Chronic pain encompasses a range of painful conditions such as migraines, fibromyalgia, or prolonged periods of pain without a specific etiological cause. In this context, a less complex, easily applicable therapy that reduces side effects and does not require a high investment from the patient becomes highly important. Currently, chronic pain treatment with topically applied endocannabinoids has been a subject of research. However, permeation studies are necessary to ensure efficacy, safety, and delivery of the active ingredient to the skin layers. Confocal Raman Spectroscopy (CRS) has emerged as an analytical method for the analysis of materials, including biological tissues. In this context, the present study brings an in vivo approach using Confocal Raman Spectroscopy to analyze in vivo the permeation of two formulations containing Cannabidiol (CBD). One formulation consists of nanoencapsulated CBD in transferosomes (TRANS), while the other is a nanolipid carrier containing nanoencapsulated CBD in transferosomes (CLN TRANS). The aim of this study is to explore the topical treatment of chronic pain using these formulations and assess their permeation characteristics. Raman data were collected from stratum corneum, epidermis and dermis at T0, T4 and T8h after topical application. It was possible to identify that both products showed to permeate to the dermis, but with different kinetics.
Visible component of sunlight has a physiologically significant effect on human skin and long-term exposure to concentrated blue light energy (sunlight, laptop, cell phones) could produce oxidative stress leading to the premature skin aging. In this work, in vivo confocal Raman spectroscopy was used to characterize biochemical changes in human skin after been irradiated with different doses of blue light. After ethical committee approval, volunteers’ phototype I and II (Fitzpatrick classification) have been selected. The River Diagnosis confocal Raman spectrometer was used, before (T0) and after 15, 30 and 60 minutes of blue light irradiation (LED 450 nm) with doses of 100 J/cm2 . It was possible to evaluate the biochemical skin damage caused in the stratum corneum and in the viable epidermis.
In the confocal Raman spectra of skin dermis, the band area in the spectral region of proline and hydroxyproline varies according to the age and health condition of the volunteers, classified as healthy young women, healthy elderly women, and diabetic elderly women. Another observation refers to the intensity variation and negative Raman shift of the amide I band. To understand these effects, we adopted a model system using the DFT/B3LYP:3-21G procedure, considering the amino acid chain formed by glycine, hydroxyproline, proline, and alanine, which interacts with two and six water molecules. Through these systems, polarizability variations were analyzed to correlate its values with the observed Raman intensities of the three groups of volunteers and to assign the vibrational spectra of the skin dermis. As a way to correlate other experimental trends, we propose a model of chemical reaction of water interchange between the bonding amino acids, in which water molecules are attached with glucose by hydrogen bonds. The theoretical results are in accordance with the observed experimental trends.
In vivo confocal Raman spectroscopy is a powerful non-invasive technique able to analyse the skin constituents. This technique was applied to transdermal perfusion studies of the vitamin A derivative in human skin. The composition of the stratum corneum (lipid bilayer) is decisive for the affinity and transport of the vitamin through skin. The vitamin A is significantly absorbed by human skin when applied with water in oil emulsion or hydro-alcoholic gel. The purpose of this study is to elucidate the behaviour of vitamin A derivative into human skin without the presence of enhancers. The results showed that the intensity band of the derivative (around 1600 cm-1), which represents the -C=O vibrational mode, was detected in different stratum corneum depths (up to 20 μm). This Raman peak of vitamin A derivative has non-coincident band with the Raman spectra of the skin epidermis, demonstrating that compound penetrated in forearm skin.
The aging process involves the reduction in the production of the major components of skin tissue. During intrinsic aging and photoaging processes, in dermis of human skin, fibroblasts become senescent and have decreased activity, which produce low levels of collagen. Moreover, there is accumulation of advanced glycation end products (AGEs). AGEs have incidence in the progression of age-related diseases, principally in diabetes mellitus and in Alzheimer's diseases. AGEs causes intracellular damage and/or apoptosis leading to an increase of the free radicals, generating a crosslink with skin proteins and oxidative stress. The aim of this study is to detect AGEs markers on human skin by in vivo Confocal Raman spectroscopy. Spectra were obtained by using a Rivers Diagnostic System, 785 nm laser excitation and a CCD detector from the skin surface down to 120 μm depth. We analyzed the confocal Raman spectra of the skin dermis of 30 women volunteers divided into 3 groups: 10 volunteers with diabetes mellitus type II, 65–80 years old (DEW); 10 young healthy women, 20–33 years old (HYW); and 10 elderly healthy women, 65–80 years old (HEW). Pentosidine and glucosepane were the principally identified AGEs in the hydroxyproline and proline Raman spectral region (1000–800 cm–1), in the 1.260–1.320 cm–1 region assignable to alpha-helical amide III modes, and in the Amide I region. Pentosidine and glucosepane calculated vibrational spectra were performed through Density Functional Theory using the B3LYP functional with 3-21G basis set. Difference between the Raman spectra of diabetic elderly women and healthy young women, and between healthy elderly women and healthy young women were also obtained with the purpose of identifying AGEs Raman bands markers. AGEs peaks and collagen changes have been identified and used to quantify the glycation process in human skin.
Raman spectroscopy has been applied to the analysis of biological samples for the last 12 years providing detection of changes occurring at the molecular level during the pathological transformation of the tissue. The potential use of this technology in cancer diagnosis has shown encouraging results for the in vivo, real-time and minimally invasive diagnosis. Confocal Raman technics has also been successfully applied in the analysis of skin aging process providing new insights in this field. In this paper it is presented the latest biomedical applications of Raman spectroscopy in our laboratory. It is shown that Raman spectroscopy (RS) has been used for biochemical and molecular characterization of thyroid tissue by micro-Raman spectroscopy and gene expression analysis. This study aimed to improve the discrimination between different thyroid pathologies by Raman analysis. A total of 35 thyroid tissues samples including normal tissue (n=10), goiter (n=10), papillary (n=10) and follicular carcinomas (n=5) were analyzed. The confocal Raman spectroscopy allowed a maximum discrimination of 91.1% between normal and tumor tissues, 84.8% between benign and malignant pathologies and 84.6% among carcinomas analyzed. It will be also report the application of in vivo confocal Raman spectroscopy as an important sensor for detecting advanced glycation products (AGEs) on human skin.
Accumulation of AGEs [Advanced Glycation End – products] occurs slowly during the human aging process. However, its formation is accelerated in the presence of diabetes mellitus. In this paper, we perform a noninvasive analysis of glycation effect on human skin by in vivo confocal Raman spectroscopy. This technique uses a laser of 785 nm as excitation source and, by the inelastic scattering of light, it is possible to obtain information about the biochemical composition of the skin. Our aim in this work was to characterize the aging process resulting from the glycation process in a group of 10 Health Elderly Women (HEW) and 10 Diabetic Elderly Women (DEW). The Raman data were collected from the dermis at a depth of 70-130 microns. Through the theory of functional density (DFT) the bands positions of hydroxyproline, proline and AGEs (pentosidine and glucosepane) were calculated by using Gaussian 0.9 software. A molecular interpretation of changes in type I collagen was performed by the changes in the vibrational modes of the proline (P) and hydroxyproline (HP). The data analysis shows that the aging effects caused by glycation of proteins degrades type I collagen differently and leads to accelerated aging process.
In this paper, we studied the phenylalanine amino acid in different systems, it means, the amino acid with different solvent molecules as water, ammonia and urea, via density functional theory (DFT). These molecules are present in the natural moisturizing factor (NMF) of skin that is found in the stratum corneum. The aim of this study is improving the understanding of the environment role and its influences. Therefore, in this paper we analyzed the vibrational modes (and compared to FT-Raman and FT-IR experimental values) and electronic properties as the charge densities (HOMO and LUMO). We also verified the hydrophobic effect and the dependence of amino acid structure on the quantities of solvent.
In this work we propose the use of experimental and theoretical reflectance anisotropy spectra (RAS) as a new tool to identify structural and dynamical aspects of the bilipid membrane and its various constituent molecules. The role of geometric details at the atomic level and macroscopic quantities, such as the membrane curvature and tilt for the different gel phases, in the theoretical RAS spectra (using Kohn-Sham density functional theory (KS-DFT)) are presented. Then the results are compared to the experimentally measured spectra taken from other techniques.
In this paper we investigated the Paracoccidoides brasiliensis fungus nanosensor by simulations of simple strand
DNA grafting on gold nanoparticle. In order to improve the knowledge of nanoparticle environment, the addiction of salt
solution was studied at the models proposed by us. Nanoparticle and DNA are represented by economic models validated
by us in this paper. In addition, the DNA grafting and salt influences are evaluated by adsorption and bond energies
calculations. This theoretical evaluation gives support to experimental diagnostics techniques of diseases.
Rapid microbiological identification and characterization are very important in dentistry and medicine. In addition to
dental diseases, pathogens are directly linked to cases of endocarditis, premature delivery, low birth weight, and loss of
organ transplants. Fourier Transform Infrared Spectroscopy (FTIR) was used to analyze oral pathogens Aggregatibacter
actinomycetemcomitans ATCC 29523, Aggregatibacter actinomycetemcomitans-JP2, and Aggregatibacter
actinomycetemcomitans which was clinically isolated from the human blood-CI. Significant spectra differences were
found among each organism allowing the identification and characterization of each bacterial species. Vibrational modes
in the regions of 3500-2800 cm-1, the 1484-1420 cm-1, and 1000-750 cm-1 were used in this differentiation. The
identification and classification of each strain were performed by cluster analysis achieving 100% separation of strains.
This study demonstrated that FTIR can be used to decrease the identification time, compared to the traditional methods,
of fastidious buccal microorganisms associated with the etiology of the manifestation of periodontitis.
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