A proof-of-concept study utilising 2D NMR spectrometry for in situ characterisation and quantitation of key biomarkers and actives in tape stripped ex vivo human skin.

The development of a semi-automated and rapid analytical technique for dermatological analysis has become a key aim of many medical and commercial entities through greater awareness of people to skin health and its importance in the 21st century. We present a proof-of-concept methodology demonstrating the use of validated non-destructive, in-situ (Nuclear Magnetic Resonance Spectroscopy) NMR techniques for characterisation and quantitation of (Natural Moisturising Factor) NMF compounds and actives from topical formulations. This quantitation is crucial for appropriate diagnosis of atopic dermatitis severity due to its association with reduced NMF abundance. This study is the first to combine diffusion NMR, semi-automated quantitation and ex-vivo skin samples to measure NMF and permeation of actives. We have shown that diffusion NMR allows for resolution between formulation components through determination of self-diffusion coefficients. We also demonstrate how the metabolomics software chenomxtm can be used to identify and quantitate individual NMF components. We show comparable results to previous literature on NMF layers in the skin, alongside reinforcing findings on permeation enhancers and heat effects on transdermal delivery of actives and formulation components. The presented methodology has shown great potential as an effective non-destructive, fast and versatile technique for dermatological analysis of physiology and actives, with future hardware and software developments in NMR making the future of dermatological analysis via NMR very promising.

Characterizing the nanomechanical properties of microcomedones after treatment with sodium salicylate ex vivo using atomic force microscopy.

OBJECTIVE: The treatment of acne presents a major clinical and dermatological challenge. Investigating the nanomechanical properties of the microcomedone precursor lesions using atomic force microscopy (AFM) may prove beneficial in understanding their softening, dissolution and prevention. Although the exact biochemical mechanism of NaSal on microcomedones is not fully understood at present, it appears to exhibit a significant exfoliation effect on the skin via corneodesmosome dissolution. METHODS: Therefore, to support this exploration, sodium salicylate (NaSal), a common ingredient employed in skin care products, is applied ex vivo to microcomedones,collected by nose strip adhesive tape, and their nanomechanical properties are assessed using AFM. Although the exact biochemical mechanism of NaSal on microcomedones is not fully understood at present, it appears to exhibit a significant exfoliation effect on the skin via corneodesmosome dissolution. RESULTS: Herein, our findings demonstrate that when microcomedones are treated with 2% NaSal, samples appeared significantly more compliant ('softer') ((1.3 ± 0.62) MPa) when compared to their pre-treated measurements ((7.2 ± 3.6) MPa; p = 0.038). Furthermore, elastic modulus maps showed that after 2% NaSal treatment, areas in the microcomedone appeared softer and swollen in some, but not in all areas, further proving the valuable impact of 2% NaSal solution in altering the biomechanical properties and morphologies in microcomedones. CONCLUSION: Our results are the first of their kind to provide qualitative and quantitative mechanobiological evidence that 2% NaSal decreases the elastic modulus of microcomedones. Therefore, this study provides evidence that NaSal can be beneficial as an active ingredient in topical treatments aimed at targeting microcomedones.

OBJECTIF: Le traitement de l'acné présente un défi clinique et dermatologique majeur. L'étude des propriétés nanomécaniques des lésions précurseurs en tant que microcomédons à l'aide de la microscopie à force atomique (AFM) peut s'avérer bénéfique pour comprendre leur ramollissement, leur dissolution et leur prévention. MÉTHODES: Par conséquent, pour soutenir cette exploration, le salicylate de sodium (NaSal), un ingrédient couramment utilisé dans les produits de soins de la peau, est appliqué ex vivo aux microcomédons et leurs propriétés nanomécaniques sont évaluées à l'aide de l'AFM. Bien que le mécanisme biochimique exact du NaSal sur les microcomédons ne soit pas entièrement compris à l'heure actuelle, il semble présenter un effet exfoliant significatif sur la peau via la dissolution des cornéodesmosomes. RÉSULTATS: Ici, nos résultats démontrent que lorsque les microcomédons sont traités avec 2% de NaSal, les échantillons semblaient significativement plus conformes ("plus doux") ((1.3 ± 0.62) MPa) par rapport à leurs mesures pré-traitées ((7.2 ± 3.6) MPa ; P = 0,03826). De plus, les cartes du module d'élasticité ont montré qu'après un traitement à 2 % de NaSal, les zones du microcomédon semblaient plus molles et gonflées dans certaines zones, mais pas dans toutes, prouvant ainsi l'impact précieux d'une solution de NaSal à 2 % dans la modification des propriétés biomécaniques et de la morphologie des microcomédons. CONCLUSION: Nos résultats sont les premiers du genre à fournir des preuves mécanobiologiques qualitatives et quantitatives que 2% de NaSal diminue le module d'élasticité des microcomédons. Par conséquent, cette étude fournit des preuves que NaSal peut être bénéfique en tant qu'ingrédient actif dans les traitements topiques visant à cibler les microcomédons.

Serum cytokine levels as markers of paralytic ileus following robotic radical prostatectomy at different pneumoperitoneum pressures.

BACKGROUND: To evaluate intraoperative and postoperative cytokines in patients who underwent robotic prostatectomy (RP) at a pressure of 12 or 15 mm Hg, and the risk of postoperative ileus. MATERIALS AND METHODS: We presented the first series evaluating intraoperative and postoperative cytokines in patients undergoing RP at a pressure of 12 or 15 mm Hg by a single surgeon. Changes in cytokine concentrations were shown to correlate with surgical outcomes and pathological states. The study investigated the changes in cytokine concentrations (interferon-γ, tumor necrosis factor-α, interleukin-1ß [IL-1ß], IL-2, IL-4, IL-6, IL-12, and IL-17) at different pneumoperitoneum pressures and their potential role in the development of postoperative ileus. RESULTS: The data on 10 consecutive patients confirmed that a lower pneumoperitoneum pressure was associated with lower cytokine levels and a lower risk of ileus. There were increased levels of postoperative interferon-γ, tumor necrosis factor-α, IL-12p70, IL-1ß, IL-2, IL-4, and IL-17a at 15 mm Hg when compared to 12 mm Hg. CONCLUSIONS: The data indicated that lower pressure RP reduced intra-/postoperative cytokine levels confirming our hypothesis. Larger patient numbers are required to further validate this but the implications of this data will benefit not only urological patients but also other speciality patients undergoing minimally invasive surgery.

Effect of allergens and irritants on levels of natural moisturizing factor and corneocyte morphology.

BACKGROUND: The irritant sodium lauryl sulfate (SLS) is known to cause a decrease in the stratum corneum level of natural moisturizing factor (NMF), which in itself is associated with changes in corneocyte surface topography. OBJECTIVE: To explore this phenomenon in allergic contact dermatitis. METHODS: Patch testing was performed on patients with previously positive patch test reactions to potassium dichromate (Cr), nickel sulfate (Ni), methylchloroisothiazolinone (MCI)/methylisothiazolinone (MI), or p-phenylenediamine. Moreover, a control (pet.) patch and an irritant (SLS) patch were applied. After 3 days, the stratum corneum from tested sites was collected, and NMF levels and corneocyte morphology, expressed as the amount of circular nanosize objects, quantified according to the Dermal Texture Index (DTI), were determined. RESULTS: Among allergens, only MCI/MI reduced NMF levels significantly, as did SLS. Furthermore, only MCI/MI caused remarkable changes at the microscopic level; the corneocytes were hexagonal-shaped with pronounced cell borders and a smoother surface. The DTI was increased after SLS exposure but not after allergen exposure. CONCLUSIONS: MCI/MI significantly decreased NMF levels, similarly to SLS. The altered corneocyte morphology suggests that skin barrier damage plays a role in the pathogenesis of MCI/MI contact allergy. The DTI seems to differentiate reactions to SLS from those to the allergens tested, as SLS was the only agent that caused a DTI increase.