structure of the PVA polymer, which resulted in an improvement in its thermal stability with an increase in the activation energy of thermal decomposition. The variation of transition temperatures with electron beam dose has been determined using DTA. The PVA thermograms were characterized by the appearance of an endothermic peak due to melting. In addition, the transmission of the PVA samples and any color changes were studied. The color intensity Δ E was greatly increased with increasing electron beam dose, and was accompanied by a significant increase in the blue color component.     

MR microscopy of water diffusion tensor in biological systems

Diffusion tensor imaging (DTI) of the control and mechanically injured spinal cord of a ratin vitro andin vivo are reported.In vitro experiments were done on a home-built 6.4 T magnetic resonance microscope. Results for formaline-fixed samples show significant differences in diffusion tensor components between gray matter and white matter of the control spinal cord. Moreover, it is shown that already 6–10 min after the injury DTI can detect changes in water diffusion in areas extending far beyond the region of primary tissue damage.In vivo experiments were performed using a 9.4 T Magnex magnet and Bruker Medspec imaging system. Good-quality DTI images, free from motion artifacts were obtained. Results from control samples confirm differences in water diffusion between white matter and gray matter, observedin vitro. In vivo experiments show that characteristic changes in water diffusion observedin vitro 6–10 min after injury are preserved 60–360 min after injury, without significant alteration during this time.     

Development of non-deterioration-type skin tissue phantom using silicone material

We developed a new type of three-layered skin tissue phantom using silicone material. This phantom maintains its optical properties in a certain period of time, typically about three months, which is much longer than about 30 min with our previous agar-type phantom. Experiments on spectral reflectance measurements and color analysis indicate the possibility of silicone-type skin tissue phantom.     

Preparation and electron microscopy of liquid crystalline condensates of DNA

Summary A brief survey of thein vitro methods of preparing DNA mesomorphic phases is presented. The significance of thesein vitro preparations for thein vivo condensed states of DNA is also discussed. The procedure of preparing a dispersion of mesomorphic condensates of unsonicated calf thymus DNA by using polyethylene glycol was investigated by the circular dichroism (CD) technique. The operational conditions able to provide at the same time good packing of the DNA chains and reproducible CD signals were searched. The morphology of the condensates thus obtained was studied by transmission electron microscopy. Work presented at the second USSR-Italy Bilateral Meeting on Liquid Crystals held in Moscow, September 15–21, 1988.     

Imaging of subcutaneous microcirculation vascular network by double correlation Optical Coherence Tomography

Spatial distribution of superficial blood vessels in human skin in vivo has been observed by using the double correlation Optical Coherence Tomography (OCT). To remove background noise, reduce the artifacts associated with patient motions and to increase the overall quality of the experimental OCT images an adaptive Wiener filtering technique has been employed. Fourier domain correlation has been subsequently applied to enhance spatial resolution of images of vascular network in human skin in vivo. Image processing has been performed on Graphics Processing Units (GPUs) utilizing Compute Unified Device Architecture (CUDA) framework in the frequency‐domain. This approach allows carrying out image processing in parallel significantly speeding up the computations. The presented results show that the double correlation method permits obtaining 2D/3D OCT images of subcutaneous microcirculation vascular network and its spatial distribution within the human skin with higher spatial resolution compare to the other OCT correlation‐based techniques developed earlier.